TW201037078A - Preparation of alpha-ketopimelic acid - Google Patents

Preparation of alpha-ketopimelic acid Download PDF

Info

Publication number
TW201037078A
TW201037078A TW099107114A TW99107114A TW201037078A TW 201037078 A TW201037078 A TW 201037078A TW 099107114 A TW099107114 A TW 099107114A TW 99107114 A TW99107114 A TW 99107114A TW 201037078 A TW201037078 A TW 201037078A
Authority
TW
Taiwan
Prior art keywords
lie
glu
ala
gly
lys
Prior art date
Application number
TW099107114A
Other languages
English (en)
Inventor
Petronella Catharina Raemakers-Franken
Martin Schurmann
Axel Christoph Trefzer
Wildeman Stefaan Marie Andre De
Original Assignee
Dsm Ip Assets Bv
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Dsm Ip Assets Bv filed Critical Dsm Ip Assets Bv
Publication of TW201037078A publication Critical patent/TW201037078A/zh

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/44—Polycarboxylic acids
    • C12P7/50—Polycarboxylic acids having keto groups, e.g. 2-ketoglutaric acid
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0008—Oxidoreductases (1.) acting on the aldehyde or oxo group of donors (1.2)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1025—Acyltransferases (2.3)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/88—Lyases (4.)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/005—Amino acids other than alpha- or beta amino acids, e.g. gamma amino acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/02—Amides, e.g. chloramphenicol or polyamides; Imides or polyimides; Urethanes, i.e. compounds comprising N-C=O structural element or polyurethanes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
    • C12P17/10—Nitrogen as only ring hetero atom
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/44—Polycarboxylic acids

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)

Description

201037078 六、發明說明： 【發明所屬之技術領域】 本發明係關於一種製備α-酮庚二酸（下文中亦稱為 「ΑΚΡ」；ΑΚΡ亦名為2-側氧基-庚二酸）之方法。本發明另 外係關於一種製備5-甲醯基戊酸（下文中亦稱為「5-FVA」） 之方法及製備6-胺基己酸（下文中亦稱為「6-ACA」）之方 法。本發明亦係關於一種製備二胺基己烷（亦名為丨，6-已二 胺）之方法。本發明另外係關於一種可用於本發明之方法 〇 中的異源細胞。本發明進一步係關於一種異源細胞在製備 ε-己内醯胺（下文中稱為「己内醯胺」）、6_胺基己酸或二 胺基己烷中之用途。 【先前技術】 一版基己烧尤其用於產生諸如耐綸6,6(nylon 6,6)之聚酿 胺。其他用途包括用作其他基本組份（例如二異氰酸伸己 醋）之起始物質及用作環氧化物之交聯劑。已知之製備方 法由丙烯腈開始經由己二腈進行。 Ο 己内醯胺為一種可用於產生例如耐綸-6或耐綸-6,12(己 内酿胺與十二内醯胺之共聚物）之聚醯胺的内醯胺。由散 裝化學品製備己内醯胺之各種方法為此項技術中所知且包 括由％己_、曱苯、苯盼、環己醇、苯或環己燒製備己内 醯胺。此等中間化合物一般由礦物油獲得。鑒於日益期望 使用更可持續性技術來製備材料，將需要提供一種由可自 千上可再生來源獲得之中間化合物或至少由可使用生 物化學方法轉化為己内醯胺之中間化合物製備己内醯胺的 146966.doc 201037078 方法。另外，將需要提供一種與習知化學方法相比，自石 化來源利用散裝化學品需要更少能量之方法。 舉例而言，如118-八6,194,572中所述，已知由6-八€八製 備己内醯胺。如WO 2005/068643中所揭示，可以生物化學 方式藉由在具有α,β-烯酸還原酶活性之酶存在下轉化6-胺 基己-2-烯酸（6-ΑΗΕ Α)來製備6-AC Α。可例如以生物化學 方式或藉由純化學合成自離胺酸製備6-AHEA。儘管藉由 WO 2005/068643中所揭示之方法可經由還原6-AHEA來製 備6-ACA，但本發明者已發現在還原反應條件下，6-AHE A可自發地且實質上不可逆地環化形成不需之副產 物，尤其為β-高脯氨酸。此環化作用可為6-ACA產生過程 中之瓶頸且可導致產率大幅度下降。 本發明者已認識到可自ΑΚΡ製備6-ACA。可以化學方 式，例如基於如 Η. JSger等人，Chem. Ber. 1959, 92, 2492-2499所述之方法，來製備ΑΚΡ。ΑΚΡ可藉由以下步驟來製 備：使用乙醇鈉作為鹼，以草酸二乙酯將環戊酮烷基化； 在強酸（2 M HC1)中使所得產物回流；及例如藉由自曱苯 中結晶來回收產物。然而，如上所指出，日益期望使用更 可持續性技術來製備材料。因此，本發明者認識到將需要 提供一種由可自生物學上可再生來源獲得之中間化合物製 備ΑΚΡ的方法。 【發明内容】 本發明之一目標在於提供製備ΑΚΡ之新穎方法，ΑΚΡ尤 其可用於製備6-ACA、二胺基己烷或另一化合物。 146966.doc 201037078 另一目標在於提供適於在製備AKP之方法中催化一或多 個反應步驟的生物催化劑。 可根據本發明解決之一或多個其他目標將由以下說明書 中得出。 本發明者已認識到可使用特定生物催化劑製備Ακρ。 因此，本發明係關於一種製備AKP之方法，其包含將α_ 酮戊二酸（AKG)轉化為α-酮己二酸（ΑΚΑ)及將心酮己二酸 轉化為α-酮庚二酸，其中使用生物催化劑（詳言之為異源 〇 生物催化劑）進行此等轉化中之至少一者。 ΑΚΡ可例如用作製備5-甲醯基戊酸（5_FVA)之中間物。 因此’本發明另外係關於一種製備5_FVA之方法，其包 含以生物催化方式使藉由本發明之方法製備之Ακρ脫去羧 基，藉此形成5-FVA。 5- FVA為例如適用於製備6_aca、己内醯胺或二胺基己 烷之中間化合物。 AKP可例如用作製備01胺基_庚二酸（AAp)之中間物。 ^ 因此，本發明另外係關於一種製備AAP之方法，其包含 以生物催化方式促使在本發明之方法中製備之Ακρ轉胺， 藉此形成AAP。 AAP為例如適用於製備6-ACA或己内醯胺之中間化合 物。 6- ACA可例如轉化為己内醯胺或二胺基己烷。 本發明另外提供一種異源細胞，其包含一或多個編碼一 或多種異源酶之異源核酸序列，該一或多種異源酶能夠在 146966.doc 201037078 由α-酮戊二酸製備α_酮庚二酸中催化至少—個反廉步驟。 該細胞可尤其用作製備至少一種選自以下之群之化合物 的方法中之生物催化劑：ΑΚΡ、5-FVA、6_ACA、二^基 己烷及己内醯胺。 1 根據本發明，當形成6-ACA及視情況存在之己内酿胺 時，未見到有關於中間產物之不當環化作用，導致產率下 降的問題。 預計本發明之方法與W0 2005/68643中所述方法相比有 相當或甚至更高之產率。預計若使用活有機體，尤盆在考 慮有機體生長及維持之方法中，則本發明之方法可尤其為 有利‘的。 另外預計在本發明 Λ ^ Τ ACA之生產力（以g/i h計）可得到提高 「Li另外指定，否則如本文中所用之術語「或」定㈣ 除非另外指定，否則如本文中所用之術語「一」定義名 |至少一」。 田提及早數形式之名詞（例如—種化合物、—種添加齊 專)時’意謂包括複數形式。因此，除非另外指定，否貝, 當提及—種特定部分（例如「化合物」）時，此意m -種」該部分’例如「至少一種化合物」。 '、非另外才日疋’否貝j當本文中提及叛酸或叛酸醋（例女 3胺基酸、5-FVA、丁二酸/丁二酸酯、乙酸/乙 時’此等術sf意謂包括質子化竣酸（游離酸）、相應寿 146966.doc 201037078 酸醋(其共軛鹼)以及其鹽。同樣，當提及胺時，此意謂包 括枭子化胺（通常為陽離子型，例如R_NH3+)及未質子化胺 (通常不帶電荷，例如r.2)。當本文中提及胺基酸（例如 6-ACA)時，此術語意謂包括呈兩性離子形式之胺基酸（其 中胺基呈質子化形式且羧酸酯基呈去質子化形式）、胺基 呈質子化形式且羧基呈其中性形式之胺基酸、及胺基呈其 中性形式且羧酸醋基呈去質子化形式之胺基酸以及其鹽f 當提及存在若干異構體（例如順式及反式異構體、尺及s 〇 對映異構體）之化合物時，化合物原則上包括可用於本發 明之特定方法中的該化合物之所有對映異構體、非對映異 構體及順式/反式異構體。 虽參考括號中之酶類別（EC)提及酶時，酶類別為基於由 國際生物化學及分子生物學聯盟命名委員會（N〇menelature Committee of the International Union of Biochemistry and M〇lecular Biology，NC_IUBMB)提供之酶命名法已將或可 將轉纟5)7類之類別，此命名法可在http://www.chem.qmul.ac.uk/ iubmb/enzyme/中找到。意欲包括（尚）未歸入指定類別中但 可歸為此類之其他適合酶。 除非另外指定，否則若本文中參考寄存編號提及蛋白質 或基因’則此編號尤其用以指具有如Unipr〇t中2〇〇8年3月 Η曰所見之序列的蛋白質或基因。 如本文中所用’術語核酸之r功能類似物」至少包括編 碼具有相同胺基酸序列之酶的其他序列及編碼該酶之同系 物的其他序列。 146966.doc 201037078 本文中所用之術語「同系物」尤其係指具有至少3〇%、 較佳至少40%、更佳至少6〇%、更佳至少65%、更佳至少 70%、更佳至少75%、更佳至少嶋、尤其至少抓、更尤 其至少90%、至少91〇/〇、至少92%、至少93%、至少94%、 至少95%、至少96%、至少97%、至少嫩或至少99%之序 列-致性的聚核苦酸或多1。被"吾同系物亦意謂包括由於 遺傳密碼簡併而不同於另一核酸序列且編碼相同多肽序列 之核酸序列（聚核苷酸序列）。 序列一致性或相似性在本文中定義為兩個或兩個以上多 肽序列或兩個或兩個以上核酸序列之間如藉由比較該等序 列所確定的關係。通常，在全長序列上比較序列一致性或 相似性，然而亦可僅比較彼此進行比對之序列之一部分。 在此項技術中’「一致性」或「相似性」亦意謂多肽序列 或核酸序列之間的序列關聯程度，視情況而定，如由該等 序列之間的匹配來測定。測定一致性或相似性之較佳方法 經設計以獲得所測試序列之間的最大匹配。在本發明之上 下文中，測疋兩種序列之間的一致性及相似性之較佳電腦 程式方法包括 BLASTP 及 BLASTN(Altschul，S. F·等人， Mol· Biol. 1990，275，403-410，由 NCBI 及其他來源
(BLAST Manual, Altschul, S.等人 ’ NCBI NLM NIH
Bethesda，MD 20894)公開獲得）。使用BLASTP供比較多肽 序列之較佳參數為間隙開口 1〇.〇，間隙擴展〇 5，Bi〇sum 62矩陣。使用blaSTN供比較核酸序列之較佳參數為間隙 開口 10.0，間隙擴展0.5，DNA全矩陣（DNA—致性矩陣）。 146966.doc 201037078 如本文中所用之異源生物催化劑（詳言之為異源細胞）為 包含異源蛋白質或異源核酸（通常作為細胞Dna或RNA之 一部分）之生物催化劑。術語「異源」在用於核酸序列 (DNA或RNA)或蛋白質時係指並非天然作為其所存在之有 機體、細胞、基因組或DNA或RNA序列之一部分存在或 在不同於其天然存在之細胞或基因組或DNA或RNA序列中 之位置的細胞或基因組或DNA或RNA序列中之位置中所發 現的核酸或蛋白質。應瞭解異源有機體中之異源DNA為玆 Ο 異源有機體之基因組的一部分。異源核酸或蛋白質對於引 入其之細胞而言非内源性的，而自另一細胞獲得或藉由合 成或重組方法產生。一般但不一定，該等核酸編碼通常不 由轉錄或表現該DNA之細胞產生的蛋白質。類似地，異源 RNA編碼通常不在存在該異源rna之細胞中表現的蛋白 質。異源核酸及蛋白質亦可稱為外來核酸或蛋白質。在本 文中術語異源核酸或蛋白質涵蓋熟習此項技術者公認為對 表現此核酸或蛋白質之細胞為異源或外來的任何核酸或蛋 ❹白質。 當提及來自特定來源之酶或另一生物催化部分時，特定 意謂包括源自於第一有機體但實際上在（經遺傳修飾之）第 一有機體中產生的重組酶或其他重組生物催化部分為來自 該第一有機體之酶或其他生物催化部分。 在本發明之一方法中’使用生物催化劑，亦即該方法中 之至少一個反應步驟由衍生自生物來源（例如有機體或自 其獲得之生物分子）之生物物質或部分催化。生物催化劑 146966.doc 201037078 可尤其包含一或多種酶。生物催化反應可包含一或多個化 學轉化反應’其中至少一個轉化反應由生物催化劑催化。 因此，「生物催化劑」可促進以下反應步驟中之化學反 應：由AKG製備AKP中之至少一個反應步驟、由AKP製備 5-FVA或AAP中之至少一個反應步驟、由5_FVA製備6_aca 中之至少一個反應步驟、由AAP製備6-AC A中之至少一個 反應步驟或由6-ACA製備己内醯胺中之至少一個反應步 驟。 生物催化劑可以任何形式使用。在一實施例中，一或多 種酶構成活有機體（諸如活的全細胞）之一部分。該等酶可 在細胞内執行催化功能。酶亦可能分泌至細胞所存在之介 質中。在一實施例中，使用自天然環境分離（自產生酶之 有機體分離）例如呈溶液、乳液、分散液、束乾細胞（懸浮 液）、溶解產物形式，或固定於支撐物上之一或多種酶。 蓉於調節反應條件之靈活性提高以使反應平衡移向所需 側’故使用自產生酶之有機體分離之酶可尤其適用。 活細胞可為生長細胞、靜止或休眠細胞（例如抱子）或處 於固定相之細胞。亦可能使用構成透性化細胞之一部分的 酶（亦即該細胞對酶受質或酶受質之前驅體可滲透）。 (用於本發明方法中之）生物催化劑原則上可為任何有機 體或可自任何有機體獲得或衍生。此有機體可為天然存在 之有機體或異源有機體。異源有機體通常為包含至少—個 編碼異源酶之核酸序列的宿主細胞，該異源酶能夠催化本 發明方法中之至少一個反應步驟。產生異源核酸序列之有 146966.doc -10- 201037078 機體可為真核或原核有機體。詳言之，該等有機體可獨立 地選自動物（包括人類）、植物、細菌、太古菌（archaea)、 酵母及真菌。 宿主細胞可為真核或原核細胞。在一實施例中，宿主細 胞係選自真菌、酵母、眼蟲類（euglenoids)、太古菌及細菌 之群。宿主細胞可尤其選自由以下組成之群：麴菌屬 、青黴菌屬黑穗病菌屬 (UstUago)、頭抱黴慝（Cephalosporitim)、髮癬蛰屬 〇 (7Wc/zc^/^ww)、擬青黴菌屬、畢赤酵母菌 屬、漢森酵母菌屬、酵母菌屬 、假絲酵母菌屬、克魯維酵母囷 屬（iir/i^verowyces)、耶氏酵母菌屬㈣⑷、芽孢桿菌 屬（5ac"/Mi)、棒狀桿菌屬狀&cierz·謂）、埃希氏菌屬 (心c/zeWc；；^)、固氮菌屬（JzoiC)kcier)、弗蘭克氏菌屬 (FranAr/a)、根瘤菌屬（及办如心·㈣）、慢生根瘤菌屬 (Bradyrhizobium)、念珠漆屬 O^abaena')、集胞漆屬 ❹ （办《ec/zocj^iz··?)、微囊滅屬（从化⑺叮*5沿）、克雷伯氏桿函屬 (尺/eWe//a)、紅細菌屬（仙〇而办acier)、假單胞菌屬 (PsewiioTnowa·?)、棲熱菌屬（TTiawws)、奇球菌屬 (Dekoeoccw)、葡糖桿菌屬（G/wco«o&acier)、甲娱:球形菌 屬（Methanosphaera)、气恢> 短择儀慝{Methanobrevibacter)、 甲炫螺菌屬、甲燒球菌屬（Mei/zanococcws)、 甲院桿菌屬（Mei/m⑽kcierz'ww)、甲烧暖球菌屬 (Mei/zawoca/i/ococcw·?)及甲炫八聯球菌屬（Mei/mwosarcMa)。 146966.doc -11 - 201037078 詳。用於本發明方法中之宿主菌株及由此之宿主細胞 了 自以下之群.大腸桿菌（五％心eW)、棕色固I 菌（上oiohciw W⑽/⑽山·〇、肺炎克雷伯氏桿菌（尺心心紿 pneumoniae)、念珠薄屬（dnabaena印·）、集跑漆屬 〇S>«d〇c少灿印·）、銅綠微囊藻（施⑽―沿 flerwgkosa)、耐幸g 射奇球菌（£)e/„〇C0CCWiS1 ra山·〇“r⑽《）、嗜 熱奇球 _ (Dehococcws geoi/zerma/z··?)、嗜熱棲熱菌 [Thermus thermophilus)、枯萆桿蛰（Bacillus subtilis)、解 殿粉芽孢桿菌（心⑼心amj/o/zXacz.ewi·)、甲酵芽孢桿菌 {Bacillus methanolicus)、麵胺酸棒狀桿菌 {Corynebacterium glutamicum)、黑麴菌（Α/^7ζ·//μ5 niger)、轰_ 专青徽 IPenicillium chrysogertum)、點青徽 (_Pew/c///z_ww 、肉色擬青徽（Pagc/Zo/Ti^^gs c<3rweM*s)、 頂頭孢黴（Cep/za/ospoWww acremom’wm)、玉米黑穗病菌 (tAsh/ago所吵山、）、曱醇酵母（Pz’c/π'α pasiork)、釀酒酵母 (Saccharomyces cerevisiae)、 莩L 酸克 魯維酵母 {Kluyveromyces /ac"、）、麥芽糖假絲酵母（CawAiia ma/ίί^α)、解脂耶氏酵母（yi3rrow/a /φο/少i/ca)、多形漢森 酵母（Hansemi!a polymorpha)、疏確礦疏化葉菌（Sulfolobus soZ/aiar/cM·?)、_嗜熱自養甲烧桿菌 thermo auto thro phi cum) ' 海沼曱烧球菌（Mei/zanococcws mizr/pa/wi/z··?)、詹氏甲烧暖球菌（似eMawocor/ciococcw·? jannashii) ' 思氏曱烧球形菌（Mei/zanos/^aera Wai/imwae)、沃氏甲烧球菌v〇"ae)、醋酸 146966.doc -12- 201037078 曱烧八聯球菌（Mei/zanosam'wa aceiz'voran^1)、巴氏甲院八 聯球菌（Mei/zawosarcz·/^ 6arA:erz·)、醋酸曱院八聯球菌、亨 氏甲烧螺菌（Μβί/2βπσ·χ7?ζ·Γ///ι^ /zMngaie/)、嗜熱甲烧鬃菌 {Methanosaeta thermophila)、史氏甲烧短桿菌 {Methanobrevibacter smithii)、萬氏曱烧球菌 {Methanococcus va«m_e///)、雜色甲烧球菌（Mei/zanococcws aeo/icws)及馬氏曱烧八聯球菌（Mei/zimosarc/wa maze/)宿主 細胞。 〇 詳言之，在AKP轉化為另一產物（例如5-FVA、AAP、二 胺基己烷或6-ACA)之一實施例中，認為宿主細胞宜為天然 能夠將AKP轉化為該產物或至少能夠催化至少一個必需反 應之有機體。舉例而言，大腸桿菌具有胺基轉移酶活性， 藉此大腸桿菌可催化由AKP形成AAP(亦參見下文）或5-FVA(若細胞亦含有適合脫羧酶，則可在細胞中形成5-FVA，亦參見下文）轉化為6-ACA之反應。 宿主細胞宜為包含催化供離胺酸生物合成之胺基己二酸 ® 途徑（亦稱為AAA途徑）之生物催化劑或其一部分的有機體 (諸如低等真核生物：真菌、酵母、眼蟲類；某些細菌， 例如棲熱菌屬、奇球菌屬；太古菌）或包含用於經由固氮 酶來固氮之生物催化劑的有機體。 在一較佳實施例中，宿主細胞為以高通量穿過AAA途徑 之有機體，諸如產黃青黴、玉米黑穗病菌或經改適較佳最 利於離胺酸產生之有機體。高通量定義為在所選產生條件 下各別有機體中為細胞蛋白質之生物合成供應離胺酸所需 146966.doc -13- 201037078 7之0。速率7至少概，更佳為至少5G%，甚至更佳為至少 7〇〇/。，最佳為至少1〇〇%。 芏夕 之二：佳:施例中’宿主細胞為產生高含量高檸檬酸酯 ==體’其可為天时在之有機體或異源有機體。此類 有機體可藉由表現在固氮酶或其同系物中所發現之必需輔 因子之形成所需的高擰檬酸合成酶而獲得。 在-實施例中，宿主細胞包含源自動物，尤其源自其一 部分’例如肝冑、胰臟、大腦、腎臟、心臟或其他器官之 異源核酸序列。動物可尤其選自哺乳動物之群，更尤其選 自兔科如〜如）、鼠科（施心e)、豬科咖㈣及^科 (Bovidae)之群。 在實施例中伯主細胞包含源自植物之異源核酸序 列。適合植物尤其包括選自以下之群的植物：鐵角蕨屬 ，葫蘆科幻尤其南瓜屬 (cwwe加），例如番南瓜（CMrcMr0加腳ah⑷（倭瓜）， 或甜瓜屬;十字花科（心似以心“⑽）’尤其芥菜 屬（々MW叩’例如擬南芥菜认Μα/ία⑽）；山靛屬 (MerwWa/⑷’例如多年生山靛⑽⑷；大 風 + 魇（Hydn〇Carpusy，及長龟 i^(Cerat〇n⑹。 在一實施例中’宿主細胞包含源自細菌之異源核酸序 列。適合細菌可尤其選自以下之群：弧菌屬、假 單胞菌屬、芽胞桿菌屬、棒狀桿菌屬、短桿菌屬 (价evzMci⑺謂）、腸球菌屬（五_r〇c〇CCMi)、鏈球菌屬 (•Sire/^ococcws)、放線卤目、克雷伯氏桿 146966.doc -14- 201037078 菌屬、乳球菌屬（Z^ci〇c〇cc⑽）、乳桿菌屬（Zaci〇hcz7/wi)、 梭菌屬、埃希氏菌屬、克雷伯氏桿菌屬、念 珠藻屬、微囊藻屬、集胞藻屬、根瘤菌屬、慢生根瘤菌 屬、棲熱菌屬、分枝桿菌屬（柳、潑酵單胞菌 屬（々m⑽⑽似）、變形桿菌屬（p⑽、土壤桿菌屬 (々rMaderhm)、土芽孢桿菌屬（Ge〇hcz7/w5)、不動桿菌 屬以〇6以如）、固氮菌屬d〇i〇6flc⑽）、雷爾氏菌屬 、紅細菌屬、副球菌屬（户arflC〇CCMi)、新鞘胺醇 〇 桿溘M [Novosphingobium、、亞硝化單胞菌屬 (iWirowmo«似）、退伍軍人桿菌屬（Zey〇„e//a)、奈瑟菌屬 (iVe心erz'a)、紅假早胞菌屬（及、葡萄球 菌屬（5Va/?/z_y/ococcwi)、奇球菌屬及沙門氏菌屬 [Salmonella、。 在一實施例中’宿主細胞包含源自太古菌之異源核酸序 列。適合太古菌可尤其選自以下之群：古丸菌屬 (^rc/iaeog/o^M·?)、氣火菌屬、鹽桿菌屬 〇 零 (Halobacterium) ' 甲烷八聯球菌屬、曱烷球菌屬、熱原體 屬（77zermo;?/a㈣α)、熱球菌屬（TTzermococcws)、火棒菌屬 (P少ro6acw/wm)、曱烷螺菌屬、火球菌屬（Pywcw)、硫 化葉菌屬（iSw//〇/〇0MS)、甲炫球菌屬、曱院球形菌屬、曱烧 火菌屬、甲烧短桿菌屬、曱烧暖球菌屬及 甲烷桿菌屬。 在一實施例中，宿主細胞包含源自真菌之異源核酸序 列。適合真菌可尤其選自以下之群：根黴菌屬 146966.doc -15- 201037078 、平革菌屬（尸/2⑽eroc/zaeie)、裸胞殼屬 (_E/Tza/ce//iz)、黑穩病達屬、脈抱徽屬、青黴 菌屬、頭孢黴屬、擬青黴菌屬、髮癣菌屬 及趟菌屬。 在一實施例中，宿主細胞包含源自酵母之異源核酸序 列。適合酵母可尤其選自以下之群：假絲酵母菌屬、漢森 酵母菌屬、克魯維酵母菌屬、耶氏酵母菌屬、裂瘦酵母菌 屬（Sc/^zoMcc/^row^c^)、畢赤酵母菌屬、耶氏酵母菌屬 及酵母菌屬。 熟習此項技術者將清楚可使用表現天然存在之生物催化 部分（諸如酶）（野生型）或在本發明之方法中具有適合活性 的天然存在之生物催化部分之突變體的生物催化劑。可藉 由熟習此項技術者已知之生物技術，例如藉由分子進化或 合理設計，來改良天然存在之生物催化部分之特性。野生 型生物催化部分之突變體可例如藉由使用熟習此項技術者 已知之突變誘發技術修飾能夠產生生物催化部分（諸如酶） 之有機體之編碼DNA來製備。此等突變誘發技術包括隨機 突變誘發、定點突變誘發、定向進化及基因重組。詳言 之，DNA可經修飾以使其編碼與野生型酶相差至少一個^ 基酸之酶’以便其編碼與野生型相比包含—❹個胺基酸 取代、缺失及/或***之酶，或以使突變體組合兩個或兩 個以上親本酶之序列，或藉由實現由此修飾之亀在適合 (宿主）細胞中表現。可藉由熟f此項技術者已知之方法， 諸如密碼子優化或密碼子對優化，例如基於如 146966.doc -16· 201037078 2008/_632中所述之方法，來達成後者。 突變生物催化劑可具有例如關於—或多個以下態樣獲改 良之特性：對受質之選擇性、活性、穩定性、溶劑耐受 !·生PH值刀布、溫度分布、受質概況、對抑制之敏感性、 輔因子利用率及受質親和力 項技術者已知之方法的適合 有改良特性之突變體。 。可藉由應用例如基於熟習此 高通量篩選或選擇法來鑑別具
G
根？本發明，由AKG製備AKPe AKG原則上可以任何方 式獲得。詳言之，可以生物催化方式藉由向異源生物催化 劑提供可轉化為AKG之適合碳源’例如藉由使該碳源醱 酵來獲仔AKG。在有利之方法中，利用碳源之全細胞生 物轉換以形成AKG，來製備AKG。 / 炭源可尤其含有至少一種選自以下之群之化合物：一元 醇夕7L醇、幾酸、二氧化碳、脂肪酸、甘油醋，包括包 含該等化合物中任一者之混合物。適合一元醇包括曱醇及 乙醇’適合多元醇包括甘油及碳水化合物。適合脂肪酸或 甘油Sa可尤其以較佳植物來源之食用油形式提供。 可尤其使用碳水化合物，因為碳水化合物通常可由生物 子上可再生之來源（諸如農產品，較佳為農業廢料）大量獲 得。較佳使用選自以下之群之碳水化合物：葡萄糖、果 糖薦糖（sucrose)、乳糖、蔗醣（sacchar〇se)、；殿粉、纖維 素及半纖維素。尤其較佳為葡萄糖、包含㈣糖之寡醣及 包含葡萄糖之多醣。 在本發明之一實施例中，使用用於將AKG轉化為aka之 146966.doc 201037078 生物催化劑’將AKG轉化為AKA，該生物催化劑之一部分 源自供離胺酸生物合成之AAA途徑。該轉化反應可包括單 個或複數個反應步驟，其步驟可藉由一或多種生物催化劑 催化。 用於催化AKG轉化為AKA之生物催化劑或其部分可為同 源或異源的。詳言之，形成供離胺酸生物合成之AAA途徑 之一部分的生物催化劑可見於選自酵母、真菌、太古菌及 細&1之群、尤其以下之群之有機體中：青黴菌屬、頭孢黴 屬、擬青黴菌屬、髮癬菌屬、麴菌屬、平革菌屬、裸胞殼 屬、黑穗病菌屬、裂殖酵母菌屬、酵母菌屬、假絲酵母菌 屬、克魯維酵母菌屬、耶氏酵母菌屬、畢赤酵母菌屬、漢 森酵母菌屬、棲熱菌屬、奇球菌屬、火球菌屬、硫化葉菌 屬、熱球菌屬、曱烷球菌屬、曱烷八聯球菌屬、曱烷暖球 菌屬、曱烷球形菌屬、甲烷火菌屬、曱烷短桿菌屬、曱烷 螺ί屬及甲烧熱桿菌屬（Methanothermobacter)。適合生物 催化劑可見於能產生高檸檬酸酯之有機體中，例如在諸如 以下之固氮菌中固氮酶複合物之生物催化劑：藍細菌 (cyan〇baCteria)(例如念珠藻屬、微囊藻屬、集胞藻屬）、根 瘤菌目（Rhizobiales)(例如根瘤菌屬、慢生根瘤菌屬）、γ-變 形菌門（γ-proteobacteria)(例如假單胞菌屬、固氮菌屬、克 雷伯氏桿菌屬）及放線菌綱（actinobacteria)(例如弗蘭克氏 菌屬）。因此’若使用基於天然包含供離胺酸生物合成之 A A A途控之宿主細胞的生物催化劑或其部分，則此系統可 為同源的。 146966.doc -18- 201037078 在本發明之一較佳實施例中，需要藉由生物催化劑實現 AKA之高生產力。含有供離胺酸生物合成之aaa途徑之生 物催化劑或其部分可藉由此項技術中已知之方法，諸如針 對此目的之突變/篩選或代謝工程改造來修飾。可藉由提 高涉及AKA形成之酶的活性及/或降低涉及其轉化為例如 胺基己二酸酯之活性，得到高含量之AKA。 涉及AKA形成之酶包括高檸檬酸合成酶（EC 2 3 3 14)、 高烏頭酸酶（EC 4·2·1·36)及高異檸檬酸脫氫酶（EC 〇 1 · 1.1.87)。此等酶在宿主細胞中之活性可藉由此項技術中 已知之諸如以下之方法來提高：（過度）表現編碼各別酶及/ 或功此同系物之基因，減輕由受質、產物或其他化合物造 成之抑制’或藉由分子進化或合理設計提高酶之催化特 性°執行定向進化之一較佳方法可基於W〇 2003/010183。 因為不希望所產生之AKA轉化為胺基己二酸（aaa)(其為 離胺酸生物合成途徑中之另一步驟），所以異源生物催化 ©劑較佳具有低活性或無活性催化此轉化反應之酶，尤其能 夠摧化此轉化反應之胺基轉移酶（諸如胺基己二酸胺基轉 移酶（EC 2.6.1.3 9))或胺基酸脫氫酶。因此，在提供生物催 化劑之佰主細胞包含編碼此類酶之基因的情況下’較佳使 該基因失活’將其剔除’或減少該基因之表現。由於此步 驟為供離胺酸產生之AAA途徑中所必需的，所以宿主細胞 且具有有限、最小之供應需要量之離胺酸用於生長及維持 的活性’但不能高程度地將AKA轉化為AAA。尤其在產黃 青黴為宿主之情況下’胺基轉移酶可具有序列ID 68之序 146966.doc •19· 201037078 列或其同系物。 可藉由若干方法使編碼不欲活性之基因失活。一種方法 為使用反義分子或RNAi分子之臨時手段（例如基於Kamath 等人20〇3. Nature 421:23卜237)。另一方法係使用可調控啟 動子系統，可使用如四環素（tetracycline)之外部觸發劑將 其關閉（例如基於 Park and Morschhauser, 2005, Eukaryot. Cell. 4:1328-1342)。又一方法係應用化學抑制劑或蛋白質 抑制劑或物理抑制劑（例如基於Tour等人2003. Nat Biotech 21:1505-1508)。一更佳之方法為移除編碼不欲活性之基因 全部或其一部分。為獲得此類突變體，可使用當前技術方 法’如單交叉重組或雙重同源重組。為此，需要構築整合 選殖載體，其可在宿主細胞染色體中之預定標靶基因座整 合。在本發明之一較佳實施例中，整合選殖載體包含與宿 主細胞基因組中之預定標靶基因座處之DNA序列同源的 DNA片段以將選殖載體之整合靶向此預定基因座。為促進 乾向整合’較佳在宿主細胞轉型之前將選殖载體線性化。 較=執行線性化以使選殖載體之至少—端但較佳每一端由 與標乾基因座同源之序列側接。側接標乾基因座之同源序 列的長度較佳為至少G.lkb，甚至較佳為至纽2kb，更佳 為至少〇.5 kb，甚至更佳為至少⑴，最佳為至少⑽。最 終最適合於實驗中之長度視有機體、標仙Μ之序列及長 較佳藉由加強宿主 體藉由同源重組靶向 細胞之同源重組能力來提 整合至宿主細胞基因組中 高核酸構築 ，亦即整合 146966.doc -20· 201037078 於預定標靶基因座中之效率。該細胞表型較佳包括如w〇 05/95624中所述之缺陷型/或wyg基因。w〇 〇5/95624 揭示一種獲得包含提高之靶向整合效率之絲狀真菌細胞的 較佳方法，其係藉由防止DNA片段非同源隨機整合至基因 組中而進行。載體系統可為單一載體或質體或兩個或兩個 以上載體或質體，其一同含有待引入宿主細胞基因組中之 全部DNA。 可藉由原生質體形成、原生質體轉型及細胞壁再生使 G Μ.細胞轉型。使真g宿主細胞轉型之適合程序描述於Ep
238023 及 Yelton 等人（1984. Proc. Nat. Acad Sci USA 81:14^474)中。使用根癌土壤桿菌（如 使絲狀真菌宿主細胞轉型之適合程序由心 G_t M.J.等人（1998. Nat. Bi〇techn〇1 16:839 842 in: Nat. Biotechnol· 1998· 16:1074)描述。亦可應用針對粗 糙脈孢菌描述之諸如電穿孔之其他方 法。 ◎使用共轉型，亦即亦使可選擇標記基因連同相關基因— 起轉型標記基因，來轉染真菌細胞。可選擇標記基因可以 物理方式連接至相Μ基因（亦即在質體上）或連接在分離片 段上。在轉染之後’針對此選擇性標記基因之存在來_選 轉型物’且隨後分析在較佳預定基因組基因座處之整合。 可選擇標記物為提供殺生物劑或病毒抗性、重金屬抗性、 自營生物原養及其類似物的產物。適用之可選擇標：物包 括（但不限於）_^乙酿胺酶）、吨（烏胺酸胺甲醯基轉移 146966.doc 21 201037078 δ#)、bar (草胺膦乙醢基轉移酶(ph〇Sphinothricinacetyltransferase))、 办沖（潮黴素碟酸轉移酶（hygromycin phosphotransferase))、 ⑴·βΖ>(硝酸還原酶）、厂yrG(乳清苷-5'-磷酸脫羧酶）、sC或 (硫酸腺苷轉移酶）、irpC(鄰胺基苯甲酸合成酶）、 ble(腐草徽素抗性蛋白（phleomycin resistance protein))以及 其等效物。最佳情況為提供一種包含第一 DNA片段之DNA 分子’該片段包含所需置換序列（亦即選擇性標記基因）在 其5'及3'側由實質上與側接標靶序列之染色體dnA序列同 源的DNA序列側接。可由第一 〇ΝΑ片段之可選擇標記物的 存在來選擇染色體DNA序列中之標靶序列經所需置換序列 置換的細胞。為提高選擇正確突變微生物菌株之相對頻 率’可將包含含有編碼選擇性標記物之基因及在真核細胞 中起作用之調節序列的表現卡匣之第二DNA片段可操作地 連接於上述片段（亦即標靶基因座之5，_側+選擇性標記基因 +標靶基因座之3'_側），且可藉由第一 DNA片段之可選擇標 記物的存在及第二選擇性標記基因之缺乏來選擇染色體 DNA序列中之標靶序列經所需置換序列置換的細胞。 在構成供離胺酸生物合成之胺基己二酸途徑之一部分的 酶系統對宿主細胞為異源的情況下，宿主細胞較佳不包括 編碼催化酮己二酸酯轉化為胺基己二酸酯之酶的基因。本 文中術語「酶系統」尤其用以指可催化特定轉化反應之單 個酶或一群酶。該轉化反應可包含一或多個含有已知或未 知中間物之化學反應，例如AKG轉化為AKA或AKA轉化為 AKP。該系統可存在於細胞内或與細胞分離。已知胺基轉 146966.doc •22- 201037078 移酶常具有廣泛的受質範圍。可需要降低存在於宿主細胞 中之該一或多種酶之活性以降低AKA轉化為AAA之活性， 同時維持其他胺基酸或細胞組份之生物合成的相關催化功 能。宿主細胞亦較佳缺乏使AKA轉化為不欲副產物之任何 其他酶活性。 在另一實施例中，利用至少一種催化AKG C!-伸長為 AKA之異源生物催化劑將AKG轉化為AKA。可使用一或多 種生物催化劑。該一或多種生物催化劑可包含一或多種源 Ο 自一或多個源有機體之酶（例如包含一種以上源自不同源 有機體之酶）。適於自AKG製備AKA之生物催化劑可尤其 選自催化α-酮戊一酸Ci-伸長為α -酮己二酸及/或α-酮己二 酸C丨-伸長為(X-酮庚二酸之生物催化劑。 此後’可利用至少一種催化ΑΚΑ伸長為ΑΚΡ之異源生物 催化劑將自AKG製備之AKA轉化為AKP。此等生物催化劑 可與催化AKG藉由C!-伸長轉化為AKA之生物催化劑相同 ©或不同。一種或一種以上生物催化劑可用於將aka轉化為 AKP。該一種或一種以上生物催化劑可包含一或多種源自 一或多個源有機體之酶（例如包含一種以上源自不同源有 機體之酶）。 已知利用Cl-伸長之生物合成途徑存在於產曱烷古生菌 (methanogenic Jrc/mea)中作為辅酶B生物合成之一部分及 生物素生物合成之一部分。認為輔酶B對此等有機體中之 甲烧產生而言為必需的’且α_酮辛二酸酯為辅酶B生物合 成中之重要中間物。在該產甲烷古生菌中，根據複數個反 146966.doc -23- 201037078 應步驟’藉由連續添加亞甲基，將α_酮戊二酸轉化成α-闕 己二酸，接著轉化成α-酮庚二酸且最終轉化成α-酮辛二酸 (亦參見圖1): a·長度為（：„之〇1-酮-酸+乙醯基-CoA今高《^寧檬酸酯+CoA-SH(圖1中之步驟！、5及9) b. 高擰檬酸酯今+高11-烏頭酸酯（由高n-檸檬酸脫水酶催 化）（圖1中之步驟2、6及10) c. 高n烏頭酸酯令今異高n-檸檬酸酯（圖1中之步驟3、7及 11) d·高n-異檸檬酸酯+NADP++長度為Cn+1之α-酮-酸+ NADPH+H++C02(圖 1 中之步驟4、8及 12) 其中η係選自1-4。 已描述產曱烧菌嗜熱甲烧八聯球菌（Mei/zimosarciwa Aermop/n’/a)及詹氏甲烷暖球菌之此重複反應順序。在諸 如以下之其他代謝途徑中之其他α-酮羧酸C!-伸長中包含類 似非迭代反應：在氧化性檸檬酸循環中草乙酸轉化為α_酮 戊二酸酯，作為得到白胺酸之異丙基蘋果酸途徑中之一部 分的α-異戊酸酯轉化為α-異己酸酯，在得到離胺酸之ααα 途徑中α-酮戊二酸醋轉化為α-酮己二酸酯，在得到異白胺 酸之丙酮酸途徑中丙酮酸酯轉化為α-酮丁酸酯，及順丁稀 二酸酯轉化為丙酮酸酯。此等反應共同定義為r Cf伸 長」。 已自詹氏曱烷暖球菌描述及表徵C!-伸長所涉及之若干 146966.doc • 24· 201037078 基因及酶。表明此等酶及編碼基因彼此相似且與其他有機 體中之C!•伸長所涉及之其他酶及其編碼基因相似。用於 使α-酮戊一酸酯迭代伸長’經α_酮己二酸酯及酮庚二酸 酉曰，形成α-酮辛二酸酯之酶的子集已藉由生物化學方式表 徵且稱作「Aks」。已在詹氏甲烷暖球菌之基因組序列中鑑 別出編碼此等酶之一些基因且已提出其他基因。 本發明者已認識到藉由將一或多個編碼^伸長所涉及之 酶系冼之核酸序列併入適合宿主細胞中，可使用匚1_伸長 Ο 在工業規模上製備AKA或AKP，使得AKA或AKP可用作製 備特殊化合物或商業產品（諸如二胺基己烧或己内酿胺）之 中間物。 用於催化C!伸長，藉此形成AK a或AKp之酶系統可尤 ^ 或多種選自以下之群的酶：高n-檸檬酸合成 酶、高η.烏頭酸酶及異_高„_檸檬酸脫氫酶，其巾η係選自卜 4 〇 棒檬酸合成酶可尤其催化Cl•伸長之「反應a」。高η_ 轉樣酸合成酶定義為能夠使鏈長為c4+n之α_酮叛二酸與乙 酿基- CoA縮合，伟溫含 p I** 使传间n-榉檬酸酯形成之酶，其中η係選 自1-4。尚η-棒檬酸合成酿7 士，甘* & 風晦了尤其為歸入或可歸入EC 2.3.3 之酶。更詳古夕 人山 ° ’、&南η_擰檬酸合成酶可選自高檸檬酸 合成酶（EC 2 3 3 14、 .3.3.14)，或可歸人 EC 2 3 3 1、2.3 3 2、 2.3.3.4或2.3.3.9。尤装齡仕达 A1 A ^ 、心4為具有高⑷檸檬酸酯活性之
AksA或其同系物。 高η-烏頭酸酶可尤其催彳 催化C1_伸長之「反應b」及/或「反 146966.doc •25- 201037078 應e」：咼烏頭酸酶定義為能夠經由高η烏頭酸酯中間物 1 個可逆随半反應（亦即高η·烏頭酸醋轉化為高n_檸 才豕酸酉曰或向n-烏頭酸醋轉化為異_高^檸檬酸酿）將高n_檸檬 酸醋轉化為異_高擰檬酸醋的酶，其中η係選自i_4。高〇_ 烏頭酸酶可尤其為歸入或可歸入EC 4.2」之酶。更詳； 之，適合高"-烏頭酸酶可選自高烏頭酸酶（EC 4.2.1.36)， 或可歸入 EC 4.2·1·3、4.2.1.33、4.2.1.79 及 4.2.1.99。尤其 較佳為選自以下具有高η·烏頭酸酶活性之群的酶：AksD、 AksE、AksD同系物及AksE同系物。 高^異檸檬酸脫氫酶可尤其催化Ciy申長之「反應d」。 異-高n-檸檬酸脫氫酶定義為能夠使異_‘_棒樣酸醋轉化」為 鏈長為C5+nia-酮-羧二酸且藉此釋放c〇2的酶，其中n係選 自"。異-高擰檬酸脫氫酶可尤其為歸入或可歸入EC 1·1·1之酶。更詳言之，適合異-高η_擰檬酸脫氫酶可選自 異-高擰檬酸脫氫酶（EC U.U7)，或可歸入Ec丨」136、 1.1.137、1_1·1.38、1丄139、1114〇、丨」丨 41、 H. 1.42、i、·1.1.82、1.1.U3、丨.1丄84、1.1丄85及 I. 1.1.286。尤其較佳為具有檬酸脫氫酶活性之 AksF或其同系物。 產甲烧菌可充當用於產生Ακρ之生物催化劑或可用作該 等生物催化劑之來源。適合生物催化劑可藉由尋找類似於 來自-伸長途徑之已知酶的蛋白質及核苷酸序列來鑑 別。可使用此項技術中熟知之生物資訊技術在序列資料庫 中有效地鑑別類似序列。可使用此項技術中已知之分子生 146966.doc -26- 201037078 物子方法（諸如南方雜交（Southern hybridization)或採用簡 併寡核苷酸之P C R技術）鑑別培養有機體及環境樣品中之類 似基因。在選殖及定序之後’該等生物催化劑可用於在異 源宿主中產生AKP。 詳§之，可使用一或多種來自選自以下之群之產甲烷菌 的用於催化（：！伸長之酶：甲烷球菌屬、甲烷螺菌屬、甲烷 暖球菌屬、甲烷八聯球菌屬、甲烷熱桿菌屬、甲烷球形菌 屬、甲烷火菌屬及甲烷短桿菌屬。更具體言之，可使用— 〇 或多種來自選自以下之群之產甲烷菌的酶：嗜熱自養甲烷 焦稈亀{Methanothermobacter thermoautotropicum)、海沿 甲烧球鹵、思氏甲烧球形菌、坎氏甲烧嗜熱菌 «印少;^ 、嗜熱曱烷八聯球菌、史氏甲烷 知·才干菌、萬氏甲烷球菌、亨氏甲烷螺菌、嗜熱甲烷鬃菌、 醋酸曱烷八聯球菌及雜色甲烷球菌。 此外，適用於催化AKG及/或AKA之〇1伸長的酶可例如 Q 見於包含經由胺基己二酸途徑催化離胺酸生物合成之酶系 統或其部分的有機體中，或含有其同系物作為其他代謝之 一部分，諸如固氮中所涉及之高檸檬酸合成酶。詳言之， 有機體係選自以下之之群：酵母及真菌，諸如青黴菌屬、 頭孢黴屬、麴菌屬、平革菌屬、裸胞殼屬、黑穗病菌屬、 擬青黴菌屬、髮癬菌屬、耶氏酵母菌屬、漢森酵母菌屬、 裂殖酵母菌屬、酵母菌屬、假絲酵母菌屬、克魯維酵母菌 屬，尤其產黃青黴、點青黴、肉色擬青黴、粉擬青黴 如·CMi)、頂頭孢黴、黑麴菌、構巢裸胞 146966.doc -27- 201037078 殼—似）、米麴菌u啊咕,加〜總）、玉 米黑穗病菌、粟酒裂殖酵母（iSc/^〇5acc/邶以 p〇m^)、釀酒酵母、解脂耶氏酵母、多形漢森酵母、白色 念珠菌（Can山而、麥芽糖假絲酵母及乳酸克魯維 酵母；細菌，諸如固氮菌屬、假單胞菌屬、克雷伯氏桿菌 屬、奇球菌屬、棲熱菌屬，尤其棕色固氮菌、施氏假單胞 菌（PwM而m⑽似、肺炎克雷伯氏桿菌、耐輻射奇 球菌、嗜熱奇球菌、嗜熱棲熱菌；及太古菌，諸如火球菌 屬、硫化葉菌屬、熱球菌屬、曱烷球菌屬、甲烷暖球菌 屬、曱烷球形菌屬、甲烷火菌屬、甲烷螺菌屬、甲烷短桿 菌屬、曱烷八聯球菌屬及曱烷熱桿菌屬，尤其超嗜熱火球 菌（P_yr〇c〇CCw /zoraoi/n·/)、硫磺礦硫化葉菌、柯氏熱球菌 (TTiermococcw·? 、海沼甲烷球菌、雜色甲烷 球菌、萬氏甲烷球菌、詹氏甲烷暖球菌、思氏甲烷球形 菌、坎氏曱炫·嗜熱菌、史氏曱烧短桿菌、嗜熱曱烧八聯球 菌、予氏曱烧螺菌、嗜熱曱烧鬃菌、醋酸曱烧八聯球菌及 嗜熱自養甲烷熱桿菌。該等酵母、真菌、細菌、太古菌或 其他有機體尤其可提供能夠催化AKG伸長為AKA及視情況 AKA伸長為APK中之「反應a」的高檸檬酸合成酶。 此外，用於催化AKP製備中之一個反應步驟之適合生物 催化劑可見於鐵角蕨屬或大風子屬中，尤其線葉鐵角蕨 (Asplenium 或泰國大風子 ，其能夠天然產生 AKP 〇 在一個較佳方法中，使用一或多種選自Aks酶及其同系 146966.doc •28- 201037078 物之群，尤其選自AksA、AksD、AksE、AksF及其同系物 之群的酶。此等Aks酶之同系物及編碼此等酶之基因的實 例示於以下頁之表中。
步驟酶名稱 1 AksA
2,3 AksD 有機體 詹氏曱烷暖球菌 嗜熱自養曱垸熱桿菌ΔΗ 海沼甲烷球菌S2 海沼甲烷球菌C5 海沼甲烷球菌C7 思氏曱烷球形菌DSM 3091 坎氏甲烷嗜熱菌AV19 史氏曱烷短桿菌ATCC35061 萬氏甲烷球菌SB 肺炎克雷伯氏桿菌 栋色固氮fe 施氏假單胞菌 雜色甲烧球菌Nankai 3_ 詹氏甲烷暖球菌 嗜熱自養甲烷熱桿菌ΔΗ 海沼甲烷球菌S2 海沼甲烷球菌C5 海沼曱烷球菌C7 思氏甲烷球形菌DSM 3091 坎氏甲烷嗜熱菌AV19 史氏甲烷短桿菌ATCC35061 萬氏甲烷球菌SB 雜色曱烷球菌Nankai3 醋酸曱烷八聯球菌 基因 蛋白質 MJ0503 NP_ _247479 MTH1630 NP_ 276742 MMP0153 NP_ _987273 MmarC5_1522 YP. _001098033 MmarC7_1153 YP. _001330370 Msp_0199 YP_ 447259 MK1209 NP 614492 Msm_0722 YP_ 001273295 Mevan—1158 YP_ _001323668 nifV P05345 nifV P05342 nifV ABP79047 Maeo_0994 YP· 001325184 MJ1003 NP_ 247997 MTH1386 NP_ _276502 Mmpl480 NP _988600 MmarC5_0098 YP_ _001096630 MmarC7_0724 YP_ _001329942 Msp_1486 YP_ _448499 MK1440 NP. _614723 Msm—0723 YP. _001273296 Mevan—0789 YP. _001323307 Maeo_0311 YP. _001324511 MA3085* NP 617978* 146966.doc -29- 201037078 亨氏曱烷螺菌JF-1 Mhun_1800* YP_503240* 嗜熱甲烷鬃菌iT Mthe—0788* ΥΡ_843217* 思氏曱烷球形菌DSM 3091 MspllOO* ΥΡ_448126* 可經由www·ncbi.nlm.nih.gov/找到基因及蛋白質參考（以 *標記之所列基因/蛋白質：在2010年3月2曰 獲得，其他所 列基因/蛋白質：在2008年4月15曰 獲得）。 步驟酶名稱有機體 基因 蛋白質 2,3 AksE 詹氏甲烷暖球菌 MJ1271 NP—248267 嗜熱自養甲烷熱桿菌ΔΗ MTH1387 NP—276503 海沼甲烷球菌S2 MMP0381 NP—987501 海沼曱烷球菌C5 MmarC5_1257 YPOO1097769 海沼甲烷球菌C7 MmarC7_1379 YP_001330593 思氏曱烷球形菌DSM 3091 Msp_1485 YP_448498 坎氏甲烷嗜熱菌AV19 MK0781 NP—614065 史氏甲烷短桿菌ATCC3 5061 Msm_0847 YP_001273420 萬氏甲烷球菌SB Mevan_1368 YP_001323877 雜色曱烷球菌Nankai3 Maeo_0652 YP_001324848 醋酸甲烷八聯球菌 MA3751* NP_618624* 亨氏曱烷螺菌JF-1 Mhun—1799* YP—503239* 思氏甲烷球形菌DSM 3091 Msp_0374* YP_447420* 嗜熱曱烷鬃菌ΡΓ Mthe 0853* YP 843282* 4 AksF 詹氏曱烷暖球菌 MJ1596 NP_248605 嗜熱自養曱烷熱桿菌ΔΗ MTH184 NP_275327 海沼甲统球菌S2 MMP0880 NP988000 海沼甲烷球菌C5 MmarC5_0688 YPOO1097214 海沼甲烷球菌C7 MmarC7_0128 YP_001329349 思氏曱烷球形菌DSM 3091 Msp_0674 YP—447715 坎氏曱烷嗜熱菌AV19 MK0782 NP_614066 史氏甲烷短桿菌ATCC35061 Msm_0373 YP001272946 萬氏曱烷球菌SB Mevan_0040 YP_001322567 雜色曱燒球菌Nankai 3 Maeo_1484 YP_001325672 醋酸曱烷八聯球菌 MA3748* NP 618621* 146966.doc -30- 201037078 亨氏甲烷螺菌 JF-l Mhun_1797* YP_503237* 思氏曱烷球形菌 DSM 3091 Msp_0674* YP_447715* 嗜熱甲烷鬃菌 Mthe_0855* YPJ43284* 史氏曱烷短桿菌 Msm 1298* YP 001273871* 可經由www.ncbi.nlm.nih.gov/找到基因及蛋白質參考 (以*標記之所列基因/蛋白質：在20 10年3月2日獲得，其他 所列基因/蛋白質：在2008年4月15曰獲得）。 詳言之，可使用由以下序列ID中任一者所表示之酶： 4、5、6、7、8、9、10、11、12、13、261、264、267、 0 273、276、279、282(AksA) ; 14、15、16、17、18、19、 20、21、22、23、186、189、192、195、225、228、 231、234(AksD) ; 24、25、26、27、28、29、30、31、 32 、 33 、 198 、 201 、 204 、 207 、 237 、 240 、 243 、 246(AksE) ; 34、35、36、37、38、39、40、41、42、 43 、 210 、 213 、 216 、 219 、 222 、 249 、 252 、 255 、 258(AksF) ; 44、45、46、47、48、49、50、51、52、 53(AksA 同系物）；54、55、56、57、58、59、60、 〇 61(AksD 同系物）；62、63、64、65、66、67(AksF 同系 物）；69、70、71、72、73、74、75、76、77、270(AksA 同系物）。 本發明方法中所製備之AKP可進一步用於製備另一化合 物，或原樣用作例如生物化學研究中之化學品，或用作pH 值缓衝化合物，例如用於諸如液相層析或毛細管電泳之製 備型或分析型分離技術中。詳言之，若需要，則可使用 AKP來製備 AAP、5-FVA、6-ACA或 α-酮辛二酸。 •31 · 146966.doc 201037078 本發明之方法中一種由AKP製備α_酮辛二酸之方法包含 使用如本文所述之生物催化劑使Ακρ經受Cl_伸長。因 此，在已藉由C!-伸長製備AKp之後’可再重複一次Ci_伸 長，藉此由(X-酮庚二酸形成辛二酸。為實現此目的， 可使用用於藉由C】-伸長自ΑΚΑ形成ΑΚΡ的如上所述之同 一組酶或其同系物。可使用與本文中針對AKp轉化為6_ AC A所述相同之原理，亦即藉由使用一或多種選自脫叛 酶、胺基轉移酶及胺基酸脫氫酶之群的能夠在本發明方法 中催化反應步驟之生物催化劑，進一步將所形成之α_酮辛 二酸轉化為7-胺基庚酸。或者，可以化學方式進行一或多 個該等後續反應步驟。以該方式製備之7_胺基庚酸可隨後 環化形成相應C7-内醯胺（亦稱為氮雜環辛_2_酮或ξ_胺基庚 内醯胺）及/或直接聚合或經由該Cf内醯胺聚合以產生耐 論-7或其共聚物。 本發明者已認識到可藉由如下方法將AKP轉化為6_ ACA，其中使第一 AKP脫羧形成5-FVA，之後可使用胺基 轉移反應由5-FVA製備6-ACA ’或使第一 AKP經受胺基轉 移反應，形成AAP，之後可藉由脫羧反應由aAP製備6_ ACA。 在用於製備6-ACA之一較佳方法中，該製備方法包含在 能夠催化α-酮酸或胺基酸（亦即包含至少一個叛酸基團及 至少一個胺基之化合物）脫去羧基之生物催化劑存在丁進 行生物催化反應。具有該催化活性之酶因此可分別稱為α 酮酸脫羧酶及胺基酸脫羧酶。 146966.doc •32· 201037078 該酸較佳為二元酸，其中該生物催化劑對緊挨著酮基或 胺基之酸基有選擇性。 一般而言’適合脫羧酶具有能夠催化AKP轉化為5-FVA 之α-酮庚二酸脫羧酶活性或能夠催化AAP轉化為6-ACA之 α-胺基庚二酸脫羧酶活性。 能夠使α-酮酸或胺基酸脫去羧基之酶可尤其選自脫羧酶 (E.C· 4.1_1)之群，較佳選自以下之群：麩胺酸脫羧酶（Ec 4.1.1.15)、二胺基庚二酸脫羧酶（EC ^丄別）、天冬胺酸 Ο 卜脫羧酶（EC 4.1丄11)、支鏈α-酮酸脫羧酶、α_酮異戊酸 脫羧酶（EC 1.2.4.4)、α·酮戊二酸脫羧酶（EC 4.1.1.71)及丙 酮酸脫羧酶（EC 4.1.1.1)。 一或多種其他適合脫叛酶可尤其選自以下之群：草酸脫 羧酶（EC 4.1.1.2)、草醯乙酸脫羧酶（EC 4.1.1>3)、乙醯乙 酸脫叛酶（EC 4.1.1.4)、纈胺酸脫緩酶/白胺酸脫羧酶（Ec 4.1 _1_ 14) ' 3-羥基麩胺酸脫羧酶（EC 4.1.1.16)、鳥胺酸脫 羧酶（EC 4.1.1.17)、離胺酸脫羧酶（EC 4.1.1.18)、精胺酸 Q 脫羧酶（EC 4.1.1.19)、2-側氧基戊二酸脫羧酶（Ec 4.1.1.71) 及二胺基丁酸脫羧酶（EC 4.1.1.86)。 脫叛酶可尤其為選自以下之群之有機體的脫羧酶：倭 瓜，育瓜，酵母；真菌，例如釀酒酵母、佛氏假絲酵母 油//⑽岣、漢森酵母菌屬（如峨ww/a )、馬克斯 克魯維酵母（尺/MJV㈣讲.⑽似）、爪哇根黴 (她力尸似㈣·㈣）、運動醱酵單胞菌（办_廳_ woh/u) ’更尤其為來自運動醱酵單胞菌及粗糙脈孢菌之 146966.doc •33· 201037078 突變體I472A ;哺乳動物，尤其來自哺乳動物大腦；及細 菌。舉例而言，可使用來自大腸桿菌之麩胺酸脫羧酶或天 冬胺酸脫羧酶，或可使用來自粗糙脈孢菌、麻瘋分枝桿菌 {Mycobacterium /e/?rae)、產氣莢膜梭菌（C/oWrM/wm 似）' 短乳桿菌、結核分枝 桿菌（A(ycc^acieHwm iwfeercw/oWs)、鏈球菌屬或乳球菌屬 之麩胺酸脫羧酶。麩胺酸脫羧酶可源自之乳球菌屬之實例 尤其包括乳酸乳球菌（jLaciococcws /acib)，諸如乳酸乳球 菌菌株B1157、乳酸乳球菌IFPL730，更尤其為乳酸乳球菌 多基因變種（lacioeoccws /ocib var. mo/i/gene·?)(以前命名 為乳酸鏈球菌多基因變種（Sire/jiococcw·? /acih var. ma/Hg叫e〇)。可尤其使用來自假單胞菌屬之草醯乙酸脫羧 酶。 在本發明之一較佳方法中，6-AC A之製備包含在能夠在 胺基供體存在下催化轉胺反應之選自胺基轉移酶（E.C. 2.6.1)之群的酶存在下進行酶促反應。 一般而言’適合胺基轉移酶具有能夠催化5-FVA轉化為 6-ACA之6-胺基己酸6-胺基轉移酶活性或能夠催化AKP轉 化為AAP之α-胺基庚二酸2-胺基轉移酶活性。 胺基轉移細可尤其選自以下之群.β -胺基異丁酸：α -類I 戊二酸胺基轉移酶、β-丙胺酸胺基轉移酶、天冬胺酸胺基 轉移酶、4-胺基-丁酸胺基轉移酶（EC 2.6.1.19)、L-離胺酸 6-胺基轉移酶（EC 2·6.1.36)、2-胺基己二酸胺基轉移酶（ec 2.6.1.39)、5-胺基戊酸胺基轉移酶（EC 2.6.1.48)、2-胺基己 146966.doc -34· 201037078 酸胺基轉移酶（EC 2.6.1.67)、離胺酸：丙酮酸6-胺基轉移 酶（EC 2.6.1.71)及芳族胺基酸胺基轉移酶（EC 2.6.1.57)。 在一實施例中，胺基轉移酶可選自以下之群：丙胺酸胺 基轉移酶（EC 2.6.1.2)、白胺酸胺基轉移酶（EC 2.6.1,6>、 丙胺酸-含氧酸胺基轉移酶（EC 2.6.1.12)、β-丙胺酸_丙酮 酸胺基轉移酶（EC 2.6.1.18)、（S)-3-胺基-2-甲基丙酸胺基 轉移i#(EC 2.6.1.22)、L，L - 一·胺基庚二酸胺基轉移酶（gc 2.6.1.83)。 © 胺基轉移酶可尤其選自來自以下之胺基轉移酶：孤菌 屬’尤其為河流孤菌（Fz’6rz_o //Μνίβ/&);假單胞菌屬，尤其 為綠膿桿菌;芽胞桿菌屬，尤其 為早氏芽胞桿菌;山散屬， 尤其為多年生山靛’更尤其為多年生山靛之芽；鐵角蕨 屬’更尤其為單邊鐵角蕨M⑴或線葉鐵 角蕨；長角豆屬，更尤其為角豆（Cerai〇m-a ⑽）；喷乳 動物；或酵母’尤其為釀酒酵母。在酶來自哺乳動物之情 況下，其可尤其源自哺乳動物腎臟、哺乳動物肝臟、哺乳 動物心臟或哺乳動物大腦。舉例而言，適合酶可選自以下 之群：來自哺乳動物腎臟之β-胺基異丁酸：α-酮戊二酸胺 基轉移酶’尤其為來自豬腎臟之ρ_胺基異丁酸：α_酮戊二 I胺基轉移酶’來自哺乳動物肝臟之丙胺酸胺基轉移 酶，尤其為來自兔肝臟之0_丙胺酸胺基轉移酶；來自哺乳 動物心臟之天冬胺酸胺基轉移酶，尤其為來自豬心臟之天 冬胺酸胺基轉移酶；來自哺乳動物肝臟之4-胺基-丁酸胺基 146966.doc -35- 201037078 轉移酶，尤其為來自豬肝臟之4-胺基-丁酸胺基轉移酶；來 自哺乳動物大腦之4-胺基-丁酸胺基轉移酶，尤其為來自人 類、豬或大鼠大腦之4-胺基丁酸胺基轉移酶；來自脈孢黴 屬之α-酮己二酸-麩胺酸胺基轉移酶，尤其為來自粗糙脈 孢菌之a-酮己二酸：麩胺酸胺基轉移酶；來自大腸桿菌之 4-胺基-丁酸胺基轉移酶；或來自棲熱菌屬之α_胺基己二酸 胺基轉移酶，尤其為來自嗜熱棲熱菌之α—胺基己二酸胺基 轉移酶；及來自梭菌屬，尤其來自胺基戊酸梭菌 麵z’„ova/en_c謂）之5_胺基戊酸胺基轉移酶。 適合2-胺基己二酸胺基轉移酶可例如由冰島火棒菌 (Pyrobaculum islandicum)氣供。 在一特定實施例中，使用包含根據序列ID 2、83、86或 任何此等序列之同系物之胺基酸序列的胺基轉移酶。 詳言之，胺基供體可為氨、銨離子、胺或胺基酸。適合 胺為一級胺及二級胺。胺基酸可具有小或^構型。胺基供 體之實例為丙胺酸、麵胺酸、異丙胺、2_胺基丁烧、2_胺 基庚烧、本基曱胺、1-苯基_1_胺基乙烧 '麵醯胺酸 '酿胺 酸、***酸、天冬胺酸、β_胺基異丁酸、卜丙胺酸、4_ 胺基丁酸及α-胺基己二酸。 在另一較佳實施例中，用於製備6-ACA之方法包含在能 夠在選自作用於供體之CH-NH2基團的氧化還原酶之群（EC 1.4)、尤其胺基酸脫氫酶（E.C.丨.^)之群的氨來源存在下 催化還原胺化反應之酶存在下進行生物催化反應。一般而 言，適合胺基酸脫氫酶具有催化5-FVA轉化為6-ACA26_ 146966.doc •36- 201037078 胺基己酸6-脫氫酶活性或具有催化AKP轉化為AAP之α-胺 基庚二酸2-脫氫酶活性。詳言之’適合胺基酸脫氫酶係選 自以下之群：二胺基庚二酸脫氫酶（EC 1.4.1.16)、離胺酸 6-脫氫酶（EC 1.4.1.18)、麵胺酸脫氫酶（ec 1.4.1.3 ; EC 1.4.1.4)及白胺酸脫氫酶（EC 1.4.1.9)。 在一實施例中’胺基酸脫氫酶可選自歸類為以下之胺基 酸脫氫酶：以NAD或NADP作為受體發揮作用之麩胺酸脫 氫酶（EC 1.4.1.3)、以NADP作為受體發揮作用之麩胺酸脫 ❹ 氫酶（EC 1.4.1.4)、白胺酸脫氫酶（Ec 1.4.1.9)、二胺基庚 二酸脫氫酶（EC 1.4.1.16)及離胺酸6-脫氫酶（EC 1.4.1.18)。 胺基酸脫氫酶可尤其源自選自以下之群之有機體：棒狀 桿菌屬，尤其為麵胺酸棒狀桿菌；變形桿菌屬，尤其為普 通變形桿菌vw/別仏）；土壤桿菌屬，尤其為根癌 土壤桿菌；土芽孢桿菌屬，尤其為嗜熱脂肪土芽孢桿菌 (Geobacillus stearothermophiluQ·，不動桿蛰餍，尤矣為不 〇 =桿菌屬（f—价L)ADP1 ;雷爾氏菌屬，尤其為 知科雷爾氏菌屬so/a«acearww);沙門氏菌屬， 尤其為鼠傷寒沙門氏菌（Sa/mcme/Za ίχρ/n.mMriMw);酵母菌 屬’尤其為釀酒酵母；短桿菌屬’尤其為黃色短桿菌 (办eWhci⑺//αν_);及芽胞桿菌屬，尤其為球形芽孢 桿菌（5⑽7/Mup/mdc似）、蠟狀芽胞桿菌（心“〜抑） 或枯草桿菌。舉例而言，適合胺基酸脫氫酶可選自來自芽 胞桿菌屬、尤其球形芽孢桿菌之二胺基庚二酸脫氫酶；來 自知·桿菌屬sp.)之二胺基庚二酸脫氫酶； 146966.doc •37· 201037078 來自棒狀桿菌屬之二胺基庚二酸脫氫酶，尤其為來自麩胺 酸棒狀桿菌之二胺基庚二酸脫氫酶；來自變形桿菌屬之二 胺基庚二酸脫氫酶，尤其為來自普通變形桿菌之二胺基庚 二酸脫氫酶；來自土壤桿菌屬、尤其根癌土壤桿菌之離胺 酸6-脫氫酶，來自土芽孢桿菌屬、尤其嗜熱脂肪土芽孢桿 菌之離胺酸6-脫氫酶；來自不動桿菌屬之以Nadh或 NADPH作為輔因子發揮作用之麩胺酸脫氫酶（ec 1.4.1.3)，尤其為來自不動桿菌ADpi之麵胺酸脫氫酶；來 自雷爾氏菌屬之麩胺酸脫氫酶（EC 1^.3)，尤其為來自茄 科雷爾氏菌屬之麩胺酸脫氫酶；來自沙門氏菌屬之以 NADPH作為輔因子發揮作用之麩胺酸脫氫酶（EC 1 _4_ 1.4) ’尤其為來自鼠傷寒沙門氏菌之麩胺酸脫氫酶； 來自酵母菌屬之麩胺酸脫氫酶（EC 143 4)，尤其為來自釀 酒酵母之楚胺酸脫氫酶；來自短桿菌屬之麩胺酸脫氫酶 (EC 1.4.1 _4) ’尤其為來自黃色短桿菌之麩胺酸脫氫酶；及 來自芽胞桿菌屬之白胺酸脫氫酶，尤其為來自躐狀芽胞桿 菌或枯草桿菌之白胺酸脫氫酶。 在一特定實施例中’在脫羧酶或其他催化AKP轉化為5-甲贐基戊酸酯（5-FVA)之生物催化劑存在下，AKP經生物 催化而轉化為5-曱醯基戊酸酯（5-FVA)。根據本發明使用 之脫羧酶可尤其選自以下之群：來自乳酸乳球菌、乳酸乳 球菌多基因變種或乳酸乳球菌乳脂亞種之α_酮酸脫羧酶； 來自乳酸乳球菌菌株Β1157或乳酸乳球菌IFPL730之支鏈α-酮酸脫叛酶；來自釀酒酵母、佛氏假絲酵母、運動醱酵單 146966.doc -38- 201037078 胞菌、漢森酵母屬、爪哇根黴、粗糙脈孢菌或馬克斯克魯 維酵母之丙酮酸脫羧酶；來自結核分枝桿菌之…酮戊二酸 脫羧酶；來自大腸桿菌、短乳桿菌、麻瘋分枝桿菌、粗糙 脈孢菌或產氣莢膜梭菌之麩胺酸脫羧酶；及來自大腸桿菌 之天冬胺酸脫羧酶。 此後，5-FVA可轉化為6-ACA。此反應可以化學方式進 行：如EP-A 628 535或DE 4 322 065中針對9-胺基壬酸（9- 胺基天竺葵酸）及12-胺基十二酸（12_胺基月桂酸）所描述， 〇 可藉由在氫化催化劑（例如SiOVAhO3支撐物上Ni)上以氨 將5-FVA還原胺化來製備6_aca，產率高。 或者，可藉由在Pt〇2上將6-肟己酸（由5_FVA與羥胺之反 應製得）氫化來獲得6-ACA。（關於同源12-胺基十二酸之合
Woods, E.O. Price, C.P. Nwaonicha J. Am. OH Chem. Soc. 1997, 74, 531-538)。 在一實施例中，可以生物催化方式在（i)胺基供體及（ii) 胺基轉移酶、胺基酸脫氫酶或能夠催化5_fVA轉化為6_ ACA之另一生物催化劑存在下將5_FVA轉化為6_acA ^詳 s之，在此類實施例中，胺基轉移酶可選自以下之群：來 自河流孤菌' 綠膿桿菌或韋氏芽胞桿菌之胺基轉移酶；來 自豬腎臟之β-胺基異丁酸：α_酮戊二酸胺基轉移酶；來自 兔肝臟之β-丙胺酸胺基轉移酶；來自多年生山靛之芽的胺 基轉移酶；來自豬肝臟或來自人類、大鼠或豬大腦之4_胺 基丁酸胺基轉移酶；來自兔肝臟之β_丙胺酸胺基轉移酶； 146966.doc -39- 201037078 及L-離胺酸： 氫酶之情況下 酮戊二酸-ε-胺基轉移酶。在使用胺基酸脫 該胺基酸脫氫酶可尤其選自來自根癌土壤 桿菌或嗜熱脂肪土芽孢桿菌 適合胺基酸脫氫酶可選自以 短桿菌屬、麩胺酸棒狀桿菌 之離胺酸6-脫氫酶之群。另一 下之群：來自球形芽孢桿菌、 或普通變形桿菌之二胺基庚二 酸脫氫酶 來自不動桿菌ADP1或茄科雷爾氏菌屬之以 NADH或NADPH作為輔因子發揮作用的麩胺酸脫氫酶⑽ 1^.3);來自鼠傷寒沙門氏菌之以NADpH作為輔因子發 揮作㈣㈣酸脫氫酶⑽來自釀酒酵母或黃色◎ 短桿菌之楚胺酸脫氫酶(EC U.M);或來自蠟狀芽胞桿菌 或枯草桿菌之白胺酸脫氫酶。 在一個特定實施例中，AKP以化學方式轉化為5_fva。 2-酮羧酸之有效化學脫羧基成相應醛可以例如基於T^ra- hedron Lett_ 1982, 幻 （4)，459_462中所述之方法使用二級 胺（例如嗎啉）’在共沸除水及同時失去c〇2下，形成中間 物烯胺而進行。中間物末端烯醯胺隨後水解為相應醛。然 後，可藉由在胺基轉移酶存在下轉胺，或藉由胺基酸脫氫◎ 酶或能夠催化轉化之另一生物催化劑之酶還原胺化，將5_
FVA以生物催化轉化為6_ACA。當描述5_FVA轉化為6_AcA 時’遠胺基轉移酶或胺基酸脫氫酶尤其可選自上述生物催 化劑。 或者’可藉由例如上述化學方法將5 — FVA轉化為6-ACA。 在一個特定實施例中，AKP以生物催化轉化為AAP係在 146966.doc 40- 201037078 ⑴胺基轉移酶，胺基酸脫氫酶，或能夠催化該轉化之另一 生物催化劑，及（ii)胺基供體存在下。根據本發明用於將 AKP轉化為AAP之該胺基轉移酶尤其可選自以下之群豬 心臟之天冬胺酸胺基轉移酶；粗糙脈孢菌或酵母之酮己 二酸：㈣酸胺基轉移酶；多年生山靛芽的胺基轉移酶； 大腸桿菌之4-胺基丁酸胺基轉移酶；嗜熱棲熱菌之&胺基 己二酸胺基轉移酶；線葉㈣蕨或單邊鐵角嚴之胺基轉移 酶；及角豆之胺基轉移酶。 適合胺基酸脫氫酶尤其可選自以下之群：不動桿菌 ADP1或祐科雷爾&菌以NADH或NADpH作為輔因子作用 之麩胺酸脫氫酶（EC山.3);鼠傷寒沙門氏菌、釀酒酵母 或黃色短桿®«NADPH作為辅因子作用之㈣酸脫氯酶 (EC 1.4.U);球形芽孢桿g、短桿菌屬、糙胺酸棒狀桿菌 或普通變形桿菌之胺基庚二酸脫氫酶。另—適合胺基酸脫 氫酶可選自以下之群：根癌土壤桿菌或嗜熱脂肪土芽抱桿 菌之離胺酸6-脫氫酶；日& 1 啤及蠟狀牙胞桿菌或枯草桿菌之白胺 酸脫氫酶。 然後，AAP可以化學方式藉由脫竣基轉化為6_aca。此 可藉由在酮《催化劑存在τ在高鴻點溶劑中加熱進行。 舉例而言，如Μ. Hashimoto, γ·邮，γ⑴喊τ iwai^ A〇ki在c/^敗ζ扣· 1986, 893_896中所描述，胺基酸在ΐ5〇_ 贿在具有ν/ν%環己_之環己醇中脫缓基產率高。 類似方法描述於Dai so之歐训直丨xh ^ ，州專利申請案1586553, 2005及 S.D. Brandt, D. Mansell ς ansell, s. Freeman, l.A. Fleet, J.F. Alder 146966.doc -41 · 201037078 J\ Pharm. Biomed. Anal. 2QQ6, 41, ST2-SS2 中。 或者，可在脫羧酶或催化AAP脫羧形成6-ACA之其他生 物催化劑存在下以生物催化方式將AAP脫羧，形成6-ACA。脫羧酶可選自能夠催化α-胺基酸脫羧之脫羧酶。詳 言之，脫羧酶可選自以下之群：來自番南瓜、黃瓜、酵母 或小牛大腦之麩胺酸脫羧酶；及二胺基庚二酸脫羧酶（EC 4.1.1.20)。二胺基庚二酸脫羧酶可例如來自能夠由二胺基 庚二酸合成離胺酸之有機體。該有機體可尤其見於細菌、 太古菌及植物中。詳言之，二胺基庚二酸脫羧酶可來自革 蘭氏陰性細菌（gram negative bacterium)，例如大腸桿菌。 在一特定實施例中，以化學方式將AKP轉化為AAP。可 藉由如針對類似化合物所述之Leuckart-Wallach催化反 應，由2-側氧基庚二酸製備AAP。使用曱酸銨之甲醇溶液 以及使用[RhCp*Cl2]2作為均相催化劑來進行此反應（M. Kitamura, D. Lee, S. Hayashi, S. Tanaka, M. Yoshimura J. Org. Chem. 2002, 67, 8685-8687)。或者，如由S. Ogo, K. Uehara及 S. Fukuzumi在 J. dm. C/zew. /Soc. 2004, 3020- 3021中所描述，可使用[IrniCp*(bpy)H20]S04作為催化劑 用曱酸銨水溶液進行Leuckart-Wallach反應。亦可藉由與 (對掌性）苄胺反應且隨後在Pd/C或Pd(OH)2/C上使中間物亞 胺氫化，將α-酮酸轉化為（對映異構性增濃）胺基酸。參看 例如 R.G. Hiskey, R.C. Northrop J· Jm. C/zew. /Soc. 1961, S3, 4798 ° 此後，可在脫羧酶或能夠將AAP脫羧形成6-ACA之另一 146966.doc -42- 201037078 生物催化劑存在下，以生物催化方式將AAP轉化為6-ACA。該脫羧酶可尤其選自上文在描述用於將AAP轉化為 6-ACA之生物催化劑時戶斤提及之生物催化劑。 或者，可藉由例如如上所述之化學方法將AAP轉化為6-ACA。 在一特定實施例中，在脫羧酶或能夠催化AKP轉化為5-FVA之其他生物催化劑存在下，以生物催化方式將AKP轉 化為5-FVA，且此後在胺基轉移酶、胺基酸脫氳酶或能夠 Ο 催化5-FVA轉化為6-ACA之其他生物催化劑存在下，將5- FVA轉化為6-ACA。適於此等反應之脫羧酶可尤其選自上 文在描述AKP經生物催化轉化為5-FVA時所提及之脫羧酶 之群。適於5 -FVA轉化之胺基轉移酶或胺基酸脫氫酶可尤 其選自上文在描述5-FVA經生物催化轉化為6-ACA時所提 及之酶。 在一特定實施例中，在胺基轉移酶、胺基酸脫氫酶或能 夠催化AKP轉化為AAP之其他生物催化劑存在下，以生物 ^ 催化方式將AKP轉化為AAP，且此後在脫羧酶存在下將 AAP轉化為6-ACA。適於此等反應之酶可尤其選自在分別 描述AKP經生物催化轉化為AAP且AAP經生物催化轉化為 6-ACA時所描述之胺基轉移酶、胺基酸脫氫酶及脫羧酶之 群。 在本發明之另一實施例中，將自根據本發明之方法製得 之AKP製備的6-AC A轉化為二胺基己烷。此反應可藉由還 原酸基，形成醛基，且使由此形成之醛基轉胺，藉此提供 146966.doc -43 - 201037078 胺基，得到二胺基己烷來實現。此反應可以化學方式或生 物催化方式實現。在本發明之一較佳方法中，該製備包含 在能夠催化酸還原形成醛基之生物催化劑存在下的生物催 化反應及/或在能夠催化該轉胺之生物催化劑存在下在胺 基供體（例如如本文中別處所述之胺基供體）存在下的生物 催化反應。 能夠催化酸基還原形成醛基之生物催化劑可尤其包含選 自以下之群的酶：氧化還原酶（EC丨.2.1)，較佳為醛脫氫 酶（EC 1.2.1.3、EC 1.2.1.4 及 EC 1.2.1.5)、乙醛脫氫酶（乙 醯化）（EC 1，2，1，1〇);天冬胺酸-半醛脫氫酶（Ec 1.2.1.11); 丙二酸-半醛脫氫酶（EC 1.2.1_15);及丁二酸-半醛脫氫酶 (EC 1.2.1.16及 EC 1.2.1.24)。 氧化還原酶原則上可獲自或源自任何有機體。該有機體 可為原核或真核有機體。詳言之，有機體可選自細菌、太 古菌、酵母、真菌、原生生物、植物及動物（包括人類）。 在一實施例中，氧化還原酶、尤其醛脫氫酶獲自或源自 選自以下之群的細菌：不動桿菌屬（尤其為鮑曼不動桿菌 及不動桿菌 NCIMB9871)、固氮螺 旋菌屬切W//_)(尤其為巴西固氮螺菌咖w//ww 、雷爾氏菌屬、包特氏菌屬、伯克 霍爾德氏菌屬（5wrA:/2〇/〇^a)、曱基桿菌屬（施咖/〇办_⑺·MW)、 黃色桿菌屬、中華根瘤菌屬WMoWn.zoMwm)、 根瘤菌屬、硝化菌屬（iWiroftacier)、布氏桿菌屬 (5rWce//a)(尤其為地中海熱布氏桿菌（反…”仏））、假單 146966.doc -44- 201037078 胞菌屬土壤桿菌屬（尤其為根癌土壤桿菌）、芽胞桿菌 屬、李氏菌屬（仏⑽⑷、產驗桿菌屬⑷⑽如叫、棒狀 桿菌屬、埃希氏菌屬及黃質細菌屬（F—cierf細p 在Λ施例中，氧化還原酶、尤其醛脫氫酶獲自或源自 &自以下之群的有機體：酵母及真菌，尤其為麴菌屬（尤 其為黑麴菌及小巢狀麴菌及青徽菌屬（尤其為 產黃青黴）。 在一貫施例中，氧化還原酶、尤其醛脫氫酶獲自或源自 〇 植物，尤其為芥菜屬，更尤其為擬南芥菜。 忐夠在轉化為二胺基己烷中催化轉胺反應之生物催化劑 可尤其包含選自胺基轉移酶（E C. 2 61)之群的酶，例如見 於如本文中別處所述之有機體中。 本發明方法中之反應條件可視用於生物催化劑（尤其為 酶）之已知條件、本文中所揭示之資訊及視情況選用之一 些常規實驗而加以選擇。 原則上’所用反應介質之pH值可在寬範圍内選擇，只要 〇 生物催化劑在該等pH條件下有活性即可。可視生物催化劑 及其他因素而定，使用鹼性、中性或酸性條件。在該方法 包括使用微生物例如以表現催化本發明方法之酶的情況 下’選擇pH值以使微生物能夠執行其預期功能。在25〇c下 之基本上水系統的情況下’ pH值可尤其在中性pH值以下 之四個pH值單位與中性pH值以上之兩個pH值單位（亦即pH 3與pH 9之間）的範圍内選擇。若水為唯一溶劑或主要溶劑 (以全部液體計’超過50重量％ ’尤其超過9〇重量。/。），則 146966.doc -45- 201037078 。為系統為水性的，其中例如少量（以全部液體計，低於 =量％’尤其低於1〇重量％)乙醇或另一溶劑可溶解，達 =成存在之微生物保持活性之濃度^其在使用酵母及/ 〆:、菌之If况下’基於25 c下之基本上水系統，酸性條件 w為車又佺詳&之，pH值可在pH 3至pH 8之範圍内。若需 =，則可使用酸及/或驗或以酸與驗之適合組合緩衝來 節pH值。 原則上’培育條件可在寬範圍内選擇，只要生物催化劑 展不足夠活性及/或生長即可。&包括好氧、 及厭氧條件。 礼 本文：厭氧條件定義為無任何氧或生物催化劑(尤其微 生物）實質上又；、、* # 、 4耗氧之條件，且通常對應於小於5 mmol/l.h之耗氡署， 重 尤其小於2.5 mm〇i/i.h或小於1 mmol/l.h之耗氧量。 好氧條件為；^供不受限生長之量的氧氣溶解於介質中 之條〗件’能夠支持至少10 一 _h，更佳大於20 〇 mmol/l.h，甚至更伟 炅佳大於50 mmol/l.h，且最佳大於1〇〇 mmol/l.h之耗氧速率。 ^乳條件定義為耗氧量受氧自氣體至液體之轉移限制之 ^厭氧條件之上限決定限氧條件之下限，亦即通常為 至少 1 min〇l/l.h ’ , 兀具為至少2.5 mmoUl.h或至少5 mmol/l.h。好氧鉻姓^ _ ’、 下限決定限氧條件之上限，亦即小 於 100 mmol/l.h、丨狄 α 5〇 mmol/l.h、小於2〇 mm〇1/l h或小 於 10 mmol/l.h。 146966.doc -46· 201037078 條件為好氧'厭氧還是限氧將視進行該方法之條件而 定，尤其為進入氣流之量及組成、所用設備之實際混合/ 質量轉移特性、所用微生物類型及微生物密度。 在本發明之一較佳方法中，至少在醱酵條件下進行Ακρ 之製備。 原則上，所用溫度並非關鍵，只要生物催化劑（尤其為 酶）展示實質上活性即可。一般而言，溫度可為至少， 尤其至少15〇c，更尤其至少2(rc ^所欲最高溫度視生物催 G 化劑而定。—般而言’該最高溫度在此項技術中已知，例 如在市售生物催化劑之情況下在產品資料表中指示，或可 基於一般常識及本文中揭示之資訊按常規確定。溫度通常 為90°C或9(TC以下，較佳為7〇。(：或7〇。(：以下，尤其為5〇^： 或5〇C以下’或更尤其為40°C或40°C以下。 詳s之，若在宿主有機體外部進行生物催化反應，則在 使用在反應介質中保留足夠活性之酶的情況下，可使用包 3有機/谷劑之尚濃度（例如大於50重量％或大於90重量〇/0) ^ 之反應介質。 可使用此項技術中本身已知之分子生物學技術構築包含 或夕種用於催化本發明方法中之反應步驟之酶的異源細 胞。舉例而言，該等技術可用於提供一種包含一或多個編 碼該一或多種生物催化劑之基因的載體。包含該一或多個 基因之載體可包含一或多個調節元件，例如一或多個啟動 子，其可操作地連接於編碼生物催化劑之基因。 如本文中所用之術語「可操作地連接」係指聚核苷酸元 146966.doc •47- 201037078 件（或編碼序列或核酸序列）呈功能關係之連接。當核酸序 列置於與另-核酸序狀功能關係切，其為「可操作地 連接」。舉例而言，若啟動子或強化子影響編碼序列之轉 錄，則其可操作地連接於編碼序列。 本文中所用之術5吾「啟動子」係指用於控制一或多個 轉錄的核^片段’其位於基因轉錄起始位點之轉錄 方向的上游’且在結構上可由以下之存在來鑑別：DNA依 賴性RNA聚合酶之結合位點、轉錄起始位點及任何其他 DNA序列’包括(但$限於)轉錄因子結合位點、抑制子及 化子蛋白結合位點及熟習此項技術者已知之直接或間接 用於調即自啟動子轉錄之量的任何其他核_酸序列。「組 成至「」啟動子為在大乡數環境及發育條件下有活性的啟動 誘V 5L」啟動子為在環境或發育調節下有活性之啟 動子。、應瞭解術語「同源」在用以指示既定（重組）核酸或 多肽刀子與既定宿主有機體或宿纟細胞之間的關係時意謂 實際上核1夂或多肽分子由相同物種(較佳相同品種或菌株) 之宿主細胞或有機體產生。 可用於實現編碼用於本發明方法之酶（尤其為諸如上文 所述之胺基轉移酶、胺基酸脫氯酶或脫叛酶）之核普酸序 列表現的啟動子可原生於編碼待表現酶之核苷酸序列，或 可異源於其可操作地連接之核苷酸序列（編碼序列）。啟動 子較佳與宿主細胞同源。 右使用異源啟動子（相對於編碼相關酶之核苷酸序列）， 則與原生於該編碼序列之啟動子相比異源啟動子較佳能 146966.doc 201037078 夠產生更高穩態水準之包含編碼序列之轉錄物（或每單位 時間能夠產生更多轉錄分子，亦即mRNA分子）。在此情況 下，適合啟動子包括熟習此項技術者所熟知之組成型與誘 導型天然啟動子以及經工程改造之啟動子。 「組成型強啟動子」為與原生宿主細胞相比使mRNA以 高頻率起始之啟動子。革蘭氏陽性微生物（Gram-positive micro-organism)中該等組成型強啟動子之實例包括8?01-26、SP01 -1 5、veg、(丙酉同酸敌化酶啟動子）及am少E。 〇 革蘭氏陽性微生物中誘導型啟動子之實例包括IPTG誘導 型Pspac啟動子、木糖誘導型PxylA啟動子。 革蘭氏陰性微生物中組成型及誘導型啟動子之實例包括 [但不 於）tac、tet、trp-tet、Ipp、lac、Ipp-lac、laclq、 Γ7、7\5、、ga/、ire、ara(尸w£>)、SP6、X-PRA X-PL。 (絲狀）真菌細胞之啟動子在此項技術中已知且可例如為 葡萄糖-6-磷酸脫氫酶即ί/Α啟動子、蛋白酶啟動子（諸如 pepA、pepB、pepC)、葡糖澱粉酶g/αΑ啟動子、澱粉酶 〇 、awyB啟動子、過氧化氫酶caiR或caiA啟動子、葡 萄糖氧化酶goxC啟動子、β -半乳糖苦酶/acA啟動子、α-葡 糖苷酶ag/A啟動子、轉譯延長因子化/Α啟動子、木糖聚糖 酶啟動子（諸如x/πΑ、、x/«C、x/«D)、纖維素酶啟動 子（諸如eg/A、eg/B、cZ>/zA)、轉錄調節因子之啟動子（諸如 areA、creA、x/nR、、priT等）或任何其他啟動子，且 可尤其見於 NCBI 網站 http://www.ncbi.nlm.nih.gov/entrez/ 中〇 146966.doc -49- 201037078 本發明亦係關於一種新穎異源細胞’其可提供一或多種 能夠在AKP之製備中及視情況在由AKP製備其他化合物（諸 如5-FVA、AAP、6-ACA、二胺基己烧或己内醯胺）中催化 至少一個反應步驟之生物催化劑。本發明亦係關於一種新 賴載體’其包含一或多個編碼一或多種能夠在AKP之製備 中及視情況在由AKP製備其他化合物（5-FVA、AAP、 ACA、二胺基己烧或己内醯胺）中催化至少一個反應步驟 之酶的基因。一或多個適合基因可尤其選自編碼如上文所 提及之酶的基因。詳言之，該等基因中之至少一者異源於 宿主有機體。 在一尤其有利之實施例中’異源細胞或載體包含 AksD、AksE、AksF及NifV基因。在另一尤其有利之實施 例中，異源細胞另外包含AksA基因。較佳AksA、AksD、 AksE及AksF基因來自詹氏甲烷暖球菌、釀酒酵母、海沼 甲烷球菌、醋酸甲烷八聯球菌、亨氏甲烷螺菌或大腸桿 菌。
NifV基因較佳來自棕色固氮菌。 在一尤其較佳實施例中，Nifv基因包含由SEQ m N〇: 149表示之序列或其功能類似物。 關於選自AksA、AksD、AksE&AksF基因之群的基因， 本發明之（所用）細胞之基因組較佳包含至少一個根據選自 以下之群的任一序列之核酸序列：SEQ 1〇 N〇丨45、丨46、 147、148，SEQ ID NO 167、168、169、170、171 ' 172、 173、174，SEQ ID NO 177、178、179、180、181、182、 146966.doc -50- 201037078 183、184 ； SEQ ID NO 224、226、236、238、248、250、 260、262 ; SEQ ID NO 227、229、239、241、251、253 ' 263、265 ; SEQ ID NO 194、196、206、208、221、223 ' 281、283 ; SEQ ID NO 188、190 ' 200、202、215、217、 272、274及其功能類似物。在一特定實施例中’細胞包含 選自序列之群的AksA、AksD、AksE及AksF基因。在另一 特定實施例中，細胞包含含有由SEQ ID NO: 149表示之序 列或其功能類似物的NifV基因、選自序列之群的AksD、 O AksE及 AksF基因。 在一尤其較佳實施例中，選自AksA、AksD、AksE及 AksF基因之群的此等基因之一、二、三或每一者包含選自 分別由SEQ ID NO: 145、146、147、148表示之序列（分別 為AksA、D、E及F)及其功能類似物的序列。在另一尤其 較佳實施例中，此等基因之一、二、三或每一者包含分別 由SEQ ID NO: 167、168、169、170表示之序列（分別為 AksA、D、E及F)及其功能類似物。 〇 在一尤其較佳實施例中，選自AksA、AksD、AksE及
AksF基因之群的此等基因之一、二、三或每一者包含選自 分別由SEQ ID NO: 260、224、236、248表示之序列（分別 為AksA、D、E及F)及其功能類似物的序列。 在一尤其較佳實施例中’選自AksA、AksD、AksE& AksF基因之群的此等基因之一、二、三或每一者包含選自 分別由SEQ ID NO: 262、226、238、250表示之序列（分別 為AksA、D、E及F)及其功能類似物的序列。 146966.doc •51 - 201037078 在一尤其較佳實施例中，選自AksA、AksD、AksE及 AksF基因之群的此等基因之一、二、三或每一者包含選自 分別由SEQ ID NO: 263、227、23 9、251表示之序列（分別 為AksA、D、E及F)及其功能類似物的序列。 在一尤其較佳實施例中，此等基因之一、二、三或每一 者包含選自分別由SEQ ID NO: 265、229、241、253表示 之序列（分別為AksA、D、E及F)及其功能類似物的序列。 在一尤其較佳實施例中，選自AksA、AksD、AksE及 AksF基因之群的此等基因之一、二、三或每一者包含選自 分別由SEQ ID NO: 281、194、206、221表示之序列（分別 為AksA、D、E及F)及其功能類似物的序列。 在一尤其較佳實施例中，選自AksA、AksD、AksE及 AksF基因之群的此等基因之一、二、三或每一者包含選自 分別由SEQ ID NO: 2 83、196、208、223表示之序歹ij (分別 為AksA、D、E及F)及其功能類似物的序列。 在一尤其較佳實施例中，選自AksA、AksD、AksE及 AksF基因之群的此等基因之一、二、三或每一者包含選自 分別由SEQ ID NO: 272、188、200、215表示之序列（分別 為AksA、D、E及F)及其功能類似物的序列。 在一尤其較佳實施例中，選自AksA、AksD、AksE及 AksF基因之群的此等基因之一、二、三或每一者包含選自 分別由SEQ ID NO: 274、190、202、217表示之序列（分別 為AksA、D、E及F)及其功能類似物的序列。 在另一尤其較佳實施例中，選自AksA、AksD、AksE及 146966.doc -52- 201037078
AksF基因之群的此等基因之一、二、三或每—者包含選自 分別由 SEQ ID NO: 161、162、163、164、165、166、 167、168、169、170、171、172、173、174表示之序列 (分別為AksA、D、E及F)及其功能類似物的序列。在另一 尤其較佳實施例中，此等基因之一、二、三或每一者包含 分別由SEQ ID NO: 177、178、179、180表示之序列（分別 為AksA、D ' E及F)及其功能類似物的序列。 在另一尤其較佳實施例中’選自AksA、AksD、AksE及 O AksF基因之群的此等基因之一、二、三或每一者包含選自 为別由SEQ ID NO: 260、224、236、248表示之序列（分別 為AksA、D、E及F)及其功能類似物的序列。 在另一尤其較佳實施例中，選自AksA、AksD、AksE及 AksF基因之群的此等基因之一、二、三或每一者包含選自 分別由SEQ ID NO·· 263、227、239、251表示之序列（分別 為AksA、D、E及F)及其功能類似物的序列。 Ο 在另一尤其較佳實施例中，選自AksA、AksD、AksE及
AksF基因之群的此等基因之一、二、三或每一者包含選自 分別由SEQ ID NO: 281、194、206、221表示之序列（分別 為AksA、D、E及F)及其功能類似物的序列。 在另一尤其較佳實施例中，此等基因之一、二、三或每 一者包含選自分別由SEq id NO: 272、188、200、215表 示之序列（分別為AksA、D、E及F)及其功能類似物的序 列。 在一尤其較佳實施例中，細胞之基因組包含由序列Π) 146966.doc •53· 201037078 145表示之核酸序列或其功能類似物、由序列ι〇 i46表示 之核酸序列或其功能類似物 '由序列⑴147表示之核酸序 列或其功能類似物、由序列ID 148表示之核酸序列或其功 能類似物、及由序列ID 149表示之核酸序列或其功能類似 物。 在一尤其較佳實施例中，細胞之基因組包含由序列ι〇 146表示之核酸序列或其功能類似物、由序列ι〇 μ?表示 之核酸序列或其功能類似物、由序列⑴148表示之核酸$ 列或其功能類似物、及由序列ID 149表示之核酸序列或其 功能類似物。 在一尤其較佳實施例中，細胞之基因組包含由序列ι〇 172表示之核酸序列或其功能類似物、由序列ι〇 表示 之核酸序列或其功能類似物、由序列出174表示之核酸序 列或其功能類似物、及由序列ID 149表示之核酸序列或其 功能類似物。 在一尤其較佳實施例中，細胞之基因組包含由序列m 224表示之核酸序列或其功能類似物、由序列m a%表示 之核酸序列或其功能類似物、由序列1〇 248表示之核酸序 列或其功能類似物、及由序列ID 149表示之核酸序列或其 功能類似物。 在一尤其較佳實施例中，細胞之基因組包含由序列id 227表示之核酸序列或其功能類似物、由序列ID 239表示 之核酸序列或其功能類似物、由序列出251表示之核酸序 列或其功能類似物、及由序列ID 149表示之核酸序列或其 146966.doc -54- 201037078 功能類似物。 在一尤其較佳實施例中，細胞之基因組包含由序列ID 194表示之核酸序列或其功能類似物、由序列id 206表示 之核酸序列或其功能類似物、由序列ID 22丨表示之核酸序 列或其功能類似物、及由序列ID 149表示之核酸序列或其 功能類似物。 在一尤其較佳實施例中，細胞之基因組包含由序列ID 188表示之核酸序列或其功能類似物、由序列m 2〇〇表示 © 之核酸序列或其功能類似物、由序列ID 215表示之核酸序 列或其功能類似物、及由序列ID 149表示之核酸序列或其 功能類似物。 在一尤其較佳實施例中，細胞之基因組包含由序列m 177表示之核酸序列或其功能類似物、由序列⑴上78表示 之核酸序列或其功能類似物、由序列1〇 179表示之核酸序 列或其功能類似物、由序列ID 18〇表示之核酸序列或其功 能類似物、及由序列ID 149表示之核酸序列或其功能類似 物。 在一尤其較佳實施例中’細胞之基因組包含由序列ID 224表示之核酸序列或其功能類似物、由序列⑴236表示 之核酸序列或其功能類似物、由序列ID 248表示之核酸序 列或其功能類似物、由序列ID 260表示之核酸序列或其功 能類似物、及由序列ID 149表示之核酸序列或其功能類似 物。 在一尤其較佳實施例中，細胞之基因組包含由序列m 146966.doc -55- 201037078 227表示之核酸序列或其功能類似物、由序列出239表示 之核酸序列或其功能類似物、由序列1〇 251表示之核酸序 列或其功能類似物、由序列ID 263表示之核酸序列或其功 能類似物、及由序⑽149表示之核酸序列或其功能類似 物。 在一尤其較佳實施例中’細胞之基因組包含由序列山 194表示之核酸序列或其功能類似物、由序列⑴表示 之核酸序列或其功能類似物、由序列1〇 221表示之核酸序 列或其功能類似物、由序列ID 281表示之核酸序列或其功 能類似物、及由序列ID 149表示之核酸序列或其功能類似 物。 在一尤其較佳實施例中，細胞之基因組包含由序列id 188表不之核酸序列或其功能類似物、由序列①2卯表示 之核酸序列或其功能類似物 '由序列山215表示之核酸序 列或其功能類似物、由序列m 272表示之核酸序列或其功 能類似物、及由序列ID 149表示之核酸序列或其功能類似 物。 已由基因組包含由SEQ ID NO: 149、167、168、169及 170表不之異源核酸序列的大腸桿菌宿主細胞實現優良社 果。 、。 已由基因組包含由SEQ ID N0: 149、168、169及17〇表 不之異源核酸序列的大腸桿菌宿主細胞實現優良結果。 已由基因組包含由序列ID 149、172、W3及W4表示之 異源核奴序列的釀酒酵母宿主細胞實現優良結果。 146966.doc 201037078 已由基因組包含由SEQ ID NO: 149、177、178、179、 180表示之異源核酸序列的大腸桿菌宿主細胞實現優良結 果。 已由基因組包含由SEQ ID NO: 149、224、236、248表 示之異源核酸序列的大腸桿菌宿主細胞實現優良結果。 已由基因組包含由SEQ ID NO: 149、227、239、251表 示之異源核酸序列的大腸桿菌宿主細胞實現優良結果。 已由基因組包含由SEQ ID NO: 149、194、206、221表 Ο 示之異源核酸序列的大腸桿菌宿主細胞實現優良結果。 已由基因組包含由SEQ ID NO: 149、188、200、251表 不之異源核酸序列的大腸桿菌宿主細胞實現優良結果。 異源細胞可尤其為如上文在描述生物催化劑時所提及之 細胞。 詳言之，本發明之異源細胞包含一或多個編碼一或多種 異源酶之異源核酸序列（其可為一或多個載體之一部分）， ❹ 該一或多種異源酶能夠在由a-酮戊二酸製備α_酮庚二酸中 或在ΑΚΡ轉化為ΑΑΡ、6-ACA、5-FVA、己内醯胺或二胺 基己烷中催化至少一個反應步驟。 在一特定實施例中，細胞包含一或多個可為同源或異源 且編碼能夠催化(X-酮戊二酸轉化為α_酮己二酸之酶系統的 核酸序列，其中該酶系統成為供離胺酸生物合成之ΑΑΑ生 物合成途徑之一部分，諸如上文更詳細描述。 異源細胞較佳不具有能夠催化…酮己二酸酯轉化為胺 基己二酸酯之胺基轉移酶活性。若此活性天然存在於細胞 146966.doc -57- 201037078 中，則可藉由使細胞DNA中編碼該等酶之基因失活、對其 進仃修飾或其缺失來移除、降低或改變活性。此活性可源 自或夕種生物催化劑。此等生物催化劑亦可例如藉由分 子進化或合理設計來修飾，以不再具有任何不欲活性但保 留任何所需活性(例如本發明上下文t之任何活性或宿主 細胞代謝所需之活性）。 異源細胞較佳不含任何可將AKP、5_FVA、AAp、6_ ACA:己内㈣或二絲己烧降解或轉〖為任何不欲副產 物之酶。若鑑別出任何該活性，例如作為己内醯胺降解途 徑之—部分，則可如上文所述來移除、降低或改變此活 性。 細胞較佳包含一或多個、編碼一或多種催化_戊二酸 C!·伸長為α-酮己二酸及/或。酮己二酸Ci_伸長為α_酮庚二 酸之酶的異源核酸序列。適合核酸序列可尤其選自諸如上 文所鑑別之編碼Aks酶或其同系物之核酸序列。 詳言之，在細胞欲用於製備AKP，Ακρ又轉化為另一產 物，諸如5-FVA或AAP ,該產物又可進一步轉化成6_ ACA、己内醯胺或二胺基己烷之情況下，異源細胞較佳包 3編碣催化該轉化之酶的核酸序列。此可為有利的，舉例 而=，因為至少一些在細胞中有活性之催化^广伸長之酶 月b夠催化AKP之不當伸長。若催化該伸長之酶原則上亦能 夠使用AKP作為受質，則藉由在細胞中表現能夠催化Ακρ 轉化為所需產物（諸如5_FVA或AAp)之酶（諸如脫羧酶或胺 基轉移酶），預期可減少或實質上避免該不當伸長。 146966.doc •58- 201037078 注意到例如詹氏甲烷暖球菌或棕色固氮菌中Ci_伸長所 涉及之一些酶具有不嚴格之受質特異性且能夠轉化不同碳 長度之欠質。已知許多酶具有不嚴格之受質特異性，從而 使其轉化非天然受質。為了提高本發明（方法中所用）異源 細胞之效率，尤其較佳提供能夠在由AKG製備AKP中催化 反應步驟且展示對AKG伸長為AKA及/或AKA伸長為AKP 具高催化活性，而對AKP之進一步伸長具低催化活性的酶 系統。可藉由諸如上述之技術來修飾能夠在由Akg製備 O AKP中催化反應步驟之一或多種酶（編碼核酸序列），以提 高關於AKG及/或AKA之伸長的反應特異性，及/或該酶（編 碼核酸序列）可經修飾，以降低AKP(作為受質）之結合親和 力，從而降低關於AKP伸長之催化活性。 該修飾可包括分子進化以產生多樣性，接著篩選所需突 變體’及/或受質結合袋之合理工程改造。改良本發明方 法中所用酶之受質特異性的技術可基於此項技術中所描述 之技術。詳言之，能夠催化C〗-伸長之「反應a」的AksA酶 或其同系物可經進化，使得關於催化AKP伸長為α-酮辛二 酸酯之催化活性相對於關於催化ΑΚΑ伸長為ΑΚΡ及/或 AKG伸長為AKA之催化活性降低。該酶較佳實質上不展示 關於催化AKP伸長為α_酮辛二酸酯之催化活性。認為催化 「反應a」之酶尤其控制藉由Cl_伸長可獲得之最大鏈長， 當然除了 AKP意欲充當α_酮辛二酸酯製備中之受質之外。 舉例而言，採用結構及序列資訊設計特定突變進行合理 工程改造已用以改良紅黴素（erythr〇mycin)聚酮合成酶之酿 146966.doc -59- 201037078 基轉移酶結構域4的受質特異性，從而接受替代性醯基供 體。已展示修飾所提出之受質結合位點可產生經修飾之能 夠容納替代性受質之結合袋，從而得到不同產物比率 (Reeves, C. D.; Murli, S.; Ashley, G. W.; Piagentini, M.; Hutchinson, C. R.; McDaniel, R. Biochemistry 2001, 40(51)，15464-15470)。合理設計與分子進化方法均已用以 改變生物催化劑BM3之受質特異性，代替或除中鏈脂肪酸 (例如肉豆蔻酸）之天然受質之外，得到大量能夠氧化多種 不同烯烴、環烯烴、芳烴及雜芳烴之突變體（Peters, M. W.; Meinhold, P.; Glieder, A.; Arnold, F. H. Journal of the
American Chemical Society 2003，125(44), 13442-13450·
Appel, D.; Lutz-Wahl, S.; Fischer, P.; Schwaneberg, U.;
Schmid, R. D. Journal of Biotechnology 2001, 88(2), 167- 171及其中參考文獻）。 在一實施例中，異源細胞包含編碼高檸檬酸合成酶之異 源核酸序列，該高檸檬酸合成酶已自接受α_酮戊二酸酯作 為受質，但α-酮己二酸酯並非適合受質之高擰檬酸合成酶 進化成亦接受ex-酮己二酸酯作為受質之高檸檬酸合成酶。 該酶可尤其為經由ΑΑΑ途徑進行離胺酸生物合成中所涉及 之例如來自以下之真菌酶或細菌酶：青黴菌屬、頭孢黴 屬、黑穗病菌屬、頭孢黴屬、擬青黴菌屬、髮癣菌屬、平 革_屬、裸胞殼屬、麵菌屬、耶氏酵母菌屬、裂殖酵母菌 屬、畢赤酵母菌屬、漢森酵母菌屬、克魯維酵母菌屬、假 絲酵母菌屬、酵母菌屬、棲熱菌屬、或奇球菌屬；或固氮 146966.doc •60- 201037078 菌’例如固氮菌屬、弗蘭克氏菌屬、集胞藻屬、念珠藻 屬、微囊藻屬、根瘤菌屬、慢生根瘤菌屬、克雷伯氏桿菌 屬或假單胞菌屬。詳言之’可使用諸如來自棕色固氮菌之 NifV之酶，經證實其對AKA具有初始活性（Zheng，L.; White, R. H.; Dean, D. R. The Journal of Bacteriology 1997，179(18)，5963-5966)。序列ID 149中展示編碼該酶之 基因。 異源細胞可尤其包含諸如上文所鑑別之編碼Aks酶或其 同系物之核酸序列，更尤其細胞可至少包含編碼Aks酶或 其同系物之核酸序列，較佳可使用編碼由序列ID 4、5、 6、7、8、9、10、11、12、13、44、45、46、47、48、 49、50、51、52、53、69、70、71、72、73、74、75、 76、77、261、264、267、270、273、276、279、282之任 一者表示的酶或其同系物之核酸序列。 在另一較佳實施例中，該細胞包含至少一個編碼由序列 ID 14、15、16、17、18、19、20、21、22、23、54、 55、56、57、58、59、60、61、186、189、192、195、 225、228、231、234之任一者表示之酶或其同系物的核酸 序列。 在另較佳實施例中，該細胞包含至少一個編碼由序列 ID 24、24、25、26、27、28、29、30、31、32、33、 198、201、204、207、237、240、243、246之任一者表示 之酶或其同系物的核酸序列。 在另一較佳實施例中，該細胞包含至少一個編碼由序列 146966.doc -61 - 201037078 ID 34、35、36、37、38 ' 39、40、41、42、43、62、 63 ' 64 、 65 ' 66 ' 67 ' 210 、 213 、 216 、 219 、 222 、 249 、 252、255、258之任一者表示之酶或其同系物的核酸序 列。 在一實施例中，異源有機體係基於具有供離胺酸生物合 成之A A A途徑的宿主細胞’其中可異源表現能夠催化c丨-伸長中之「反應a」的高檸檬酸合成酶（諸如AksA或其同系 物）。該高檸檬酸合成酶較佳能夠選擇性催化AKG及/或 AKA之伸長（反應a)中的反應步驟，而實質上不催化AKP之 伸長。在該情況下’宜缺失任何内源性高棒檬酸合成酶， 尤其在其能夠催化AKP之伸長反應中之「反應&」的情況 下。此類宿主細胞可隨後有效地含有一或多種在AKG C!-伸長為AKA及/或AKA C〗-伸長為AKP中具有功能活性的高 檸檬酸合成酶。隨後可由内源性酶（諸如涉及胺基己二酸 途徑之酶）催化實現AKG及/或AKA伸長之其他反應。 在一實施例中，異源細胞包含編碼具有α_酮庚二酸胺基 轉移酶活性之酶的核酸序列及/或編碼具有α_胺基庚二酸脫 羧酶活性之酶的核酸序列（含其之重組載體）。 在一較佳實施例中，本發明之異源細胞包含編碼具有 ΑΚΡ脫羧活性之_的核酸序列及/或編碼具有5_FVA胺基 轉移酶活性之酶的核酸序列。在一較佳實施例中，本發明 之異源細胞包含編碼具有α-胺基庚二酸2_脫氫酶或AKp胺 基轉移酶活性之酶的核酸序列及/或編碼具有α_胺基庚二酸 脫羧酶活性之酶的核酸序列。 146966‘doc 62· 201037078 在一較佳實施例中，本發明之異源細胞包含編碼具有6-胺基己酸6-脫氫酶活性之酶的核酸序列及視情況存在之編 碼具有α-酮庚二酸脫羧酶活性之酶的核酸序列。 【實施方式】 現將由下列實例來說明本發明。 實例 實例1 : 一般方法 分子及遺傳技術 〇 標準遺傳及分子生物學技術一般在此項技術中已知且早 先已描述（Maniatis 等人 1982「Molecular cloning: a laboratory manual」· Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y. ; Miller 1972「Experiments in molecular genetics」，Cold Spring Harbor Laboratory, Cold Spring Harbor ; Sambrook 及 Russell 2001「Molecular cloning: a laboratory manual」（第 3 版），Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press » Q ^ r F. Ausubel 等人編，1 Current protocols in molecular biology」，Green Publishing and Wiley Interscience, New York 1987)。 質體及菌株 早先已描述pMS470(Balzer，D_; Ziegelin, G_; Pansegrau, W.; Kruft, V.; Lanka, E. Nucleic Acids Research 1992, 20(8)，185 1-1858)及 pBBRlMCS(Kovach ME, Phillips RW， Elzer PH, Roop RM 2nd, Peterson KM. Biotechniques. 1994 146966.doc -63- 201037078 年5月；16(5):800-2. pBBRIMCS: —種宿主範圍廣泛之選 殖載體）。大腸桿菌菌株TOP10及DH10B(Invitrogen， Carlsbad, CA，USA)用於所有選殖程序。大腸桿菌菌株 BL21 Al(Invitrogen, Carlsbad, CA, USA)及 BL21(Novagen (EMD/Merck)，Nottingham, UK)用於蛋白質表現。 pRS414、pRS415 及 pRS416(Sikorski,R.S.及 Hieter，P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae Genetics 122 (1)，19-27 (1989) ; Christianson，T.W.，Sikorski，R.S·， Dante，M·，Shero,J.H及 Hieter，P· Multifunctional yeast high-copy-number shuttle vectors. Gene 110 (1), 119-122 (1992)) 用於在釀酒酵母中表現。釀酒酵母菌株€丑1^?1^113-6B(wra3 ' trpl ' leu2 ' MATa) ' CEN.PK \\2>-SO{ura3 ' MATa)、CEN.PK 102-3A〇ra3、/ew2、MATa)及 CEN.PK 113-9D(wra3、、MATa)用於蛋白質表現。 培養基
2><ΤΥ 培養基（16 g/1 騰蛋白脒（tryptopeptone)、10 gM酵母 提取物、5 g/1 NaCl)用於大腸桿菌生長。補充抗生素（100 pg/ml 胺苄青黴素（ampicillin)、50-100 pg/ml 新黴素 (neomycin))以在大腸桿菌中維持質體。為誘導大腸桿菌中 基因表現，使用0.02%(***糖）及0.2 mM(IPTG)最終濃 度之***糖（對於BL21-AI衍生物）及IPTG(對於 卩]^470、？86111]^8衍生物）。在^19基本培養基（12.8 g/L Na2HP04.7H20 > 3 g/L ΚΗ2Ρ04、0.5 g/L NaCl、1 g/L 146966.doc -64- 201037078
Na2HP04.7H20、3 g/L ΚΗ2Ρ04、0.5 g/L NaCl、1 g/L NH4C1、2 mM MgS〇4、0.1 mM CaCl2)中，以葡萄糖（Ια%)或甘油 （1-4%)作 為碳源 ，由 大腸桿 菌產生 AKP ， 如下 文進一步說明。 具有4%半乳糖之Verduyn培養基用於釀酒酵母生長。 鑑別質體
藉由此項技術中一般已知之遺傳學、生物化學及/或表 型方式，諸如轉型子對抗生素之抗性、轉型子或質體DNA 〇 純化之PCR診斷分析、純化之質體DNA之限制性分析或 DNA序列分析，來鑑別攜帶不同基因之質體。藉由限制性 消化及若包括PCR步驟，則另外藉由定序來證實所述所有 新構築體之完整性。 用於測定α-酮酸、6-ACA、AAP、5-FVA及高⑷檸檬酸酯 之UPLC-MS/MS分析法 1.8 μιη，1 00 mmx2.1 mm之 Waters HSS Τ3 管柱用於分離 α-酮酸、6-ACA、AAP、5-FVA及高⑷檸檬酸酯，其中如表 〇 W 1中所述梯度溶離。溶離劑A由含有0.1 %甲酸之LC/MS級水 組成，且溶離劑B由含有0.1%甲酸之乙腈組成。流速為 0.25 ml/min且管柱溫度保持在40°C下不變。 表1 :用於分離α-酮酸、6-ACA、5-FVA、AAP及高⑷檸檬 酸酯之梯度溶離方案 時間(分鐘） 0 5.0 5.5 10 10.5 15 %A 100 85 20 20 100 100 %B 0 15 80 80 0 0 使用多反應監測（MRM)，Waters micromass Quattro 146966.doc ·65· 201037078 micro API用於正離子或負離子電離模式之電噴霧中，視所 分析化合物而定。在500 L/hr之流速下，離子源溫度保持 在130°C下，而去溶劑化溫度為350°C。 對於AKG、AKA、AKP、5-FVA、高擰檬酸酯及高2-檸 檬酸酯，以10-14 eV使去質子化分子成碎片，得到損失了 例如H20、C0及C02之特定碎片。 對於6-ACA及AAP，以13 eV使質子化分子成碎片，得到 損失了Η20、ΝΗ3及CO之特定碎片。 為測定濃度，合成製備之化合物的外標校準曲線用以計 算各別離子之響應因子。該響應因子用以計算樣品中之濃 度。用溶離劑A適當（2-10倍）稀釋樣品以克服離子抑制及 基質效應。
實例2:由大勝桿菌產生AKP 構築AKP生物合成途徑 自資料庫擷取以下各物之蛋白質序列：詹氏曱烷球菌蛋 白高檸檬酸合成酶（AksA，MJ0503，[序列ID 4])、高烏頭 酸酶小次單元（AksE，MJ1271，[序列ID 24])、高烏頭酸酶 大次單元（AksD，MJ1003，[序列ID 14])及高異檸檬酸脫 氫酶（AksF，MJ1596，[序列ID 34])、來自海沼曱烷球菌 C5之其同系物（高檸檬酸合成酶（AksA，MmarC5_1522， [序列ID 7])、高烏頭酸酶小次單元（AksE，MmarC5 1257， [序列ID 27])、高烏頭酸酶大次單元（AksD，MmarC5 0098，[序列ID 17])及高異檸檬酸脫氫酶（AksF，MmarC5 0688，[序列ID 37]))及棕色固氮菌高檸檬酸合成酶NifV[序 146966.doc •66· 201037078 列 ID 75]。 (使用W008000632中所述之方法）針對大腸桿菌，對詹 氏甲烷球菌及海沼曱烷球菌之基因進行密碼子對優化且以 合成方式構築（Geneart, Regensburg, Germany)。在優化程 序中，避免内部限制性位點且在起始及終止處引入常見限 制性位點以便在表現載體中次選殖。亦添加來自pMS47〇 之tac啟動子序列在AksD上游。各ORF在共同核糖體結合 位點及前導序列之前以驅動PMS470中之轉譯。亦添加來 Ο 自PMS470之tac啟動子序列在AksD上游。將合成AksA[詹 氏甲烷暖球菌序列ID 167，海沼曱烷球菌序列ID 177]/ AksF[詹氏甲烷暖球菌序列ID 168，海沼曱烷球菌序列ID 178]卡匣以Ndel/Xbal切割且將合成AksD[詹氏甲烷暖球菌 序列ID 169，海沼曱烷球菌序列ID 179]/AksE[詹氏曱烷暖 球菌序列ID 170，海沼曱烷球菌序列ID 180]卡匣以 Xbal/Hindlll切割。將來自詹氏曱烷暖球菌之含有Aks基因 之片段***pMS470之Ndel/Hindlll位點以獲得載體pAKP- 〇 W 180。將含有來自海沼曱烷球菌之Aks基因之片段*** pMS470之Ndel/Hindlll位點以獲得載體pAKP-182。
自 D. Dean(Zheng L，White RH, Dean DR. Purification of the Azotobacter vinelandii nifV-encoded homocitrate synthase. J Bacteriol. 1997年 9月；179(18):5963_6)獲得含 有來自棕色固氮菌之NifV[序列ID 149]的大腸桿菌表現構 築體（pDB555)。使用 phusion DNA聚合酶（Finnzymes)，使 用引子 Avine-WT-R-BamHI[序列 ID 150]及 Avine-WT-F-SacI 146966.doc -67- 201037078 [序列ID 151]，自此載體對基因進行PCR擴增，且在具 有BamHI/SacI之ρΑΚΡ-1 80中AksA上游選殖，得到載體 pAKP-281 □。亦使用引子Avine-WT-R-HindIII[序列ID 152] 及 Avine-WT-F-Hindlll[序列 ID 153]對 m/F基因進行 PCR擴 增且在具有Hindlll之pAKP-180及pAKP-182中AksE[序列ID 170]下游選殖，分別得到載體pAKP-279及pAKP-280。 為使pAKP279及pAKP28 1中之ahJ基因失活，分別以 BamHI及5g/II消化質體，得到三個片段（566bps、1134bps 及7776bps)。自瓊脂糖凝膠中分離1134bps及7776bps大小 〇 之片段且彼此接合。在轉型至大腸桿菌之後，檢查質體之 位向且選擇兩個片段以與在原始質體PAKP279及pAKP281 中相同之方式定向的質體，分別得到pAKP322及 PAKP323 ° 大腸桿菌中蛋白質表現及代謝物產生 將質體pAKP-279、pAKP-280、pAKP-281、pAKP-322及 pAKP-323轉型至大腸桿菌BL21中進行表現。起始培養物 在具有10 ml 2 χΤΥ培養基之管中生長隔夜。將200 μΐ培養 ◎ 物轉移至含有20 ml 2χΤΥ培養基之搖瓶中。在迴旋式震盪 器中在30°C及280 rpm下培育燒瓶。4 h後，添加0.2 mM最 終濃度之IPTG且在30°C及280 rpm下培育燒瓶4-16 h。藉由 離心自20 ml培養物收集細胞且在24孔板中將其再懸浮於4 ml具有適合碳源之M9培養基中。在30-37°C及210 rpm下培 育24-72 h後，藉由離心收集細胞，且將離心塊與上清液分 離，且儲存在-20°C下以供分析。 146966.doc • 68 · 201037078 藉由離心收穫來自小規模生長（參見前一段落）之細胞。 將細胞離心塊再懸浮於1 ml 1 00%乙醇中且劇烈渦動。將 細胞懸浮液在9 5 °C下加熱2分鐘且藉由離心移除細胞碎 片。在真空乾燥器中蒸發上清液且將所得離心塊溶解於 200 μΐ去離子水中。藉由離心移除剩餘碎片且將上清液儲 存在-20°C下。 分析上清液及細胞提取物 在進行UPLC-MS/MS分析之前，將來自細胞部分之上清 〇 液及提取物以水稀釋5倍。結果清楚地表明重組菌株中存 在AKP及AAP。預期大腸桿菌中存在之天然胺基轉移酶催 化AKP轉化為AAP。
表2 :以葡萄糖或甘油作為碳源產生AKP 質體 部分 碳源 AKP [mg/1] AAP [mg/1] pAKP-279 上清液 葡萄糖 3 n.d. pAKP-279 細胞 葡萄糖 2 n.d. pAKP-281 上清液 葡萄糖 3 n.d. pAKP-281 細胞 葡萄糖 2 n.d. pAKP-280 上清液 葡萄糖 2 n.d. pAKP-322 上清液 葡萄糖 10 3 pAKP-322 細胞 葡萄糖 8 12 pAKP-323 上清液 葡萄糖 7 3 pAKP-323 細胞 葡萄糖 7 1 _ 上清液 葡萄糖 n.d. n.d. - 細胞 葡萄糖 n.d. n.d. pAKP-281 上清液 甘油 12 1 pAKP-281 細胞 甘油 6 4 pAKP-322 上清液 甘油 57 5 pAKP-322 細胞 甘油 8 12 146966.doc •69· 201037078 ρΑΚΡ-323 上清液 甘油 47 4 ρΑΚΡ-323 細胞 甘油 4 7 上清液 甘油 n.d. n.d. 細胞 甘油 n.d. n.d. η. d.=未偵測到 結果清楚地表明重組菌株中存在AKP及AAP。預期大腸 桿菌中存在之天然胺基轉移酶催化AKP轉化為AAP。自構 築體移除AksA對所產生之AKP及AAP之量具有積極影響。 實例3:由釀酒酵母產生AKP 構築AKP生物合成途徑 (使用W008000632中所述之方法）針對釀酒酵母’對詹 氏曱烷球菌基因進行密碼子對優化。自釀酒酵母基因組資 料庫（www.yeastgenome.org，如在3 1/3/08獲得）操取啟動子 及終止子序列。將tpil啟動子中位置-5處之T變成A以產生 釀酒酵母之共同kozak序列。如表3中所示，以合成方式 (Geneart, Regensburg, Germany)製造啟動子-基因-終止子 卡g。 表3 :啟動子-基因-終止子卡匡 啟動子 基因 終止子 tdhl MJ0503 [序列Π) Hi] tdhl tpil MJ1003 [序列 ID 172] tpil enol MJ1271 [序列 ID 173] enol tdh3 MJ1596 [序列id 1741 tdh3 在優化程序中’避免内部限制性位點且在起始及末端處 引入常見限制性位點以便在表現載體中次選殖。 以Sall/EcoRI切割合成AksA卡匣且以EcoRI/Xbal切割合 成AksF卡匣’且將兩個片段接合至pRS4 1 5以獲得ρΑΚΡ- 146966.doc -70· 201037078 136。類似地’將合成AksD及AksE卡匣***pRS416中以獲 得 pAKP-146。將來自 pAKP-136 之 AksA-AksF 卡匣以 Xhol/Kpnl消化且***pAKP-146中，得到pAKP-141。以合 成方式製造具有編碼線粒體信號肽（mtSP)且N-末端融合至 MJ0503、MJ1271、MJ1003 及 MJ1596 之 207bp序列[序列 ID 158]之類似構築體（pfanner n, Neupert W. Distinct steps in the import of ADP/ATP carrier into mitochondria. J Biol Chem. 1987年 6 月 5 日；262(16):7528-36)。將由啟動子-mtSP-基因-終止子組成之合成片段組合於pRS4l6中以獲得 pAKP-140。使用 phusi〇n DNA 聚合酶，以引子 AksA-Avine-F[序列 ID 1 54]及 AksA-Avine-Rl [序列 ID 1 55]，自 pDB555對m/K進行PCR擴增。使用phusion DNA聚合酶， 以引子 Pgal2-F2[序列 ID 156]及 Pgal2-R[序列 ID 157]，自 pAKP-47對抑/2啟動子進行擴增。藉由PCR，使用Phusion DNA 聚合酶及引子 pgal2-F2[序列 ID 1 53]及 AksA-Avine-R1[序列ID 155]，來融合兩個PCR片段，且在具有 Hpal/AscI之pAKP-47中選殖所得融合產物，得到pPgal2-nifV_Ttdhl。藉由KpnI/Spel 自此構築體中移除pPgal2-nifV-Ttdhl卡匣且將其***經KpnI/Spel消化之pAKP-140及 pAKP-141 中，置換 MJ0503(AksA)[序列 ID 167]，且分別得 到構築體pAKP-3 05及pAKP-306。 構築產生AKP之釀酒酵母菌株 根據如Gietz及Woods所述之方法（Gietz, R.D.及Woods, R.A. (2002).Transformation of yeast by the Liac/SS carrier 146966.doc -71· 201037078 DNA/PEG method. Methods in Enzymology 350: 87-96)以 1 μg pAKP-305或pAKP_306質體DNA使釀酒酵母菌株 CEN.PK113-5D轉型。將細胞塗於具有lx無胺基酸酵母氮 源基礎及2%葡萄糖之瓊脂板上。
以釀酒酵母產生AKP 為產生AKP，在30°C及280 rpm下，將起始培養物在10 ml含有具有4%半乳糖之Verduyn培養基之管中好氧生長隔 夜。用25 ml具有4%半乳糖之新鮮Verduyn培養基稀釋培養 物至0.5 OD且在30°C及280 rpm下厭氧及好氧培育2及5天 (好氧培養）及4天（厭氧培養）。藉由離心收穫細胞，且如實 例2中針對大腸桿菌所述來製備上清液及細胞部分樣品， 以供UPLC-MS/MS分析。 表4 :結果 質體 部分 AKP [mg/11 PAKP305 上清液 1 PAKP305 細胞 2 PAKP306 上清液 1 實例4:選殖胺基轉移酶及脫羧酶之標靶基因 設計表現構築體 添加αίβ位點至核糖體結合位點及起始密碼子之上游及 終止密碼子下游的所有基因中以便於使用Gateway技術 (Invitrogen，Carlsbad, CA, USA)進行選瘦。 基因合成及質體構築 由DNA2.0獲得合成基因且根據DNA2.0之標準程序進行 146966.doc -72- 201037078 密碼子優化，以在大.腸桿菌中表現。對來自河流孤菌 JS17[SEQ ID No. 1]及韋氏芽胞桿菌 KBAB4[SEQ ID No. 82]之分別編碼河流孤菌JS 17 ω-胺基轉移酶[SEQ ID No. 2] 及韋氏芽胞桿菌KBAB4胺基轉移酶（ZP_01186960)[SEQ ID No. 83]之胺基酸序列的胺基轉移酶基因進行密碼子優化， 且藉由DNA合成獲得所得序列[SEQ ID No. 3]及[SEQ ID No. 85] ° 亦對來自大腸桿菌[SEQ ID No. 105]、釀酒酵母[SEQ ID 〇 No. 108]、運動醱酵單胞菌[SEQ ID No. Ill]、乳酸乳球菌 [SEQ ID No. 114]、[SEQ ID No_ 117]、及結核分枝桿菌 [SEQ ID No. 120]之分別編碼河流弧菌JS17 ω-胺基轉移酶 [SEQ ID No. 3]、韋氏芽孢桿菌ΚΒΑΒ4胺基轉移酶 (ZP_01186960)[SEQ ID No. 84]、大腸桿菌二胺基庚二酸 脫羧酶LysA[SEQ ID No. 106]、釀酒酵母丙酮酸脫羧酶 Pdc[SEQ ID No. 109]、運動礙酵單胞菌丙嗣酸脫叛酶 PdcI472A[SEQ ID No. 112]、乳酸乳球菌支鏈α-酮酸脫羧
Q 酶KdcA[SEQ ID No. 11 5]及α-酮異戊酸脫羧酶KivD[SEQ 10 1^〇.118]及結核分枝桿菌〇1-酮戊二酸脫羧酶反§(1[3£(^10 No. 12 1 ]之胺基酸序列的基因進行密碼子優化，且藉由 DNA合成分別獲得所得序列[SEQ ID No. 107]、[SEQ ID No. 110]、[SEQ ID No. 63]、[SEQ ID No. 116]、[SEQ ID No. 119]及[SEQ ID No. 122]。 使用Gateway技術（Invitrogen)，如製造商方案 (www.invitrogen.com)中所述，經由引入之attB位點及 146966.doc -73- 201037078 pDONR201(Invitrogen)作為進入載體，將基因構築體選殖 至pBAD/M_yc-His-DEST表現載體中。以此方式分別獲得表 現載體pBAD-K//_AT、pBAD-5w_ATpBAD-L>^、pBAD-尸心、pBAD-PdcI472A、pBAD-kdcA 及 pBAD-kivD。藉由 以各別pBAD-表現載體轉型化學感受態大腸桿菌 TOPlO(Invitrogen)來獲得相應表現菌株。 藉由PCR進行選殖 藉由 PCR ，使用 PCR Supermix High Fidelity (Invitrogen) ’根據製造商說明書’使用如下表中所列之引 子’自基因組DNA擴增編碼生物催化劑之各種基因。 表5:用於各種基因之引子综述 基因來源 L基因序列ID 酶序列ID 引子序列ID 綠膿桿菌 85 86 87 及 88 綠膿桿菌 μοι 102 135及136 綠膿桿菌 141 142 147及148 綠膿桿菌 143 144 149及150 枯草桿菌 89 90 123及124 枯草桿菌 91 92 125及126 枯草桿菌 139 140 145及146 類球紅細菌(及/奶而如故r sphaeroides) 93 94 127及128 嗜肺性退伍軍人桿菌(IegioweZ/α pneumovhilia) 95 96 129及130 歐洲亞硝化單胞菌(MYmyo was europaea) 97 98 131及132 淋病奈瑟菌(A/e/aeria gcworr/ioeae) 99 100 133及134 沼澤紅假單胞菌 (Rhodopseudomonas palustris) 103 104 137及138 146966.doc -74· 201037078 藉由瓊脂糖凝膠電泳分析PCR反應且使用QIAquick PCR 純化套組（Qiagen，Hilden, Germany)自凝膠溶離正確尺寸 之PCR產物。使用Gateway技術（Invitrogen)，如製造商方 案中所述，經由引入之位點及pDONR-zeo(Invitrogen) 作為進入載體，將經純化PCR產物選殖至pBAD/A/>c?-His-DEST表現載體中。藉由DNA定序來驗證由PCR選殖之基 因的序列。以此方式獲得表現載體pBAD-Pae-_gi9946143_AT 、 pB AO-Bsu_gil 6078032_AT 、 pBAD- ❹ Bsu_gil6080075_AT ' pBAD-5i«_gil6077991_AT ' pBAD-
Rsp AT ' pBAD-L/?w_AT ' pBAO-Neu_AT ' pBAD-iV^o_AT ' pB AD-P<3e_gi995 1299_AT 、pBAD-Pae_gi995 1 072 AT 、 pBAD-P<ae_gi995 163 0_AT 及 pBAD-i?/><3_AT。藉由以 pBAD 構築體轉型化學感受態大腸桿菌TOPlO(Invitrogen)，獲得 相應表現痛株。 實例5:大腸桿菌之生長以供蛋白質表現 在具有940 μΐ含0_02%(w/v)L-***糖之培養基的96深 〇 孔板中進行小規模生長。藉由以96孔印模（Kiihner， Birsfelden, Switzerland)自冷;東儲備培養物轉移細胞來接 種。在25°C下，在迴旋式震盪器（300 rpm，5 cm振幅）上培 育培養板48 h。通常達到2-4之OD62〇nm。 實例6 :製備細胞溶解產物 製備溶解緩衝液 溶解緩衝液含有以下成份： 表6 :溶解緩衝液 146966.doc -75- 201037078 1M MOPS pH 7.5 5 ml DNAse I 級 II(Roche) 10 mg 溶菌酶 200 mg MgS04.7H20 123.2 me 二硫蘇糖醇(DTT) 154.2 me H20(Milli〇) 補足至100 ml 在即將使用之前製備新鮮溶液。 藉由溶解製備無細胞提取液 藉由離心收穫來自小規模生長（參見前一段）之細胞且棄 去上清液。將在離心期間形成之細胞離心塊在_ 2 〇 下冷 凍至少16 h且隨後在冰上解凍。將5〇〇卜丨新鮮製備之溶解 緩衝液添加至各孔中且藉由使培養板劇烈渦動2 _ 5 m丨n使細 胞再懸浮。為實現溶解’將培養板在室溫下培育3〇分鐘。 為移除細胞碎片，將培養板在4〇C及6000 g下離心20分鐘。 將上清液轉移至新鮮培養板上且保持在冰上，直至再次使 用。 藉由音波處理製備無細胞提取液 藉由離心收穫來自中等規模生長（參見前一段）之細胞且 棄去上清液。將1 ml磷酸鉀緩衝液（pH 7)添加至0.5 g濕細 胞離心塊中且藉由劇烈渦動使細胞再懸浮。為實現溶解， 將細胞音波處理20分鐘。為移除細胞碎片，將溶解產物在 4°C及6000 g下離心20分鐘。將上清液轉移至新鮮管中，且 在-20°C下冷凍，直至再次使用。 實例7:藉由5_甲釀基戊酸甲酯化學水解來製備5甲酿 146966.doc -76· 201037078 基戊酸 如下藉由5-甲醯基戊酸甲酯化學水解來製備胺基轉移酶 反應之受質（亦即5-甲醯基戊酸）：用NaOH使100/〇(w/v)5-曱 醯基戊酸甲酯水溶液之PH值為14.1。在201：下培育24 h之 後，用HC1使pH值為7.1。 實例8: 5-甲醢基戊酸轉化為6_ACA之酶促反應 除非另外規定，否則製備包含1 〇 mM 5-甲醯基戊酸、20 mM外消旋a-曱基苄胺及2〇〇 μΜ 5，-填酸吡哆搭於50 mM磷 酸鉀緩衝液（pH 7.0)中之反應混合物。將丨〇〇 μι反應混合物 分配至孔板之各孔中。為開始反應，將20 μΐ無細胞提取液 添加至各孔中。在37°C下在震盪器上培育反應混合物24 h。此外，在相同條件下培育化學空白混合物（不含無細胞 提取液）及生物空白（含有pBAD/M_yc-His C之大腸桿菌 TOP 1 〇)。藉由HPLC-MS分析樣品。結果概述於下表中。
表7:在胺基轉移酶存在下由5-FVA形成6-ACA 生物催化劑 6-ACA 濃度[mg/kg] 大腸桿菌 ΤΟΡ10/ρΒΑϋ-β7_ΑΤ 43* 大腸桿菌 TOP 10/pBAD-Pae_AT 930 大腸桿菌 TOPlO/pBAD-Pae AT 25* 大腸桿菌 TOP10/pBAD-5we AT 24* 大腸桿菌 TOP 10/pB AD-&w_gi 16077991 AT 288 大腸桿菌TOPlO/pBAD·/^ gi9951072 AT 1087 大腸桿菌TOPlO/pBAD-尸從 gi9951630 AT 92 含有pBAD/柳c-His C之大腸桿菌TOPI 0(生物空白） 0.6 無(化學空白） n.d. n-d.:未偵測到 *方法不同之處在於：使用10 μ丨無細胞提取液而非20 μΐ， 5'-磷酸吡哆醛濃度為50 μΜ而非200 μΜ，且孔中反應混合 146966.doc -77- 201037078 物之體積為190 μΐ而非100 μΐ。 表明在胺基轉移酶存在下由5-FVA形成6-ACA。 實例9 : ΑΚΡ轉化為5-甲醯基戊酸之酶促反應 製備包含50 mM ΑΚΡ、5 mM氯化鎂、100 μΜ 5·-磷酸吡 哆醛（對於LysA)或1 mM二磷酸噻胺（對所有其他酶）於100 mM填酸鉀缓衝液（pH 6.5)中之反應混合物。將4 ml反應混 合物分配至反應容器中。為開始反應，將1 ml藉由音波處 理獲得之無細胞提取液添加至各孔。在商用草乙酸脫羧酶 (Sigma-Aldrich產品編號04878)情況下，使用50 U。在37°C Ο 下以磁性攪拌器培育反應混合物48 h。此外，在相同條件 下培育化學空白混合物（不含無細胞提取液）及生物空白（含 有 pBAD/M_yc-His C之大腸桿菌 TOP10)。藉由 HPLC-MS 分 析反應期間來自不同時間點之樣品。結果概述於下表中。
表8:在脫羧酶存在下由AKP形成5-FVA 生物催化劑 5-FVA 濃度[mg/kg] 3h 18h 48 h 大腸桿菌 TOPlO/pBAD-LysA 150 590 720 大腸桿菌 TOP 10/pBAD-Pdc 1600 1700 1300 大腸桿菌 TOP 10/pBAD-PdcI472A 2000 2000 1600 大腸桿菌 T〇Pl〇/pBAD-KdcA 3300 2300 2200 大腸桿菌 TOP 10/pBAD-KivD 820 1400 1500 草乙酸脫羧酶 n.d. 6 10 含有pBAD/Mjc-His C之大腸桿菌TOP 10(生物空白） n.d. n.d. n.d. 無G匕學空白） n.d. n.d. n.d.
n.d.:未偵測到 表明在脫羧酶存在下由AKP形成5-FVA。 146966.doc • 78 · 201037078 表明在脫羧酶存在下由AKP形成5_FVA。 實例1〇:在重組脫缓酶存在下Ακρ轉化為6_ACA之梅促 反應 製備包含50 mM AKP、5 mM氯化鎂、1〇〇 5,_填酸吡 哆醛（對於LysA)或1 mM二磷酸噻胺（對所有其他測試生物 催化劑）於100 mM磷酸鉀緩衝液（pH 6.5)中之反應混合物。 將4 ml反應混合物分配至反應容器中。為開始反應，將i ml無細胞提取液添加至各孔中。在37它下將反應混合物以 磁性搜掉器培育4 8 h。此外，在相同條件下培育化學空白 混合物（不含無細胞提取液）及生物空白（含有pB ad/儿/yc_ His C之大腸桿菌TOP 10)。藉由HPLC-MS分析反應期間來 自不同時間點之樣品。結果概述於下表中。
表9 :在脫羧酶存在下由AKP形成6-ACA 星物催化劑 6-AC A 濃度[mg/kgl 3h 18h 48 h 大腸桿菌 TOPlO/pBAD-LysA n.a. 0.01 0 大腸桿菌 TOPlO/pBAD-Pdc 0.1 0.3 n.a. 大腸桿菌 TOP10/pBAD-PdcI472A 0.03 0.1 0.2 A 腸桿菌 TOP 10/pB AD-KdcA 0.04 0.1 0.3 大腸桿菌丁 OP 10/pBAD-KivD n.a. 0.3 0.6 含有pBAD/柳c-His C之大腸桿菌TOP10(生物空白） n.d. n.d. n.d. 無(化學空白） n.d· n.d. n.d. n.a. =未分析 n.d. =未偵測到 表明在脫羧酶存在下由AKP形成6-ACA。預期大腸桿菌 含有天然5-FVA胺基轉移酶活性。 實例11 :在重組脫羧酶及重組胺基轉移酶存在下AKP轉 146966.doc -79· 201037078 化為6-ACA之酶促反應 製備包含50 mM AKP、5 mM氯化鎂、100 μΜ 5’-嗔酸。比 哆醛、1 mM二磷酸噻胺及50 mM外消旋α-曱基苄胺於ι〇〇 mM磷酸鉀緩衝液（pH 6.5)中之反應混合物。將1.6 ml反應 混合物分配至反應容器中。為開始反應，將0.2 ml含有脫 羧酶之無細胞提取液及0.2 ml含有胺基轉移酶之無細胞提 取液添加至各反應容器中。在37°C下以磁性攪拌器培育反 應混合物48 h。此外，在相同條件下培育化學空白混合物 (不含無細胞提取液）及生物空白（含有pBAD/M_yc-His C之 大腸桿菌TOP10)。藉由HPLC-MS分析反應期間來自不同 時間點之樣品。結果概述於下表中。
表1 0 :在重組脫敌酶及重組胺基轉移酶存在下由AKP形成 6-ACA 48小 時後之6-ACA澧廑fme/kel ’ π AT DC 大腸桿菌 TOP10/pBAD-Vfl- AT 大腸#菌 TOP 10/pB AD-B we-AT 大运每！ TOPlO/pBAD-PAE 09946143 AT 大腸桿菌 TOPlO/pBAD-Pdc 183.4 248.9 117.9 大腸桿菌 TOPlO/pBAD- PdcI472A 458.5 471.6 170.3 大腸桿菌 TOPlO/pBAD-KdcA 497.8 497.8 275.1 大腸桿菌 TOPlO/pBAD-KivD 510.9 510.9 314.4 AT=胺基轉移酶 DC=脫羧酶 在化學空白及生物空白中未偵測到6_aca。 此外，結果展示與宿主細胞僅具有重組脫羧酶（且無重 146966.doc •80· 201037078 實例12 :在大腸桿菌中產生6-ACA 製備使胺基轉移酶與脫羧酶共表現之構築體 如實例4中所述，構築含有編碼將AKP轉化為5-甲醯基 戊酸（5-FVA)及5-FVA轉化為6-ACA之酶之基因的質體。為 使胺基轉移酶與脫羧酶共表現，使用Phusion DNA聚合酶 及弓1 子 pF113-F-NsiI(aaattatgcatACAGCATGGCCTGCAACG) 及 pF113-R-AgeI(aaattaccggtCAGGGTTATTGTCTCATGAG)， 自 pF113(pJF119EH之衍生物（Ftirste，J. Ρ·, W. Pansegrau，R. Ο Frank, Η· Blocker, Ρ. Scholz, Μ· Bagdasarian及 Ε. Lanka. 1986. Molecular cloning of the plasmid RP4 primase region in a multi-host-range tacP expression vector. Gene 48:119-131)，其在位置515及5176處分別含有兩個Notl位點，其中 tac啟動子為編號開始）對tac啟動子卡匣進行PCR擴增，且 將所得PCR片段融合至經Nsil/Agel消化之pBBRIMCS (Kovach ME, Phillips RW, Elzer PH, Roop RM 2nd, Peterson KM. Biotechniques· 1994 年 5 月；16(5):800-2. Ο 冒 pBBRIMCS: a broad-host-range cloning vector)，得到 pBBR-lac。對來自河流孤菌JS17(Seq ID NO:l)之胺基轉移 酶基因進行密碼子優化（Seq ID NO: 3)。根據製造商說明 書，分別使用引子對 AT-Vfl_for_Ec(AAATTT GGTACC GCTAGGAGGAATTAACCATG)+AT-Vfl_rev_Ec(AAATTT ACTAGT AAGCTGGGTTTACGCGACTTC)及 AT-Pa01_for一
Ec(AAATTT GGTACC GCTAGGAGGAATTAACCATG)+AT-Pa01_rev—Ec(AAATTT ACTAGTACAAGAAAGCTGGGTTCAAG)， 146966.doc • 81 - 201037078 使用 Phusion DNA聚合酶，自 pBAD/M_yc-His-DEST-AT-Vfl 及pBAD/Mjc-his-DEST-PAOl對此密碼子優化基因及來自 綠膿桿菌PA01之分別編碼AT-Vfl及AT-PA01(Seq ID 85)之 基因進行PCR擴增。 根據製造商說明書，使用Phusion DNA聚合酶且使用引 子 Kdc_for_Ec(AAATTT ACTAGT GGCTAGGAGGAATTACATATG) 及 Kdc一rev一Ec (AAATTT AAGCTT ATTACTTGTTCTGCTCCGCAAAC) ，自pBAD/Myc-His-DEST-DC-KdcA對來自乳酸乳球菌之 編碼乳酸乳球菌支鏈α-酮酸脫羧酶KdcA(Seq ID NO:116)之 脫羧酶基因進行PCR擴增。以KpnI/Spel消化胺基轉移酶片 段且以Spel/Hindlll消化脫羧酶片段。將兩個片段分別接 合至經KpnI/Hindlll消化之pBBR-lac以獲得pAKP-94(含有 編碼AT-PA01及KdcA之基因）及pAKP-96(含有編碼AT-Vfl 及KdcA之基因）。 大腸桿菌中蛋白質表現及代謝物產生 使質體pAKP-323(在實例2中所述）與pAKP96共轉型至大 腸桿菌BL21中進行表現。如實例2中所述使培養物生長。 如實例2中所述製備樣品以供分析且如實例1中所述藉由 LC-MS-MS進行分析。 表11 : 質體1 質體2 碳源 培育 培養基 培養條件 6-ACA Γμ§/11 pAKP-323 pAKP-96 葡萄糖 24 h 2χΤΥ 24 孔 ΜΤΡ 18 pAKP-323 pAKP-96 甘油 24 h 2χΤΥ 24 孔 ΜΤΡ 109 pAKP-323 pAKP-96 葡萄糖 24 h Μ9 24 孔 ΜΤΡ 8 pAKP-323 pAKP-96 甘油 24 h Μ9 24 孔 ΜΤΡ 107 pAKP-323 pAKP-96 葡萄糖 24 h Μ9 搖瓶 1120 146966.doc -82- 201037078 以質體ρΑΚΡ-322(菌株eAKP233)、質體pAKp96(菌株 eAKP 71)或以質體PAKP94(菌株eAKP70)使大腸桿菌BL21 轉型。使培養物在具有10 ml 2χΤΥ培養基之管中生長隔 夜。將200 μΐ培養物轉移至含有20 ml 2 χΤΥ培養基之搖瓶 中。在30°C及280 rpm下，於迴旋式震盪器中培育燒瓶。4 h後’添加0.1 mM最終濃度之IPTG，且在30°C及280 rpm下 培育燒瓶4 h。藉由離心自1 〇 ml培養物收集細胞且將其再 懸浮於10 ml含有0.4%葡萄糖之M9培養基中。在24孔板中 〇 將兩種培養物以各種比率混合，且在37°c及210 rpm下培 育48-72 h之後，藉由離心收集上清液且儲存於-20°C下以 供分析。如實例2中所述來製備樣品以供LC-MS-MS分析且 如實例1中所述進行分析。 表12 : 菌株 eAKP233 菌株eAKP71 菌株eAKP70 培育 6-ACA|"pg/ll 5 ml 0 ml Oml 48 h 0 4 ml 1 ml Oml 48 h 0 2.5 ml 2.5 ml Oml 48 h 26 1 ml 4 ml Oml 48 h 40 0 ml 5 ml Oml 48 h 0 5 ml 0 ml Oml 72 h 0 4 ml 0 ml 1ml 72 h 68 2.5 ml Oml 2.5 ml 72 h 71 1 ml Oml 4 ml 72 h 85 0 ml Oml 5 ml 72 h 0 實例13 :構築其他太古菌之AKP生物合成途徑 自資料庫擷取以下各物之蛋白質序列：醋酸甲烷八聯球 菌高烏頭酸酶小次單元（AksE，MA3751 ’ [序列ID 225])、 146966.doc -83 - 201037078 高烏頭酸酶大次單元（AksD，MA3085，[序列ID 237])及高 異檸檬酸脫氫酶（AksF，MA3748，[序列ID 249])、來自亨 氏曱烷螺菌高烏頭酸酶小次單元（AksE ， Mhun_1799，[序列id 228])、高烏頭酸酶大次單元 (AksD ’ Mhun_1800，[序列ID 240])及高異檸檬酸脫氫酶 (AksF，Mhun—1797，[序列ID 252])之其同系物、來自海 沼甲烷球菌幻高烏頭酸酶小次單元（AksE，MMP0381，[序 列ID 207])、高烏頭酸酶大次單元（AksD，MMP1480，[序 列ID 195])及高異擰檬酸脫氫酶（AksF，[序列ID 222])之其 同系物、來自萬氏曱烷球菌高烏頭酸酶小次單元 (AksE，Mevan_13 68，[序列ID 201])、高烏頭酸酶大次單 元（AksD ’ Mevan_07 89，[序列ID 189])及高異檸檬酸脫氫 酶（AksF，Mevan_0040，[序列ID 216])之其同系物、及棕 色固氮菌高檸檬酸合成酶NifV[序列ID 75]。 表13 : 質體ID 供體有機體 NifV Seq ID AksD Seq ID AksE Seq ID AksF Seq ID pAKP-358 醋酸曱烷八聯球菌及棕色固氮菌 mm 149 236 224 248 pAKP-359 亨氏曱烷螺菌JF-1及棕色固氮菌 (NifV) 149 239 227 251 pAKP376 萬氏曱烷球菌及棕色固氮菌 (NifV) 149 188 200 215 pAKP378 海沼曱烷球菌*52及棕色固氮菌 _(綱 149 194 206 221 (使用W008000632中所述方法）針對大腸桿菌，對編碼 146966.doc • 84· 201037078 PAKP378 海沼曱烷球菌及棕色固氮菌 149 194 206 221 (NifV) —-—— (使用W008000632中所述方法）針對大腸桿菌，對編碼 高烏頭酸酶小次單元（AksE)、高烏頭酸酶大次單元（AksD) 及高異檸檬酸脫氫酶（AksF)之基因進行密碼子對優化（表 13)。以合成方式（Geneart, Regensburg, Germany)製備含有 優化基因連同野生型nifV基因（Seq ID 149)之構築體。在優 化程序中，避免内部限制性位點，且在起始及停止處引入 常見限制性位點以便在表現載體中次選殖。亦添加來自 pMS470之tac啟動子序列在AksD上游。各ORF在共同核糖 體結合位點及前導序列之前以驅動pMS470中之轉譯。亦 添加來自pMS470之tac啟動子序列在AksD上游。以 Ndel/Xbal切割合成AksA/AksF卡匣且以Xbal/Hindlll切割 合成AksD/AksE卡匣。將含有Aks基因之片段***pMS470 之 Ndel/Hindlll位點中以獲得載體 pAKP-358、pAKP359、 pAKP376及pAKP378 ° 大腸桿菌中蛋白質表現及代謝物產生 將質體轉型至大腸桿菌BL21中進行表現。使起始培養物 在具有10 ml 2χΤΥ培養基之管中生長隔夜。將200 μΐ培養 物轉移至含有20 ml 2xTY培養基之搖瓶中。在30°C及280 rpm下，在迴旋式震盪器中培育燒瓶。4 h後，添加0.2 mM 最終濃度之IPTG，且在30°C及280 rpm下培育燒瓶4-16 h。 藉由離心自20 ml培養物收集細胞且在24孔板中將其再懸 浮於4 ml具有適合碳源之M9培養基中。在30-37°C及210 146966.doc -85- 201037078 rpm下培育24-72 h之後，藉由離心收集細胞，且將離心塊 與上清液分離且儲存於-20°C下以供分析。 製備細胞部分以供分析 藉由離心收穫來自小規模生長（參見前一段）之細胞。將 細胞離心塊再懸浮於1 ml 1 00°/。乙醇中且劇烈渦動。在 95°C下加熱細胞懸浮液2分鐘且藉由離心移除細胞碎片。 在真空乾燥器中蒸發上清液且將所得離心塊溶解於200 μΐ 去離子水中。藉由離心移除剩餘碎片且將上清液儲存於 -20。。下。 分析上清液及細胞提取物 在進行UPLC-MS/MS分析之前，將來自細胞部分之上清 液及提取物以水稀釋5倍。表14中所示之結果清楚地表明 重組菌株中存在ΑΚΡ及ΑΑΡ。預期大腸桿菌中存在之天然 胺基轉移酶催化ΑΚΡ轉化為ΑΑΡ。 表14 :以甘油作為碳源產生ΑΚΡ 質體 部分 碳源 AKP[mg/ll AAP[mg/l] _ 上清液 甘油 n.d. n.d. _ 細胞 甘油 n.d. n.d. PAKP358 上清液 甘油 21 24 PAKP359 上清液 甘油 19 27 PAKP376 上清液 甘油 3 0.4 PAKP378 上清液 甘油 650 134 n. d.=未偵測到 結果清楚地表明重組菌株中存在AKP及AAP。預期大腸 桿菌中存在之天然胺基轉移酶催化AKP轉化為AAP。 146966.doc -86- 201037078 (5-FVA)及5-FVA轉化為6-ACA之酶的質體，而質體 pAKP94及pAKP96描述於實例12中。對於用運動醱酵單胞 菌丙酮酸脫缓酶PdcI472A[SEQ ID No. 112]及α-酮異戊酸 脫羧酶KivD[SEQ ID No. 118]更換存在於ρΑΚΡ 94及 ρΑΚΡ9ό中之乳酸乳球菌支鍵α_酉同酸脫缓酶KdeA[SEQ ID No. 115]，分別以Ndel及HinD3消化質體pBAD-kivD及 pBAD-PdcI472A。將含有脫羧酶基因之1，6 kb片段分離且 接合至經Ndel/HinD3消化之載體pAKP94中，分別得到 Ο ρΑΚΡ 326 及 ΡΑΚΡ327。將來自 pBAD-kivD 之 1.6 kb Ndel/HinD3片段選殖至 pAKP96中，得到pAKP330。 大腸桿菌中蛋白質表現及代謝物產生 將質體轉型至大腸桿菌BL21中進行表現。使起始培養物 在具有10 ml 2xTY培養基之管中生長隔夜。將200 μΐ培養 物轉移至含有20 ml 2χΤΥ培養基之搖瓶中。在30°C及280 rpm下，在迴旋式震盪器中培育燒瓶。4 h後，添加0.2 mM 最終濃度之IPTG，且在30°C及280 rpm下培育燒瓶4 h。藉 〇 W 由離心自20 ml培養物收集細胞且在24孔板中將其再懸浮 於4 ml具有1%甘油及500 mgd AKP之2xTY培養基中。在 3 0°C及210 rpm下培育48 h後，藉由離心收集細胞，且將離 心塊與上清液分離且儲存於-20°C下以供分析。
表15:在大腸桿菌中產生6-ACA 質體 胺基轉移酶 脫羧酶 Mg/1AAP Mg/16-ACA PAKP326 PA01 kivD 34 21 PAKP327 PA01 pdcI472A 25 1 PAKP330 Vfl kivD 18 18 146966.doc -87· 201037078 結果清楚地表明重組菌株中存在6-ACA及AAP。預期 腸桿菌中存在之天然胺基轉移酶催化AKP轉化為AAP。 146966.doc •88· 序列表 201037078 <11〇> 荷蘭商DSM智財資產公司 <120> α·酮庚二酸之製備 <140> 099107114 <141> 2010-03-11 <150> 09154840.4; 09170092.2; 09180441.9 <151> 2009-03-11; 2009-09-11; 2009-12-22 <210> 1 <211> 1362 <212> DNA <213> 河流孤菌 <210>
<211> CDS <212> (1)..(1362) <400> 1 48 atg aac aaa ccg caa age tgg gaa gee egg gee gag acc tat teg etc Met Asn Lys Pro Gin Ser Trp Glu Ala Arg Ala Glu Thr Tyr Ser Leu 1 5 10 15 o tat ggt ttc acc gac atg cct teg ctg cat cag ege ggc aeg gtc gtc Tyr Gly Phe Thr Asp Met Pro Ser Leu His Gin Arg Gly Thr Val Val 20 25 30 gtg acc cat ggc gag gga ccc tat ate gtc gat gtg aat ggc egg cgt Val Thr His Gly Glu Gly Pro Tyr lie Val Asp Val Asn Gly Arg Arg 35 40 45 96 144 tat ctg gac gee aac teg ggc ctg tgg aac atg gtc geg ggc ttt gac Tyr Leu Asp Ala Asn Ser Gly Leu Trp Asn Met Val Ala Gly Phe Asp 50 55 60 192 cac aag ggg ctg ate gac gee gee aag gee caa tac gag cgt ttt ccc His Lys Gly Leu lie Asp Ala Ala Lys Ala Gin Tyr Glu Arg Phe Pro 65 70 75 80 240 ggt tat cac gee ttt ttc ggc ege atg tee gat cag aeg gta atg ctg Gly Tyr His Ala Phe Phe Gly Arg Met Ser Asp Gin Thr Val Met Leu 85 90 95 teg gaa aag ctg gtc gag gtg teg ccc ttt gat teg ggc egg gtg ttc Ser Glu Lys Leu Val Glu Val Ser Pro Phe Asp Ser Gly Arg Val Phe 100 105 110 tat aca aac teg ggg tee gag geg aat gac acc atg gtc aag atg eta Tyr Thr Asn Ser Gly Ser Glu Ala Asn Asp Thr Met Val Lys Met Leu 115 120 125 tgg ttc ctg cat gca gee gag ggc aaa ccg caa aag ege aag ate ctg Trp Phe Leu His Ala Ala Glu Gly Lys Pro Gin Lys Arg Lys lie Leu 130 135 140 288 336 384 432 acc ege tgg aac gee tat cac ggc gtg acc gee gtt teg gee age atg Thr Arg Trp Asn Ala Tyr His Gly Val Thr Ala Val Ser Ala Ser Met 145 150 155 160 acc ggc aag ccc tat aat teg gtc ttt ggc ctg ccg ctg ccg ggc ttt Thr Gly Lys Pro Tyr Asn Ser Val Phe Gly Leu Pro Leu Pro Gly Phe 165 170 175 gtg cat ctg acc tgc ccg cat tac tgg ege tat ggc gaa gag ggc gaa Val His Leu Thr Cys Pro His Tyr Trp Arg Tyr Gly Glu Glu Gly Glu 180 185 190 480 528 146966·序列表.doe 576 201037078 acc gaa gag cag ttc gtc gcc cgc etc gee ege gag ctg gag gaa aeg 624
Thr Glu Glu Gin Phe Val Ala Arg Leu Ala Arg Glu Leu Glu Glu Thr 195 200 205 ate cag cgc gag ggc gcc gac acc ate gcc ggt ttc ttt gcc gaa ccg 672 lie Gin Arg Glu Gly Ala Asp Thr lie Ala Gly Phe Phe Ala Glu Pro 210 215 220 gtg atg ggc geg ggc ggc gtg att ccc ccg gcc aag ggc tat ttc cag 720
Val Met Gly Ala Gly Gly Val lie Pro Pro Ala Lys Gly Tyr Phe Gin 225 230 235 240 geg ate ctg cca ate ctg cgc aaa tat gac ate ccg gtc ate teg gac 768
Ala lie Leu Pro lie Leu Arg Lys Tyr Asp lie Pro Val lie Ser Asp 245 250 255 gag gtg ate tgc ggt ttc gga cgc acc ggt aac acc tgg ggc tgc gtg 816
Glu Val lie Cys Gly Phe Gly Arg Thr Gly Asn Thr Trp Gly Cys Val 260 265 270 acc tat gac ttt aca ccc gat gca ate ate teg tee aag aat ett aca 864
Thr Tyr Asp Phe Thr Pro Asp Ala lie lie Ser Ser Lys Asn Leu Thr 275 280 285 geg ggc ttt ttc ccc atg ggg geg gtg ate ett ggc ccg gaa ett tee 912
Ala Gly Phe Phe Pro Met Gly Ala Val lie Leu Gly Pro Glu Leu Ser 290 295 300 aaa egg ctg gaa acc gca ate gag geg ate gag gaa ttc ccc cat ggc 960
Lys Arg Leu Glu Thr Ala lie Glu Ala lie Glu Glu Phe Pro His Gly 305 310 315 320 ttt acc gcc teg ggc cat ccg gtc ggc tgt get att geg ctg aaa gca 1008
Phe Thr Ala Ser Gly His Pro Val Gly Cys Ala lie Ala Leu Lys Ala 325 330 335 ate gac gtg gtg atg aat gaa ggg ctg get gag aac gtc cgc cgc ett 1056 lie Asp Val Val Met Asn Glu Gly Leu Ala Glu Asn Val Arg Arg Leu 340 345 350 gee ccc cgt ttc gag gaa agg ctg aaa cat ate gcc gag cgc ccg aac 1104
Ala Pro Arg Phe Glu Glu Arg Leu Lys His lie Ala Glu Arg Pro Asn 355 360 365 ate ggt gaa tat cgc ggc ate ggc ttc atg tgg geg ctg gag get gtc 1152 lie Gly Glu Tyr Arg Gly lie Gly Phe Met Trp Ala Leu Glu Ala Val 370 375 380 aag gac aag gca age aag aeg ccg ttc gac ggc aac ctg teg gtc age 1200
Lys Asp Lys Ala Ser Lys Thr Pro Phe Asp Gly Asn Leu Ser Val Ser 385 390 395 400 gag cgt ate gcc aat acc tgc acc gat ctg ggg ctg att tgc egg ccg 1248
Glu Arg lie Ala Asn Thr Cys Thr Asp Leu Gly Leu lie Cys Arg Pro 405 410 415 ett ggt cag tee gtc gtc ett tgt ccg ccc ttt ate ctg acc gag geg 1296
Leu Gly Gin Ser Val Val Leu Cys Pro Pro Phe lie Leu Thr Glu Ala 420 425 430 146966-序列表.doe 1344 201037078 cag atg gat gag atg ttc gat aaa etc gaa aaa gee ett gat aag gtc Gin Met Asp Glu Met Phe Asp Lys Leu Glu Lys Ala Leu Asp Lys Val 435 440 445 ttt gee gag gtt gee tga 1362
Phe Ala Glu Val Ala 450 <210> 2 <211> 453 <212> PRT <213>河流孤菌 <400> 2
Met Asn Lys Pro Gin Ser Trp Glu Ala Arg Ala Glu Thr Tyr Ser Leu 15 10 15
Tyr Gly Phe Thr Asp Met Pro Ser Leu His Gin Arg Gly Thr Val Val 20 25 30
Val Thr His Gly Glu Gly Pro Tyr lie Val Asp Val Asn Gly Arg Arg 35 40 45
Tyr Leu Asp Ala Asn Ser Gly Leu Trp Asn Met Val Ala Gly Phe Asp 50 55 60
His Lys Gly Leu lie Asp Ala Ala Lys Ala Gin Tyr Glu Arg Phe Pro 65 70 75 80
Gly Tyr His Ala Phe Phe Gly Arg Met Ser Asp Gin Thr Val Met Leu 85 90 95
Ser Glu Lys Leu Val Glu Val Ser Pro Phe Asp Ser Gly Arg Val Phe 100 105 110
Tyr Thr Asn Ser Gly Ser Glu Ala Asn Asp Thr Met Val Lys Met Leu 115 120 125
Trp Phe Leu His Ala Ala Glu Gly Lys Pro Gin Lys Arg Lys lie Leu 130 135 140
Thr Arg Trp Asn Ala Tyr His Gly Val Thr Ala Val Ser Ala Ser Met 145 150 155 160
Thr Gly Lys Pro Tyr Asn Ser Val Phe Gly Leu Pro Leu Pro Gly Phe 165 170 175
Val His Leu Thr Cys Pro His Tyr Trp Arg Tyr Gly Glu Glu Gly Glu 146966-序列表.doc 201037078 180 185 190
Thr Glu Glu Gin Phe Val Ala Arg Leu Ala Arg Glu Leu Glu Glu Thr 195 200 205 lie Gin Arg Glu Gly Ala Asp Thr lie Ala Gly Phe Phe Ala Glu Pro 210 215 220
Val Met Gly Ala Gly Gly Val lie Pro Pro Ala Lys Gly Tyr Phe Gin 225 230 235 240
Ala lie Leu Pro lie Leu Arg Lys Tyr Asp lie Pro Val lie Ser Asp 245 250 255
Glu Val lie Cys Gly Phe Gly Arg Thr Gly Asn Thr Trp Gly Cys Val 260 265 270
Thr Tyr Asp Phe Thr Pro Asp Ala lie lie Ser Ser Lys Asn Leu Thr 275 280 285
Ala Gly Phe Phe Pro Met Gly Ala Val lie Leu Gly Pro Glu Leu Ser 290 295 300
Lys Arg Leu Glu Thr Ala lie Glu Ala lie Glu Glu Phe Pro His Gly 305 310 315 320
Phe Thr Ala Ser Gly His Pro Val Gly Cys Ala lie Ala Leu Lys Ala 325 330 335 lie Asp Val Val Met Asn Glu Gly Leu TVla Glu Asn Val Arg Arg Leu 340 345 350
Ala Pro Arg Phe Glu Glu Arg Leu Lys His lie Ala Glu Arg Pro Asn 355 360 365 lie Gly Glu Tyr Arg Gly lie Gly Phe Met Trp Ala Leu Glu Ala Val 370 375 380
Lys Asp Lys Ala Ser Lys Thr Pro Phe Asp Gly Asn Leu Ser Val Ser 385 390 395 400
Glu Arg lie Ala Asn Thr Cys Thr Asp Leu Gly Leu lie Cys Arg Pro 405 410 415
Leu Gly Gin Ser Val Val Leu Cys Pro Pro Phe lie Leu Thr Glu Ala 420 425 430 -4 146966-序列表.doc 201037078
Gin Met Asp Glu Met Phe Asp Lys Leu Glu Lys Ala Leu Asp Lys Val 435 440 445
Phe Ala Glu Val Ala 450 o
<210> 3 <211> 1362 <212> DNA <213> 人工 <220> <223>河流孤菌JS17(〇-胺基轉移酶密碼子優化基因 <400> 3 atgaataaac cacagtcttg ggaagctcgt gctgaaacct atagcctgta cggctttacc gatatgccgt ctctgcacca gcgtggtact gtagtggtaa cgcacggtga gggcccgtac atcgtggacg ttaatggccg gcgggcttcg accacaaagg ggttatcacg cgttctttgg gttgaagtgt ccccgttcga aacgatacca tggttaagat cgtaaaattc tgacccgttg accggtaaac cgtataacag tgcccgcact actggcgtta ctggctcgtg aactggaaga tttgcggagc ctgttatggg gcaatcctgc cgatcctgcg ggcttcggcc gcaccggtaa atcattagct ctaaaaacct ccagaactgt ctaagcgcct ttcactgcta gcggccatcc atgaacgagg gcctggcgga aaacacattg ctgagcgccc ctggaagcag ttaaagataa gaacgtatcg ctaatacctg ccgttacctg gatgcaaaca cctgatcgat gccgcaaaag ccgtatgagc gaccagactg tagcggtcgt gtcttttaca gctgtggttc ctgcacgcag gaacgcttat cacggtgtga cgtgttcggt ctgccgctgc tggtgaggaa ggcgaaactg aaccattcaa cgcgaaggtg tgccggcggt gtgattccgc caagtacgac attccggtta cacctggggc tgcgtgacgt gactgcgggt ttcttcccta ggaaaccgcc atcgaggcaa ggtaggctgc gcaatcgcgc aaacgtgcgc cgcctggcgc gaacattggc gaatatcgcg agctagcaag accccgttcg tacggacctg ggtctgatct gcggcctgtg gaacatggtt cgcagtacga acgcttcccg tgatgctgag cgaaaaactg ctaactctgg cagcgaggct cggaaggcaa acctcagaaa ctgctgtttc cgcatctatg ctggcttcgt gcatctgacc aggaacagtt cgtggcgcgt cagatactat cgcgggcttc cggcgaaggg ctatttccag tttctgacga agtgatctgc atgacttcac tccggacgca tgggcgccgt aatcctgggc tcgaagagtt cccgcacggt tgaaggcgat cgatgttgtc cgcgttttga agaacgtctg gcatcggttt catgtgggcc acggcaacct gtccgtgagc gccgtccgct gggtcagtcc 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 146966·序列表.doc 1260 201037078 gtagttctgt gcccaccatt tatcctgacc gaagcgcaga tggatgaaat gttcgataaa 1320 ctggagaaag ctctggataa agtgttcgct gaagtcgcgt aa 1362 <210> 4 <211> 406
<212> PRT <213>詹氏甲烷暖球菌DSM2661 <400> 4
Met Thr Lys Val Leu Val Met Phe Met Asp Phe Leu Phe Glu Asn Ser 15 10 15
Trp Lys Ala Val Cys Pro Tyr Asn Pro Lys Leu Asp Leu Lys Asp lie 20 25 30
Tyr lie Tyr Asp Thr Thr Leu Arg Asp Gly Glu Gin Thr Pro Gly Val 35 40 45
Cys Phe Thr Lys Glu Gin Lys Leu Glu lie Ala Arg Lys Leu Asp Glu 50 55 60
Leu Gly Leu Lys Gin lie Glu Ala Gly Phe Pro lie Val Ser Glu Arg 65 70 75 80
Glu Ala Asp lie Val Lys Thr lie Ala Asn Glu Gly Leu Asn Ala Asp 85 90 95 lie Leu Ala Leu Cys Arg Ala Leu Lys Lys Asp lie Asp Lys Ala lie 100 105 110
Glu Cys Asp Val Asp Gly lie lie Thr Phe lie Ala Thr Ser Pro Leu 115 120 125
His Leu Lys Tyr Lys Phe Asn Asn Lys Ser Leu Asp Glu lie Leu Glu 130 135 140
Met Gly Val Glu Ala Val Glu Tyr Ala Lys Glu His Gly Leu Phe Val 145 150 155 160
Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Pro lie Glu Asp Leu lie 165 170 175
Lys Val His Lys Ala Ala Glu Glu Ala Gly Ala Asp Arg Val His lie 180 185 190
Ala Asp Thr Thr Gly Cys Ala Thr Pro Gin Ser Met Glu Phe lie Cys -6- 146966-序列表.doc 201037078 195 200 205
Lys Thr Leu Lys Glu Asn Leu Lys Lys Ala His lie Gly Val His Cys 210 215 220
His Asn Asp Phe Gly Phe Ala Val lie Asn Ser lie Tyr Gly Leu lie 225 230 235 240
Gly Gly Ala Lys Ala Val Ser Thr Thr Val Asn Gly lie Gly Glu Arg 245 250 255
Ala Gly Asn Ala Ala Leu Glu Glu Leu lie Met Ala Leu Thr Val Leu 260 265 270
Tyr Asp Val Asp Leu Gly Leu Asn Leu Glu Val Leu Pro Glu Leu Cys 275 280 285 〇
Arg Met Val Glu Glu Tyr Ser Gly lie Lys Met Pro Lys Asn Lys Pro 290 295 300 lie Val Gly Glu Leu Val Phe Ala His Glu Ser Gly lie His Val Asp 305 310 315 320
Ala Val lie Glu Asn Pro Leu Thr Tyr Glu Pro Phe Leu Pro Glu Lys 325 330 335 lie Gly Leu Lys Arg Asn lie Leu Leu Gly Lys His Ser Gly Cys Arg 340 345 350
Ala Val Ala Tyr Lys Leu Lys Leu Met Gly lie Asp Tyr Asp Arg Glu 355 360 365 o
Met Leu Cys Glu lie Val Lys Lys Val Lys Glu lie Arg Glu Glu Gly 370 375 380
Lys Phe lie Thr Asp Glu Val Phe Lys Glu He Val Glu Glu Val Leu 385 390 395 400
Arg Lys Arg Asn Lys Asn 405
<210> 5 <211> 391 <212> PRT <213>耐熱曱烷熱桿菌DH <400> 5 146966-序列表.doc 201037078
Met Arg Tyr Phe Val Ser Pro Phe Asn Lys Glu Ala Glu Leu Lys Phe 1 5 10 15
Pro Asp Arg lie Thr lie Tyr Asp Thr Thr Leu Arg Asp Gly Glu Gin 20 25 30
Thr Pro Gly Val Cys Leu Gly Thr Glu Glu Lys Leu Glu lie Ala Arg 35 40 45
Lys Leu Asp Glu Leu Gly lie His Gin lie Glu Ser Gly Phe Pro Val 50 55 60
Val Ser Glu Gin Glu Arg Val Ser Val Lys Ser lie Ala Asn Glu Gly 65 70 75 80
Leu Asn Ala Glu lie Leu Ala Leu Cys Arg Thr Lys Lys Asp Asp lie 85 90 95
Asp Ala Ala lie Asp Cys Asp Val Asp Gly Val lie Thr Phe Met Ala 100 105 110
Thr Ser Asp Leu His Leu Lys His Lys Leu Lys Leu Thr Arg Glu Glu 115 120 125
Ala Leu Asn Val Cys Met Asn Ser lie Glu Tyr Ala Lys Asp His Gly 130 135 140
Leu Phe Leu Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Asp Leu Asp 145 150 155 160
Phe Leu Lys Gin lie Tyr Arg Lys Ala Glu Asn Tyr Gly Ala Asp Arg 165 170 175
Val His 工le Ala Asp Thr Val Gly Ala lie Ser Pro Gin Gly Met Asp 180 185 190
Tyr Leu Val Arg Glu Leu Arg Arg Asp lie Lys Val Asp 工le Ala Leu 195 200 205
His Cys His Asn Asp Phe Gly Met Ala Leu Ser Asn Ser lie Ala Gly 210 215 220
Leu Leu Ala Gly Gly Thr Ala Val Ser Thr Thr Val Asn Gly lie Gly 225 230 235 240 M6966·序列表.doc 201037078
Glu Arg Ala Gly Asn Thr Ser Leu Glu Glu Leu lie Met Ala Leu Arg 245 250 255 lie 工le Tyr Glu Val Asp Leu Gly Phe Asn lie Gly Val Leu Tyr Glu 260 265 270
Leu Ser Arg Leu Val Glu Lys His Thr Arg Met Lys Val Pro Glu Asn 275 280 285
Lys Pro lie Val Gly Arg Asn Val Phe Arg His Glu Ser Gly lie His 290 295 300
Val Asp Ala Val lie Glu Glu Pro Leu Thr Tyr Glu Pro Phe Leu Pro 305 310 315 320
Glu Met lie Gly His Gin Arg Lys lie Val Leu Gly Lys His Ser Gly 325 330 335
Cys Arg Ala Val Lys Ala Lys Leu Glu Glu Tyr Gly lie Asp Val Thr 340 345 350
Arg Asp Glu Leu Cys Arg lie Val Glu Glu Val Lys Lys Asn Arg Glu 355 360 365
Lys Gly Lys Tyr lie Asn Asp Glu Leu Phe Tyr Arg lie Val Lys Ser 370 375 380
Val Arg Gly Pro Val Asp Phe 385 390 <210> 6 <211> 386
<212> PRT <213> 海沼曱烷球菌S2 <400> 6
Met Asp Trp Lys Ala Val Ser Pro Tyr Asn Pro Lys Leu Asp Leu Lys 15 10 15
Asp Cys Tyr Leu Tyr Asp Thr Thr Leu Arg Asp Gly Glu Gin Thr Pro 20 25 30
Gly Val Cys Phe Ala Gly Asp Gin Lys Leu Glu lie Ala Lys Lys Leu 35 40 45
Asp Glu Leu Lys lie Lys Gin lie Glu Ala Gly Phe Pro lie Val Ser 50 55 60 146966-序列表.doc -9- 201037078
Glu Asn Glu Arg Lys Ala lie Lys Ser lie Thr Gly Glu Gly Leu Asn 65 70 75 80
Ala Gin lie Leu Ala Leu Ser Arg Val Leu Lys Glu Asp lie Asp Lys 85 90 95
Ala lie Glu Cys Asp Val Asp Gly lie lie Thr Phe lie Ala Thr Ser 100 105 110
Pro Met His Leu Lys Tyr Lys Leu His Lys Asn Leu Asp Glu Val Glu 115 120 125
Glu Met Gly Met Lys Ala Val Glu Tyr Ala Lys Asp His Gly Leu Phe 130 135 140
Val Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Pro Leu Glu Asp lie 145 150 155 160 lie Arg lie His Lys Asn Ala Glu Glu His Gly Ala Asp Arg Val His 165 170 175 lie Ala Asp Thr Leu Gly Cys Ala Thr Pro Gin Ala Met Tyr His lie 180 185 190
Cys Ser Glu Leu Ser Lys His Leu Lys Lys Ala His lie Gly Val His 195 200 205
Cys His Asn Asp Phe Gly Phe Ala Val lie Asn Ser lie Tyr Gly Leu 210 215 220 lie Gly Gly Ala Lys Ala Val Ser Thr Thr Val Asn Gly lie Gly Glu 225 230 235 240
Arg Ala Gly Asn Ala Ala lie Glu Glu lie Ala Met Ala Leu Lys Val 245 250 255
Leu Tyr Asp His Asp Met Gly Leu Asn Thr Glu lie Leu Thr Glu lie 260 265 270
Ser Lys Leu Val Glu Asn Tyr Ser Lys lie Lys lie Pro Glu Asn Lys 275 280 285
Pro Leu Val Gly Glu Met Val Phe Tyr His Glu Ser Gly lie His Val 290 295 300 -10- 146966-序列表.doc 201037078
Asp Ala Val Leu Glu Asn Pro Leu Thr Tyr Glu Pro Phe Leu Pro Glu 305 310 315 320
Lys lie Gly Gin Lys Arg Lys lie lie Leu Gly Lys His Ser Gly Cys 325 330 335
Arg Ala Val Ala His Arg Leu Gin Glu Leu Gly Leu Glu Ala Ser Arg 340 345 350
Asp Glu Leu Trp Glu lie Val Lys Lys Thr Lys Glu Thr Arg Glu Asp 355 360 365
Gly Thr Glu lie Ser Asp Glu Val Phe Lys Asn lie Ala Glu Lys lie 370 375 380
lie Lys 385 <210> . 7 <211> 386
<212> PRT <213> 海沼曱烷球菌C5 <400> 7
Met Asp Trp Lys Ala Val Ser Pro Tyr Asn Pro Lys Leu Asn Leu Lys 1 5 10 15
Asp Cys Tyr Leu Tyr Asp Thr Thr Leu Arg Asp Gly Glu Gin Thr Pro 20 25 30
Gly Val Cys Phe Thr His Asp Gin Lys Leu Glu lie Ala Lys Lys Leu 35 40 45 ❹
Asp Glu Leu Lys lie Lys Gin lie Glu Ala Gly Phe Pro lie Val Ser 50 55 60
Glu Asn Glu Arg Lys Ala lie Lys Ser lie Thr Gly Glu Gly Leu Asn 65 70 75 80
Ala Gin lie Leu Ala Leu Ser Arg Val Leu Lys Glu Asp lie Asp Lys 85 90 95
Ala lie Glu Cys Asp Val Asp Gly lie lie Thr Phe lie Ala Ala Ser 100 105 110
Pro Met His Leu Lys Tyr Lys Leu His Lys Ser Leu Asp Glu Val Glu -11 - 146966-序列表.doc 201037078 115 120 125
Glu Met Gly Met Lys Ala Val Glu Tyr Ala Lys Asp His Gly Leu Phe 130 135 140
Val Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Pro Val Glu Asp Leu 145 150 155 160 lie Arg lie His Lys Asn Ala Glu Glu His Gly Ala Asn Arg Val His 165 170 175 lie Ala Asp Thr Leu Gly Cys Ala Thr Pro Gin Ala Met Tyr His lie 180 185 190
Cys Ser Glu Leu Ser Ser Asn Leu Lys Lys Ala His lie Gly Val His 195 200 205
Cys His Asn Asp Phe Gly Phe Ala Val lie Asn Ser lie Tyr Gly Leu 210 215 220 工le Gly Gly Ala Lys Ala Val Ser Thr Thr Val Asn Gly lie Gly Glu 225 230 235 240
Arg Ala Gly Asn Ala Ala lie Glu Glu lie Val Met Ala Leu Lys Val 245 250 255
Leu Tyr Asp His Asp Met Gly Leu Asn Thr Glu lie Leu Thr Glu lie 260 265 270
Ser Lys Leu Val Glu Asn Tyr Ser Lys lie Arg lie Pro Glu Asn Lys 275 280 285
Pro Leu Val Gly Glu Met Ala Phe Tyr His Glu Ser Gly lie His Val 290 295 300
Asp Ala Val Leu Glu Asn Pro Leu Thr Tyr Glu Pro Phe Leu Pro Glu 305 310 315 320
Lys lie Gly Gin Lys Arg Lys lie lie Leu Gly Lys His Ser Gly Cys 325 330 335
Arg Ala Val Ala His Arg Leu Gin Glu Leu Gly Leu Glu Ala Ser Arg 340 345 350
Glu Glu Leu Trp Glu lie Val Lys Lys Thr Lys Glu Thr Arg Glu Glu 355 360 365 -12- 146966-序列表.doc 201037078
Gly Thr Glu lie Ser Asp Glu Val Phe Lys Asn lie Val Asp Lys lie 370 375 380 lie Lys 385
> > > > 0 12 3 1111 2 2 2 2 V < < V 8 386 PRT 海沼甲烷球菌C7 <400> 8 Met Asp Trp Lys Ala Val Ser Pro Tyr Asn Pro Lys Leu Asp Leu Lys 15 10 15 Ο
Asp Cys Tyr Leu Tyr Asp Thr Thr Leu Arg Asp Gly Glu Gin Thr Pro 20 25 30
Gly Val Cys Phe Thr His Asp Gin Lys Leu Glu lie Ala Lys Lys Leu 35 40 45
Asp Glu Leu Lys lie Lys Gin lie Glu Ala Gly Phe Pro lie Val Ser 50 55 60
Glu Asn Glu Arg Lys Cys lie Lys Ser lie Thr Gly Glu Gly Leu Asn 65 70 75 80
Ala Gin lie Leu Ala Leu Ser Arg Val Leu Lys Glu Asp lie Asp Lys 85 90 95 ο
Ala lie Glu Cys Asp Val Asp Gly lie lie Thr Phe lie Ala Ala Ser 100 105 110
Pro Met His Leu Lys Tyr Lys Leu His Lys Ser Leu Asp Glu Val Glu 115 120 125
Glu Met Gly Met Lys Ala Val Glu Tyr Ala Lys Asp His Gly Leu Phe 130 135 140
Val Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Pro lie Glu Asp lie 145 150 155 160 lie Arg lie His Lys Asn Ala Glu Glu His Gly Ala Asp Arg Val His 165 170 175 146966·序列表.doc 13· 201037078 lie Ala Asp Thr Leu Gly Cys Ala Thr Pro Gin Ser Met Tyr Tyr lie 180 185 190
Cys Ser Glu Leu Ser Lys His Leu Lys Lys Ala His lie Gly Val His 195 200 205
Cys His Asn Asp Phe Gly Phe Ala Val lie Asn Ser lie Tyr Gly Leu 210 215 220
Leu Gly Gly Ala Lys Ala Val Ser Thr Thr Val Asn Gly lie Gly Glu 225 230 235 240
Arg Ala Gly Asn Ala Ala lie Glu Glu lie Val Met Ala Leu Lys Val 245 250 255
Leu Tyr Asp Tyr Asp Met Gly Leu Asn Thr Glu lie Leu Thr Glu Met 260 265 270
Ser Lys Leu Val Glu Lys Tyr Ser Lys lie Arg lie Pro Glu Asn Lys 275 280 285
Pro Leu Val Gly Glu Met Ala Phe Tyr His Glu Ser Gly lie His Val 290 295 300
Asp Ala Val Leu Glu Asn Pro Leu Thr Tyr Glu Pro Phe Leu Pro Glu 305 310 315 320
Lys lie Gly Gin Lys Arg Lys lie He Leu Gly Lys His Ser Gly Cys 325 330 335
Arg Ala Val Ala His Arg Leu Gin Glu Leu Gly Leu Glu Thr Ser Arg 340 345 350
Asn Glu Leu Trp Glu lie Val Lys Lys Thr Lys Glu Thr Arg Glu Glu 355 360 365
Gly Thr Glu lie Ser Asp Glu Val Phe Lys Asn lie Val Asp Lys lie 370 375 380 lie Lys 385 <210> 9
<211> 279 <212> PRT <213>思氏甲烷球形菌DSM 3091 146966-序列表.doc -14- 201037078 <400> 9
Met Gly Leu Ser Asp Leu His Leu Glu Val Lys lie Asn Lys Pro Arg 15 10 15
Asp Val Val Asn Gin lie Cys Met Asp Ala 工le Asp Tyr Gly Lys Asp 20 25 30
His Gly Leu Phe Val Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Glu 35 40 45
Leu Pro Lys Leu Leu Asp Val Tyr Lys Gin Ala Gin Asp His Gly Ala 50 55 60
Asp Arg lie His lie Ala Asp Thr Thr Gly Ser lie Asn Pro Tyr Ala 65 70 75 80
Thr Gin Tyr Leu Val Lys Asn lie Lys Lys Glu lie Asp Thr Glu lie 85 90 95
Ala Leu His Cys His Asn Asp Phe Gly Phe Ala Val Ala Asn Ser lie 100 105 110
Ala Gly Leu Phe Glu Gly Ala Thr Ala lie Ser Thr Thr Val Asn Gly 115 120 125 lie Gly Glu Arg Ala Gly Asn Ala Ser Leu Glu Glu Leu lie Met Ser 130 135 140
Leu Lys Leu Leu Tyr Asn Lys Asp Leu Gly Phe Lys Thr Glu Val lie 145 150 155 160
Tyr Glu Leu Ser Gin Leu Val Ser Lys Tyr Ser Lys lie Pro lie Ser 165 170 175
Asp Ser Lys Ala lie Val Gly Asn Asn Val Phe Arg His Glu Ser Gly 180 185 190 lie His Val Asp Ala lie Val Lys Asn Pro Leu Ala Tyr Glu Pro Phe 195 200 205 lie Pro Glu Met lie Gly Thr Lys Arg Gin lie Val Leu Gly Lys His 210 215 220
Ser Gly Lys Ser Ala Val lie Glu Lys Leu Asp Thr Leu Asn lie Lys 225 230 235 240 15- 146966-序列表.doc 201037078
Val Asp Asp Thr Gin Leu Ser Gin lie Val Ser Leu Val Lys Gin Glu 245 250 255
Arg Glu Arg Gly Glu Glu lie Thr Asn Asn Lys Phe Asp Glu lie Leu 260 265 270
Glu Lys Val Asn lie Lys Arg 275 <210> 10 <211> 397
<212> PRT <213>坎氏曱烷嗜熱菌AV19 <400> 10
Met Gin Ser Pro Tyr Val Arg Glu Ala Val Arg Glu Met Asp Leu Pro 15 10 15
Asp Glu Val lie Val Tyr Asp Thr Thr Leu Arg Asp Gly Glu Gin Thr 20 25 30
Pro Gly Val Ser Phe Thr Pro Glu Gin Lys Leu Glu 工le Ala His Leu 35 40 45
Leu Asp Glu Leu Gly Val Gin Gin He Glu Ala Gly Phe Pro Val Val 50 55 60
Ser Glu Gly Glu Arg Asp Ala Val Arg Arg lie Ala His Glu Gly Leu 65 70 75 80
Asn Ala Asp lie Leu Cys Leu Ala Arg Thr Leu Arg Gly Asp Val Asp 85 90 95
Ala Ala Leu Asp Cys Asp Val Asp Gly Val lie Thr Phe lie Ala Thr 100 105 110
Ser Glu Leu His Leu Lys His Lys Leu Arg Met Ser Arg Glu Glu Val 115 120 125
Leu Glu Arg lie Ala Asp Thr Val Glu Tyr Ala Lys Asp His Gly Leu 130 135 140
Trp Val Ala Phe Ser Ala Glu Asp Gly Thr Arg Thr Glu Phe Glu Phe 145 150 155 160
Leu Glu Arg Val Tyr Arg Thr Ala Glu Glu Cys Gly Ala Asp Arg Val •16- 146966-序列表.doc 201037078 165 170 175
His Ala Thr Asp Thr Val Gly Val Met lie Pro Ala Ala Met Arg Leu 180 185 190
Phe Val Ala Lys lie Arg Glu Val Val Asp Leu Pro lie Gly Val His 195 200 205
Cys His Asp Asp Phe Gly Met Ala Val Ala Asn Ser Leu Ala Ala Val 210 215 220
Glu Ala Gly Ala Gin Ala lie Ser Thr Thr Val Asn Gly lie Gly Glu 225 230 235 240
Arg Ala Gly Asn Ala Ala Leu Glu Glu Val lie Met Ala Leu Lys Glu 245 250 255
O
Leu Tyr Gly He Asp Pro Gly Phe Asn Thr Glu Val Leu Ala Glu Leu 260 265 270
Ser Arg Lys Val Ser Glu Tyr Ser Gly He Asp Val Pro Pro Asn Lys 275 280 285
Ala Val Val Gly Glu Asn Ala Phe Arg His Glu Ser Gly lie His Val 290 295 300
Ala Ala Val Leu Glu Glu Pro Arg Thr Tyr Glu Pro lie Asp Pro Lys 305 310 315 320
Glu Val Gly Met Asn Arg Lys He Val Leu Gly Lys His Thr Gly Arg 325 330 335 〇
Lys Ala Val Val Ala Lys Leu Glu Glu Leu Gly Val Glu Pro Glu Glu 340 345 350
Glu lie Val Glu Glu Val Leu Lys Arg lie Lys Ala Leu Gly Asp Arg 355 360 365
Arg Val Arg Val Thr Asp Ser Lys Leu Glu Glu lie Val Arg Asn Val 370 375 380
Leu Glu Ser Arg Gly Asp Arg Asp Asp Pro Gly Ser Arg 385 390 395 <210> 11 <211> 390 17· 146966-序列表.doc 201037078
<212> PRT <213>史氏甲烷短桿菌ATCC35061 <400> 11
Met Gin Tyr Tyr lie Ser His Tyr Asn Lys Glu Pro Glu Leu Asn Phe 15 10 15
Pro Asp Glu lie Thr Val Tyr Asp Thr Thr Leu Arg Asp Gly Glu Gin 20 25 30
Thr Pro Gly Val Cys Phe Ser Pro Glu Glu Lys Leu Glu 工le Ala Lys 35 40 45
Lys Leu Asp Glu Val Lys lie Lys Gin lie Glu Ala Gly Phe Pro lie 50 55 60
Val Ser Lys Lys Glu Gin Glu Ser Val Lys Ala lie Thr Ser Glu Gly 65 70 75 80
Leu Asn Ala Gin lie lie Ser Leu Ser Arg Thr Lys Lys Glu Asp lie 85 90 95
Asp Ala Ala Leu Asp Cys Asp Val Asp Gly Val 工le Thr Phe Met Gly 100 105 110
Thr Ser Asp 工le His Leu Glu His Lys Met His lie Gly Arg Gin Glu 115 120 125
Ala Leu Asn Thr Cys Met Asn Ala lie Glu Tyr Ala Lys Asp His Gly 130 135 140
Leu Phe Val Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Asp Leu Asp 145 150 155 160
Phe Leu Lys Arg lie Tyr Asn Lys Ala Glu Ser Tyr Gly Ala Asp Arg 165 170 175
Val His lie Ala Asp Thr Thr Gly Ala lie Thr Pro Gin Gly lie Thr 180 185 190
Tyr Leu Val Lys Glu Leu Lys Lys Asp Val Asn lie Asp 工le Ala Leu 195 200 205
His Cys His Asn Asp Phe Gly Leu Ala Val lie Asn Ser lie Ser Gly 210 215 220 146966-序列表.doc - 18 - 201037078
Val Leu Ala Gly Ala Asn Gly lie Ser Thr Thr Val Asn Gly lie Gly 225 230 _ 235 240
Glu Arg Ala Gly Asn Ala Ser Leu Glu Glu Val lie Met Ser Leu Lys 245 250 255
Leu Leu Tyr Gly Lys Asp Leu Gly Phe Lys Thr Lys His lie Lys Glu 260 265 270
Leu Ser Glu Leu Val Ser Lys Ala Ser Gly Leu Pro Val Pro Tyr Asn 275 280 285
Lys Pro Val Val Gly Asn Asn Val Phe Arg His Glu Ser Gly lie His 290 295 300 o
Val Asp Ala Val lie Glu Glu Pro Leu Cys Tyr Glu Pro Tyr lie Pro 305 310 315 320
Glu Leu Val Gly Gin Lys Arg Gin Leu Val Leu Gly Lys His Ser Gly 325 330 335
Cys Arg Ala Val Arg Ala Lys Leu Asn Glu Cys Asp Leu Asp Val Ser 340 345 350
Asp Asp Thr Leu lie Glu lie Val Lys Lys Val Lys Lys Ser Arg Glu 355 360 365
Glu Gly Thr Tyr lie Asn Asp Asp Val Phe Lys Glu 工le Val Lys Ser 370 375 380
Cys Asn Tyr Lys Lys Glu 385 390 <210> 12 <211> 386
<212> PRT
<213>萬氏甲炫球菌SB <400> 12
Met Asp Trp Lys Glu Val Ser Gin Tyr Asn Pro Lys Leu Asp Leu Lys 15 10 15
Glu Cys Tyr Val Tyr Asp Thr Thr Leu Arg Asp Gly Glu Gin Thr Pro 20 25 30
Gly Val Cys Phe Thr Gly Asn Gin Lys Leu Glu lie Ala Lys Lys Leu 35 40 45 -19- 146966-序列表.doc 201037078
Asp Asp Leu Gly lie Lys Gin lie 50 55
Glu Ala Gly Phe Pro Thr Val Ser 60
Glu Asn Glu Arg Lys Cys lie Lys Ser lie Ser Ser Glu Gly Leu Asn 65 70 75 80
Ala Asp lie Leu Ala Leu Ser Arg Val Leu Lys Glu Asp lie Asp Arg 85 90 95
Ala lie Glu Cys Asp Val Asp Gly lie lie Thr Phe Val Ala Thr Ser 100 105 110
Pro Met His Leu Lys Tyr Lys Leu His Lys Ser Phe Glu Glu Val Glu 115 120 125
Glu Met Gly Met Lys Ala lie Glu Tyr Ala Lys Asp His Gly Leu Phe 130 135 140
Val Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Ser lie Glu Asn lie 145 150 155 160 lie Lys 工le His Lys Asn Ala Glu Asp Tyr Gly Ala Asp Arg Val His 165 170 175 lie Ala Asp Thr Leu Gly Cys Ala Thr Pro Gin Ser Met Tyr Gin lie 180 185 190
Cys Ser Glu Leu Asn Lys Ser Leu Lys Lys Ala His lie Gly Val His 195 200 205
Cys His Asn Asp 210
Phe Gly Phe Ala Ala lie Asn Ser lie Tyr Gly Leu 215 220
Met Gly Gly Ala Lys Ala Val Ser Thr Thr Val Asn Gly lie Gly Glu 225 230 235 240
Arg Ala Gly Asn Ala Ala Leu Glu Glu Val Val Met Ala Leu Lys Val 245 250 255
Leu Tyr Asn Tyr Asp Met Gly Leu Asn Thr Glu Leu lie Met Glu Thr 260 265 270
Ser Lys Leu Val Glu Thr Tyr Ser Lys lie Lys Val Pro Glu Asn Lys 275 280 285 20- 146966、序列表.doc 201037078
Pro Leu Val Gly Glu Met Val Phe Tyr His Glu Ser Gly lie His Val 290 295 300
Asp Ala Val Leu Glu Asn Pro Leu Thr Tyr Glu Pro Phe Leu Pro Glu 305 310 315 320
Lys lie Gly Gin Lys Arg Lys lie Val Leu Gly Lys His Ser Gly Cys 325 330 335
Arg Ala Val Ala Tyr Arg Leu Asn Glu Leu Gly Phe Glu Ala Thr Arg 340 345 350
Asp Glu Leu Trp Glu lie Val Lys Lys Thr Lys Glu Thr Arg Glu Gin 355 360 365 ❹
Gly Thr Glu lie Ser Asp Glu Val Phe Lys Asn lie Val Thr His lie 370 375 380
Leu Asn 385 <210> 13 <211> 387
<212> PRT <213>雜色甲烷球菌Nankai3 <400> 13
Met Asn Trp Lys Glu Val Cys Gin Tyr Asn Pro Lys Leu Asn Leu Glu 15 10 15
Asp Cys Tyr lie Tyr Asp Thr Thr Leu Arg Asp Gly Glu Gin Thr Pro 20 25 30 〇
Gly Val Cys Phe Ser Met Glu Gin Lys Leu Asp lie Ala Lys Lys Leu 35 40 45
Asp Glu Leu Gly Val Lys Gin lie Glu Ala Gly Phe Pro Ala Val Ser 50 55 60
Lys Ser Glu lie Glu Asn Val Lys Lys lie Ala Asn Glu Gly Leu Asn 65 70 Ί5 80
Ala Glu lie Leu Ala Leu Ser Arg Ala Leu Gin Gly Asp lie Asp Lys 85 90 95
Ala Leu Ser Cys Asp Val Asp Gly lie lie Thr Phe lie Ala Ala Ser -21 - 146966-序列表.doc 201037078 100 105 110
Pro Leu His Leu Lys Tyr Lys Leu His Lys Ser lie Glu Glu Val Glu 115 120 125
Glu Met Gly Met Lys Ala Val Glu Tyr Ala Lys Asp His Gly Leu Phe 130 135 140
Val Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Pro lie Glu Asp Leu 145 150 155 160
Val Arg lie His Lys Asn Ala Glu Glu His Gly Ala Asp Arg Val His 165 170 175 lie Ala Asp Thr Thr Gly Cys Gly Thr Pro Gin Ser lie Gin Tyr lie 180 185 190
Cys Ser Glu Leu Ser Asn Asn Leu Lys Lys Ala His lie Gly Val His 195 200 205
Cys His Asn Asp Phe Gly Leu Ala Val lie Asn Ser lie Tyr Gly Leu 210 215 220
Leu Gly Gly Ala Lys Ala Ala Ser Thr Thr Val Asn Gly lie Gly Glu 225 230 235 240
Arg Ala Gly Asn Ala Pro Leu Glu Glu Leu Leu Leu Thr Met Asn Val 245 250 255
Leu Tyr Asp Val Lys Thr Asp Leu Asn lie Ser lie lie Lys Glu Leu 260 265 270
Ser Thr Met Val Glu Asn Tyr Ser Gly lie Lys lie Pro Val Asn Lys 275 280 285
Pro lie Val Gly Asp Lys Val Phe Tyr His Glu Ser Gly lie His Val 290 295 300
Asp Ala Val lie Glu Asn Pro Leu Thr Tyr Glu Pro Phe Leu Pro Glu 305 310 315 320
Arg lie Gly Gin Lys Arg Glu lie Val Leu Gly Lys His Ser Gly Cys 325 330 335
Ser Ala Val Glu Ser Lys Leu Lys Glu Leu Gly Leu Glu Val Pro Lys 340 345 350 -22· 146966·序列表.doc 201037078
Asp Arg lie Trp Asp Leu Val Lys Lys Val Lys Thr Thr Arg Glu Gly 355 360 365
Gly Glu Asp lie Asp Asp Glu Met Phe lie Lys lie Val Asp lie lie 370 375 380
Asn Lys Gin 385 <210> 14 <211> 420
<212> PRT <213> 詹氏甲烷暖球菌DSM2661 <400> 14
Met Thr Leu Val Glu Lys lie Leu Ser Lys Lys Val Gly Tyr Glu Val
Cys Ala Gly Asp Ser lie Glu Val Glu Val Asp Leu Ala Met Thr His 20 25 30
Asp Gly Thr Thr Pro Leu Ala Tyr Lys Ala Leu Lys Glu Met Ser Asp 35 40 45
Ser Val Trp Asn Pro Asp Lys lie Val Val Ala Phe Asp His Asn Val 50 55 60
Pro Pro Asn Thr Val Lys Ala Ala Glu Met Gin Lys Leu Ala Leu Glu 65 70 75 80
Phe Val Lys Arg Phe Gly lie Lys Asn Phe His Lys Gly Gly Glu Gly 85 90 95 lie Cys His Gin lie Leu Ala Glu Asn Tyr Val Leu Pro Asn Met Phe 100 105 110
Val Ala Gly Gly Asp Ser His Thr Cys Thr His Gly Ala Phe Gly Ala 115 120 125
Phe Ala Thr Gly Phe Gly Ala Thr Asp Met Ala Tyr lie Tyr Ala Thr 130 135 140
Gly Glu Thr Trp lie Lys Val Pro Lys Thr lie Arg Val Asp lie Val 145 150 155 160 23· 146966-序列表.doc 201037078
Gly Lys Asn Glu Asn Val Ser Ala Lys Asp lie Val Leu Arg Val Cys 165 170 175
Lys Glu lie Gly Arg Arg Gly Ala Thr Tyr Met Ala lie Glu Tyr Gly 180 185 190
Gly Glu Val Val Lys Asn Met Asp Met Asp Gly Arg Leu Thr Leu Cys 195 200 205
Asn Met Ala lie Glu Met Gly Gly Lys Thr Gly Val lie Glu Ala Asp 210 215 220
Glu lie Thr Tyr Asp Tyr Leu Lys Lys Glu Arg Gly Leu Ser Asp Glu 225 230 235 240
Asp lie Ala Lys Leu Lys Lys Glu Arg lie Thr Val Asn Arg Asp Glu 245 250 255
Ala Asn Tyr Tyr Lys Glu lie Glu lie Asp lie Thr Asp Met Glu Glu 260 265 270
Gin Val Ala Val Pro His His Pro Asp Asn Val Lys Pro lie Ser Asp 275 280 285
Val Glu Gly Thr Glu lie Asn Gin Val Phe lie Gly Ser Cys Thr Asn 290 295 300
Gly Arg Leu Ser Asp Leu Arg Glu Ala Ala Lys Tyr Leu Lys Gly Arg 305 310 315 320
Glu Val His Lys Asp Val Lys Leu lie Val lie Pro Ala Ser Lys Lys 325 330 335
Val Phe Leu Gin Ala Leu Lys Glu Gly lie lie Asp lie Phe Val Lys 340 345 350
Ala Gly Ala Met lie Cys Thr Pro 355 360
His Gin Gly Val Leu Ala Glu Gly 370 375
Arg Asn Phe Lys Gly Arg Met Gly 385 390
Gly Cys Gly Pro Cys Leu Gly Ala 365 Glu lie Cys Leu Ser Thr Thr Asn 380 His lie Asn Ser Tyr lie Tyr Leu 395 400
Ala Ser Pro Lys lie Ala Ala lie Ser Ala Val Lys Gly Tyr lie Thr -24- 146966-序列表 _doc 201037078 405 410 415
Asn Lys Leu Asp 420 <210> 15 <211> 428
<212> PRT
<213>耐熱甲烷熱桿菌DH <400> 15
Met Val Lys Met Asn Met Thr Glu Lys lie Leu Ala Glu Ala Ala Gly 15 10 15
Leu Arg Glu Val Thr Pro Gly Glu lie lie Glu Ala Arg Val Asp Leu 20 25 30
Ala Met Thr His Asp Gly Thr Ser Pro Pro Thr lie Arg Thr Phe Arg 35 40 45
Asp lie Ala Ser Arg Gly Gly Pro Ala Arg Val Trp Asp Pro Glu Arg 50 55 60 lie Val Met Val Phe Asp His Asn Val Pro Ala Asn Thr lie Gly Ala 65 70 75 80
Ala Glu Phe Gin Arg Val Thr Arg Glu Phe Ala Arg Glu Gin Gly lie 85 90 95
Val Asn lie Phe Gin Asn Ala Ala Gly lie Cys His Gin Val Leu Pro 100 105 110
Glu Arg Gly Phe Val Arg Pro Gly Met Val 工le Val Gly Ala Asp Ser 115 120 125
His Thr Cys Thr Tyr Gly Ala Phe Gly Ala Phe Ala Thr Gly Met Gly 130 135 140
Ala Thr Asp Met Ala Met Val Phe Ala Thr Gly Lys Thr Trp Phe Met 145 150 155 160
Val Pro Glu Ala Met Arg lie Glu Val Thr Gly Glu Pro Glu Gly His 165 170 175
Val Tyr Ala Lys Asp Val lie Leu His lie lie Gly Glu lie Gly Val 180 185 190 -25· 146966-序列表.doc 201037078
Asp Gly Ala Thr Tyr Arg Ser Val Glu Phe Thr Gly Asp Thr lie Glu 195 200 205
Ser Met Asp Val Ser Gly Arg Met Thr lie Cys Asn Met Ala Val Glu 210 215 220
Met Gly Ala Lys Asn Gly lie Met Glu Pro Asn Arg Gin Thr Leu Asp 225 230 235 240
Tyr Val Arg Ala Arg Thr Gly Arg Glu Phe Arg Val Tyr Ser Ser Asp 245 250 255
Glu Asp Ser Gin Tyr Leu Glu Asp His His Phe Asp Val Ser Asp Leu 260 265 270
Glu Pro Gin Val Ala Cys Pro Asp 275 280
His Arg Val Glu Gly Thr His lie 290 295
Thr Asn Gly Arg Tyr Glu Asp Leu 305 310
Asp Val Asp Asn Val Tyr Pro Val 285 Asp Glu Ala Phe Leu Gly Ser Cys 300 Lys lie Ala Ala Glu Val lie Gly 315 320
Asp Arg Arg Val His Glu Asp Val Arg Phe lie Val Ser Pro Ala Ser 325 330 335
Arg Glu lie Tyr Leu Lys Ala Leu Glu Asp Gly lie lie Glu Thr Phe 340 345 350 lie Arg Ala Gly Ala lie Val Cys Asn Pro Gly Cys Gly Pro Cys Leu 355 360 365
Gly Ala His Met Gly Val Leu Ala Pro Gly Glu Val Ser lie Ala Thr 370 375 380
Thr Asn Arg Asn Phe Arg Gly Arg Met Gly Asp Pro Ala Ser Ser Val 385 390 395 400
Tyr Leu Ala Asn Pro Ala Val Val Ala Glu Ser Ala lie Glu Gly Val 405 410 415 lie Ser Ala Pro Gin Gin Glu Ala Gly Asn Gly Cys 420 425 -26· 146966-序列表.doc 201037078 <210> 16 <211> 418
<212> PRT <213>海沼甲烷球菌S2 <400> 16
Met Thr Leu Ala Glu Lys lie lie Ser Lys Asn Val Gly Lys Asn Val 1 5 10 15
Tyr Ala Lys Asp Ser Val Glu lie Ser Val Asp lie Ala Met Thr His 20 25 30
Asp Gly Thr Thr Pro Leu Thr Val Lys Ala Phe Glu Gin lie Ser Asp 35 40 45
Lys Val Trp Asp Asn Glu Lys lie Val lie lie Phe Asp His Asn lie 50 55 60 o
Pro Ala Asn Thr Ser Lys Ala Ala Asn Met Gin Val lie Thr Arg Glu 65 70 75 80
Phe lie Lys Lys Gin Gly lie Lys Asn Tyr Tyr Leu Asp Gly Glu Gly 85 90 95 工le Cys His Gin Val Leu Pro Glu Lys Gly His Val Lys Pro Asn Met 100 105 110 lie lie Ala Gly Ala Asp Ser His Thr Cys Thr His Gly Ala Phe Gly 115 120 125
Ala Phe Ala Thr Gly Phe Gly Ala Thr Asp Met Gly Tyr Val Tyr Ala 130 135 140 〇
Thr Gly Lys Thr Trp Leu Arg Val Pro Glu Thr lie Arg Val Asn Val 145 150 155 160
Thr Gly Glu Asn Glu Asn lie Ser Gly Lys Asp lie lie Leu Lys Thr 165 170 175
Cys Lys Glu Val Gly Arg Arg Gly Ala Thr Tyr Met Ser Leu Glu Tyr 180 185 190
Gly Gly Asn Ala Val His Asn Leu Ser Met Asp Glu Arg Met Val Leu 195 200 205
Ser Asn Met Ala lie Glu Met Gly Gly Lys Ala Gly lie lie Glu Ala 210 215 220 27- 146%6·序列表.doc 201037078
Asp Asp Thr Thr Tyr Arg Tyr Leu Glu Asn Ala Gly Val Ser Arg Glu 225 230 235 240
Glu lie Leu Glu Leu Lys Lys Asn Lys lie Thr Val Asp Glu Ser Glu 245 250 255
Glu Asp Tyr Tyr Lys Thr lie Glu Phe Asp lie Thr Gly Met Glu Glu 260 265 270
Gin Val Ala Cys Pro His His Pro Asp Asn Val Lys Gly Val Ser Glu 275 280 285
Val Glu Gly Thr Glu Leu Asn Gin Val Phe lie Gly Ser Cys Thr Asn 290 295 300
Gly Arg Leu Asn Asp Leu Arg lie Ala Ala Lys Tyr Leu Lys Gly Lys 305 310 315 320
Lys Val Asn Glu Asn Thr Arg Leu lie Val lie Pro Ala Ser Lys Ser 325 330 335 lie Phe Lys Glu Ala Leu Asn Glu Gly Leu lie Asp lie Phe Val Asp 340 345 350
Ser Gly Ala Leu lie Cys Thr Pro Gly Cys Gly Pro Cys Leu Gly Ala 355 360 365
His Gin Gly Val Leu Gly Asp Gly Glu Val Cys Leu Ala Thr Thr Asn 370 375 380
Arg Asn Phe Lys Gly Arg Met Gly Asn Thr Asn Ala Gin Val Tyr Leu 385 390 395 400
Ser Ser Pro Lys lie Ala Ala Lys Ser Ala Val Lys Gly Tyr 工le Thr 405 410 415
Asn Glu <210> 17 <211> 418
<212> PRT <213>海沼甲烷球菌C5 <400> 17 -28- 146966-序列表.doc 201037078
Met Thr Leu Ala Glu Lys lie lie Ser Lys Asn Val Gly Lys Asn Val 15 10 15
Tyr Ala Gly Asp Ser Val Glu lie Asp Val Asp Val Ala Met Thr His 20 25 30
Asp Gly Thr Thr Pro Leu Thr Val Lys Ala Phe Glu Gin lie Ser Asp 35 40 45
Lys Val Trp Asp Asn Glu Lys lie Val lie lie Phe Asp His Asn lie 50 55 60
Pro Ala Asn Thr Ser Lys Ala Ala Asn Met Gin Val lie Thr Arg Glu 65 70 75 80
Phe lie Lys Lys Gin Gly lie Lys Asn Tyr Tyr Leu Asp Gly Glu Gly 85 90 95 lie Cys His Gin Val Leu Pro Glu Lys Gly His Val Lys Pro Asn Met 100 105 110 lie lie Ala Gly Ala Asp Ser His Thr Cys Thr His Gly Ala Phe Gly 115 120 125
Ala Phe Ala Thr Gly Phe Gly Ala Thr Asp Met Gly Tyr Val Tyr Ala 130 135 140
Thr Gly Lys Thr Trp Leu Arg Val Pro Glu Thr lie Gin Val Asn Val 145 150 155 160
Thr Gly Glu Asn Glu Asn lie Ser Gly Lys Asp lie lie Leu Lys Thr 165 170 175
Cys Lys Glu Val Gly Arg Arg Gly Ala Thr Tyr Leu Ser Leu Glu Tyr 180 185 190
Gly Gly Asn Ala Val Gin Asn Leu Asp Met Asp Glu Arg Met Val Leu 195 200 205
Ser Asa Met Ala lie Glu Met Gly Gly Lys Ala Gly lie lie Glu Ala 210 215 220
Asp Asp Thr Thr Tyr Lys Tyr Leu Glu Asn Ala Gly Val Ser Arg Glu 225 230 235 240
Glu lie Leu Asn Leu Lys Lys Asn Lys lie Lys Val Asn Glu Ser Glu -29- 146966-序列表.doc 201037078 245 250 255
Glu Asn Tyr Tyr Lys Thr Phe Glu Phe Asp lie Thr Asp Met Glu Glu 260 265 270
Gin lie Ala Cys Pro His His Pro Asp Asn Val Lys Gly Val Ser Glu 275 280 285
Val Ser Gly lie Glu Leu Asp Gin Val Phe lie Gly Ser Cys Thr Asn 290 295 300
Gly Arg Leu Asn Asp Leu Arg lie Ala Ala Lys His Leu Lys Gly Lys 305 310 315 320
Lys Val Asn Glu Ser Thr Arg Leu lie Val lie Pro Ala Ser Lys Ser 325 330 335 lie Phe Lys Glu Ala Leu Lys Glu Gly Leu lie Asp Thr Phe Val Asp 340 345 350
Ser Gly Ala Leu lie Cys Thr Pro Gly Cys Gly Pro Cys Leu Gly Ala 355 360 365
His Gin Gly Val Leu Gly Asp Gly Glu Val Cys Leu Ala Thr Thr Asn 370 375 380
Arg Asn Phe Lys Gly Arg Met Gly Asn Thr Lys Ser Glu Val Tyr Leu 385 390 395 400
Ser Ser Pro Ala lie Ala Ala Lys Ser Ala Val Lys Gly Tyr lie Thr 405 410 415
Asn Glu <210> 18 <211> 418
<212> PRT <213> 海沼曱烷球菌C7 <400> 18
Met Thr Leu Ala Glu Lys lie lie Ser Lys Asn Val Gly Lys Asn Val 15 10 15
Tyr Ala Gly Asp Ser Val Glu lie Asp Val Asp lie Ala Met Thr His 20 25 30 •30 146966-序列表.doc 201037078
Asp Gly Thr Thr Pro Leu Thr Val Lys Ala Phe Glu Gin lie Ser Asp 35 40 45
Lys Val Trp Asp Asn Glu Lys lie Val lie lie Phe Asp His Asn lie 50 55 60
Pro Ala Asn Thr Ser Lys Ala Ala Asn Met Gin Val lie Thr Arg Glu 65 70 75 80
Phe lie Lys Lys His Gly lie Lys Asn Tyr Tyr Leu Asp Gly Glu Gly 85 90 95 lie Cys His Gin Val Leu Pro Glu Lys Gly His Val Lys Pro Asn Met 100 105 110 o lie lie Ala Gly Ala Asp Ser His Thr Cys Thr His Gly Ala Phe Gly 115 120 125
Ala Phe Ala Thr Gly Phe Gly Ala Thr Asp Met Gly Phe Val Tyr Ala 130 135 140
Thr Gly Lys Thr Trp Leu Arg Val Pro Glu Thr lie Arg Val Asn Val 145 150 155 160
Thr Gly Glu Asn Glu Asn lie Ser Gly Lys Asp lie lie Leu Lys Thr 165 170 175
Cys Lys Glu Val Gly Arg Ser Gly Ala Thr Tyr Met Ser Leu Glu Tyr 180 185 190
Gly Gly Asn Ala Val Gin Asn Leu Glu Met Asn Glu Arg Met Val Leu 195 200 205
Ser Asn Met Ala lie Glu Met Gly Gly Lys Ala Gly lie lie Glu Ala 210 215 220
Asp Asp Thr Thr Tyr Lys Tyr Leu Glu Asn Ala Gly Val Ser Arg Glu 225 230 235 240
Glu lie Leu Asn Leu Lys Lys Asn Lys lie Thr Val Asn Glu Ser Glu 245 250 255
Glu Asn Tyr Tyr Lys Thr lie Glu Phe Asp lie Thr Asp Met Glu Glu 260 265 270 -31 · 146966-序列表.doc 201037078
Gin lie Ala Cys Pro His Asn Pro Asp Asn Val Lys Gly Val Ser Glu 275 280 285
Val Ser Gly Thr Glu Leu Asp Gin Val Phe lie Gly Ser Cys Thr Asn 290 295 300
Gly Arg Leu Asn Asp Leu Arg lie Ala Ala Lys Tyr Leu Lys Gly Lys 305 310 315 320
Lys Val Asn Glu Asn Thr Arg Leu lie Val lie Pro Ala Ser Lys Ser 325 330 335 lie Phe Ala Gly Ala Leu Lys Glu Gly Leu lie Asp lie Phe Val Glu 340 345 350
Ser Gly Ala Leu lie Cys Thr Pro Gly Cys Gly Pro Cys Leu Gly Ala 355 360 365
His Gin Gly Val Leu Gly Asp Gly Glu Val Cys Leu Ala Thr Thr Asn 370 375 380
Arg Asn Phe Lys Gly Arg Met Gly Asn Thr Lys Ala Glu Val Tyr Leu 385 390 395 400
Ser Ser Pro Lys lie Ala Ala Lys Ser Ala Val Lys Gly Tyr lie Thr 405 410 415
Asn Glu <210> 19 <211> 415
<212> PRT <213> 思氏甲烷球形菌DSM3091 <400> 19
Met Asn lie Ser Glu Lys lie Leu Ala Lys Ala Ser Asn Lys Glu Glu 15 10 15
Val Ser Pro Gly Asp Thr lie Thr Ala Asn lie Asp Val Ala Met Ser 20 25 30
His Asp Gly Thr Ser Pro Pro Thr 工le Lys Val Phe Glu Lys 工le Ala 35 40 45
Asp Lys Val Trp Asp Pro Glu Lys 工le Val Leu Val Phe Asp His Val 50 55 60 -32- 146966·序列表.doc 201037078 lie Pro Ala Asn Thr lie Gly Ser Ala Glu Phe Gin Gin Val Val Arg 65 70 75 80
Glu Phe Gly Lys Lys Gin Lys lie Pro Asn Met Tyr lie Gin Gly Glu 85 90 95
Gly Val Cys His Glu Val Leu Pro Asp Tyr Gly His Val Lys Pro Ser 100 105 110
Thr Val lie Val Gly Ala Asp Ser His Thr Cys Thr Tyr Gly Ala Phe 115 120 125
Gly Ala Phe Ser Thr Gly Leu Gly Ala Thr Asp Leu Ala Met Val Tyr 130 135 140
Ala Thr Gly Gin Thr Trp Phe Asn Val Pro Glu Ser Leu Lys lie Asn
145 150 155 160
Val Asn Gly Thr Leu Asn Glu Asn Val Tyr Ser Lys Asp Val lie Leu 165 170 175
Lys lie lie Lys Glu Leu Gly Ala Tyr Gly Ala Thr Tyr Lys Ser Leu 180 185 190
Glu Phe His Gly Asp Thr lie Asp Asn Met Ser Val Ala Ser Arg Leu 195 200 205
Thr Met Thr Asn Met Ala lie Glu Cys Gly Ala Lys Asn Gly lie Met 210 215 220
Val Pro Asn Lys Gin Thr Lys Glu Tyr Leu Ser Gin Arg Gly lie Thr 225 230 235 240
Asp Tyr Thr lie Thr Thr Ala Ser Lys Asp Ala Glu Tyr Glu Lys lie 245 250 255
Tyr Asp Phe Asp Val Asp Asp Leu Gin Pro Gin lie Ala Cys Pro His 260 265 270
Asn Val Asp Asn Val Glu Asp lie Asp Lys Val Ala Gly Thr His lie 275 280 285
Asp Gin Ala Val Leu Gly Ser Cys Thr Asn Gly Arg Tyr Glu Asp Leu 290 295 300 •33· 146966-序列表.doc 201037078
Leu Gin Ala Ala Glu Val lie Glu Gly His Lys lie His Glu Asp Val 305 310 315 320
Glu Leu Leu Val Phe Pro Ala Ser Arg His Val Tyr Glu Lys Ala lie 325 330 335
Glu Thr Gly Val lie Gin Thr Leu Leu Lys Ser Asn Ala lie lie Cys 340 345 350
Asn Pro Gly Cys Gly Pro Cys Leu Gly Ala His Met Gly Val Met Thr 355 360 365
Asp Asp Met Thr Cys lie Ser Thr Thr Asn Arg Asn Phe Leu Gly Arg 370 375 380
Met Gly Ser Ala Lys Ser Tyr Val Tyr Leu Ser Asn Pro Ala Val Val 385 390 395 400
Ala Ala Ser Ala lie Lys Gly Glu lie Thr Asn Pro Ser Glu lie 405 410 415 <210> 20 <211> 418
<212> PRT <213>坎氏甲烷嗜熱菌AV19 <400> 20
Met Gly Lys Thr Met Ala Glu Lys lie Leu Ser Arg Ala Ser Gly Glu 15 10 15
Asp Ala Glu Ala Gly Asp lie Val Val Ala Asn lie Asp Val Ala Met 20 25 30
Val His Asp lie Thr Gly Pro lie Thr Val Gin Arg Leu Glu Glu Net 35 40 45
Gly Val Glu Arg Val Trp Asp Pro Ser Lys lie Val Val Leu Phe Asp 50 55 60
His Gin Val Pro Ala Asp Ser Val Glu Ala Ala Glu Asn His Lys lie 65 70 75 80
Met Arg Glu Phe Val 85
Glu Glu Gin Gly lie Glu His Phe Tyr Asp Val 90 95
Arg Glu Gly Val Cys His Gin Val Leu Pro Glu Lys Gly His Val Arg 34· 146966·序列表.doc 201037078 100 105 110
Pro Gly Asp Val lie Val Gly Ala Asp Ser His Thr Cys Thr His Gly 115 120 125
Ala Leu Gly Ala Phe Ala Thr Gly lie Gly Ser Thr Asp Met Ala Ala 130 135 140
Val Phe Ala Thr Gly Lys Leu Trp Phe Arg Val Pro Glu Thr Tyr Arg 145 150 155 160
Val Glu lie Thr Gly Glu Leu Pro Glu Gly Val Tyr Ala Lys Asp Val 165 170 175
Val Leu Lys Val Thr Gly Glu lie Gly Ala Asp Gly Ala Thr Tyr Met ▲ 180 185 190 ❹
Ala lie Glu Tyr His Gly Glu Val Val Arg Glu Met Ser Val Ser Asp 195 200 205
Arg Met Cys Leu Cys Asn Met Ala lie Glu Met Gly Ala Lys Thr Gly 210 215 220
Met Val Pro Pro Asp Glu Lys Thr Leu Glu Tyr Val Lys Lys Arg Ala 225 230 235 240
Gly Thr Glu Gly Arg Pro Val Glu Pro Asp Pro Asp Ala Arg Tyr Glu 245 250 255
Ala Glu Leu Thr Leu Asp Val Ser Asp Leu Glu Pro Gin Val Ala Lys 260 265 270
G
Pro Phe Ser Pro Asp Asn Val Val Pro Val Gly Glu Val Glu Gly lie 275 280 285
Ala lie Asp Gin Val Phe lie Gly Ser Cys Thr Asn Gly Arg Tyr Glu 290 295 300
Asp Leu Lys Val Ala Ala Glu Val Leu Glu Gly Glu Glu Val His Asp 305 310 315 320
Asp Val Arg Leu lie Val lie Pro Ala Ser Arg Glu Val Tyr His Arg 325 330 335
Thr Leu Lys Asp Gly Val Leu Glu Val Leu His Glu Ala Gly Ala Leu 340 345 350 -35· 146966-序列表.doc 201037078 lie Cys Pro Pro Asn Cys Gly Pro Cys Leu Gly Gly His Met Gly Val 355 360 365
Leu Ala Glu Gly Glu Arg Cys Val Ala Thr Ser Asn Arg Asn Phe Pro 370 375 380
Gly Arg Met Gly His Arg Glu Ser Glu Val Tyr Leu Ala Ser Pro Ala 385 390 395 400
Thr Ala Ala Ala Ser Ala lie Glu Gly Glu lie Thr Asp Pro Arg Pro 405 410 415
Tyr Leu <210> 21 <211> 417
<212> PRT <213> 史氏甲烷短桿菌ATCC35061 <400> 21
Met Asn lie Thr Glu Lys lie Leu Ser Ala Lys Ala Lys Lys Glu Val 15 10 15
Thr Pro Gly Glu lie lie Glu lie Pro Val Asp Leu Ala Met Ser His 20 25 30
Asp Gly Thr Ser Pro Pro Ala lie Lys Thr Phe Glu Lys Val Ala Thr 35 40 45
Lys Val Trp Asp Asn Glu Lys lie Ala lie Val Phe Asp His Asn Val 50 55 60
Pro Ala Asn Thr lie Gly Ser Ala Glu Phe Gin Lys Val Cys Arg Asp 65 70 75 80
Phe lie Lys Lys Gin Lys lie Thr Lys Asn Tyr lie His Gly Asp Gly 85 90 95 lie Cys His Gin Val Leu Pro Glu Lys Gly Leu Val Glu Pro Gly Lys 100 105 110
Val lie Val Gly Ala Asp Ser His Thr Cys Thr Tyr Gly Ala Tyr Gly 115 120 125 -36- 146966-序列表.doc 201037078
Ala Phe Ser Thr Gly Met Gly Ala Thr Asp Leu Ala Met Val Tyr Ala 130 135 140
Thr Gly Lys Thr Trp Phe Met Val Pro Glu Ala lie Lys Met Glu Val 145 150 155 160
Ser Gly Glu Leu Asn Ser Tyr Thr Ala Pro Lys Asp lie lie Leu Lys 165 170 175 lie lie Gly Glu Val Gly lie Ala Gly Ala Thr Tyr Lys Thr Ala Glu 180 185 190
Phe Cys Gly Glu Thr lie Glu Lys Met Gly Val Glu Gly Arg Ala Thr 195 200 205
工le Cys Asn Met Ala lie Glu Met Gly Ala Lys Asn Gly lie Met Glu 210 215 220
Pro Asn Lys Glu Val lie Gin Tyr Val Ser Gin Arg Thr Gly Lys Lys 225 230 235 240
Glu Ser Glu Leu Asn lie Val Lys Ser Asp Glu Asp Ala Gin Tyr Ser 245 250 255
Glu Glu Met His Phe Asp lie Thr Asp Met Glu Pro Gin lie Ala Cys 260 265 270
Pro Asn Asp Val Asp Asn Val Lys Asp lie Ser Lys Val Glu Gly Thr 275 280 285
Ala Val Asp Gin Cys Leu lie Gly Ser Cys Thr Asn Gly Arg Leu Ser 290 295 300
Asp Leu Lys Asp Ala Tyr Glu lie Leu Lys Asp Asn Glu lie Asn Asn 305 310 315 320
Asp Thr Arg Leu Leu lie Leu Pro Ala Ser Ala Glu lie Tyr Lys Gin 325 330 335
Ala lie His Glu Gly Tyr 工le Asp Ala Phe lie Asp Ala Gly Ala lie 340 345 350 lie Cys Asn Pro Gly Cys Gly Pro Cys Leu Gly Gly His Met Gly Val 355 360 365
Leu Ser Glu Gly Glu Thr Cys Leu Ser Thr Thr Asn Arg Asn Phe Lys -37- 146966-序列表.doc 201037078 370 375 380
Gly Arg Met Gly Asp Pro Lys Ser Ser Val Tyr Leu Ala Asn Ser Lys 385 390 395 400
Val Val Ala Ala Ser Ala lie Glu Gly Val lie Thr Asn Pro Lys Asp 405 410 415
Leu <210> 22 <211> 418
<212> PRT
<213>萬氏甲:¾球菌SB <400> 22
Met Thr Leu Ala Glu Ala lie Leu Ser Lys Lys Leu Gly Lys Asn Val 15 10 15
Tyr Ala Lys Asp Ser Val Glu lie Asp Val Asp Leu Ala Met Thr His 20 25 30
Asp Gly Thr Thr Pro Leu Thr Val Lys Ala Phe Glu Glu lie Ser Asp 35 40 45
Arg Val Phe Asp Asn Lys Lys lie Val lie Val Phe Asp His Asn lie 50 55 60
Pro Ala Asn Thr Ser Lys Ala Ala Asn Met Gin lie lie Thr Arg Asp 65 70 75 80
Phe lie Lys Lys His Asp lie Lys Asn Tyr Tyr Leu Asp Gly Glu Gly 85 90 95 lie Cys His Gin lie Leu Pro Glu Lys Gly His Val Lys Pro Asn Met 100 105 110
Val lie Val Gly Ala Asp Ser His Thr Cys Thr His Gly Ala Phe Gly 115 120 125
Ala Phe Ala Thr Gly Phe Gly Ala Ser Asp Met Gly Tyr Val Tyr Ala 130 135 140
Thr Gly Lys Thr Trp Phe Arg Val Pro Glu Thr lie Arg Val Asn Val 145 150 155 160 •38- 146966-序列表.doc 201037078
Thr Gly Lys Asn Glu Asn lie Ser Gly Lys Asp lie Val Leu Lys Thr 165 170 175
Cys Lys Glu Val Gly Arg Ser Gly Ala Thr Tyr Met Ala Leu Glu Tyr 180 185 190
Gly Gly Ser Ala Val Lys Ala Leu Asn Met Asp Glu Arg Met Val Leu 195 200 205
Cys Asn Met Ala 工le Glu Met Gly Gly Lys Val Gly Leu lie Glu Ala 210 215 220
Asp His Thr Thr Tyr Asp Tyr Leu Lys Asn Ala Gly Val Ser Asn Gin 225 230 235 240 ❹
Glu lie Ala Glu Leu Gin Arg Asn Lys lie Ser lie Thr Glu Asn Glu 245 250 255
Glu Thr Tyr Phe Lys Thr Val Glu Phe Asp lie Thr Asp Met Glu Glu 260 265 270
Gin Val Ala Cys Pro His His Pro Asp Asn Val Lys Gly lie Ser Glu 275 280 285
Val Leu Gly Thr Pro lie Asp Gin lie Phe lie Gly Ser Cys Thr Asn 290 295 300
Gly His lie Gly Asp Leu Arg lie Ala Ala Lys lie Leu Lys Gly Lys 305 310 315 320
Ser lie Asn Lys Asn Thr Arg Leu lie Val lie Pro Ala Ser Lys Ser 325 330 335 lie Leu Lys Gin Ala Leu Asn Glu Gly Leu lie Asp lie Phe Val Asp 340 345 350
Phe Gly Ala Leu lie Cys Ala Pro Gly Cys Gly Pro Cys Leu Gly Ala 355 360 365
His Glu Gly Val Leu Gly Asp Gly Glu Val Cys Leu Ala Thr Thr Asn 370 375 380
Arg Asn Phe Lys Gly Arg Met Gly Asn lie Asn Ser Glu Val Tyr Leu 385 390 395 400 39- 146966-序列表.doc 201037078
Ser Ser Pro Ala He Ala Ala Lys Ser Ala lie Lys Gly His lie Thr 405 410 415
Asn Glu <210> 23 <211> 421
<212> PRT <213〉雜色甲烷球菌Nankai3 <400> 23
Met Thr Leu Ala Glu Glu lie Leu Ser Lys Lys Val Gly Lys Lys Val 15 10 15
Lys Ala Gly Asp Val Val Glu He Asp lie Asp Leu Ala Met Thr His 20 25 30
Asp Gly Thr Thr Pro Leu Ser Ala Lys Ala Phe Lys Gin lie Thr Asp 35 40 45
Lys Val Trp Asp Asn Lys Lys lie Val lie Val Phe Asp His Asn Val 50 55 60
Pro Ala Asn Thr Leu Lys Ala Ala Asn Met Gin Lys lie Thr Arg Glu 65 70 75 80
Phe lie Lys Glu Gin Asn lie lie Asn His Tyr Leu Asp Gly Glu Gly 85 90 95
Val Cys His Gin Val Leu Pro Glu Asn Gly His lie Gin Pro Asn Met 100 105 110
Val lie Ala Gly Gly Asp Ser His Thr Cys Thr Tyr Gly Ala Phe Gly 115 120 125
Ala Phe Ala Thr Gly Phe Gly Ala Thr Asp Met Gly Asn lie Tyr Ala 130 135 140
Thr Gly Lys Thr Trp Leu Lys Val Pro Lys Thr lie Arg lie Asn Val 145 150 155 160
Asn Gly Glu Asn Asp Lys lie Thr Gly Lys Asp lie lie Leu Lys lie 165 170 175
Cys Lys Glu Val Gly Arg Ser Gly Ala Thr Tyr Met Ala Leu Glu Tyr 180 185 190 -40 146966·序列表.doc 201037078
Gly Gly Glu Ala lie Lys Lys Leu Ser Met Asp Glu Arg Met Val Leu 195 200 205
Ser Asn Met Ala lie Glu Met Gly Gly Lys Val Gly Leu lie Glu Ala 210 215 220
Asp Glu Thr Thr Tyr Asn Tyr Leu Arg Asn Val Gly lie Ser Glu Glu 225 230 235 240
Lys lie Leu Glu Leu Lys Lys Asn Gin lie Thr lie Asp Glu Asn Asn 245 250 255 lie Asp Asn Asp Asn Tyr Tyr Lys lie lie Asn lie Asp lie Thr Asp 260 265 270 o
Met Glu Glu Gin Val Ala Cys Pro His His Pro Asp Asn Val Lys Asn 275 280 285 lie Ser Glu Val Lys Gly Ala Pro lie Asn Gin Val Phe lie Gly Ser 290 295 300
Cys Thr Asn Gly Arg Leu Asn Asp Leu Arg lie Ala Ser Lys Tyr Leu 305 310 315 320
Lys Gly Lys Lys Val His Asn Asp Val Arg Leu lie Val lie Pro Ala 325 330 335
Ser Lys Ser lie Phe Lys Gin Ala Leu Lys Glu Gly Leu lie Asp lie 340 345 350 〇
Phe Val Asp Ala Gly Ala Leu lie Cys Thr Pro Gly Cys Gly Pro Cys 355 360 365
Leu Gly Ala His Gin Gly Val Leu Gly Asp Gly Glu Val Cys Leu Ala 370 375 380
Thr Thr Asn Arg Asn Phe Lys Gly Arg Met Gly Asn Thr Thr Ala Glu 385 390 395 400 工le Tyr Leu Ser Ser Pro Ala lie Ala Ala Lys Ser Ala lie Lys Gly 405 410 415
Tyr lie Thr Asn Glu 420 146966-序列表.doc -41 - 201037078 <210> 24 <211> 170
<212> PRT <213> 詹氏甲烷暖球菌DSM2661 <400> 24
Met lie lie Lys Gly Arg Ala His Lys Phe Gly Asp Asp Val Asp Thr 15 10 15
Asp Ala lie lie Pro Gly Pro Tyr Leu Arg Thr Thr Asp Pro Tyr Glu 20 25 30
Leu Ala Ser His Cys Met Ala Gly lie Asp Glu Asn Phe Pro Lys Lys 35 40 45
Val Lys Glu Gly Asp Val lie Val Ala Gly Glu Asn Phe Gly Cys Gly 50 55 60
Ser Ser Arg Glu Gin Ala Val lie Ala lie Lys Tyr Cys Gly lie Lys 65 70 75 80
Ala Val lie Ala Lys Ser Phe Ala Arg lie Phe Tyr Arg Asn Ala lie 85 90 95
Asn Val Gly Leu lie Pro lie lie Ala Asn Thr Asp Glu lie Lys Asp 100 105 110
Gly Asp lie Val Glu lie Asp Leu Asp Lys Glu Glu lie Val lie Thr 115 120 125
Asn Lys Asn Lys Thr lie Lys Cys Glu Thr Pro Lys Gly Leu Glu Arg 130 135 140
Glu lie Leu Ala Ala Gly Gly Leu Val Asn Tyr Leu Lys Lys Arg Lys 145 150 155 160
Leu lie Gin Ser Lys Lys Gly Val Lys Thr 165 170 <210> 25 <211〉 170
<212> PRT <213>耐熱甲烷熱桿菌DH <400> 25
Met Glu Gly lie lie Arg Gly Arg Val Trp Arg Phe Gly Asp Asn Val 15 10 15 -42 146966-序列表.doc 201037078
Asp Thr Asp Met lie lie Pro Gly Arg Tyr Leu Arg Thr Phe Ser Leu 20 25 30
Asp Glu Leu Ala Ser His Val Met Glu Gly Ala Arg Pro Glu Phe Ala 35 40 45
Ser Gin Val Arg Lys Gly Asp lie lie Val Ala Gly Arg Asn Phe Gly 50 55 60
Cys Gly Ser Ser Arg Glu Gin Ala Pro Val Ala Leu Lys His Ala Gly 65 70 75 80
Val Val Ala lie lie Ala Glu Ser Phe Ala Arg lie Phe Tyr Arg Asn 85 90 95
Ala lie Asn lie Gly Leu Pro Val lie Met Ala Lys Val Asp Ala Asp 100 105 110
Asp Gly Asp Glu Val Ser lie Asp Leu Arg Ser Gly Gin lie Arg Asn 115 120 125
Leu Thr Ala Gly Ser Glu Tyr Arg Met Lys Pro Phe Asn Asp Tyr Met 130 135 140
Leu Ser lie Leu Glu Asp Gly Gly Leu Val Asn His Tyr Leu Lys Thr 145 150 155 160 lie Asp Thr Gly lie Ser Gly Asp Glu Gly 165 170 <210> 26 <211> 161 <212> PRT <213> 海沼甲烷球菌S2 <400> 26
G
Met Lys lie Thr Gly Lys Val His Leu Phe Gly Asp Asp He Asp Thr 1 5 10 15
Asp Ala lie lie Pro Gly Ala Tyr Leu Lys Thr Thr Asp Glu Tyr Glu 20 25 30
Leu Ala Ser His 35
Cys Met Ala Gly lie Asp Glu Asn Phe Pro Glu Arg 40 45 -43 · 146966-序列表.doc 201037078
Val Glu Asp Gly Asp Phe Leu Val Ala Gly Glu Asn Phe Gly Cys Gly 50 55 60
Ser Ser Arg Glu Gin Ala Pro lie Ala lie Lys Tyr Cys Gly lie Lys 65 70 75 80
Ala lie lie Val Glu Ser Phe Ala Arg lie Phe Tyr Arg Asn Cys lie 85 90 95
Asn Leu Gly Val Phe Pro lie Glu Cys Lys Gly lie Ser Lys His Val 100 105 110
Lys Asp Gly Asp Val lie Glu Leu Asp Leu Glu Glu Lys Lys Val lie 115 120 125
Leu Lys Asp Thr Val Leu Asp Cys Asn Leu Pro Thr Gly Thr Ala Lys 130 135 140
Asp lie Met Asp Glu Gly Gly Leu lie Asn Tyr Ala Lys Lys Gin Lys 145 150 155 160
Asn <210> 27 <211> 161 <212> PRT <213> 海沼曱烷球菌C5 <400> 27
Met Lys lie Thr Gly Lys Val His Val Phe Gly Asp Asp lie Asp Thr 15 10 15
Asp Ala lie lie Pro Gly Ala Tyr Leu Lys Thr Thr Asp Glu Tyr Glu 20 25 30
Leu Ala Ser His Cys Met Ala Gly lie Asp Glu Asp Phe Pro Glu Met 35 40 45
Val Lys Glu Gly Asp Phe Leu Val Ala Gly Glu Asn Phe Gly Cys Gly 50 55 60
Ser Ser Arg Glu Gin Ala Pro lie Ala lie Lys Tyr Cys Gly lie Lys 65 70 75 80
Ala lie lie Val Glu Ser Phe Ala Arg lie Phe Tyr Arg Asn Cys lie 85 90 95 -44 146966-序列表.doc 201037078
Asn Leu Gly Val Phe Pro lie Glu Cys Lys Gly lie Ser Lys His Val 100 105 110
Lys Asp Gly Asp Leu lie Glu Leu Asp Leu Glu Asn Lys Lys Val lie 115 120 125
Leu Lys Asp Lys Val Leu Asp Cys His lie Pro Thr Gly Thr Ala Lys 130 135 140
Asp lie Met Asp Glu Gly Gly Leu lie Asn Tyr Ala Lys Lys Gin Lys 145 150 155 160
Asn <210>
<211> <212> <213> 28 161
PRT 海沼曱烷球菌C7 <400> 28
Met Lys lie Thr Gly Lys Val His Leu Phe Gly Asp Asp Val Asp Thr 15 10 15
Asp Ala lie lie Pro Gly Ala Tyr Leu Lys Thr Thr Asp Glu Tyr Glu 20 25 30
Leu Ala Ser His Cys Met Ala Gly lie Asp Glu Asp Phe Pro Glu Met 35 40 45
Val Glu Glu Gly Asp Phe Leu Val Ala Gly Glu Asn Phe Gly Cys Gly 50 55 60
Ser Ser Arg Glu Gin Ala Pro lie Ala lie Lys Tyr Cys Gly lie Lys 65 70 75 80
Ala lie lie Val Glu Ser Phe Ala Arg lie Phe Tyr Arg Asn Cys lie 85 90 95
Asn Leu Gly Val Phe Pro lie Glu Cys Lys Gly lie Ser Lys His Val 100 105 110
Lys Asp Gly Asp Ser lie Glu Leu Asp Leu Glu Asn Lys Lys Val lie 115 120 125 -45- 146966·序列表.doc 201037078
Leu Lys Asp Thr Val Leu Asn Cys His Leu Pro Thr Gly Thr Ala Lys 130 135 140
Glu lie Met Asp Glu Gly Gly Leu lie Asn Tyr Ala Lys Lys His Lys 145 150 155 160
Asn <210> 29 <211> 163
<212> PRT <213〉 思氏甲烷球形菌DSM 3091 <400> 29
Met Asp Ser Met Lys Gly Lys Val Trp Thr Phe Arg Asp Cys lie Asp 15 10 15
Thr Asp Val lie lie Ala Gly Arg Tyr Leu Arg Thr Phe Asn Pro Glu 20 25 30
Asp Leu Ala Ala His Val Met Glu Ala Glu Asp Pro Glu Phe Ser Ser 35 40 45
Lys Val Gly Lys Gly Asp lie lie Val Gly Gly Trp Asn Phe Gly Cys 50 55 60
Gly Ser Ser Arg Glu Gin Ala Pro Val Ala lie Lys Thr Ala Gly Val 65 70 75 80
Ser Ala Val lie Ala Lys Ser Phe Ala Arg lie Phe Tyr Arg Asn Ala 85 90 95 lie Asn lie Gly Leu Pro Val lie Thr Ala Asp lie Glu Val Asp Glu 100 105 110
Gly Asp lie Leu Glu Val Asn lie Glu Asp Gly lie lie lie Asn Glu 115 120 125
Thr Thr Lys Lys Thr Phe Lys lie Lys Pro Phe Asp Ala Glu Met Leu 130 135 140
Asp lie Leu Glu Asn Gly Gly Leu Val Asn Gin Tyr Leu Lys Asn Lys 145 150 155 160
Lys Glu Val 46- 146966-序列表.doc 201037078 <210> 30
<211> 170 <212> PRT <213>坎氏甲烷嗜熱菌AV19 <400> 30
Met Arg Asp Val lie Arg Gly Arg Ala Trp Val Phe Gly Asp Asp lie 15 10 15
Asp Thr Asp Gin lie lie Pro Gly Arg Tyr Leu Thr Thr Gin Asp Pro 20 25 30
Glu Glu Leu Ala Lys His Val Met Glu Gly Ala Asp Pro Glu Phe Pro 35 40 45
Glu Lys Val Arg Glu Gly Asp Val lie Val Ala Gly Lys Asn Phe Gly 50 55 60 o
Cys Gly Ser Ser Arg Glu His Ala Pro lie Ala Leu Lys Ala Ala Gly 65 70 75 80 lie Ala Cys Val Val Thr Arg Ser Phe Ala Arg lie Phe Tyr Arg Asn 85 90 95
Ala lie Asn Leu Gly Leu Pro Leu Val Val Cys Pro Gly Val Asp Asp 100 105 110
Ala Phe Glu Asp Gly Gin Gly lie Glu Val Asn Leu Arg Glu Gly Tyr 115 120 125
Val Arg Asn Leu Asp Thr Gly Glu Glu Leu Glu Ala Lys Pro Leu Pro 130 135 140 ❹
Asp Phe Met Met Arg lie Leu Glu Ala Gly Gly Leu Val Glu Leu lie 145 150 155 160
Lys Arg Glu Gly Pro Arg Ala Phe Glu Gly 165 170 <210> 31
<211> 161 <212> PRT <213>史氏甲烷短桿菌ATCC35061 <400> 31
Met Asp lie lie Lys Gly Lys Thr Trp Thr Phe Gly Glu Asn lie Asp 47- 146966-序列表.doc 201037078 1 5 10 15
Thr Asp Val lie lie Pro Gly Arg Tyr Leu Arg Thr Phe Asn Pro Gin 20 25 30
Asp Leu Ala Asp His Val Leu Glu Gly Glu Arg Pro Asp Phe Thr Lys 35 40 45
Asn Val Lys Lys Gly Asp lie lie Val Ala Asp Glu Asn Phe Gly Cys 50 55 60
Gly Ser Ser Arg Glu Gin Ala Pro Val Ala lie Lys Thr Ala Gly Val 65 70 75 80
Asp Ala lie Val Ala Lys Ser Phe Ala Arg lie Phe Tyr Arg Asn Ala 85 90 95 lie Asn lie Gly Leu Pro Val lie Val Cys Asp lie Gin Ala Lys Asp 100 105 110
Gly Asp lie lie Asn lie Asp Leu Ser Lys Gly lie Leu Thr Asn Glu 115 120 125
Thr Thr Gly Glu Ser Val Thr Phe Glu Pro Phe Lys Glu Phe Met Leu 130 135 140
Asp lie Leu Glu Asp Asn Gly Leu Val Asn His Tyr Leu Lys Glu Lys 145 150 155 160
Gin <210> 32 <211> 161
<212> PRT
<213>萬氏甲烧球菌SB <400> 32
Met Lys Leu Lys Gly Lys Ala His Val Phe Ser Asp Asp Val Asp Thr 15 10 15
Asp Ala lie lie Pro Gly Ala Tyr Leu Arg Thr Thr Asp Val Tyr Glu 20 25 30
Leu Ala Ser His Cys Met Ala Gly lie Asp Glu Asn Phe Pro Lys Lys 35 40 45 -48- 146966-序列表.doc 201037078
Val Asn Leu Gly Asp Phe lie Val Ala Gly Glu Asn Phe Gly Cys Gly 50 55 60
Ser Ser Arg Glu Gin Ala Pro lie Ser lie Lys Tyr Leu Gly lie Ser 65 70 75 80
Ala lie lie Ala Glu Ser Phe Ala Arg lie Phe Tyr Arg Asn Ser lie 85 90 95
Asn Leu Gly Val lie Pro lie Glu Cys Lys Asn lie Ser Lys His Val 100 105 110
Lys Thr Gly Asp Leu lie Glu Leu Asp Leu Glu Asn Lys Lys lie lie 115 120 125
Leu Lys Asp lie Val Leu Glu Cys Thr Val Pro Thr Gly Lys Ala Lys 130 135 140
Glu lie lie Asp Leu Gly Gly Leu lie Asn Tyr Ala Lys Ala Gin Met 145 150 155 160
Gly <210> 33 <211> 165
<212> PRT <213>雜色甲烷球菌Nankai3 <400> 33
Met lie lie Lys Gly Asn lie His Leu Phe Gly Asp Asp lie Asp Thr 15 10 15
G
Asp Ala lie lie Pro Gly Ala Tyr Leu Lys Thr Thr Asp Pro Lys Glu 20 25 30
Leu Ala Ser His Cys Met Ala Gly lie Asp Glu Lys Phe Ser Thr Lys 35 40 45
Val Lys Asp Gly Asp lie lie Val Ala Gly Glu Asn Phe Gly Cys Gly 50 55 60
Ser Ser Arg Glu Gin Ala Pro lie Ser lie Lys His Thr Gly lie Lys 65 70 75 80
Ala Val Val Ala Glu Ser Phe Ala Arg lie Phe Tyr Arg Asn Cys lie 49- 146966-序列表.doc 201037078 85 90 95
Asn lie Gly Leu lie Pro lie Thr Cys Glu Gly lie Asn Glu Gin lie 100 105 110
Gin Asn Leu Lys Asp Gly Asp Thr lie Glu lie Asp Leu Gin Asn Glu 115 120 125
Thr lie Lys lie Asn Ser Met Met Leu Asn Cys Gly Ala Pro Lys Gly 130 135 140 lie Glu Lys Glu lie Leu Asp Ala Gly Gly Leu Val Gin Tyr Thr Lys 145 150 155 160
Asn Lys Leu Lys Lys 165 <210> 34 <211> 347
<212> PRT <213>詹氏曱烷暖球菌DSM2661 <400> 34
Met Met Lys Val Cys Val lie Glu Gly Asp Gly lie Gly Lys Glu Val 15 10 15 工le Pro Glu Ala lie Lys lie Leu Asn Glu Leu Gly Glu Phe Glu lie 20 25 30 lie Lys Gly Glu Ala Gly Leu Glu Cys Leu Lys Lys Tyr Gly Asn Ala 35 40 45
Leu Pro Glu Asp Thr lie Glu Lys Ala Lys Glu Ala Asp lie 工le Leu 50 55 60
Phe Gly Ala lie Thr Ser Pro Lys Pro Gly Glu Val Gin Asn Tyr Lys 65 70 75 80
Ser Pro lie lie Thr Leu Arg Lys Met Phe His Leu Tyr Ala Asn Val 85 90 95
Arg Pro lie Asn Asn Phe Gly lie Gly Gin Leu lie Gly Lys lie Ala 100 105 110
Asp Tyr Glu Phe Leu Asn Ala Lys Asn lie Asp lie Val lie lie Arg 115 120 125 •50- 146966-序列表.doc 201037078
Glu Asn Thr Glu Asp Leu Tyr Val Gly Arg Glu Arg Leu Glu Asn Asp 130 135 140
Thr Ala lie Ala Glu Arg Val lie Thr Arg Lys Gly Ser Glu Arg lie 145 150 155 160 lie Arg Phe Ala Phe Glu Tyr Ala lie Lys Asn Asn Arg Lys Lys Val 165 170 175
Ser Cys lie His Lys Ala Asn Val Leu Arg lie Thr Asp Gly Leu Phe 180 185 190
Leu Glu Val Phe Asn Glu lie Lys Lys His Tyr Asn lie Glu Ala Asp 195 200 205 o
Asp Tyr Leu Val Asp Ser Thr Ala Met Asn Leu lie Lys His Pro Glu 210 215 220
Lys Phe Asp Val lie Val Thr Thr Asn Met Phe Gly Asp lie Leu Ser 225 230 235 240
Asp Glu Ala Ser Ala Leu lie Gly Gly Leu Gly Leu Ala Pro Ser Ala 245 250 255
Asn lie Gly Asp Asp Lys Ala Leu Phe Glu Pro Val His Gly Ser Ala 260 265 270
Pro Asp lie Ala Gly Lys Gly lie Ala Asn Pro Met Ala Ser lie Leu 275 280 285
Ser lie Ala Met Leu Phe Asp Tyr lie Gly Glu Lys Glu Lys Gly Asp 290 295 300
Leu lie Arg Glu Ala Val Lys Tyr Cys Leu lie Asn Lys Lys Val Thr 305 310 315 320
Pro Asp Leu Gly Gly Asp Leu Lys Thr Lys Asp Val Gly Asp Glu lie 325 330 335
Leu Asn Tyr lie Arg Lys Lys Leu Lys Gly Tyr 340 345 <210> 35 <211> 331
<212> PRT
<213>耐熱甲烷熱桿菌DH -51 - 146966-序列表.doc 201037078 <400> 35
Met Tyr Arg lie Thr Val lie Pro Gly Asp Gly lie Gly Val Glu Val 15 10 15
Met Glu Ala Ala Leu His Val Leu Gin Ala Leu Glu lie Glu Phe Glu 20 25 30
Phe Thr His Ala Glu Ala Gly Asn Glu Cys Phe Arg Arg Cys Gly Asp 35 40 45
Thr Leu Pro Glu Glu Thr Leu Lys Leu Val Arg Lys Ala Asp Ala Thr 50 55 60
Leu Phe Gly Ala Val Thr Thr Val Pro Gly Gin Lys Ser Ala lie lie 65 70 75 80
Thr Leu Arg Arg Glu Leu Asp Leu Phe Ala Asn Leu Arg Pro Val Lys 85 90 95
Ser Leu Pro Gly Val Pro Cys Leu Tyr Pro Asp Leu Asp Phe Val lie 100 105 110
Val Arg Glu Asn Thr Glu Asp Leu Tyr Val Gly Asp Glu Glu Tyr Thr 115 120 125
Pro Glu Gly Ala Val Ala Lys Arg lie lie Thr Arg Thr Ala Ser Arg 130 135 140
Arg lie Ser Gin Phe Ala Phe Gin Tyr Ala Gin Lys Glu Gly Met Gin 145 150 155 160
Lys Val Thr Ala Val His Lys Ala Asn Val Leu Lys Lys Thr Asp Gly 165 170 175 lie Phe Arg Asp Glu Phe Tyr Lys Val Ala Ser Glu Tyr Pro Gin Met 180 185 190
Glu Ala Asn Asp Tyr Tyr Val Asp Ala Thr Ala Met Tyr Leu lie Thr 195 200 205
Gin Pro Gin Glu Phe Gin Thr lie Val Thr Thr Asn Leu Phe Gly Asp 210 215 220 lie Leu Ser Asp Glu Ala Ala Gly Leu lie Gly Gly Leu Gly Leu Ala 225 230 235 240 52- 146966-序列表.doc 201037078
Pro Ser Ala Asn lie Gly Glu Lys Asn Ala Leu Phe Glu Pro Val His 245 250 255
Gly Ser Ala Pro Gin lie Ala Gly Lys Asn 工le Ala Asn Pro Thr Ala 260 265 270
Met lie Leu Thr Thr Thr Leu Met Leu Lys His Leu Asn Lys Lys Gin 275 280 285
Glu Ala Gin Lys lie Glu Lys Ala Leu Gin Lys Thr Leu Met Arg Gly 290 295 300 lie Met Thr Pro Asp Leu Gly Gly Thr Ala Ser Thr Met Glu Met Ala 305 310 315 320
Glu Ala lie Lys Glu Glu He Val Lys Gly Glu 325 330 <210> 36 <211> 339
<212> PRT <213> 海沼甲烷球菌S2 <400> 36
Met Arg Asn Thr Pro Lys lie Cys Val lie Asn Gly Asp Gly lie Gly 15 10 15
Asn Glu Val Val Pro Glu Thr Val Arg Val Leu Asn Glu Leu Gly Asp 20 25 30
Phe Glu Phe lie His Ala His Ala Gly Tyr Glu Cys Phe Lys Arg Cys 35 40 45 ❹
Gly Asp Ala lie Pro Glu Asn Thr lie Glu lie Ala Lys Glu Ser Asp 50 55 60
Cys lie Leu Phe Gly Ser Val Thr Thr Pro Lys Pro Thr Glu Leu Lys 65 70 75 80
Asn Lys Ser Tyr Arg Ser Pro lie Leu Thr Leu Arg Lys Glu Leu Asp 85 90 95
Leu Tyr Ala Asn lie Arg Pro Thr Tyr Asn Phe Asp Asn Leu Asp Phe 100 105 110 -53- 146966·序列表.doc 201037078
Val lie lie Arg Glu Asn Thr Glu Gly Leu Tyr Val Lys Lys Glu Tyr 115 120 125
Tyr Asp Glu Lys Asn Glu Val Ala lie Ala Glu Arg lie lie Ser Lys 130 135 140
Phe Gly Ser Ser Arg lie Val Lys Phe Ala Phe Asp Tyr Ala Val Gin 145 150 155 160
Asn Asn Arg Lys Lys Val Ser Cys lie His Lys Ala Asn Val Leu Arg 165 170 175
Val Thr Asp Gly Leu Phe Leu Glu Val Phe Glu Glu Met Ser Lys His 180 185 190
Tyr Glu Lys Leu Gly lie Lys Ser Asp Asp Tyr Leu lie Asp Ala Thr 195 200 205
Ala Met Tyr Leu lie Arg Asn Pro Gin Met Phe Asp Val Leu Val Thr 210 215 220
Thr Asn Leu Phe Gly Asp lie Leu Ser Asp Glu Ala Ala Gly Leu lie 225 230 235 240
Gly Gly Leu Gly Met Ser Pro Ser Ala Asn lie Gly Asp Lys Asn Gly 245 250 255
Leu Phe Glu Pro Val His Gly Ser Ala Pro Asp lie Ala Gly Lys Gly 260 265 270 lie Ser Asn Pro lie Ala Thr lie Leu Ser Ala Ala Met Met Leu Asp 275 280 285
His Leu Lys Met Asn Lys Glu Ala Glu Tyr lie Arg Lys Ala Val Lys 290 295 300
Lys Thr Val Glu Cys Lys Tyr Leu Thr Pro Asp Leu Gly Gly Asn Leu 305 310 315 320
Lys Thr Phe Glu Val Thr Glu Lys lie lie Glu Ser lie Arg Ser Gin 325 330 335
Met lie Gin <210> 37 •54- 146966-序列表.doc 201037078 <211> 339 <212> PRT <213> 海沼曱烷球菌C5 <400> 37
Met Arg Asn Thr Pro Lys lie Cys Val lie Asn Gly Asp Gly lie Gly 15 10 15
Asn Glu Val lie Pro Glu Thr Val Arg Val Leu Asn Glu lie Gly Asp 20 25 30
Phe Glu Phe lie Glu Thr His Ala Gly Tyr Glu Cys Phe Lys Arg Cys 35 40 45
Gly Asp Ala lie Pro Glu Lys Thr lie Glu lie Ala Lys Glu Ser Asp 50 55 60
Ser lie Leu Phe Gly Ser Val Thr Thr Pro Lys Pro Thr Glu Leu Lys 65 70 75 80
Asn Lys Pro Tyr Arg Ser Pro lie Leu Thr Leu Arg Lys Glu Leu Asp 85 90 95
Leu Tyr Ala Asn lie Arg Pro Thr Phe Asn Phe Lys Asn Leu Asp Phe 100 105 110
Val lie lie Arg Glu Asn Thr Glu Gly Leu Tyr Val Lys Lys Glu Tyr 115 120 125
Tyr Asp Glu Lys Asn Glu Val Ala Thr Ala Glu Arg lie lie Ser Lys 130 135 140
Phe Gly Ser Ser Arg lie Val Lys Phe Ala Phe Asp Tyr Ala Leu Gin 145 150 155 160
Asn Asn Arg Lys Lys Val Ser Cys lie His Lys Ala Asn Val Leu Arg 165 170 175 lie Thr Asp Gly Leu Phe Leu Gly Val Phe Glu Glu lie Ser Lys Lys 180 185 190
Tyr Glu Lys Leu Gly lie Val Ser Asp Asp Tyr Leu lie Asp Ala Thr 195 200 205
Ala Met Tyr Leu lie Arg Asn Pro Gin Met Phe Asp Val Met Val Thr 210 215 220 55- 146966·序列表.doc 201037078
Thr Asn Leu Phe Gly Asp lie Leu Ser Asp Glu Ala Ala Gly Leu lie 225 230 235 240
Gly Gly Leu Gly Met Ser Pro Ser Ala Asn lie Gly Asp Lys Asn Gly 245 250 255
Leu Phe Glu Pro Val His Gly Ser Ala Pro Asp lie Ala Gly Lys Gly 260 265 270 lie Ser Asn Pro lie Ala Thr lie Leu Ser Ala Ala Met Met Leu Asp 275 280 285
His Leu Lys lie Asn Lys Glu Ala Glu Tyr lie Arg Asn Ala Val Lys 290 295 300
Lys Thr Val Glu Cys Lys Tyr Leu Thr Pro Asp Leu Gly Gly His Leu 305 310 315 320
Lys Thr Ser Glu Val Thr Glu Lys lie lie Glu Ser lie Lys Ser Gin 325 330 335
Met lie Gin <210> 38 <211> 339
<212> PRT <213> 海沼甲烷球菌C7 <400> 38
Met Arg Asn Thr Pro Lys lie Cys Val lie Asn Gly Asp Gly lie Gly 15 10 15
Asn Glu Val 工le Pro Glu Thr Val Arg Val Leu Ser Glu lie Gly Asp 20 25 30
Phe Glu Phe lie Glu Thr His Ala Gly Tyr Glu Cys Phe Lys Arg Cys 35 40 45
Gly Asp Ala lie Pro Glu Lys Thr lie Glu lie Ala Lys Glu Ser Asp 50 55 60
Ser lie Leu Phe Gly Ser Val Thr Thr Pro Lys Pro Thr Glu Leu Lys 65 70 75 80
Asn Lys Pro Tyr Arg Ser Pro lie Leu Thr Leu Arg Lys Glu Leu Asp 56- 146966-序列表.doc 201037078 85 90 95
Leu Tyr Ala Asn lie Arg Pro Thr Phe Asn Phe Lys Asp Leu Asp Phe 100 105 110
Val lie lie Arg Glu Asn Thr Glu Gly Leu Tyr Val Lys Lys Glu Tyr 115 120 125
Tyr Asp Glu Lys Asn Glu Val Ala lie Ala Glu Arg Val lie Ser Lys 130 135 140
Phe Gly Ser Ser Arg lie Val Lys Tyr Ala Phe Asp Tyr Ala Leu Gin 145 150 155 160
Asn Asn Arg Lys Lys Val Ser Cys lie His Lys Ala Asn Val Leu Arg 165 170 175 o lie Thr Asp Gly Leu Phe Leu Glu Val Phe Glu Glu lie Ser Lys Lys 180 185 190
Tyr Glu Lys Leu Gly lie Ala Ser Asp Asp Tyr Leu lie Asp Ala Thr 195 200 205
Ala Met Tyr Leu lie Arg Asn Pro Gin Met Phe Asp Val Met Val Thr 210 215 220
Thr Asn Leu Phe Gly Asp lie Leu Ser Asp Glu Ala Ala Gly Leu lie 225 230 235 240
Gly Gly Leu Gly Met Ser Pro Ser Ala Asn lie Gly Asp Lys Asn Gly 245 250 255 ❹
Leu Phe Glu Pro Val His Gly Ser Ala Pro Asp 工le Ala Gly Lys Gly 260 265 270 lie Ser Asn Pro lie Ala Ser lie Leu Ser Ala Ala Met Met Leu Asp 275 280 285
His Leu Asn Met Asn Lys Glu Ala Glu Cys lie Arg Asn Ala Val Lys 290 295 300
Lys Ala Val Glu Cys Lys Tyr Leu Thr Pro Asp Leu Gly Gly Asn Leu 305 310 315 320
Lys Thr Ser Glu Val Thr Asp Lys lie lie Glu Ser lie Lys Ser Gin 325 330 335 -57 146966-序列表.doc 201037078
Met Val Gin <210> 39 <211> 323
<212> PRT <213> 思氏曱烷球形菌DSM 3091 <400> 39
Met Tyr Lys lie Thr Val lie Pro Gly Asp Gly lie Gly Gin Glu Val 15 10 15
Met Gin Pro Thr lie Asp lie Leu Glu Thr Leu Asn Ser Lys Phe Glu 20 25 30
Phe lie Pro Lys Glu Ala Gly Lys Glu Cys Tyr Gin Lys Tyr Asp Thr 35 40 45
Asn Leu Pro Glu Glu Thr lie Val Gin Cys Arg Glu Ser Asp Ser Thr 50 55 60
Leu Phe Gly Ala Val Thr Ser lie Pro Gin Gin Lys Ser Ala lie Val 65 70 75 80
Thr Leu Arg Lys Glu Leu Asp Leu Tyr Val Asn Gin Arg Pro lie His 85 90 95
Ser Tyr Thr Asn Pro Asp lie Asp Phe Thr lie lie Arg Glu Asn Ser 100 105 110
Glu Gly Leu Tyr Ser His lie Glu Glu Ser Thr Gly Asp Glu Ala lie 115 120 125
Ala lie Arg Lys lie Thr Tyr Lys Ala Ser Glu Arg lie lie Asn Tyr 130 135 140
Ala Phe Asn Tyr Ala Leu Lys Thr Glu Lys Ser Lys Val Thr Ala Ser 145 150 155 160
His Lys Ala Asn Val Leu Pro Val Thr Asp Gly lie Phe Lys Asn Thr 165 170 175
Phe Tyr Lys Val Ala Ser Asn Tyr Pro Thr lie Lys Ser Asn Asp Tyr 180 185 190 •58 146966-序列表.doc 201037078
Tyr lie Asp Ala Met Ala Met Tyr Leu lie Thr Asn Pro Ala Gin Phe 195 200 205
Asp lie lie Val Thr Thr Asn Leu Phe Gly Asp lie Leu Ser Asp Glu 210 215 220
Gly Gly Gly Leu Val Gly Thr Leu Gly Leu lie Pro Ser Ala Asn lie 225 230 235 240
Gly Asp Lys Thr Gly Leu Phe Glu Pro Val His Gly Ser Ala Pro Asp 245 250 255 lie Ala Gly Leu Asn Lys Ala Asn Pro lie Ala Met lie Leu Ser Ser 260 265 270
Cys Leu Met Leu Glu Tyr Leu Gly Leu Tyr Asp Asp Ala Lys Arg lie 275 280 285
Gin Asn Ala Val Glu Glu Thr lie Ser Glu Ser Lys Val Lys Thr Pro 290 295 300
Asp Met Gly Gly His Asn Asn Thr Gin Asp Val Ala Asn Asn lie Leu 305 310 315 320
His Arg Leu <210> <211> <212> <213> 40 335 PRT 坎氏曱烷嗜熱菌AV19 ❹ <400> 40
Met Ala Tyr Lys lie Ala Val lie Pro Gly Asp Gly lie Gly Pro Glu 15 10 15
Val lie Glu Ala Ala Leu His Val lie Glu Pro Leu lie Asp Ala Glu 20 25 30
Phe Val Glu Gly Glu Ala Gly Asp Glu Cys Ala Glu Lys His Gly Asp 35 40 45
Pro Leu Pro Glu Asp Thr Leu Glu Leu Cys His Glu Ala Asp Ala lie 50 55 60
Leu Phe Gly Ala Ala Gly Glu Thr Ala Ala Asp Val lie Val Arg Leu 65 70 75 80 59- 146966-序列表.doc 201037078
Arg Gin Glu Leu Asp Leu Tyr Ala Asn lie Arg Pro Val Arg Gly Phe 85 90 95
Pro Gly Leu Arg Glu Leu Thr Gly Glu Pro Tyr Val Arg Asp Asp Val 100 105 110
Asp Phe Val lie Val Arg Glu Asn Thr Glu Gly Leu Tyr Ser Gly lie 115 120 125
Glu Gly Arg Phe Arg Asp Thr Ala Tyr Thr Leu Arg lie lie Thr Glu 130 135 140
Glu Gly Thr Arg Arg lie Ala Glu Val Ala Cys Asp Leu Ala Glu Glu 145 150 155 160
Arg Gly Ser Asn Thr Val Thr Cys Val His Lys Ala Asn Val Met Arg 165 170 175
Glu Thr Cys Gly Leu Phe Arg Glu Val Cys Lys Glu Val Val Glu Ser 180 185 190
Arg Gly Leu Glu Phe Glu Glu Tyr Tyr Val Asp Ala Ala Ala Met Phe 195 200 205
Met lie Thr Glu Pro Glu Arg Phe Asp Val Val Val Thr Pro Asn Met 210 215 220
Phe Gly Asp lie Leu Ser Asp Glu Ala Ala Ala Leu Val Gly Gly Leu 225 230 235 240
Gly Leu Ala Pro Ser Gly Asn Val Gly Asp Arg His Gly Leu Phe Glu 245 250 255
Pro Val His Gly Ser Ala Pro Asp lie Ala Gly Lys Gly lie Ala Asn 260 265 270
Pro Phe Ala Thr lie Leu Ser Ala Val Met Met Leu Glu Trp Leu Gly 275 280 285
Glu Asp Glu Ala Ala Glu Ala Val Arg Glu Ala Val Gly Glu Ala lie 290 295 300
Arg Glu Gly Val Val Thr Pro Asp Leu Gly Gly Asp Lys Lys Thr Met 305 310 315 320 -60- 146966·序列表.doc 201037078
Glu Val Ala Glu Phe Val Arg Glu Ala Ala Leu Asn Arg Val Gin 325 330 335 <210> 41 <211> 336
<212> PRT <213> 史氏曱烷短桿菌ATCC35061 <400> 41
Met Ser Thr Ser Asn Lys Lys Asp Asn Lys Tyr Gin lie Ala Val lie 15 10 15
Pro Gly Asp Gly lie Gly Lys Glu Val Met Glu Ala Thr lie Ser Val 20 25 30
Leu Asp Glu Leu Asp Val Asp Phe Asp Tyr lie Tyr Gly lie Ala Gly 35 40 45
Asp Glu Cys Asn Glu Glu His Gly Thr Pro Leu Pro Gin Glu Thr lie 50 55 60
Asp lie Val Arg Asp Ser Asp Ala Cys Leu Phe Gly Ala Ala Gly Glu 65 70 75 80
Thr Ala Ala Asp Val lie Val Lys lie Arg Gin Glu Met Lys Met Phe 85 90 95
Ala Asn Leu Arg Pro Val Lys Ser Tyr Pro Asn Thr Lys Ser Leu Phe 100 105 110
Glu Asn Val Asp Phe Met lie Val Arg Glu Asn Thr Glu Gly Leu Tyr 115 120 125 〇 工le Ala Asp Gin Glu Glu Glu Thr Glu Asp Gly Ala 工le Ala Lys Arg 130 135 140
Val lie Thr Arg Glu Ala Glu Glu Arg lie lie Asp Tyr Ala Phe Gin 145 150 155 160
Tyr Ala Lys Asp Asn Asn Arg Thr Lys Val Thr Ala Val His Lys Ala 165 170 175
Asn Val Leu Lys Lys Thr Asp Gly Leu Phe Lys Lys lie Phe Tyr Glu 180 185 190
Val Gly Glu Lys Tyr Pro Asp lie Asp Thr Glu Asp Phe Tyr Val Asp -61 - 146966·序列表.doc 201037078 195 200 205
Ala Thr Ala Met Tyr Leu Val Thr Gin Pro Gin Glu Phe Gin Val Val 210 215 220
Val Thr Thr Asn Leu Phe Gly Asp lie Leu Ser Asp Glu Gly Ala Gly 225 230 235 240
Leu Val Gly Gly Leu Gly Leu lie Pro Ser Ala Asn lie Gly Ala Asp 245 250 255
Gly Ala Leu Phe Glu Pro Val His Gly Ser Ala Pro Asp lie Ala Gly 260 265 270
Gin Gin Lys Ala Asn Pro lie Ala Met Met Leu Ser Ala lie Met Met 275 280 285
Leu Arg Tyr Leu Gly Glu Asn Asp Ala Ala Asp Lys Phe Asp Ala Ala 290 295 300 lie Leu Lys Val Leu Ser Glu Gly Lys Thr Leu Thr Gly Asp Leu Gly 305 310 315 320
Gly Ser Ala Thr Thr Met Glu Val Ala Gin Ala Val·Lys Asn Ala Leu 325 330 335 <210> 42 <211> 337
<212> PRT <213> 萬氏曱烷球菌SB <400> 42
Met Gly Tyr Met Pro Lys lie Cys Val lie Thr Gly Asp Gly lie Gly 15 10 15
Lys Glu Val Val Pro Glu Thr Leu Arg Val Leu Asn Glu Val His Asp 20 25 30
Phe Glu Tyr lie Glu Ala His Ala Gly Tyr Glu Cys Phe Lys Arg Cys 35 40 45
Gly Glu Ser lie Pro Glu Ser Thr lie Gin Thr Ala Lys Asn Ser Asp 50 55 60
Ser lie Leu Phe Gly Ser Val Thr Thr Pro Lys Pro Thr Glu Leu Lys 65 70 75 80 62- 146966-序列表.doc 201037078
Asn Lys Pro Tyr Arg Ser Pro lie Leu Thr Leu Arg Gin Glu Leu Asp 85 90 95
Leu Tyr Ala Asn lie Arg Pro Thr Tyr Asn Phe Lys Asp Leu Asp Phe 100 105 no
Val lie lie Arg Glu Asn Thr Glu Cys Leu Tyr Val Lys Arg Glu Tyr 115 120 125
Tyr Asp Glu He Asn Glu Val Ala He Ala Glu Arg He He Ser Lys 130 135 140
Lys Gly Ser Glu Arg lie lie Lys Phe Ala Phe Glu Tyr Ala Arg Leu 145 150 155 160
Asn Asn Arg Lys Lys Val Ser Cys He His Lys Ala Asn Val Leu Arg 165 170 175
Val Thr Asp Gly Leu Phe Leu Glu lie Phe Glu Lys lie Ala Lys Leu 180 185 190
Tyr Glu Asn Phe Gly lie Ser Ser Asn Asp Tyr Lau lie Asp Ala Thr 195 200 205
Ala Met Tyr Leu lie Lys Asn Pro Tyr Met Phe Asp Val Met Val Thr 210 215 220
Thr Asn Leu Phe Gly Asp lie Leu Ser Asp Glu Ala Ala Gly Leu lie 225 230 235 240
Gly Gly Leu Gly Met Ser Pro Ser Ala Asn lie Gly Asp Asn Leu Gly 245 250 255
Leu Phe Glu Pro Val His Gly Ser Ala Pro Asp He Ala Gly Lys Gly 260 265 270 lie Ser Asn Pro lie Ala Thr lie Leu Ser Ala Ser Met Met Leu Asp 275 280 285
His Leu Lys Met Asn Lys Lys Ala Glu lie lie Arg Asn Ala Val Lys 290 295 300
Lys Thr lie Asn Asn Gly Tyr Leu Thr Pro Asp Leu Gly Gly Ser Leu 305 310. 315 320 •63- 146966·序列表.doc 201037078
Lys Thr Ser Glu Val Val Asn Lys Val lie Glu Phe lie Arg Asp Glu 325 330 335 lie <210> 43 <211> 343
<212> PRT <213> 雜色曱烧球菌Nankai3 <400> 43
Met Lys lie Pro Lys lie Cys Val lie Glu Gly Asp Gly lie Gly Lys 15 10 15
Glu Val lie Pro Glu Thr Val Arg lie Leu Lys Glu lie Gly Asp Phe 20 25 30
Glu Phe lie Tyr Glu His Ala Gly Tyr Glu Cys Phe Lys Arg Cys Gly 35 40 45
Asp Ala lie Pro Glu Lys Thr Leu Lys Thr Ala Lys Glu Cys Asp Ala 50 55 60 lie Leu Phe Gly Ala Val Ser Thr Pro Lys Leu Asp Glu Thr Glu Arg 65 70 75 80
Lys Pro Tyr Lys Ser Pro lie Leu Thr Leu Arg Lys Glu Leu Asp Leu 85 90 95
Tyr Ala Asn Val Arg Pro lie His Lys Leu Asp Asn Ser Asp Ser Ser 100 105 110
Asn Asn lie Asp Phe lie lie lie Arg Glu Asn Thr Glu Gly Leu Tyr 115 120 125
Ser Gly Val Glu Tyr Tyr Asp Glu Glu Lys Glu Leu Ala lie Ser Glu 130 135 140
Arg His lie Ser Lys Lys Gly Ser Lys Arg lie lie Lys Phe Ala Phe 145 150 155 160
Glu Tyr Ala Val Lys His His Arg Lys Lys Val Ser Cys lie His Lys 165 170 175
Ser Asn lie Leu Arg lie Thr Asp Gly Leu Phe Leu Asn lie Phe Asn 180 185 190 •64 146966-序列表.doc 201037078
Glu Phe Lys Glu Lys Tyr Lys Asn Glu Tyr Asn lie Glu Gly Asn Asp 195 200 205
Tyr Leu Val Asp Ala Thr Ala Met Tyr lie Leu Lys Ser Pro Gin Met 210 215 220
Phe Asp Val lie Val Thr Thr Asn Leu Phe Gly Asp lie Leu Ser Asp 225 230 235 240
Glu Ala Ser Gly Leu Leu Gly Gly Leu Gly Leu Ala Pro Ser Ala Asn 245 250 255 lie Gly Asp Asn Tyr Gly Leu Phe Glu Pro Val His Gly Ser Ala Pro 260 265 270
Asp lie Ala Gly Lys Gly Val Ala Asn Pro lie Ala Ala Val Leu Ser 275 280 285
Ala Ser Met Met Leu Tyr Tyr Leu Asp Met Lys Glu Lys Ser Arg Leu 290 295 300
Leu Lys Asp Ala Val Lys Gin Val Leu Ala His Lys Asp lie Thr Pro 305 310 315 320
Asp Leu Gly Gly Asn Leu Lys Thr Lys Glu Val Ser Asp Lys lie lie 325 330 335
Glu Glu Leu Arg Lys lie Ser 340 <210> 44 <211> 440 <212> PRT <213> 醸酒酵母 <400> 44
Met Ser Glu Asn Asn Glu Phe Gin Ser Val Thr Glu Ser Thr Thr Ala 15 10 15
Pro Thr Thr Ser Asn Pro Tyr Gly Pro Asn Pro Ala Asp Tyr Leu Ser 20 25 30
Asn Val Lys Asn Phe Gin Leu lie Asp Ser Thr Leu Arg Glu Gly Glu 35 40 45 -65- 146966·序列表.doc 201037078
Gin Phe Ala Asn Ala Phe Phe Asp Thr Glu Lys Lys lie Glu lie Ala 50 55 60
Arg Ala Leu Asp Asp Phe Gly Val Asp Tyr lie Glu Leu Thr Ser Pro 65 70 75 80
Val Ala Ser Glu Gin Ser Arg Lys Asp Cys Glu Ala lie Cys Lys Leu 85 90 95
Gly Leu Lys Ala Lys lie Leu Thr His lie Arg Cys His Met Asp Asp 100 105 110
Ala Arg Val Ala Val Glu Thr Gly Val Asp Gly Val Asp Val Val lie 115 120 125
Gly Thr Ser Lys Phe Leu Arg Gin Tyr Ser His Gly Lys Asp Met Asn 130 135 140
Tyr 工le Ala Lys Ser Ala Val Glu Val lie Glu Phe Val Lys Ser Lys 145 150 155 160
Gly lie Glu lie Arg Phe Ser Ser Glu Asp Ser Phe Arg Ser Asp Leu 165 170 175
Val Asp Leu Leu Asn lie Tyr Lys Thr Val Asp Lys lie Gly Val Asn 180 185 190
Arg Val Gly 工le Ala Asp Thr Val Gly Cys Ala Asn Pro Arg Gin Val 195 200 205
Tyr Glu Leu lie Arg Thr Leu Lys Ser Val Val Ser Cys Asp lie Glu 210 215 220
Cys His Phe His Asn Asp Thr Gly Cys Ala lie Ala Asn Ala Tyr Thr 225 230 235 240
Ala Leu Glu Gly Gly Ala Arg Leu lie Asp Val Ser Val Leu Gly lie 245 250 255
Gly Glu Arg Asn Gly lie Thr Pro Leu Gly Gly Leu Met Ala Arg Met 260 265 270 lie Val Ala Ala Pro Asp Tyr Val Arg Ser Lys Tyr Lys Leu His Lys 275 280 285 lie Arg Asp lie Glu Asn Leu Val Ala Asp Ala Val Glu Val Asn lie 66- 146966-序列表.doc 201037078 290 295 300
Pro Phe Asn Asn Pro lie Thr Gly Phe Cys Ala Phe Thr His Lys Ala 305 310 315 320
Gly lie His Ala Lys Ala lie Leu Ala Asn Pro Ser Thr Tyr Glu lie 325 330 335
Leu Asp Pro His Asp Phe Gly Met Lys Arg Tyr lie His Phe Ala Asn 340 345 350
Arg Leu Thr Gly Trp Asn Ala lie Lys Ser Arg Val Asp Gin Leu Asn 355 360 365
Leu Asn Leu Thr Asp Asp Gin lie Lys Glu Val Thr Ala Lys lie Lys 370 375 380
Lys Leu Gly Asp Val Arg Pro Leu Asn lie Asp Asp Val Asp Ser lie 385 390 395 400 lie Lys Asp Phe His Ala Glu Leu Ser Thr Pro Leu Leu Lys Pro Val 405 410 415
Asn Lys Gly Thr Asp Asp Asp Asn lie Asp lie Ser Asn Gly His Val 420 425 430
Ser Lys Lys Ala Lys Val Thr Lys 435 440 <210> 45 <211> 428 <212> PRT <213>釀酒酵母 <400> 45
Met Thr Ala Ala Lys Pro Asn Pro Tyr Ala Ala Lys Pro Gly Asp Tyr 15 10 15
Leu Ser Asn Val Asn Asn Phe Gin Leu lie Asp Ser Thr Leu Arg Glu 20 25 30
Gly Glu Gin Phe Ala Asn Ala Phe Phe Asp Thr Glu Lys Lys lie Glu 35 40 45 lie Ala Arg Ala Leu Asp Asp Phe Gly Val Asp Tyr lie Glu Leu Thr 50 55 60 -67- 146966-序列表.doc 201037078
Ser Pro Val Ala Ser Glu Gin Ser Arg Lys Asp Cys Glu Ala lie Cys 65 70 75 80
Lys Leu Gly Leu Lys Ala Lys lie Leu Thr His lie Arg Cys His Met 85 90 95
Asp Asp Ala Lys Val Ala Val Glu Thr Gly Val Asp Gly Val Asp Val 100 105 110
Val lie Gly Thr Ser Lys Phe Leu Arg Gin Tyr Ser His Gly Lys Asp 115 120 125
Met Asn Tyr lie Ala Lys Ser Ala Val Glu Val lie Glu Phe Val Lys 130 135 140
Ser Lys Gly lie Glu lie Arg Phe Ser Ser Glu Asp Ser Phe Arg Ser 145 150 155 160
Asp Leu Val Asp Leu Leu Asn lie Tyr Lys Thr Val Asp Lys lie Gly 165 170 175
Val Asn Arg Val Gly lie Ala Asp Thr Val Gly Cys Ala Asn Pro Arg 180 185 190
Gin Val Tyr Glu Leu lie Arg Thr Leu Lys Ser Val Val Ser Cys Asp 195 200 205 lie Glu Cys His Phe His Asn Asp Thr Gly Cys Ala lie Ala Asn Ala 210 215 220
Tyr Thr Ala Leu Glu Gly Gly Ala Arg Leu lie Asp Val Ser Val Leu 225 230 235 240
Gly lie Gly Glu Arg Asn Gly lie Thr Pro Leu Gly Gly Leu Met Ala 245 250 255
Arg Met lie Val Ala Ala Pro Asp Tyr Val Lys Ser Lys Tyr Lys Leu 260 265 270
His Lys lie Arg Asp lie Glu Asn Leu Val Ala Asp Ala Val Glu Val 275 280 285
Asn lie Pro Phe Asn Asn Pro lie Thr Gly Phe Cys Ala Phe Thr His 290 295 300 -68 - 146966·序列表.doc 201037078
Lys Ala Gly lie His Ala Lys Ala lie Leu Ala Asn Pro Ser Thr Tyr 305 310 315 320
Glu lie Leu Asp Pro His Asp Phe Gly Met Lys Arg Tyr lie His Phe 325 330 335
Ala Asn Arg Leu Thr Gly Trp Asn Ala lie Lys Ala Arg Val Asp Gin 340 345 350
Leu Asn Leu Asn Leu Thr Asp Asp Gin lie Lys Glu Val Thr Ala Lys 355 360 365 lie Lys Lys Leu Gly Asp Val Arg Ser Leu Asn lie Asp Asp Val Asp 370 375 380
O
Ser lie lie Lys Asn Phe His Ala Glu Val Ser Thr Pro Gin Val Leu 385 390 395 400
Ser Ala Lys Lys Asn Lys Lys Asn Asp Ser Asp Val Pro Glu Leu Ala 405 410 415
Thr lie Pro Ala Ala Lys Arg Thr Lys Pro Ser Ala 420 425 <210> 46 <211> 393
<212> PRT <213>乳酸克魯維酵母 <400> 46
Met Ser Val Asn Ser Asn Pro Tyr Ala Pro Ser Pro Asn Asp Leu Leu 15 10 15
Ser Asn Val Cys Asn Phe Gin Leu lie Glu Ser Thr Leu Arg Glu Gly 20 25 30
Glu Gin Phe Ala Ser Ala Phe Phe Ser Thr Glu Lys Lys lie Glu lie 35 40 45
Ala Lys Ala Leu Asp Asp Phe Gly Val Asp Tyr lie Glu Leu Thr Ser 50 55 60
Pro Val Ala Ser Glu Gin Ser Arg Ser Asp Cys Glu Ala lie Cys Lys 65 70 75 80
Leu Gly Leu Lys Ala Lys lie Leu Thr His lie Arg Cys His Met Asp 85 90 95 -69- 146966·序列表.doc 201037078
Asp Ala Arg Val Ala Val Glu Thr 100 105 lie Gly Thr Ser Lys Phe Leu Arg 115 120
Asn Tyr lie Ala Lys Ser Ala lie 130 135
Gly Val Asp Gly Val Asp Val Val 110
Glu Tyr Ser His Gly Lys Asp Met 125
Glu Val lie Glu Phe Val Lys Ser 140
Lys Gly Leu Glu lie Arg Phe Ser Ser Glu Asp Ser Phe Arg Ser Asp 145 150 155 160 lie Val Asp Leu Leu Asn lie Tyr Lys Thr Val Asp Lys lie Gly Val 165 170 175
Asn Arg Val Gly lie Ala Asp Thr Val Gly Cys Ala Asn Pro Arg Gin 180 185 190
Val Tyr Glu Leu Val Arg Thr Leu Lys Ser Val Val Ser Cys Asp lie 195 200 205
Glu Cys His Phe His Asp Asp Thr Gly Cys Ala lie Gly Asn Ser Tyr 210 215 220
Ser Ala Leu Glu Ala Gly Ala Arg Leu lie Asp Val Ser Val Leu Gly 225 230 235 240 lie Gly Glu Arg Asn Gly lie Thr Ser Leu Gly Gly Leu Met Ala Arg 245 250 255
Met lie Val Ser Ala Pro Glu Tyr Val Lys Ser Lys Tyr Lys Leu His 260 265 270
Lys Leu Arg Asp Leu Glu Asn Leu Val Ala Asp Ala Val Ser Val Asn 275 280 , 285
Val Pro Phe Asn Asn Pro lie Thr Gly Phe Cys Ala Phe Thr His Lys 290 295 300
Ala Gly lie His Ala Lys Ala lie Leu Ala Asn Pro Ser Thr Tyr Glu 305 310 315 320 lie Leu Asn Pro Glu Asp Phe Gly Met Lys Arg Tyr lie His Phe Ala 325 330 335 70· 146966-序列表.doc 201037078
Asn Arg Leu Thr Gly Trp Asn Ala lie Lys Ser Arg Val Glu Gin Leu 340 345 350
Asn Leu His Leu Ser Asp Asp Gin lie Lys Glu Val Thr Ser Lys lie 355 360 365
Lys Gin lie Gly Asp Val Arg Gin Leu Ser lie Glu Asp Val Asp Thr 370 375 380 lie lie Lys Asp Tyr His Ser Glu Leu 385 390 <210> 47 <211> 490
<212> PRT <213>黃孢原毛平革菌
<221> misc—feature <222> (62)7. (62) <223> Xaa可為任何天然存在之胺基酸 <400> 47
Leu Ser lie Leu Val Ala lie Gin Lys Leu Glu Pro Cys Cys Lys Met 1 5 10 15
Cys Pro His Ala Asn Gly Asp Ser Thr Pro Asn Asp Pro Ser Gin Met 20 25 30
Val Pro Val Asp Leu Ser Asn Gly Thr Ser His Gin Ala Ser Val Gin 35 40 45
Ser Asn Ser Asn Gly His Ala Ala Thr Asn Gly Ala Ala Xaa Asn Pro 50 55 60
Tyr Ala Pro Arg Ala Ser Asp Phe Leu Ser Asn Val Ser Asn Phe Lys 65 70 75 80 lie lie Glu Ser Thr Leu Arg Glu Gly Glu Gin Phe Ala Asn Ala Phe 85 90 95
Phe Asp Thr Lys Thr Lys lie Ala lie Ala Lys Ala Leu Asp Ala Phe 100 105 110
Gly Val Glu Tyr He Glu Leu Thr Ser Pro Ala Ala Ser Glu Gin Ser 115 120 125 -71 - 146966-序列表.doc 201037078
Arg Arg Asp Cys Glu Ala lie Cys Lys Leu Gly Leu Lys Ala Lys lie 130 135 140
Leu Thr His lie Arg Cys His Met Asp Asp Ala Arg lie Ala Val Glu 145 150 155 160
Thr Gly Val Asp Gly Val Asp Val Val 工le Gly Thr Ser Ser Phe Leu 165 170 175
Arg Glu Phe Ser His Gly Lys Asp Met Ala Tyr 工le Thr Lys Thr Ala 180 185 190 lie Glu Val lie Glu Phe Val Lys Ser Lys Gly lie Glu Val Arg Phe 195 200 205
Ser Ser Glu Asp Ser Phe Arg Ser Asp Leu Val Asp Leu Leu Ser lie 210 215 220
Tyr Gin Thr Val Asp Lys lie Gly Val Asn Arg Val Gly lie Ala Asp 225 230 235 240
Thr Val Gly Cys Ala Asn Pro Arg Gin Val Tyr Asp Leu Val Arg Thr 245 250 255
Leu Arg Gly Val Val Lys Cys Asp lie Glu lie His Leu His Asn Asp 260 265 270
Thr Gly Met Ala lie Ala Asn Ala Tyr Thr Ala Leu Glu Ala Gly Ala 275 280 285
Thr His 工le Asp Thr Ser Val Leu Gly lie Gly Glu Arg Val Gly lie 290 295 300
Thr Pro Leu Gly Gly Leu Val Ala Cys Leu Tyr Ala Ala Asn Pro Glu 305 310 315 320
Tyr Val Lys Ser Lys Tyr Asn Leu Pro Met Leu Arg Glu lie Glu Asn 325 330 335
Leu Val Ala Glu Ala Val Glu Val Asn lie Pro Phe Met Asn Pro lie 340 345 350
Thr Gly Tyr Cys Ala Phe Thr His Lys Ala Gly lie His Ala Lys Ala 355 360 365 72· 146966-序列表.doc 201037078 lie Leu Asn Asn Pro Ser Thr Tyr Glu lie Leu Lys Pro Glu Asp Phe 370 375 380
Gly Leu Thr Arg Tyr Val Ser lie Gly His Arg Leu Thr Gly Trp Asn 385 390 395 400
Ala Val Lys Ser Arg Val Glu Gin Leu Gly Leu Lys Leu Thr Asp Glu 405 410 415
Glu lie Lys Asp Val Thr Ala Lys lie Lys Glu Leu Ala Asp Val Arg 420 425 430
Thr Gin Ser Met Asp Asp Val Asp Thr Leu Leu Arg Val Tyr His Ser 435 440 445 o
Gly lie Gin Ser Gly Glu Leu Ala Ala Gly Gin Arg Glu Ala Leu Asp 450 455 460
Arg Leu Leu Arg Lys His Arg Glu Gly Thr Met Ser Arg Glu Pro Ser 465 470 475 480
Val Ser Arg Pro Ser Thr Pro Thr Gin Ala 485 490 <210> 48 <211> 441
<212> PRT <213>乳酸克魯維酵母 <400> 48
Met Ser Ser Asn Gin Asp Phe Gin Pro Val Thr Glu Ser Ala Ser Ser 15 10 15 ❹
Val Thr Lys Phe Gin Gin Asn Pro Tyr Gly Pro Asn Pro Ala Asp Tyr 20 25 30
Leu Ser Asn Val Asn Asn Tyr Gin Leu lie Asp Ser Thr Leu Arg Glu 35 40 45
Gly Glu Gin Phe Ala Asn Ala Phe Phe Asp Thr Glu Lys Lys lie Glu 50 55 60 lie Ala Lys Ala Leu Asp Asp Phe Gly Val Asp Tyr lie Glu Leu Thr 65 70 75 80
Ser Pro Val Ala Sar Glu Gin Ser Arg Arg Asp Cys Glu Ala lie Cys 85 90 95 -73 146966-序列表.doc 201037078
Lys Leu Gly Leu Lys Ala Lys lie Leu Thr His lie Arg Cys His Met 100 105 110
Asp Asp Ala Arg Val Ala Val Glu Thr Gly Val Asp Gly Val Asp Val 115 120 125
Val lie Gly Thr Ser Lys Phe Leu Arg Gin Tyr Ser His Gly Lys Asp 130 135 140
Met Asn Tyr lie Ala Lys Ser Ala lie Glu Val lie Glu Phe Val Lys 145 150 155 160
Ser Lys Gly lie Glu lie Arg Phe Ser Ser Glu Asp Ser Phe Arg Ser 165 170 175
Asp Leu Val Asp Leu Leu Asn lie Tyr Lys Thr Val Asp Lys lie Gly 180 185 190
Val Asn Arg Val Gly lie Ala Asp Thr Val Gly Cys Ala Asn Pro Arg 195 200 205
Gin Val Tyr Glu Leu Val Arg Thr Leu Lys Ser Val Val Ser Cys Asp 210 215 220 lie Glu Cys His Phe His Asn Asp Thr Gly Cys Ala lie Ala Asn Ala 225 230 235 240
Tyr Thr Ala Leu Glu Gly Gly Ala Arg Leu lie Asp Val Ala Val Leu 245 250 255
Gly lie Gly Glu Arg Asn Gly lie Thr Pro Leu Gly Gly Leu Met Ala 260 265 270
Arg Met lie Val Ala Ala Pro Glu Tyr Thr Lys Ser Lys Tyr Lys Leu 275 280 285
His Lys lie Arg Asp lie Glu Asn Leu lie Ala Glu Ala Val Glu Val 290 295 300
Asn lie Pro Phe Asn Asn Pro lie Thr Gly Phe Cys Ala Phe Thr His 305 310 315 320
Lys Ala Gly lie His Ala Lys Ala lie Leu Ala Asn Pro Ser Thr Tyr 325 330 335 •74- 146966·序列表.doc 201037078
Glu lie Leu Asp Pro His Asp Phe Gly Met Lys Arg Tyr lie His Phe 340 345 350
Ala Asn Arg Leu Thr Gly Trp Asn Ala lie Lys Ser Arg Val Asp Gin 355 360 365
Leu Asn Leu Asn Leu Thr Asp Asp Gin Val Lys Glu Val Thr Ala Lys 370 375 380 lie Lys Lys Leu Gly Asp lie Arg Pro Leu Asn lie Asp Asp Val Asp 385 390 395 400
Ser lie lie Lys Asp Phe His Ala Glu Val Ser Thr Pro Gin Leu Arg 405 410 415
Ala Val Arg Arg Asp Asp Asn Asp Val Asn Asp lie Asp lie Gin Glu 420 425 430
Pro Ser Asn Lys Lys Thr Lys Val Glu 435 440 <210> 49 <211> 418 <212> PRT <213>粟酒裂殖酵母 <400> 49
Met Ser Val Ser Glu Ala Asn Gly Thr Glu Thr lie Lys Pro Pro Met 15 10 15
Asn Gly Asn Pro Tyr Gly Pro Asn Pro Ser Asp Phe Leu Ser Arg Val 20 25 30 〇
Asn Asn Phe Ser lie lie Glu Ser Thr Leu Arg Glu Gly Glu Gin Phe 35 40 45
Ala Asn Ala Phe Phe Asp Thr Glu Lys Lys lie Gin lie Ala Lys Ala 50 55 60
Leu Asp Asn Phe Gly Val Asp Tyr lie Glu Leu Thr Ser Pro Val Ala 65 70 75 80
Ser Glu Gin Ser Arg Gin Asp Cys Glu Ala lie Cys Lys Leu Gly Leu 85 90 95
Lys Cys Lys lie Leu Thr His 工le Arg Cys His Met Asp Asp Ala Arg 75· 146966·序列表.doc 201037078 100 105 110
Val Ala Val Glu Thr Gly Val Asp 115 120
Ser Gin Tyr Leu Arg Lys Tyr Ser 130 135 lie Asp Ser Ala Thr Glu Val lie 145 150
Gly Val Asp Val Val He Gly Thr 125 His Gly Lys Asp Met Thr Tyr lie 140 Asn Phe Val Lys Ser Lys Gly lie 155 160
Glu Val Arg Phe Ser Ser Glu Asp Ser Phe Arg Ser Asp Leu Val Asp 165 170 175
Leu Leu Ser Leu Tyr Lys Ala Val Asp Lys lie Gly Val Asn Arg Val 180 185 190
Gly lie Ala Asp Thr Val Gly Cys Ala Thr Pro Arg Gin Val Tyr Asp 195 200 205
Leu lie Arg Thr Leu Arg Gly Val Val Ser Cys Asp lie Glu Cys His 210 215 220
Phe His Asn Asp Thr Gly Met Ala lie Ala Asn Ala Tyr Cys Ala Leu 225 230 235 240
Glu Ala Gly Ala Thr His lie Asp Thr Ser lie Leu Gly lie Gly Glu 245 250 255
Arg Asn Gly lie Thr Pro Leu Gly Ala Leu Leu 7Vla Arg Met Tyr Val 260 265 270
Thr Asp Arg Glu Tyr lie Thr His Lys Tyr Lys Leu Asn Gin Leu Arg 275 280 285
Glu Leu Glu Asn Leu Val Ala Asp Ala Val Glu Val Gin lie Pro Phe 290 295 300
Asn Asn Tyr lie Thr Gly Met Cys Ala Phe Thr His Lys Ala Gly lie 305 310 315 320
His Ala Lys Ala lie Leu Ala Asn Pro Ser Thr Tyr Glu lie Leu Lys 325 330 335
Pro Glu Asp Phe Gly Met Ser Arg Tyr Val His Val Gly Ser Arg Leu 340 345 350 -76- 146966·序列表.doc 201037078
Thr Gly Trp Asn Ala lie Lys Ser Arg Ala Glu Gin Leu Asn Leu His 355 360 365
Leu Thr Asp Ala Gin Ala Lys Glu Leu Thr Val Arg lie Lys Lys Leu 370 375 380
Ala Asp Val Arg Thr Leu Ala Met Asp Asp Val Asp Arg Val Leu Arg 385 390 395 400
Glu Tyr His Ala Asp Leu Ser Asp Ala Asp Arg lie Thr Lys Glu Ala 405 410 415
Ser Ala <210> 50 ^ <211> 465 <212> PRT <213>黑麴菌 <400> 50
Met Cys Pro Gly Ala Asp His Glu Pro Asn Gly Gin Ala Asn Val Ala 15 10 15
Asn Gly Asn Gly Asn Asn Gly Glu His Pro Gly Phe Thr Ala Val Glu 20 25 30
Thr Arg Gin Asn Pro His Pro Ser Val Ser Arg Asn Pro Tyr Gly His 35 40 45
Asn Val Gly Val Thr Asp Phe Leu Ser Asn Val Ser Arg Phe Gin lie
lie Glu Ser Thr Leu Arg Glu Gly Glu Gin Phe Ala Asn Ala Phe Phe 65 70 75 80
Asp Thr Glu Lys Lys lie Glu lie Ala Lys Ala Leu Asp Glu Phe Gly 85 90 95
Val Asp Tyr lie Glu Leu Thr Ser Pro Cys Ala Ser Glu Gin Ser Arg 100 105 110
Lys Asp Cys Glu Ala lie Cys Lys Leu Gly Leu Lys Ala Lys lie Leu 115 120 125 -77· 146966-序列表.doc 201037078
Thr His lie Arg Cys His Met Asp Asp Ala Arg lie Ala Val Glu Thr 130 135 140
Gly Val Asp Gly Val Asp Val Val lie Gly Thr Ser Ser Tyr Leu Arg 145 150 155 160
Glu His Ser His Gly Lys Asp Met Thr Tyr lie Lys Asn Thr Ala lie 165 170 175
Glu Val lie Glu Phe Val Lys Ser Lys Gly lie Glu lie Arg Phe Ser 180 185 190
Ser Glu Asp Ser Phe Arg Ser Asp Leu Val Asp Leu Leu Ser lie Tyr 195 200 205
Ser Ala Val Asp Lys Val Gly Val Asn Arg Val Gly lie Ala Asp Thr 210 215 220
Val Gly Cys Ala Ser Pro Arg Gin Val Tyr Glu Leu Val Arg Val Leu 225 230 235 240
Arg Gly Val Val Ser Cys Asp lie Glu Thr His Phe His Asn Asp Thr 245 250 255
Gly Cys Ala lie Ala Asn Ala Tyr Cys Ala Leu Glu Ala Gly Ala Thr 260 265 270
His 工le Asp Thr Ser Val Leu Gly lie Gly Glu Arg Asn Gly lie Thr 275 280 285
Pro Leu Gly Gly Leu Met Ala Arg Met Met Val Ala Asp Pro Glu Tyr 290 295 300
Val Lys Ser Lys Tyr Arg Leu Glu Lys Leu Lys Asp lie Glu Asp Leu 305 310 315 320
Val Ala Glu Ala Val Glu Val Asn lie Pro Phe Asn Asn Tyr lie Thr 325 330 335
Gly Phe Cys Ala Phe Thr His Lys Ala Gly lie His Ala Lys Ala lie 340 345 350
Leu Asn Asn Pro Ser Thr Tyr Glu lie lie Asn Pro Ala Asp Phe Gly 355 360 365
Met Ser Arg Tyr Val His Phe Ala Ser Arg Leu Thr Gly Trp Asn Ala 78· 146966·序列表.doc 201037078 370 375 380 lie Lys Ser Arg Ala Gin Gin Leu Lys lie Glu Met Thr Asp Asp Gin 385 390 395 400
Tyr Lys Glu Cys Thr Ala Lys lie Lys Ala Leu Ala Asp lie Arg Pro 405 410 415 lie Ala Val Asp Asp Ala Asp Ser lie lie Arg Ala Tyr Tyr Arg Asn 420 425 430
Leu Lys Leu Gly Glu Asn Lys Pro Leu Leu Asp Leu Thr Ala Asp Glu 435 440 445 o
Gin Ala Gin Phe Ala Ala Lys Glu Lys Glu Leu Ala Ala Gin Ala Ser 450 455 460
Ala 465 <210> 51 <211> 445 <212> PRT <213>構巢裸胞殼 <400> 51
Met Cys Pro Gly Asp His Pro Gly Phe Thr Ala Val Gin Thr Arg Gin 1 5 10 15
Asn Pro His Pro Ser Arg Asn Pro Tyr Gly His Asn Val Gly Val Thr 20 25 30
Asp Phe Leu Ser Asn Val Ser Arg Phe Lys lie lie Glu Ser Thr Leu 35 40 45
Arg Glu Gly Glu Gin Phe Ala Asn Ala Phe Phe Asp Thr Gin Lys Lys 50 55 60
He Glu lie Ala Lys Ala Leu Asp Glu Phe Gly Val Asp Tyr lie Glu 65 70 75 80
Leu Thr Ser Pro Cys Ala Ser Glu Gin Ser Arg Leu Asp Cys Glu Ala 85 90 95
lie Cys Lys Leu Gly Leu Lys Ala Lys lie Leu Thr His lie Arg Cys 100 105 HO -79 146966-序列表.doc 201037078
His Met Asp Asp Ala Arg Val Ala Val Glu Thr Gly Val Asp Gly Val 115 120 125
Asp Val Val lie Gly Thr Ser Ser Tyr Leu Arg Glu His Ser His Gly 130 135 140
Lys Asp Met Thr Tyr lie Lys Asn Thr Ala lie Glu Val lie Glu Phe 145 150 155 160
Val Lys Ser Lys Gly lie Glu lie Arg Phe Ser Ser Glu Asp Ser Phe 165 170 175
Arg Ser Asp Leu Val Asp Leu Leu Ser lie Tyr Ser Ala Val Asp Gin 180 185 190
Val Gly Val Asn Arg Val Gly lie Ala Asp Thr Val Gly Cys Ala Ser 195 200 205
Pro Arg Gin Val Tyr Glu Leu lie Arg Val Leu Arg Gly Val Val Ser 210 215 220
Cys Asp lie Glu Thr His Phe His Asn Asp Thr Gly Cys Ala 工le Ala 225 230 235 240
Asn Ala Tyr Cys Ala Leu Glu Ala Gly Ala Thr His lie Asp Thr Ser 245 250 255
Val Leu Gly lie Gly Glu Arg Asn Gly lie Thr Pro Leu Gly Gly Leu 260 265 270
Met Ala Arg Met Met Val Ala Asp Pro Gin Tyr Val Lys Ser Lys Tyr 275 280 285
Lys Leu Glu Lys Leu Lys Asp lie Glu Asp Leu Val Ala Glu Ala Val 290 295 300
Glu Val Asn lie Pro Phe Asn Asn Tyr lie Thr Gly Phe Cys Ala Phe 305 310 315 320
Thr His Lys Ala Gly lie His Ala Lys Ala lie Leu Asn Asn Pro Ser 325 330 335
Thr Tyr Glu lie lie Asn Pro Ala Asp Phe Gly Met Ser Arg Tyr Val 340 345 350 •80· 146966-序列表.doc 201037078
His Phe Ala Ser Arg Leu Thr Gly Trp Asn Ala lie Lys Ser Arg Ala 355 360 365
Gin Gin Leu Asn Val His Met Thr Asp Asp Gin Tyr Lys Glu Cys Thr 370 375 380
Ala Lys lie Lys Ala Leu Ala Asp lie Arg Pro lie Ala lie Asp Asp 385 390 395 400
Ala Asp Ser lie lie Arg Ala Tyr Tyr Arg Asn Leu Ser Ser Gly Glu 405 410 415
Asn Lys Pro Leu Met Asp Leu Thr Ala Asp Glu His Ala Gin Phe Leu 420 425 430
Ala Lys Glu Lys Glu Leu Thr Glu Ser Gly Thr Ala Leu 435 440 445 <210> 52 <211> 474 <212> PRT <213>產黃青黴 <400> 52
Met Val Leu Leu Pro Pro Ser Leu Pro Val Cys Gin Leu Lys Val Thr 15 10 15
Ala Pro Glu Phe Pro Ser Asn Phe Tyr Leu Asp Gly Asp His Ser Gly 20 25 30
Phe Val Gly lie Glu Thr Arg Gin Asn Pro His Pro Ser Ala Ser Arg 35 40 45 o
Asn Pro Tyr Gly His Asp Ala Gly Val Thr Asp Phe Leu Ser Asn Val 50 55 60
Ser Arg Phe Gin lie lie Glu Ser Thr Leu Arg Glu Gly Glu Gin Phe 65 70 75 80
Ala Asn Ala Phe Phe Asp Thr Ala Lys Lys lie Glu lie Ala Lys Ala 85 90 95
Leu Asp Asp Phe Gly Val Asp Tyr lie Glu Leu Thr Ser Pro Cys Ala 100 105 110
Ser Glu Gin Ser Arg Ala Asp Cys Glu Ala lie Cys Lys Leu Gly Leu 115 120 125 •81 146966·序列表.doc 201037078
Lys Ala Lys lie Leu Thr His lie Arg Cys His Met Asp Asp Ala Arg 130 135 140 lie Ala Val Glu Thr Gly Val Asp Gly Val Asp Val Val lie Gly Thr 145 150 155 160
Ser Ser Tyr Leu Arg Glu His Ser His Gly Lys Asp Met Thr Tyr lie 165 170 175
Lys Asn Ala Ala lie Glu Val lie Glu Phe Val Lys Ser Lys Gly lie 180 185 190
Glu lie Arg Phe Ser Ser Glu Asp Ser Phe Arg Ser Asp Leu Val Asp 195 200 205
Leu Leu Ser lie Tyr Ser Ala Val Asp Lys Val Gly Val Asn Arg Val 210 215 220
Gly lie Ala Asp Thr Val Gly Cys Ala Ser Pro Arg Gin Val Tyr Glu 225 230 235 240
Leu Val Arg Val Leu Arg Gly Val Val Gly Cys Asp lie Glu Thr His 245 250 255
Phe His Asn Asp Thr Gly Cys Ala lie Ala Asn Ala Phe Cys Ala Leu 260 265 270
Glu Ala Gly Ala Thr His lie Asp Thr Ser Val Leu Gly lie Gly Glu 275 280 285
Arg Asn Gly lie Thr Pro Leu Gly Gly Leu Met Ala Arg Met Met Val 290 295 300
Ala Asp Arg Glu Tyr Val Lys Ser Lys Tyr Lys Leu Glu Lys Leu Lys 305 310 315 320
Glu lie Glu Asp Leu Val Ala Glu Ala Val Glu Val Asn lie Pro Phe 325 330 335
Asn Asn Tyr lie Thr Gly Phe Cys Ala Phe Thr His Lys Ala Gly lie 340 345 350
His Ala Lys Ala 工le Leu Asn Asn Pro Ser Thr Tyr Glu lie lie Asn 355 360 365 -82- 146966-序列表.doc 201037078
Pro Ala Asp Phe Gly Met Ser Arg Tyr Val His Phe Ala Ser Arg Leu 370 375 380
Thr Gly Trp Asn Ala lie Lys Ser Arg Ala Gin Gin Leu Lys Leu Glu 385 390 395 400
Met Thr Asp Thr Gin Tyr Lys Glu Cys Thr Ala Lys lie Lys Ala Met 405 410 415
Ala Asp lie Arg Pro lie Ala Val Asp Asp Ala Asp Ser lie lie Arg 420 425 430
Ala Tyr His Arg Asn Leu Lys Ser Gly Glu Asn Lys Pro Leu Leu Asp 435 440 445 o
Leu Thr Ala Glu Glu Gin Ala Ala Phe Ala Ala Lys Glu Lys Glu Leu 450 455 460
Leu Glu Ala Gin Ala Ala Gly Leu Pro Val 465 470 <210> 53 <211> 446 <212> PRT <213>解脂耶氏酵母 <400> 53 Met Cys Ala Thr Asp Asn Ala Pro Ala Ala Asn Ala Ala Pro Glu Lys 15 10 15
Pro Ser Asn Val Gly Val Glu Val Gly His Thr Gly Glu Gin Thr Asn 20 25 30 〇
Pro Tyr Gly Ala Asn Pro Ala Asp Phe Leu Ser Asn Val Ser Lys Phe 35 40 45
Gin Leu lie Glu Ser Thr Leu Arg Glu Gly Glu Gin Phe Ala Ser Ala 50 55 60
Phe Phe Asp Thr Glu Thr Lys lie Glu lie Ala Lys Ala Leu Asp Asp 65 70 75 80
Phe Gly Val Asp Tyr lie Glu Leu Thr Ser 85 90
Pro Ala Ala Ser Glu Gin 95
Ser Arg Ser Asp Cys Glu Ala lie Cys Lys Leu Gly Leu Lys Ala Lys 146966-序列表.doc -83 - 201037078 100 105 110 lie Leu Thr His lie Arg Cys His Met Asp Asp Ala Arg Leu Ala Val 115 120 125
Ser Thr Gly Val Asp Gly Val Asp Val Val lie Gly Thr Ser Gin Phe 130 135 140
Leu Arg Gin Tyr Ser His Gly Lys Asp Met Asn Tyr lie Ala Gin Ser 145 150 155 160
Ala Val Glu Val lie Glu Phe Val Lys Ser His Gly lie Glu lie Arg 165 170 175
Phe Ser Ser Glu Asp Ser Phe Arg Ser Asp Leu Val Asp Leu Leu Asn 180 185 190 lie Tyr Arg Thr Val Asp Lys lie Gly Val Asn Arg Val Gly lie Ala 195 200 205
Asp Thr Val Gly Cys Ala Asn Pro Arg Gin Val Tyr Glu Leu Val Arg 210 215 220
Thr Leu Lys Ser Val Val Ser Cys Asp lie Glu Cys His Phe His Asn 225 230 235 240
Asp Thr Gly Cys Ala 工le Ala Asn Ala Tyr Thr Ala Leu Glu Ala Gly 245 250 255
Ala Asn Leu lie Asp Val Ser Val Leu Gly lie Gly Glu Arg Asn Gly 260 265 270 lie Thr Ser Leu Gly Gly Leu Met Ala Arg Met lie Ala Ala Asp Arg 275 280 285
Asp Tyr Val Leu Ser Lys Tyr Lys Leu His Lys Leu Arg Asp Leu Glu 290 295 300
Asn Leu Val Ala Asp Ala Val Gin Val Asn lie Pro Phe Asn Asn Pro 305 310 315 320 lie Thr Gly Phe Cys Ala Phe Thr His Lys Ala Gly lie His Ala Lys 325 330 335
Ala lie Leu Ala Asn Pro Ser Thr Tyr Glu lie Leu Asn Pro Ala Asp 340 345 350 -84 146966-序列表.doc 201037078
Phe Gly Len Thr Arg Tyr lie His Phe Ala Asn Arg Leu Thr Gly Trp 355 360 365
Asn Ala 工le Lys Ser Arg Val Asp Gin Leu Asn Leu His Leu Thr Asp 370 375 380
Ala Gin Cys Lys Asp Val Thr Ala Lys lie Lys Lys Leu Gly Asp Val 385 390 395 400
Arg Ser Leu Asn lie Asp Asp Val Asp Ser lie lie Arg Glu Phe His 405 410 415
Ala Asp Val Thr Ser Thr Pro Thr Val Ala Ala Thr Glu Gly Pro Ala 420 425 430 o
Val Glu Asp Glu Pro Ala Ala Lys Lys Ala Lys Thr Glu Glu 435 440 445 <210> 54 <211> 687
<212> PRT <213>黃孢原毛平革菌 <400> 54 lie Pro Gin Thr Val He Glu Lys Val Val Gin Lys Tyr Ala Val Gly 1 5 10 15
Leu Pro Gly Asp Lys Val Val Lys Ala Gly Asp Tyr Val Met lie Arg 20 25 30 .
Pro Glu His Val Met Thr His Asp Asn Thr Gly Pro Val lie Ser Lys 35 40 45 ❹
Phe Lys Ser lie Gly Ala Thr Arg lie Tyr Asn Pro Lys Gin Val Val 50 55 60
Phe Thr Leu Asp His Asp Val Gin Asn Lys Ser Glu Lys Asn Leu Lys 65 70 75 80
Lys Tyr Ala Thr He Glu Ala Phe Ala Arg Thr His Gly He Asp Phe 85 90 95
Tyr Pro Ala Gly Arg Gly He Gly His Gin Val Leu Val Glu Glu Gly 100 105 110 85- 146966-序列表.doc 201037078
Tyr Ala Phe Pro His Thr Leu Thr Val Ala Ser Asp Ser His Ser Asn 115 120 125
Met Tyr Gly Gly Val Gly Cys Val Gly Thr Pro lie Val Arg Thr Asp 130 135 140
Ala Ala Ala Leu Trp Ala Thr Gly Gin Thr Trp Trp Gin Val Pro Arg 145 150 155 160
Met Val Lys Val Glu Phe Lys Gly Arg Leu Ala Pro Gly Val Ser Gly 165 170 175
Lys Asp Val lie Val Ala Leu Cys Gly Ser Phe Asn Lys Asp Glu Val 180 185 190
Leu Asn Ala Ala lie Glu Phe Ser Gly Glu Gly Val Gin His Leu Thr 195 200 205
Val Asp Glu Arg Leu Thr lie Ala Asn Met Thr Thr Glu Trp Gly Ala 210 215 220
Leu Val Gly Val Phe Pro Val Asp Asp Val Thr Leu Ser Trp Tyr Glu 225 230 235 240
Arg Met Leu Lys Lys Leu Glu Leu Arg Thr Phe Ser Thr Pro Ala Leu 245 250 255
Gly Ser Ser lie Pro Pro Pro Pro Glu His Pro Arg lie Asn Arg Ala 260 265 270
Arg Leu Asp Ala Leu Arg Ala Asn Asn Leu Arg Ser Asp Ala Asp Ala 275 280 285
Glu Tyr Ser Ser His Leu Val Phe Asp Leu Ser Thr Leu Val Pro Tyr 290 295 300
Val Ser Gly Pro Asn Ser Val Lys Val Ala Asn Pro Leu Pro Lys Leu 305 310 315 320
Glu Glu Ala Lys lie Lys 工le Asn Lys Ala Tyr Leu Leu Ser Cys Thr 325 330 335
Asn Ala Arg Ala Ser Asp lie Ala Ala Ala Ala Ala Val lie Lys Gly 340 345 350
His Lys Val His Pro Asp Val Gin Phe Tyr Phe Ala Pro Ala Ser Ser •86· 146966-序列表.doc 201037078 355 360 365
Glu Val Gin Arg Glu Ala Glu Gin Ser Gly Asp Trp Glu Thr Leu lie 370 375 380
Gly Ala Gly Ala Lys Pro Leu Pro Ala Gly Cys Gly Pro Cys lie Gly 385 390 395 400
Leu Gly Thr Gly Leu Leu Glu Glu Gly Glu Val Gly lie Ser Ala Thr 405 410 415
Asn Arg Asn Tyr Lys Gly Arg Met Gly His Pro Leu Ala Gin Ala Tyr 420 425 430
Leu Ala Ser Pro Ala Val Val Ala Ala Ser Ala lie Lys Gly Tyr lie 435 440 445
Ala Gly Pro Asp Ser Leu Asp Pro Ser Lys Leu Pro Pro Ala Gly Ala 450 455 460
Pro Thr Phe Ser lie Val Asn Ser Pro Ser Ser Gly Ala Lys Ala Ser 465 470 475 480
Gin Lys Glu Pro Val Leu Val Gly Phe Pro Glu Thr Phe Ala Gly Pro 485 490 495
Leu Leu Phe Ala Pro Gin Asp Asn Leu Asn Thr Asp Gly lie Tyr Pro 500 505 510
Gly Lys Tyr Thr Tyr Gin Asp Asp lie Thr Leu Glu Arg Gin Ala Glu 515 520 525 〇
Val Val Met Glu Asn Tyr Asp Pro Thr Phe Ala Gin Leu Asp Ala His 530 535 540
Thr Lys Arg Gly Val Val Leu Val Ser Gly Tyr Asn Phe Gly Thr Gly 545 550 555 560
Ser Ser Arg Glu Gin Ala Ala Thr Ala Leu Lys Ser Ala Gly lie Pro 565 570 575 lie Val lie Ala Gly Ser Phe Gly Asp lie Phe Lys Arg Asn Ala lie 580 585 590
Asn Asn Gly Leu Val Cys Val Glu Ser Pro Glu Leu Val Ala Asp Leu 595 600 605 • 87 - 146966-序列表.doc 201037078
Thr Ala Gin Phe Ala Lys Asp Gly Lys Arg Gly Ala Gly Gly Lys Glu 610 615 620
Gly Glu Leu Thr Val Asn Lys Gly Leu Ser Ala Glu Val Lys Val Val 625 630 635 640
Asp Gly Ala Leu His Val Thr Phe Pro Asp Gly Lys Thr Lys Thr Tyr 645 650 655
Thr lie Gin Pro Val Gly Ala Ser Val Gin Glu Leu Trp Leu Cys Gly 660 665 670
Gly Leu Glu Gly Tyr Val Leu Lys Ala lie Gin Ala Glu Asn Phe 675 680 685 <210> 55 <211> 721 <212> PRT <213>粟酒裂殖酵母 <400> 55
Met Asp Ser Gly Glu Met His His Pro Tyr Gin Ala Phe Ser Lys Val 15 10 15
Gly Lys Cys Glu lie Ser Gin Thr Asn Pro Ser Phe Ser Ser Gly Met 20 25 30
Arg Cys Leu Val Arg Ser Ala Asp lie Gin Phe Lys Gly lie Cys Gly 35 40 45
Leu Thr Arg Gly Phe Ala Ser Phe Asn Lys Pro Pro Gin Thr lie Thr 50 55 60
Glu Lys lie Val Gin Lys Phe Ala Gin Asn lie Pro Glu Asn Lys Tyr 65 70 75 80
Val Arg Ser Gly Asp Tyr Val Thr lie Lys Pro Lys His Cys Met Ser 85 90 95
His Asp Asn Ser Trp Pro Val Ala Leu Lys Phe Met Gly lie Gly Ala 100 105 110
Lys Lys Val Phe Asp Asn Arg Gin lie Val Cys Thr Leu Asp His Asp 115 120 125 88· 146966-序列表.doc 201037078
Val Gin Asn Lys Ser Glu Ala Asn Leu Arg Lys Tyr Lys Asn lie Glu 130 135 140
Ser Phe Ala Lys Gly Gin Gly lie Asp Phe Tyr Pro Ala Gly Arg Gly 145 150 155 160 lie Gly His Gin lie Met Val Glu Gin Gly Tyr Ala Met Pro Gly Ser 165 170 175
Met Ala Val Ala Ser Asp Ser His Ser Asn Thr Tyr Gly Gly Val Gly 180 185 190
Cys Leu Gly Thr Pro lie Val Arg Thr Asp Ala Ala Ala lie Trp Ala 195 200 205
o
Thr Gly Gin Thr Trp Trp Gin lie Pro Pro lie Ala Arg Val Asn Leu 210 215 220
Val Gly Gin Leu Pro Lys Gly Leu Ser Gly Lys Asp lie lie Val Ser 225 230 235 240
Leu Cys Gly Ala Phe Asn His Asp Glu Val Leu Asn His Ala lie Glu 245 250 255
Phe Tyr Gly Glu Gly Leu Asn Ser Leu Ser lie Glu Ser Arg Leu Thr 260 265 270 lie Ala Asn Met Thr Thr Glu Trp Gly Ala Leu Ser Gly Leu Phe Pro 275 280 285
Thr Asp Glu Lys Leu Leu Ala Trp Tyr Glu Asp Arg Leu Lys Phe Leu 290 295 300
Gly Pro Asn His Pro Arg Val Asn Arg Glu Thr Leu Asp Ala lie Lys 305 310 315 320
Ala Ser Pro lie Leu Ala Asp Glu Gly Ala Phe Tyr Ala Lys His Leu 325 330 335 lie Leu Asp Leu Ser Thr Leu Ser Pro Ala Val Ser Gly Pro Asn Ser 340 345 350
Val Lys Val Tyr Asn Ser Ala Ala Thr Leu Glu Lys Lys Asp lie Leu 355 360 365 lie Lys Lys Ala Tyr Leu Val Ser Cys Thr Asn Gly Arg Leu Ser Asp -89- 146966_ 序列表.doc 201037078 370 375 380 lie His Asp Ala Ala Glu Thr Val Lys Gly Lys Lys Val Ala Asp Gly 385 390 395 400
Val Glu Phe Tyr Val Gly Ala Ala Ser Ser Glu Val Glu Ala Ala Ala 405 410 415
Gin Lys Asn Gly Asp Trp Gin Thr Leu lie Asp Ser Gly Ala Arg Thr 420 425 430
Leu Pro Ala Gly Cys Gly Pro Cys lie Gly Leu Gly Thr Gly Leu Leu 435 440 445
Lys Asp Gly Glu Val Gly lie Ser Ala Thr Asn Arg Asn Phe Lys Gly 450 455 460
Arg Met Gly Ser Arg Glu Ala Leu Ala Tyr Leu Ala Ser Pro Ala Val 465 470 475 480
Val Ala Ala Ser Ala lie Ala Gly Lys lie Val Ala Pro Glu Gly Phe 485 490 495
Lys Asn Ala Val Ser Leu Val Ser Ala Val Asp lie Thr Asp Lys Val 500 505 510
Asn Lys Gin Thr Ala Ser Lys Ser Ser Thr Glu Ala Val Asp Ser Glu 515 520 525
Thr Ala lie lie Asp Gly Phe Pro Ser lie Val Ala Gly Glu 工le Val 530 535 540
Phe Cys Asp Ala Asp Asn Leu Asn Thr Asp Gly lie Tyr Pro Gly Arg 545 550 555 560
Tyr Thr Tyr Arg Asp Asp lie Thr Lys Glu Glu Met Ala Lys Val Cys 565 570 575
Met Glu Asn Tyr Asp Ser Glu Phe Gly Lys Lys Thr Lys Lys Asp Asp 580 585 590 lie Leu Val Ser Gly Phe Asn Phe Gly Thr Gly Ser Ser Arg Glu Gin 595 600 605
Ala Ala Thr Ala lie Leu Ser Arg Gly lie Pro Leu Val Val Gly Gly 610 615 620 90- 146966-序列表.doc 201037078
Ser Phe Ser Asp lie Phe Lys Arg Asn Ser lie Asn Asn Ala Leu Leu 625 630 635 640
Ala lie Gin Leu Pro Asp Leu Val Gin Lys Leu Arg Thr Ala Phe Ala 645 650 655
Asn Glu Ser Lys Glu Leu Thr Arg Arg Thr Gly Trp His Leu Lys Trp 660 665 670
Asp Val Arg Lys Ser Thr Val Thr Val Thr Thr Ser Asp Asn Lys Glu 675 680 685 Met Ser Trp Lys lie Gly Glu Leu Gly Asn Ser Val Gin Ser Leu Phe 690 695 700 o
Val Arg Gly Gly Leu Glu Gly Trp Val Lys His Glu lie Ser Lys Ser 705 710 715 720
Asn <210> <211> <212> <213>
56 693 PRT 乳酸克魯維酵母 <400> 56
Met Phe Arg Val Gin Arg Leu Arg Met Phe Ser Thr Ser Arg Ala Leu 15 10 15
Tyr Ala Gly Gin Asn Met Thr Glu Lys 工le Val Gin Arg His Ala Val 20 25 30 o
Gly Leu Pro Glu Gly Lys Thr Val Val Ser Gly Asp Tyr Val Ser lie 35 40 45
Lys Pro Ala His Cys Met Ser His Asp Asn Ser Trp Pro Val Ala Leu 50 55 60
Lys Phe Met Gly Leu Gly Ala Ser Thr lie Lys Asn Pro Arg Gin Val 65 70 75 80
Val Asn Thr Leu Asp His Asp Val Gin Asn Lys Ser Glu Lys Asn Leu 85 90 95 146966-序列表.doc -91 - 201037078
Thr Lys Tyr Lys Asn lie Glu Asn Phe Ala Lys Lys His Gly lie Asp 100 105 110
Phe Tyr Pro Ala Gly Arg Gly lie Gly His Gin lie Met lie Glu Glu 115 120 125
Gly Tyr Ala Phe Pro Leu Thr Met Thr Val Ala Ser Asp Ser His Ser 130 135 140
Asn Thr Tyr Gly Gly lie Gly Ala Leu Gly Thr Pro lie Val Arg Thr 145 150 155 160
Asp Ala Ala Ala lie Trp Ala Thr Gly Gin Thr Trp Trp Gin lie Pro 165 170 175
Pro Val Ala Gin Val Glu Leu Lys Gly Glu Leu Pro Ala Gly lie Ser 180 185 190
Gly Lys Asp lie lie Val Ala Leu Cys Gly Val Phe Asn Gin Asp Gin 195 200 205
Val Leu Asn His Ala lie Glu Phe Thr Gly Asp Ser Leu Asp Lys lie 210 215 220
Pro lie Asp Tyr Arg Leu Thr lie Ala Asn Met Thr Thr Glu Trp Gly 225 230 235 240
Ala Leu Ser Gly Leu Phe Pro Val Asp Asn Val Leu Leu Asp Phe Tyr 245 250 255
Arg Asn Arg Leu Thr Lys Val Gly Asn Asn His Pro Arg lie Asn Glu 260 265 270
Ala Arg lie Asn Glu Leu Gin Ala Lys Ser Asp Ser Leu Gin Ala Asp 275 280 285
Pro Asp Ala Lys Tyr Ala Lys Lys Leu lie lie Asp Leu Ser Thr Leu 290 295 300
Thr His Tyr Val Ser Gly Pro Asn Ser Val Lys lie Ser Ser Thr Val 305 310 315 320
Asp Asp Leu Ser Lys Gin Asp lie Lys Val Asn Lys Ala Tyr Leu Val 325 330 335
Ser Cys Thr Asn Ser Arg Leu Ser Asp Leu Glu Ser Ala Ala Asn Val -92- 146966-序列表.doc 201037078 340 345 350
Val Cys Pro Ser Gly Asp 工le Asn Gin Val His Lys Val Ala Glu Gly 355 360 365
Val Glu Phe Tyr lie Ala Ala Ala Ser Ser Glu Val Glu Ala Glu Ala 370 375 380
Arg Ala Thr Gly Ala Trp Gin Lys Leu Leu Asn Ala Gly Cys Leu Pro 385 390 395 400
Leu Pro Ala Gly Cys Gly Pro Cys lie Gly Leu Gly Thr Gly Leu Leu 405 410 415
Glu Glu Gly Gin Val Gly lie Ser Ala Thr Asn Arg Asn Phe Lys Gly 420 425 430
Arg Met Gly Ser Lys Asp Ala Leu Ala Tyr Leu Ala Ser Pro Ser Val 435 440 445
Val Ala Ala Ser Ala lie Leu Gly Lys lie Gly Ser Pro Ala Glu Val 450 455 460
Leu Gly Thr Lys Asp Pro Asn Phe Thr Gly Val Val Ala Thr Val Glu 465 470 475 480
Asp Ala Pro Ala Thr Ser Ala Asp Gly Lys Asp Val Ala Asp Glu Ser 485 490 495
Gly Ala Ser Gly Ser Val Glu lie Leu Glu Gly Phe Pro Ser Glu lie _ 500 505 510 〇
Ser Gly Glu Leu Val Leu Cys Asp Ala Asp Asn lie Asn Thr Asp Gly 515 520 525 lie Tyr Pro Gly Lys Tyr Thr Tyr Gin Asp Asp Val Pro Lys Glu Thr 530 535 540
Met Ala Lys Val Cys Met Glu Asn Tyr Asp Pro Asp Phe Gin Thr Lys 545 550 555 560
Ala Asn Pro Gly Asp lie Leu lie Ser Gly Phe Asn Phe Gly Thr Gly 565 570 575
Ser Ser Arg Glu Gin Ala Ala Thr Ala lie Leu Ala Lys Gly lie Lys 580 585 590 -93- 146966-序列表.doc 201037078
Leu Val Val Ser Gly Ser Phe Gly Asn lie Phe Phe Arg Asn Ser lie 595 600 605
Asn Asn Ala Leu Leu Thr Leu Glu lie Pro Ala Leu lie Asn Met Leu 610 615 620
Arg Asp Arg Tyr Lys Asp Ala Pro Lys Glu Leu Thr Arg Arg Thr Gly 625 630 635 640
Trp Phe Leu Lys Trp Asp Val Ser Gin Ala Lys Val Tyr Val Thr Glu 645 650 655
Gly Ser Val Asn Gly Pro lie Val Leu Glu Gin Lys Val Gly Glu Leu 660 665 670
Gly Lys Asn Leu Gin Glu lie lie Val Lys Gly Gly Leu Glu Ser Trp 675 680 685
Val Lys Ser Gin Leu 690 <210> 57 <211> 693 <212> PRT <213> 釀酒酵母 <400> 57
Met Leu Arg Ser Thr Thr Phe Thr Arg Ser Phe His Ser Ser Arg Ala 15 10 15
Trp Leu Lys Gly Gin Asn Leu Thr Glu Lys lie Val Gin Ser Tyr Ala 20 25 30
Val Asn Leu Pro Glu Gly Lys Val Val His Ser Gly Asp Tyr Val Ser 35 40 45 lie Lys Pro Ala His Cys Met Ser His Asp Asn Ser Trp Pro Val Ala 50 55 60
Leu Lys Phe Met Gly Leu Gly Ala Thr Lys lie Lys Asn Pro Ser Gin 65 70 75 80 lie Val Thr Thr Leu Asp His Asp lie Gin Asn Lys Ser Glu Lys Asn 85 90 95 -94- 146966-序列表.doc 201037078
Leu Thr Lys Tyr Lys Asn lie Glu Asn Phe Ala Lys Lys His His lie 100 105 110
Asp His Tyr Pro Ala Gly Arg Gly lie Gly His Gin lie Met lie Glu 115 120 125
Glu Gly Tyr Ala Phe Pro Leu Asn Met Thr Val Ala Ser Asp Ser His 130 135 140
Ser Asn Thr Tyr Gly Gly Leu Gly Ser Leu Gly Thr Pro lie Val Arg 145 150 155 160
Thr Asp Ala Ala Ala lie Trp Ala Thr Gly Gin Thr Trp Trp Gin lie 165 170 175
Pro Pro Val Ala Gin Val Glu Leu Lys Gly Gin Leu Pro Gin Gly Val 180 185 190
Ser Gly Lys Asp lie lie Val Ala Leu Cys Gly Leu Phe Asn Asn Asp 195 200 205
Gin Val Leu Asn His Ala lie Glu Phe Thr Gly Asp Ser Leu Asn Ala 210 215 220
Leu Pro lie Asp His Arg Leu Thr lie Ala Asn Met Thr Thr Glu Trp 225 230 235 240
Gly Ala Leu Ser Gly Leu Phe Pro Val Asp Lys Thr Leu lie Asp Trp 245 250 255
Tyr Lys Asn Arg Leu Gin Lys Leu Gly Thr Asn Asn His Pro Arg lie 260 265 270
Asn Pro Lys Thr lie Arg Ala Leu Glu Glu Lys Ala Lys lie Pro Lys 275 280 285
Ala Asp Lys Asp Ala His Tyr Ala Lys Lys Leu lie lie Asp Leu Ala 290 295 300
Thr Leu Thr His Tyr Val Ser Gly Pro Asn Ser Val Lys Val Ser Asn 305 310 315 320
Thr Val Gin Asp Leu Ser Gin Gin Asp lie Lys lie Asn Lys Ala Tyr 325 330 335
Leu Val Ser Cys Thr Asn Ser Arg Leu Ser Asp Leu Gin Ser Ala Ala -95- 146966-序列表.doc 201037078 340 345 350
Asp Val Val Cys Pro Thr Gly Asp Leu Asn Lys Val Asn Lys Val Ala 355 360 365
Pro Gly Val Glu Phe Tyr Val Ala Ala Ala Ser Ser Glu lie Glu Ala 370 375 380
Asp Ala Arg Lys Ser Gly Ala Trp Glu Lys Leu Leu Lys Ala Gly Cys 385 390 395 400 lie Pro Leu Pro Ser Gly Cys Gly Pro Cys lie Gly Leu Gly Ala Gly 405 410 415
Leu Leu Glu Pro Gly Glu Val Gly lie Ser Ala Thr Asn Arg Asn Phe 420 425 430
Lys Gly Arg Met Gly Ser Lys Asp Ala Leu Ala Tyr Leu Ala Ser Pro 435 440 445
Ala Val Val Ala Ala Ser Ala Val Leu Gly Lys lie Ser Ser Pro Ala 450 455 460
Glu Val Leu Ser Thr Ser Glu lie Pro Phe Ser Gly Val Lys Thr Glu 465 470 475 480 lie lie Glu Asn Pro Val Val Glu Glu Glu Val Asn Ala Gin Thr Glu 485 490 495
Ala Pro Lys Gin Ser Val Glu lie Leu Glu Gly Phe Pro Arg Glu Phe 500 505 510
Ser Gly Glu Leu Val Leu Cys Asp Ala Asp Asn lie Asn Thr Asp Gly 515 520 525 lie Tyr Pro Gly Lys Tyr Thr Tyr Gin Asp Asp Val Pro Lys Glu Lys 530 535 540
Met Ala Gin Val Cys Met Glu Asn Tyr Asp Ala Glu Phe Arg Thr Lys 545 550 555 560
Val His Pro Gly Asp lie Val Val Ser Gly Phe Asn Phe Gly Thr Gly 565 570 575
Ser Ser Arg Glu Gin Ala Ala Thr Ala Leu Leu Ala Lys Gly lie Asn 580 585 590 -96 146966-序列表.doc 201037078
Leu Val Val Ser Gly Ser Phe Gly Asn lie Phe Ser Arg Asn Ser lie 595 600 605
Asn Asn Ala Leu Leu Thr Leu Glu lie Pro Ala Leu lie Lys Lys Leu 610 615 620
Arg Glu Lys Tyr Gin Gly Ala Pro Lys Glu Leu Thr Arg Arg Thr Gly 625 630 635 640
Trp Phe Leu Lys Trp Asp Val Ala Asp Ala Lys Val Val Val Thr Glu 645 650 655
Gly Ser Leu Asp Gly Pro Val lie Leu Glu Gin Lys Val Gly Glu Leu 660 665 670
Gly Lys Asn Leu Gin Glu lie lie Val Lys Gly Gly Leu Glu Gly Trp 675 680 685
Val Lys Ser Gin Leu 690 <210> 58 <211> 769 <212> PRT <213>黑麴菌 <400> 58
Met Gin Ser Arg Leu Leu Pro Ser Gly Pro Gly Arg Arg Trp lie Ser 15 10 15
Leu Arg Val Pro Asn Thr Pro Gin Arg Arg Ala Phe Ala Ser Thr Arg 20 25 30 o
Phe Leu Phe Gin Asp Val Phe Gin Ser Gin Leu Asp Asp Pro Ser Ser 35 40 45
Ala Ala Leu Phe Ser Ser Leu Gin Ser Ser Arg Ala Val Pro Gin Thr 50 55 60
Leu Thr Glu Lys lie Val Gin Lys Tyr Ala Val Gly Leu Pro Asp Gly 65 70 75 80
Lys Phe Val Lys Ser Gly Asp Tyr Val Thr lie Ala Pro His Arg lie 85 90 95 •97· 146966-序列表.doc 201037078
Met Thr His Asp Asn Ser Trp Pro Val Ala Leu Lys Phe Met Ser lie 100 105 110
Gly Ala Ser Lys Met His Asp Pro Asn Gin Val Val Met Thr Leu Asp 115 120 125
His Asp Val Gin Asn Lys Thr Glu Lys Asn Leu Gin Lys Tyr Arg Gin 130 135 140 lie Glu Glu Phe Ala Lys Gin His Gly Val Glu Phe Tyr Pro Ala Gly 145 150 155 160
Arg Gly lie Gly His Gin lie Met Val Glu Glu Gly Phe Ala Trp Pro 165 170 175
Gly Thr Leu Val Val Ala Ser Asp Ser His Ser Asn Thr Tyr Gly Ala 180 185 190
Val Ala Ser Val Gly Thr Pro lie Val Arg Thr Asp Ala Ala Ser lie 195 200 205
Trp Ala Thr Gly Lys Thr Trp Trp Gin He Pro Pro Val Ala Lys Val 210 215 220
Thr Phe Thr Gly lie Leu Pro Pro Gly Val Thr Gly Lys Asp Val lie 225 230 235 240
Val Ala Leu Cys Gly Leu Phe Asp Lys Asp Asp Val Leu Asn His Ala 245 250 255 lie Glu Phe Thr Gly Ser Glu Glu Thr Met Arg Ser Leu Pro Met Asp 260 265 270
Ser Arg Leu Thr lie Ala Asn Met Thr Thr Glu Trp Gly Ala Leu Ser 275 280 285
Gly Leu Phe Pro Met Asp Gly Val Leu Lys Gly Trp Leu Lys Gly Lys 290 295 300
Ala Thr Thr Ala Ala Met Gly Leu Ala Asp Gly Pro Phe Lys Thr Leu 305 310 315 320
Ala Ala Arg Asn Phe Thr His Pro Ala lie Glu Gin Leu Phe Val Asn 325 330 335
Pro Leu Thr Ala Asp Lys Gly Ala Lys Tyr Ala Lys Glu Leu Phe Leu -98 146966-序列表.doc 201037078 340 345 350
Asp Leu Ser Thr Leu Ser Pro Tyr Val Ser Gly Pro Asn Ser Val Lys 355 360 365 lie Ala Thr Pro Leu Lys Glu Leu Glu Ala Gin Asp lie Lys Val Asp 370 375 380
Lys Ala Tyr Leu Val Ser Cys Thr Asn Ser Arg Ala Ser Asp lie Ala 385 390 395 400
Ala Ala Ala Lys Val Phe Lys Asp Ala Ala Glu Lys Asn Gly Gly Lys 405 410 415
Val Pro Lys lie Ala Asp Gly Val Lys Phe Tyr lie Ala Ala Ala Ser 420 425 430 o lie Pro Glu Gin Leu Ala Ala Glu Gly Ala Gly Asp Trp Gin Thr Leu 435 440 445
Leu Glu Ala Gly Ala Thr Ala Leu Pro Ala Gly Cys Gly Pro Cys lie 450 455 460
Gly Leu Gly Thr Gly Leu Leu Glu Pro Gly Glu Val Gly lie Ser Ala 465 470 475 480
Ser Asn Arg Asn Phe Lys Gly Arg Met Gly Ser Thr Glu Ala Lys Ala 485 490 495
Tyr Leu Gly Ser Pro Glu lie Val Ala Ala Ser Ala Leu Ser Gly Lys 500 505 510 o
Leu Ser Gly Pro Gly Trp Tyr Gin Pro Pro Glu Gly Trp Thr Glu Val 515 520 525
Val Arg Gly Glu Gly Asp Gly lie Arg Glu Glu Asp Arg Met Leu Asn 530 535 540
Thr Glu Gin Ala Leu Glu Lys Leu Leu Gly Gin Leu Asp Asp Leu Val 545 550 555 560
Ala Asp Gly Glu Lys Arg Phe Ala Pro Glu Glu Lys Val Glu Glu Glu 565 570 575
Gly Gly Leu Thr Glu Val Tyr Pro Gly Phe Pro Glu Arg Val Ser Gly 580 585 590 -99 146966-序列表.doc 201037078
Glu lie Val Phe Cys Asp Ala Asp Asn Leu Asn Thr Asp Ala lie Tyr 595 600 605
Pro Gly Tyr Trp Thr Tyr Gin Asp Asn Val Pro Val Glu Lys Met Ala 610 615 620
Glu Val Cys Met Ser Asn Tyr Asp Lys Glu Phe Ala Ser lie Ala Lys 625 630 635 640
Glu Gly Asp lie Leu Val Val Gly Tyr Asn Phe Gly Cys Gly Ser Ser 645 650 655
Arg Glu Gin Ala Ala Thr Ala Leu Leu Ala Lys Gin lie Pro Leu Val 660 665 670
Val Ser Gly Ser Phe Gly Asn lie Phe Ser Arg Asn Ser lie Asn Asn 675 680 685
Ala Leu Met Gly Leu Glu Val Pro Arg Leu Val Ser Arg Leu Arg Glu 690 695 700
Glu Phe Gly Asp Lys Gin Leu Thr Arg Arg Thr Gly Trp Thr Leu Thr 705 710 715 720
Trp Asp Val Arg Arg Ser Gin lie Glu 工le Gin Glu Gly Gin Asn Gly 725 730 735
Pro Lys Trp Thr His Lys Val Gly Glu Leu Pro Pro Asn Val Gin Glu 740 745 750 lie lie Ala Lys Gly Gly Leu Glu Lys Trp Val Lys Asn Ala lie Glu 755 760 765
Ala <210> 59 <211> 776 <212> PRT <213>構巢裸胞殼 <400> 59
Met Gin Ser Arg Leu Val Ser Gin Ser Gly Leu Gly Arg Arg Trp Ala 15 10 15 -100- 146966-序列表.doc 201037078
Val Leu Arg Cys Ala Leu Ser Lys Thr Tyr Gin Arg Arg Thr Leu Thr 20 25 30
Ser Thr Arg Arg Gin Phe Gin Asp 35 40
Pro Thr Ser Ala Ala Leu Phe Ser 50 55
Pro Gin Thr Leu Thr Glu Lys lie 65 70
Pro Gin Gly Lys Phe Val Lys Ser 85 9
Val Phe Gin Ser Gin Leu Glu Asp 45 Ala Leu Asn Ser Ser Lys Ala Val 60 Val Gin Lys Tyr Ser Val Gly Leu 75 80 Gly Asp Tyr Val Thr lie Gin Pro D 95
His Arg Cys Met Thr His Asp Asn Ser Trp Pro Cys Ala Leu Lys Phe 100 105 110
Met Ser lie Gly Ala Ser Arg Leu His Asn Pro Asp Gin lie Val Met 115 120 125
Thr Leu Asp His Asp Val Gin Asn Lys Ser Asp Lys Asn Leu Lys Lys 130 135 140
Tyr Arg Gin lie Glu Glu Phe Ala Thr Gin His Gly Val Glu Phe Tyr 145 150 155 160
Pro Ala Gly Arg Gly lie Gly His Gin lie Met 工le Glu Glu Gly Phe 165 170 175
Ala Trp Pro Gly Thr Leu Ala Val Ala Ser Asp Ser His Ser Asn Met 180 185 190
Tyr Gly Gly Val Gly Cys Leu Gly Thr Pro lie Val Arg Thr Asp Ala 195 200 205
Ala Ser Val Trp Ala Thr Gly Lys Thr Trp Trp Gin lie Pro Pro Val 210 215 220
Ala Lys Val Thr Phe Lys Gly Val Leu Pro Pro Gly Val Thr Gly Lys 225 230 235 240
Asp Val lie Val Ala Leu Cys Gly Leu Phe Asn Lys Asp Asp Val Leu 245 250 255
Asn His Ala lie Glu Phe Thr Gly Ser Glu Glu Thr Met Arg Ser Leu 101 - 146966-序列表.doc 201037078 260 265 270
Ser Val Asp Thr Arg Leu Thr lie Ala Asn Met Thr Thr Glu Trp Gly 275 280 285
Ala Leu Ser Gly Leu Phe Pro lie Asp Ser Val Leu Lys Gly Trp Leu 290 295 300
Arg Gly Lys Ala Thr Thr Ala Ala Met Gly Leu Ala Asp Gly Pro Phe 305 310 315 320
Lys Thr Arg Ala Ala Glu Arg Phe Thr His Pro Leu Leu Glu Gin Leu 325 330 335
Phe Glu Asn Pro Leu Thr Ala Asp Lys Gly Ala Lys Tyr Ala Lys Glu 340 345 350
Leu Phe Leu Asp Leu Ser Ser Leu Ser Pro Tyr Val Ser Gly Pro Asn 355 360 365
Ser Val Lys Val Ala Thr Pro Leu Lys Glu Leu Glu Ala Gin Asn lie 370 375 380
Lys Val Asp Lys Ala Tyr Leu Val Ser Cys Thr Asn Ser Arg Ala Ser 385 390 395 400
Asp lie Ala Ala Ala Ala Lys Val Phe Lys Glu Ala Ala Glu Lys Asn 405 410 415
Lys Phe Tyr lie Ala 430
Gly Gly Lys lie Pro Lys lie Ala Asp Gly Val 420 425
Ala Ala Ser lie Pro Glu Gin Leu Ala Ala Glu Gly Asn Gly Asp Trp 435 440 445
Gin Thr Leu Leu Glu Ala Gly Ala Thr Gin Leu Pro Ala Gly Cys Gly 450 455 460
Pro Cys lie Gly Met Gly Gin Gly Leu Leu Glu Pro Gly Glu Val Gly 465 470 475 480 lie Ser Ala Ser Asn Arg Asn Phe Lys Gly Arg Met Gly Ser Thr Glu 485 490 495
Ala Lys Ala Tyr Leu Gly Ser Pro Glu Val Val Ala Ala Ser Ala Leu 500 505 510 •102 146966-序列表.doc 201037078
Ser Gly Lys Leu Ser Gly Pro Gly Trp Tyr Gin Thr Pro Glu Gly Trp 515 520 525
Thr Glu Val lie Arg Gly Glu Gly Asp Gly lie Arg Glu Glu Asp Arg 530 535 540
Met Leu Thr Asn Glu Glu Ala Leu Glu Lys lie lie Gly Gin Leu Asp 545 550 555 560
Asp Leu Val Ala Asp Gly Glu Lys Arg Phe Ala Ser Glu Thr Pro Ala 565 570 575
Val Glu Glu Ser Glu Gin Gly Leu Thr Glu lie Tyr Pro Gly Phe Pro 580 585 590
Glu Arg Val Ser Gly Glu Leu Val Phe Cys Asp Ala Asp Asn Val Asn 595 600 605
Thr Asp Gly lie Tyr Pro Gly Lys Tyr Thr Tyr Gin Asp Asp Val Pro 610 615 620
Pro Glu Thr Met Ala Arg Val Cys Met Glu Asn Tyr Asp Pro Glu Phe 625 630 635 640
Ser Thr Thr Ala Lys Glu Gly Asp lie Leu Val Ser Gly Phe Asn Phe 645 650 655
Gly Cys Gly Ser Ser Arg Glu Gin Ala Ala Thr Ala lie Leu Ala Lys 660 665 670
Lys lie Pro Leu Val Val Ser Gly Ser Phe Gly Asn lie Phe Ser Arg 675 680 685
Asn Ser lie Asn Asn Ala Leu Met Gly Leu Glu Val Pro Arg Leu Val 690 695 700
Asn Arg Leu Arg Glu Thr Phe Gly Ser Gly Asp Lys Val Leu Thr Arg 705 710 715 720
Arg Thr Gly Trp Thr Leu Thr Trp Asp Val Arg Lys Ser Gin lie Glu 725 730 735
Val Gin Glu Gly Pro Gly Gly Pro Lys Trp Thr His Lys Val Gly Glu 740 745 750 • 103- 146966-序列表.doc 201037078
Leu Pro Pro Asn Val Gin Glu lie lie Ala Lys Gly Gly Leu Glu Lys 755 760 765
Trp Val Lys Asn Ala lie Gly Ala 770 775 <210> 60 <211> 774 <212> PRT <213>產黃青黴 <400> 60
Met Pro Ser Ala Glu Ser Gly Pro Lys Thr Leu Tyr Asp Lys Val Phe 15 10 15
Gin Asp His lie Val Asn Glu Gin Glu Asp Gly Thr Cys Leu lie Tyr 20 25 30 lie Asp Arg His Leu Val His Glu Val Thr Ser Pro Gin Ala Phe Glu 35 40 45
Gly Leu Lys Asn Ala Ser Arg Gin Val Arg Arg Pro Asp Cys Thr Leu 50 55 60
Ala Thr Val Asp His Asn lie Pro Thr Ser Ser Arg Lys Asn Phe Lys 65 70 75 80
Asn Ala Ala Asp Phe lie Lys Glu Asn Asp Ser Arg Leu Gin Cys Thr 85 90 95
Thr Leu Glu Glu Asn Val Lys Asp Phe Gly Leu Thr Tyr Phe Gly Met 100 105 110
Gly Asp Lys Arg Gin Gly lie Val His lie lie Gly Pro Glu Gin Gly 115 120 125
Phe Thr Leu Pro Gly Thr Thr Val Val Cys Gly Asp Ser His Thr Ser 130 135 140
Thr His Gly Ala Phe Gly Ala Leu Ala Phe Gly lie Gly Thr Ser Glu 145 150 155 160
Val Glu His Val Leu Ala Thr Gin Thr Leu lie Thr Arg Arg Ser Lys 165 170 175
Asn Met Arg lie Gin Val Asp Gly Glu Leu Pro Ala Gly Val Thr Ser 104· 146966-序列表.doc 201037078 180 185 190
Lys Asp Val Val Leu His lie lie Gly Val lie Gly Thr Ala Gly Gly 195 200 205
Asn Gly Ala Val lie Glu Phe Cys Gly Ser Val lie Arg Gly Leu Ser 210 215 220
Met Glu Ala Arg Met Ser Met Cys Asn Met Ser lie Glu Gly Gly Ala 225 230 235 240
Arg Ala Gly Met lie Ala Pro Asp Glu lie Thr Phe Glu Tyr Leu Lys 245 250 255
Gly Arg Pro Leu Ala Pro Lys Tyr Gly Ser Ala Glu Trp Asn Lys Ala 260 265 270 o
Thr Ser Tyr Trp Ser Ser Leu Lys Ser Asp Ala Gly Ala Lys Tyr Asp 275 280 285
Ser Glu Val Phe lie Asp Gly Lys Asp lie lie Pro Thr lie Ser Trp 290 295 300
Gly Thr Ser Pro Gin Asp Val Val Pro lie Thr Gly Val Val Pro Ser 305 310 315 320
Pro Asp Asp Phe Glu Asp Glu Asn Arg Lys Ala Ser Cys Lys Arg Ala 325 330 335
Leu Glu Tyr Met Gly Leu Val Ser Gly Thr Pro Met Lys Asp Val Val 340 345 350 〇
Val Asp Lys Val Phe lie Gly Ser Cys Thr Asn Ala Arg lie Glu Asp 355 360 365
Leu Arg Ala Ala Ala Lys Val Val Asn Gly Arg Lys Val Ala Ser Asn 370 375 380 lie Lys Arg Ala Met lie Val Pro Gly Ser Gly Leu Val Lys Glu Gin 385 390 395 400
Ala Glu Ser Glu Gly Leu Asp Lys Val Phe Thr Asp Ala Gly Phe Glu 405 410 415
Trp Arg Glu Ala Gly Cys Ser Met Cys Leu Gly Met Asn Pro Asp lie 420 425 430 -105- 146966-序列表.doc 201037078
Leu Ser Pro Lys Glu Arg Cys Ala Ser Thr Ser Asn Arg Asn Phe Glu 435 440 445
Gly Arg Gin Gly Ala Gin Gly Arg Thr His Leu Met Ser Pro Ala Met 450 455 460
Ala Ala Thr Ala Ala lie Val Gly Lys Leu Ala Asp Val Arg Glu His 465 470 475 480
Val Val Ala Ser Pro Val Leu Gly Lys Ala Ser Pro Lys lie Asp Val 485 490 495
Gin Pro Val Phe Glu Ser Pro Glu Thr Glu Asp Glu Leu Asp Arg Val 500 505 510
Leu Asp Arg Pro Ala Asp Asn Glu Pro His Thr Asn Ser Ser Ala Pro 515 520 525
Ala Ser Gly Gly Gly Lys Ser Thr Gly Leu Pro Thr Phe Thr Thr Leu 530 535 540
Lys Gly lie Ala Ala Pro Leu Asp Arg Ala Asn Val Asp Thr Asp Ala 545 550 555 560 lie lie Pro Lys Gin Phe Leu Lys Thr lie Lys Arg Thr Gly Leu Gly 565 570 575
Thr Ala Leu Phe Tyr Glu Leu Arg Tyr Thr Asp Asp Lys Glu Asn Pro 580 585 590
Asp Phe Val Leu Asn Gin Gly lie Tyr Arg Asp Ser Lys lie Leu Val 595 600 605
Val Thr Gly Pro Asn Phe Gly Cys Gly Ser Ser Arg Glu His Ala Pro 610 615 620
Trp Ala Leu Leu Asp Phe Gly lie Lys Cys lie lie Ala Pro Ser Phe 625 630 635 640
Ala Asp lie Phe Phe Asn Asn Thr Phe Lys Asn Gly Met Leu Pro Val 645 650 655
Val Val Ser Asp Glu Val Ala Leu Gin Lys lie Ala Asp Glu Ala Arg 660 665 670 •106- 146966-序列表.doc 201037078
Ala Gly Arg Glu Val Glu Val Asp Leu Val Asn Gin Glu lie Lys Asp 675 680 685
Ala Gin Gly Asn Lys lie Thr Ser Phe Glu Val Glu Ala Phe Arg Lys 690 695 700
His Cys Leu lie Asn Gly Leu Asp Asp lie Gly Leu Thr Leu Gin Met 705 710 715 720
Glu Ser Lys lie Arg Ser Phe Glu Ser Lys Arg Thr Leu Asp Thr Pro 725 730 735
Trp Leu Asp Gly Ser Ala Tyr Leu Arg Arg Asp Arg Arg Gly Ala Thr 740 745 750
Met Val Glu Ala Ala Pro Val Pro Lys Thr Asn Arg Gly Asp Val Lys 755 760 765
Asn Glu Pro Leu Glu Trp 770 <210> 61 <211> 785 <212> PRT <213>產黃青黴 <400> 61
Met Ser Pro Cys Ser Met Leu Leu Lys Arg Val Ala Arg Pro Pro Val 1 5 10 15
Ser Thr Thr Cys Arg Leu Val Arg Pro Arg Trp Ala Pro Ser Phe Gly 20 25 30 ❹
Val Pro Ser Arg Thr lie His His Pro Leu Arg Ser Val Ser Lys Ser 35 40 45
Leu Ser Thr Arg Ala Leu Ser Thr Thr Ala Pro Ala Arg Val Glu Gly 50 55 60
Phe His Ser Gin His Glu Asn Ala Ser lie Pro Phe Ser Glu Thr Pro 65 70 75 80
Ser Glu Lys Arg Thr Pro Gin Thr Leu Thr Glu Lys lie Val Gin Arg 85 90 95
Tyr Ala Val Gly Leu Pro Glu Gly Lys Leu Val Arg Ser Gly Asp Tyr 107- 146966·序列表.doc 201037078 100 105 110 lie Ser Leu Ala Pro Gly Tyr Cys Met Thr His Asp Asn Ser Trp Pro 115 120 125
Val Ala Leu Lys Phe Met Ser Met Gly Ala Thr Lys lie His Arg Pro 130 135 140
Glu Gin lie Val Met Thr Leu Asp His Asp Val Gin Asn Thr Ser Ala 145 150 155 160
Ala Asn Leu Lys Lys Tyr Glu Gin lie Glu Thr Phe Ala Gly Gin His 165 170 175
Gly lie Asp Phe Tyr Pro Ala Gly Arg Gly lie Gly His Gin Val Met 180 185 190
Val Glu Glu Gly Tyr Ala Trp Pro Gly Thr Met Ala Val Ala Ser Asp 195 200 205
Ser His Ser Asn His Tyr Gly Gly Val Gly Cys Leu Gly Thr Ala Val 210 215 220
Val Arg Thr Asp Ala Ala Ser lie Trp Ala Thr Ser Arg Thr Trp Trp 225 230 235 240
Gin lie Pro Pro Val Ala Arg Val Thr Phe Thr Gly Thr Leu Pro Ala 245 250 255
Gly Val Thr Gly Lys Asp Val lie Val Ala Leu Cys Gly Leu Phe Asn 260 265 270
Ser Asp Val Leu Asn His Ala lie Glu Phe Thr Gly Ser Glu Glu Thr 275 280 285
Met Glu Ser Leu Leu Val Asp Ser Arg Leu Thr lie Ala Asn Met Thr 290 295 300
Thr Glu Trp Gly Ala Leu Thr Gly Leu Phe Pro lie Asp Arg Thr Leu 305 310 315 320
Lys Arg Trp Leu Arg Tyr Lys Ala Thr Glu Ala Ala Met Ser Glu Asp 325 330 335
Arg Thr Thr Arg Gin Arg lie Thr His Glu Arg lie Asp Glu Leu Phe 340 345 350 _ 108 146966-序列表.doc 201037078
Ala Asn Pro Leu Thr Ala Asp Pro Asp Ala Gin Tyr Ala Lys Gin Leu 355 360 365
Tyr Leu Asn Leu Ser Thr Leu Ser Pro Tyr Val Ser Gly Pro Asn Ser 370 375 380
Val Lys Val Ala Thr Pro Leu Asn Glu Leu Ala Gin Gin Asn lie Lys 385 390 395 400
Val Asn Arg Ala Tyr lie Val Ser Cys Thr Asn Ser Arg Ala Ser Asp 405 410 415
Leu Ala Ala Ala Ala Lys Val Phe Lys Asp Ala Ala Lys Ala Asn Pro 420 425 430 ❹
Gly Thr Thr Pro 435
Lys lie Ala Asp Gly Val Lys Leu Tyr lie Ala Ala 440 445
Ala Ser Ala Pro 450
Glu Gin Glu Ala Ala Glu Ser Thr Gly Asp Trp Gin 455 460
Ala Leu Leu Asp Ala Gly Ala Gin Pro Leu Pro Ala Gly Cys Gly Pro 465 470 475 480
Cys lie Gly Leu Gly Thr Gly Leu Leu Glu Pro Gly Glu Val Gly lie 485 490 495
Ser Ala Ser Asn Arg Asn Phe Lys Gly Arg Met Gly Ser Arg Asp Ala 500 505 510
Leu Ala Tyr Leu Ala Ser Pro Glu Val Val Ala Ala Ser Ala Leu Ser 515 520 525
Gly Val lie Ser Gly Pro Gly Ala Tyr Gin Val Pro Glu Asn Trp Ser 530 535 540
Gly Val Glu His Gly Phe Gly Thr Gly Leu Pro Pro Thr Thr Glu Asn 545 550 555 560
Glu Leu Thr Asn Leu Leu Gin Gin Met Glu Ser Leu lie Asp Arg Val 565 570 575
Glu Ser Ala Gly Glu Asp Ser Lys Pro Ala Thr Glu lie Leu Pro Gly 580 585 590 -109 146966-序列表.doc 201037078
Phe Pro Glu Arg lie Ser Gly Glu lie Val Phe Leu Asp Ala Asp Asn 595 600 605
Leu Asp Thr Asp Asn lie Tyr Pro Gly Lys Leu Thr Tyr Gin Asp Asn 610 615 620
Val Ser Lys Asp Asp Met Ala Ala Ala Cys Met Gin Asn Tyr Asp Pro 625 630 635 640
Glu Phe Lys Gly lie Ala Lys Pro Ser Asp lie Leu Val Ala Gly Phe 645 650 655
Asn Phe Gly Cys Gly Ser Ser Arg Glu Gin Ala Ala Thr Ala lie Leu 660 665 670
Ala Lys Gin lie Pro Leu Val Val Ala Gly Ser Phe Gly Asn lie Phe 675 680 685
Ser Arg Asn Ser lie Asn Asn Ala Leu Met Gly Leu Glu Val Pro Arg 690 695 700
Leu lie Glu Arg Leu Arg Ala Ser Phe Ala Gin Pro Pro Pro Gly Asp 705 710 715 720
Ala Gly Arg Gin Leu Thr Arg Arg Thr Gly Trp Thr Leu Thr Trp Asp 725 730 735
Val Lys Arg Ser Val Val Glu Val Lys Glu Gly Glu Ser Gly Glu Ser 740 745 750
Trp Thr Glu Gin Val Gly Glu Leu Pro Ala Asn Val Gin Glu lie lie 755 760 765
Ala Glu Gly Gly Leu Glu Ala Trp Val Lys Gly Lys Val Ala Lys Ser 770 775 780
Glu 785 <210> 62 <211> 360
<212> PRT <213>黃孢原毛平革菌 <400> 62
Met Ala Phe Arg Leu Pro Leu Arg Arg Ala Leu Ser Thr Ala Ala Ala 110· 146966-序列表.doc 201037078 5 10 15
Ser Arg Ser Ser Leu Lys lie Gly Leu Val Pro Ala Asp Gly lie Gly 20 25 30
Arg Glu Val lie Pro Ala Ala Arg Gin Ala lie Glu Ala Leu Gly Ser 35 40 45
Asp lie Pro Lys Pro Glu Phe Val Asp Leu Leu Ala Gly Phe Glu Leu 50 55 60
Phe Thr Arg Thr Gly Thr Ala Leu Pro Glu Glu Thr Val Gin Ala Leu 65 70 75 80
Lys Glu Cys Asp Cys Ala Leu Phe Gly Ala Val Ser Ser Pro Ser Arg 85 90 95 o
Arg Val Thr Gly Tyr Ser Ser Pro lie Val Ala Leu Arg Lys lie Leu 100 105 110
Asp Leu Tyr Ala Asn Val Arg Pro Val Val Ala Pro Thr Pro Glu Glu 115 120 125
Lys Pro Asn Val Asp Leu lie Val Val Arg Glu Asn Thr Glu Cys Leu 130 135 140
Tyr Val Lys Gin Glu Gin Met Thr Pro Thr Glu Asn Gly Arg Glu Ala 145 150 155 160
Arg Ala Thr Arg Val He Thr Glu Arg Ala Ser Arg Arg lie Gly Gin 165 170 175 ❹
Met Ala Phe Glu Leu Ala Lys Ala Arg Pro Arg Lys His Val Thr lie 180 185 190 lie His Lys Ser Asn Val Leu Ser lie Thr Asp Gly Leu Phe Arg Glu 195 200 205
Thr Val Arg Ser Val Pro Arg Leu Asn Glu Gly Lys Tyr Asp Asp Val 210 215 220
Glu lie Ala Glu Gin Leu Val Asp Ser Ala Val Tyr Arg Leu Phe Arg 225 230 235 240
Glu Pro His lie Tyr Asp Val Met Val Ala Pro Asn Leu Tyr Gly Asp 245 250 255 •111 146966-序列表.doc 201037078 lie lie Ser Asp Ala Ala Ala Ala Leu Val Gly Ser Leu Gly Leu ‘Val 260 265 270
Pro Ser Val Asn Ala Gly Asp Asn Phe Val Met Gly Glu Pro Val His 275 280 285
Gly Ser Ala Pro Asp He Ala Gly Gin Gly lie Ala Asn Pro He Ala 290 295 300
Ser He Arg Ser Ala Ala Leu Met Leu Arg His Leu Gly Tyr Gly Ala 305 310 315 320
Pro Ala Asp Arg Leu Asp Lys Ala Val Asp Glu Val lie Arg Glu Gly 325 330 335
Gin He Leu Thr Pro Asp Leu Gly Gly Lys Ser Lys Thr Gin Asp Val 340 345 350
Val Asp Ala Val Leu Lys Arg lie 355 360 <210> 63 <211> 362 <212> PRT <213>粟酒裂殖酵母 <400> 63
Met Ser Ala Thr Arg Arg lie Val Leu Gly Leu lie Pro Ala Asp Gly 1 5 10 15 lie Gly Lys Glu Val Val Pro Ala Ala Arg Arg Leu Met Glu Asn Leu 20 25 30
Pro Ala Lys His Lys Leu Lys Phe Asp Phe lie Asp Leu Asp Ala Gly 35 40 45
Trp Gly Thr Phe Glu Arg Thr Gly Lys Ala Leu Pro Glu Arg Thr Val 50 55 60
Glu Arg Leu Lys Thr Glu Cys Asn Ala Ala Leu Phe Gly Ala Val Gin 65 70 75 80
Ser Pro Thr His Lys Val Ala Gly Tyr Ser Ser Pro lie Val Ala Leu 85 90 95 •112- 146966-序列表.doc 201037078
Arg Lys Lys Met Gly Leu Tyr Ala Asn Val Arg Pro Val Lys Ser Leu 100 105 HO
Asp Gly Ala Lys Gly Lys Pro Val Asp Leu Val lie Val Arg Glu Asn 115 120 125
Thr Glu Cys Leu Tyr Val Lys Glu Glu Arg Met Val Gin Asn Thr Pro 130 135 140
Gly Lys Arg Val Ala Glu Ala lie Arg Arg lie Ser Glu Glu Ala Ser 145 150 155 160
Thr Lys lie Gly Lys Met Ala Phe Glu lie Ala Lys Ser Arg Gin Lys 165 170 175
o lie Arg Glu Ser Gly Thr Tyr Ser lie His Lys Lys Pro Leu Val Thr 180 185 190 lie lie His Lys Ser Asn Val Met Ser Val Thr Asp Gly Leu Phe Arg 195 200 205
Glu Ser Cys Arg His Ala Gin Ser Leu Asp Pro Ser Tyr Ala Ser lie 210 215 220
Asn Val Asp Glu Gin lie Val Asp Ser Met Val Tyr Arg Leu Phe Arg 225 230 235 240
Glu Pro Glu Cys Phe Asp Val Val Val Ala Pro Asn Leu Tyr Gly Asp 245 250 255 lie Leu Ser Asp Gly Ala Ala Ser Leu lie Gly Ser Leu Gly Leu Val 260 265 270
Pro Ser Ala Asn Val Gly Asp Asn Phe Val Met Ser Glu Pro Val His 275 280 285
Gly Ser Ala Pro Asp lie Ala Gly Arg Gly lie Ala Asn Pro Val Ala 290 295 300 ,
Thr Phe Arg Ser Val Ala Leu Met Leu Glu Phe Met Gly His Gin Asp 305 310 315 320
Ala Ala Ala Asp lie Tyr Thr Ala Val Asp Lys Val Leu Thr Glu Gly 325 330 335
Lys Val Leu Thr Pro Asp Leu Gly Gly Lys Ser Gly Thr Asn Glu lie 113- 146966-序列表.doc 201037078 340 345 350
Thr Asp Ala Val Leu Ala Asn lie His Asn 355 360 <210> 64 <211> 360 <212> PRT <213>構巢裸胞殼 <400> 64
Met Ala Ala Ala Arg Thr Leu Arg lie Gly Leu lie Pro Gly Asp Gly 15 10 15 工le Gly Arg Glu Val lie Pro Ala Gly Arg Arg lie Leu Glu Ala Leu 20 25 30
Pro Ala Ser Leu Asn Leu Lys Phe Asn Phe Val Asp Leu Asp Ala Gly 35 40 45
Tyr Asp Cys Phe Lys Arg Thr Gly Thr Ala Leu Pro Asp Lys Thr Val 50 55 60
Glu Val Leu Lys Lys Glu Cys Asp Gly Ala Leu Phe Gly Ala Val Ser 65 70 75 80
Ser Pro Ser Thr Lys Val Ala Gly Tyr Ser Ser Pro lie Val Ala Leu 85 90 95
Arg Lys Lys Leu Asp Leu Phe Ala Asn Val Arg Pro Val Lys Thr Thr 100 105 110
Ala Gly Thr Ser Ala Gly Lys Pro lie Asp Leu Val lie Val Arg Glu 115 120 125
Asn Thr Glu Asp Leu Tyr Val Lys Glu Glu Ser Thr Glu Glu Thr Pro 130 135 140
Asn Gly Lys Val Ala Arg Ala lie Lys Gin lie Ser Glu Arg Ala Ser 145 150 155 160
Ser Arg lie Ala Thr lie Ala Gly Glu lie Ala Leu Arg Arg Gin Asn 165 170 175 lie Arg Asp Gly Ala Ala Ala Ser Gly Leu Arg Thr Lys Pro Met Val 180 185 190 -114 146966·序列表.doc 201037078
Thr lie Thr His Lys Ser Asn Val Leu Ser Gin Thr Asp Gly Leu Phe 195 200 205
Arg Glu Thr Ala Arg Ala Ala Leu Ala Ala Gin Lys Phe Ser Ser Val 210 215 220
Glu Val Glu Glu Gin lie Val Asp Ser Met Val Tyr Lys Leu Phe Arg 225 230 235 240
Gin Pro Glu Tyr Tyr Asp Val lie Val Ala Pro Asn Leu Tyr Gly Asp 245 250 255 lie Leu Ser Asp Gly Ala Ala Ala Leu Val Gly Ser Leu Gly Leu Val 260 265 270
Pro Ser Ala Asn Val Gly Asp Asn Phe Ala lie Gly Glu Pro Cys His 0 275 280 285
Gly Ser Ala Pro Asp lie Glu Gly Lys Asn lie Ala Asn Pro lie Ala 290 295 300
Thr Leu Arg Ser Val Ala Leu Met Leu Glu Phe Leu Gly Glu Glu Gin 305 310 315 320
Ala Ala Ala Lys lie Tyr Ala Ala Val Asp Gly Asn Leu Asp Glu Gly 325 330 335
Lys Tyr Leu Ser Pro Asp Met Gly Gly Lys Ala Thr Thr Thr Glu Val 340 345 350
Leu Glu Asp Val Leu Lys Arg Leu 355 360 <210> 65 <211> 359 <212> PRT <213>產黃青黴 <400> 65
Met Ala Ala Ala Arg Thr Leu Arg lie Gly Leu lie Pro Gly Asp Gly 15 10 15 lie Gly Arg Glu Val lie Pro Ala Gly Arg Arg lie Leu Glu Ser Leu 20 25 30
Pro Ser Ser Leu Asn Leu Lys Phe Ser Phe Val Asp Leu Asp Ala Gly 115- 146966-序列表.doc 201037078 35 40 45
Tyr Glu Thr Phe Gin Lys Thr Gly Thr Ala Leu Pro Asp Lys Thr Val 50 55 60
Asp Thr Leu Lys Lys Glu Cys Asp Gly Ala Leu Phe Gly Ala Val Ser 65 70 75 80
Ser Pro Ser Thr Lys Val Ala Gly Tyr Ser Ser Pro lie Val Ala Leu 85 90 95
Arg Lys Lys Leu Asp Leu Tyr Ala Asn Val Arg Pro Val Lys Thr Thr 100 105 110
Ala Gly Asn Ser Asn Gly Lys Pro lie Asp Leu Val lie Val Arg Glu 115 120 125
Asn Thr Glu Asp Leu Tyr Val Lys Glu Glu Arg Thr lie Glu Gly Pro 130 135 140
Asn Gly Lys Val Ala Glu Ala lie Lys Arg lie Ser Glu Lys Ala Ser 145 150 155 160
Phe Arg lie Ser Asn lie Ala Gly Glu lie Ala Leu Arg Arg Gin Asn 165 170 175 工le Arg Ala Ala Ser Pro Thr Ser Thr Arg Asp Gin Pro Met Val Thr 180 185 190 lie Thr His Lys Ser Asn Val Leu Ser Gin Thr Asp Gly Leu Phe Arg 195 200 205
Glu Thr Ala Arg Arg Ala Leu Ser Ala Glu Lys Phe Ser Ser Val Phe 210 215 220
Val Glu Glu Gin lie Val Asp Ser Met Val Tyr Lys Leu Phe Arg Gin 225 230 235 240
Pro Glu Phe Tyr Asp Val lie Val Ala Pro Asn Leu Tyr Gly Asp lie 245 250 255
Leu Ser Asp Gly Ala Ala Ala Leu Val Gly Ser Leu Gly Leu Val Pro 260 265 270
Ser Ala Asn Val Gly Asp Gly Phe Ala lie Gly Glu Pro Cys His Gly 275 280 285 -116· 146966-序列表.doc 201037078
Ser Ala Pro Asp lie Glu Gly Lys Gly lie Ser Asn Pro lie Ala Thr 290 295 300 lie Arg Ser Val Ala Leu Met Leu Glu Phe Leu Gly Glu Glu Lys Ala 305 310 315 320
Ala Ala Gin lie Tyr Ala Ala Val Asp Gly Asn Leu Asp Ala Ala Gin 325 330 335
Phe Leu Thr Pro Asp Met Gly Gly Lys Ala Thr Thr Gin Gin Val Leu 340 345 350
Asp Asp Val Leu Lys Arg Leu 355
<210> 66 <211> 371 <212> PRT <213>釀酒酵母 <400> 66
Met Phe Arg Ser Val Ala Thr Arg Leu Ser Ala Cys Arg Gly Leu Ala 15 10 15
Ser Asn Ala Ala Arg Lys Ser Leu Thr lie Gly Leu lie Pro Gly Asp 20 25 30
Gly lie Gly Lys Glu Val lie Pro Ala Gly Lys Gin Val Leu Glu Asn 35 40 45
Leu Asn Ser Lys His Gly Leu Ser Phe Asn Phe lie Asp Leu Tyr Ala 50 55 60
G
Gly Phe Gin Thr Phe Gin Glu Thr Gly Lys Ala Leu Pro Asp Glu Thr 65 70 75 80
Val Lys Val Leu Lys Glu Gin Cys Gin Gly Ala Leu Phe Gly Ala Val 85 90 95
Gin Ser Pro Thr Thr Lys Val Glu Gly Tyr Ser Ser Pro lie Val Ala 100 105 110
Leu Arg Arg Glu Met Gly Leu Phe Ala Asn Val Arg Pro Val Lys Ser 115 120 125 -117- 146966-序列表.doc 201037078
Val Glu Gly Glu Lys Gly Lys Pro lie Asp Met Val lie Val Arg Glu 130 135 140
Asn Thr Glu Asp Leu Tyr lie Lys lie Glu Lys Thr Tyr lie Asp Lys 145 150 155 160
Ala Thr Gly Thr Arg Val Ala Asp Ala Thr Lys Arg lie Ser Glu lie 165 170 175
Ala Thr Arg Arg lie Ala Thr lie Ala Leu Asp lie Ala Leu Lys Arg 180 185 190
Leu Gin Thr Arg Gly Gin Ala Thr Leu Thr Val Thr His Lys Ser Asn 195 200 205
Val Leu Ser Gin Ser Asp Gly Leu Phe Arg Glu lie Cys Lys Glu Val 210 215 220
Tyr Glu Ser Asn Lys Asp Lys Tyr Gly Gin lie Lys Tyr Asn Glu Gin 225 230 235 240 lie Val Asp Ser Met Val Tyr Arg Leu Phe Arg Glu Pro Gin Cys Phe 245 250 255
Asp Val lie Val Ala Pro Asn Leu Tyr Gly Asp lie Leu Ser Asp Gly 260 265 270
Ala Ala Ala Leu Val Gly Ser Leu Gly Val Val Pro Ser Ala Asn Val 275 280 285
Gly Pro Glu lie Val lie Gly Glu Pro Cys His Gly Ser Ala Pro Asp 290 295 300 lie Ala Gly Lys Gly lie Ala Asn Pro lie Ala Thr lie Arg Ser Thr 305 310 315 320
Ala Leu Met Leu Glu Phe Leu Gly His Asn Glu Ala Ala Gin Asp lie 325 330 335
Tyr Lys Ala Val Asp Ala Asn Leu Arg Glu Gly Ser lie Lys Thr Pro 340 345 350
Asp Leu Gly Gly Lys Ala Ser Thr Gin Gin Val Val Asp Asp Val Leu 355 360 365
Ser Arg Leu -118- 146966·序列表.doc 201037078 370 <210> 67 <211> 369
<212> PRT <213>乳酸克魯維酵母 <400> 67
Met Met Arg Thr Arg Phe lie Gin Leu Ser Arg Arg Ala Tyr Ala Ser 15 10 15
Asn Ala Lys Asn Leu Thr lie Gly Leu lie Pro Gly Asp Gly lie Gly 20 25 30
Lys Glu Val lie Pro Ala Gly Lys Lys lie Leu Glu Ser Leu Asn Pro 35 40 45
Lys Tyr Gly Leu Ser Phe Lys Phe lie Asp Leu Gin Ala Gly Trp Glu 50 55 60
Thr Phe Gin Asn Thr Gly Lys Ala Leu Pro Asp Glu Thr lie Asp lie 65 70 75 80
Leu Lys Asn Gin Cys Glu Gly Ala Leu Phe Gly Ala Val Gin Ser Pro 85 90 95
Thr Thr Lys Val Glu Gly Tyr Ser Ser Pro lie Val Ala Leu Arg Lys 100 105 110
Asn Leu Gly Leu Phe Ala Asn Val Arg Pro Val Lys Ser Val Asp Gly 115 120 125
Thr Lys Asp Arg Lys Val Asp Leu Val lie Val Arg Glu Asn Thr Glu 130 135 140
Asp Leu Tyr lie Lys Leu Glu Lys Ser Tyr lie Asp Glu Ala Thr Gly 145 150 155 160
Thr Arg Val Ala Asp Ala Thr Lys Arg lie Thr Glu lie Ala Thr Lys 165 170 175
Asn lie Ala Thr lie Ala Leu Gin lie Ala Gin Gin Arg Leu Glu Gin 180 185 190
Asn Gly His Ala Thr Leu Thr Val Thr His Lys Ser Asn Val Leu Ser 195 200 205 -119- 146966·序列表.doc 201037078
Gin Ser Asp Gly Leu Phe Arg Glu Val Cys Arg Glu Thr Tyr Glu Ala 210 215 220
Asn Lys Asp Lys Tyr Gly Gly Val Gin Tyr Asn Glu Gin He Val Asp 225 230 235 240
Ser Met Val Tyr Arg Met Phe Arg Glu Pro Glu Cys Phe Asp Val Val 245 250 255
Val Ala Pro Asn Leu Tyr Gly Asp lie Leu Ser Asp Gly Ala Ala Ala 260 265 270
Leu Val Gly Ser Leu Gly Val Val Pro Ser Ala Asn Val Gly Pro Asn 275 280 285 lie Val lie Gly Glu Pro Cys His Gly Ser Ala Pro Asp lie Ala Gly 290 295 300
Lys Gly lie Ser Asn Pro lie Ala Thr lie Arg Ser Thr Ala Leu Met 305 310 315 320
Leu Glu Phe Leu Gly Tyr Pro Glu Pro Ala Lys Asp lie His Lys Ala 325 330 335
Val Asp Ala Asn lie Arg Glu Gly Lys Tyr Leu Thr Pro Asp Leu Gly 340 345 350
Gly Asn Ser Thr Thr Gin Gin Val Leu Glu Asp Val Leu Ser Lys Leu 355 360 365
Asp <210> 68 <211> 536 <212> PRT <213>產黃青黴 <400> 68
Met Ser Pro Pro Thr Ala Leu Asp Val Asn Leu Val Gly Val Thr Asp 15 10 15
Thr Ser Thr Val Pro Val Pro Glu Pro Leu Thr Val Asn Gly Val Ser 20 25 30
Ala Trp Arg Glu Lys Thr Ala Lys Val Pro Thr Gly Val Ala Ala Ala •120· 146966-序列表.doc 201037078 35 40 45
Cys Asn Ser Asp Met Phe Lys Ser Pro lie Cys Tyr Thr Lys Pro Lys 50 55 60
Ala Lys Gin Phe Glu His Arg Phe Ser Leu Glu Ala Lys Ser Arg Lys 65 70 7 5 80
Ala Ser Thr Leu Lys Thr Ala Ala Arg Tyr Leu Lys Thr Pro Gly Leu 85 90 95
He Ser Leu Gly Gly Gly Leu Pro Ser Pro Glu Tyr Phe Pro Phe Glu 100 105 110 o
His Leu Asp lie Lys Val Pro Thr Ala Pro Gly Phe Ser Pro Glu Ala 115 120 125
Thr Arg Glu Ser Gly Thr Val Leu Arg Ala Gly Lys His Asp lie Gin 130 135 140
Glu Gly Thr Ser Thr Tyr Asp Leu Glu lie Ala Leu Asn Tyr Gly Gin 145 150 155 160
Ala Thr Gly Ala Ala Pro Leu Leu Arg Phe Val Thr Glu His Thr Glu 165 170 175 lie lie His Ser Pro Pro Tyr Ser Asp Trp Gin Cys Thr Leu Thr Ala 180 185 190
Gly Ser Thr Tyr Ala Trp Asp Thr Ala Leu Arg Val Phe Cys Glu Arg 195 200 205 〇
Gly Asp Tyr lie Leu Met Glu Glu Tyr Thr Phe Ala Ser Ala Ala Glu 210 215 220
Thr Ala Phe Pro Leu Gly lie Lys Val Ala Gly lie Pro Met Asp Glu 225 230 235 240
Gin Gly Leu lie Pro Glu Ala Met Asp Lys lie Leu Gly Asp Trp Asp 245 250 255
Val Ala Ala Arg Gly Ala Arg Lys Pro His Val Leu Tyr Thr lie Pro 260 265 270
Thr Gly Gin Asn Pro Thr Gly Ala Thr Gin Ser Ala Glu Arg Arg His 275 280 285 146966-序列表.doc -121 - 201037078
Ala Val Tyr Lys Val Ala Gin Lys His Asp Leu lie lie Val Glu Asp 290 295 300
Glu Pro Tyr Tyr Phe Leu Gin Met Gin Pro Tyr Thr Ser Gly Asp Ala 305 310 315 320
Ser Pro Val Pro Pro Pro Ser Ser His Glu Glu Phe lie Asn Ser Leu 325 330 335
Val Pro Ser Phe Leu Ser Met Asp Thr Asp Gly Arg Val Val Arg Leu 340 345 350
Glu Ser Phe Ser Lys Val lie Ser Pro Gly Ser Arg Val Gly Trp lie 355 360 365
Val Ala Ser Glu Gin lie lie Glu Arg Phe lie Arg Asn Phe Glu Val 370 375 380
Ser Ser Gin Asn Pro Ser Gly lie Ala Gin lie Ala Leu Phe Lys Leu 385 390 395 400
Leu Asp Glu His Trp Gly His Ser Gly Tyr Leu Asp Trp Leu lie Asn 405 410 415
Leu Arg Met Ser Tyr Thr Ala Arg Arg Asp Ser Leu Val His Ala Cys 420 425 430
Glu Lys His Leu Pro Arg Glu lie Val His Trp Glu Ala Pro Ala Ala 435 440 445
Gly Met Phe Gin Trp Met Ser lie Asp Trp Arg Lys His Pro Gly lie 450 455 460
Ala Ala Gly Lys Thr His Ala Asp lie Glu Glu Glu lie Phe Leu Ser 465 470 475 480
Ala Val Asn Gly Gly Val Leu Leu Ser Arg Gly Ser Trp Phe Lys Pro 485 490 495
Asp His Asp Thr Val Glu Glu Lys Met Phe Phe Arg Ala Thr Phe Ala 500 505 510
Ala Ala Ser Ser Glu Lys lie Asp Glu Ala lie Ser Arg Phe Ala Gin 515 520 525 •122· 146966·序列表.d〇c 201037078
Ser Leu Arg Ala Gin Phe Gly Leu 530 535 <210> 69 <211> 376 <212> PRT <213>嗜熱棲熱菌 <400> 69
Met Arg Glu Trp Lys lie lie Asp Ser Thr Leu Arg Glu Gly Glu Gin 15 10 15
Phe Glu Lys Ala Asn Phe Ser Thr Gin Asp Lys Val Glu lie Ala Lys 20 25 30
Ala Leu Asp Glu Phe Gly lie Glu Tyr lie Glu Val Thr Thr Pro Val 35 40 45 o
Ala Ser Pro Gin Ser Arg Lys Asp Ala Glu Val Leu Ala Ser Leu Gly 50 55 60
Leu Lys Ala Lys Val Val Thr His lie Gin Cys Arg Leu Asp Ala Ala 65 70 75 80
Lys Val Ala Val Glu Thr Gly Val Gin Gly lie Asp Leu Leu Phe Gly 85 90 95
Thr Ser Lys Tyr Leu Arg Ala Ala His Gly Arg Asp lie Pro Arg lie 100 105 110 lie Glu Glu Ala Lys Glu Val lie Ala Tyr lie Arg Glu Ala Ala Pro 115 120 125 〇
His Val Glu Val Arg Phe Ser Ala Glu Asp Thr Phe Arg Ser Glu Glu 130 135 140
Gin Asp Leu Leu Ala Val Tyr Glu Ala Val Ala Pro Tyr Val Asp Arg 145 150 155 160
Val Gly Leu Ala Asp Thr Val Gly Val Ala Thr Pro Arg Gin Val Tyr 165 170 175
Ala Leu Val Arg Glu Val Arg Arg Val Val Gly Pro Arg Val Asp lie 180 185 190
Glu Phe His Gly His Asn Asp Thr Gly Cys Ala He Ala Asn Ala Tyr 123- 146966-序列表.doc 201037078 195 200 205
Glu Ala 工le Glu Ala Gly Ala Thr His Val Asp Thr Thr lie Leu Gly 210 215 220 工le Gly Glu Arg Asn Gly lie Thr Pro Leu Gly Gly Phe Leu Ala Arg 225 230 235 240
Met Tyr Thr Leu Gin Pro Glu Tyr Val Arg Arg Lys Tyr Lys Leu Glu 245 250 255
Met Leu Pro Glu Leu Asp Arg Met Val Ala Arg Met Val Gly Val Glu 260 265 270 lie Pro Phe Asn Asn Tyr lie Thr Gly Glu Thr Ala Phe Ser His Lys 275 280 285
Ala Gly Met His Leu Lys Ala lie Tyr lie Asn Pro Glu Ala Tyr Glu 290 295 300
Pro Tyr Pro Pro Glu Val Phe Gly Val Lys Arg Lys Leu lie lie Ala 305 310 315 320
Ser Arg Leu Thr Gly Arg His Ala lie Lys Ala Arg Ala Glu Glu Leu 325 330 335
Gly Leu His Tyr Gly Glu Glu Glu Leu His Arg Val Thr Gin His lie 340 345 350
Lys Ala Leu Ala Asp Arg Gly Gin Leu Thr Leu Glu Glu Leu Asp Arg 355 360 365 lie Leu Arg Glu Trp lie Thr Ala 370 375 <210> 70 <211> 393
<212> PRT <213>耐輻射奇球菌 <400> 70
Met Ala Gly lie Phe Met Thr Asp Ala Pro Pro Pro Leu lie Pro Ala 15 10 15
Arg Ser Trp Ala lie lie Asp Ser Thr Leu Arg Glu Gly Glu Gin Phe 20 25 30 124- 146966-序列表 _doc 201037078
Ala Arg Gly Asn Phe Gly Thr Asp Asp Lys Val Glu lie Ala Arg Ala 35 40 45
Leu Asp Ala Phe Gly Ala Glu Tyr lie Glu Val Thr Thr Pro Met Val 50 55 60
Ser Glu Gin Thr Arg Gin Asp lie Arg Lys Leu Thr Gly Leu Gly Leu 65 70 75 80
Arg Ala Lys Phe Leu Thr His Val Arg Cys His Met Glu Asp Val Gin 85 90 95
Arg Ala Val Asp Thr Gly Val Asp Gly Leu Asp Leu Leu Phe Gly Thr 100 105 110
Ser Ser Phe Leu Arg Glu Phe Ser His Gly Lys Ser lie Ala Gin lie 115 120 125 lie Asp Thr Ala Gly Glu Val lie Gly Trp lie Lys Thr His His Pro 130 135 140
Glu Leu Glu He Arg Phe Ser Ala Glu Asp Thr Phe Arg Ser Glu Glu 145 150 155 160
Ala Asp Leu Met Ala Val Tyr Ser Ala Val Ser Glu Leu Gly Val His 165 170 175
Arg Val Gly Leu Ala Asp Thr Val Gly Val Ala Thr Pro Arg Gin Val 180 185 190
Tyr Thr Leu Val Arg Glu Val Arg Lys Val lie His Glu Gly Cys Gly 195 200 205 lie Glu Phe His Gly His Asn Asp Thr Gly Cys Ala Val Ser Asn Ala 210 215 220
Tyr Glu Ala lie Glu Ala Gly Ala Thr His lie Asp Thr Thr lie Leu 225 230 235 240
Gly lie Gly Glu Arg Asn Gly lie Thr Pro Leu Gly Gly Leu Leu Ala 245 250 255
Arg Met Phe Thr Phe Asp Pro Gin Gly Leu lie Asp Lys Tyr Asn Leu 260 265 270 • 125· 146966-序列表.doc 201037078
Glu Leu Leu Pro Glu Leu Asp Arg Met lie Ala Arg Met Val Asp Leu 275 280 285
Pro Val Pro Trp Asn Asn Tyr Leu Thr Gly Glu Phe Ala Tyr Asn His 290 295 300
Lys Ala Gly Met His Leu Lys Ala lie Tyr Leu Asn Pro Gly Ala Tyr 305 310 315 320
Glu Ala lie Pro Pro Gly Val Phe Gly Val Gly Arg Arg lie Gin Ala 325 330 335
Ala Ser Lys Val Thr Gly Lys His Ala 工le Ala Tyr Lys Ala Arg Glu 340 345 350
Leu Gly Leu His Tyr Gly Glu Asp Ala Leu Arg Arg Val Thr Asp His 355 360 365 lie Lys Ser Leu Ala Glu Gin Asp Glu Leu Asp Asp Ala His Leu Glu 370 375 380
Gin Val Leu Arg Glu Trp Val Ser Ala 385 390 <210> 71 <211> 389 <212> PRT <213>嗜熱奇球菌 <400> 71
Met Thr Pro Asp Ser Ser Thr Pro Leu lie Pro Ala Arg Ser Trp Ala 15 10 15 lie lie Asp Ser Thr Leu Arg Glu Gly Glu Gin Phe Ala Arg Gly Asn 20 25 30
Phe Lys Thr Gly Asp Lys lie Glu lie Ala Arg Leu Leu Asp Ala Phe 35 40 45
Gly Ala Glu Phe Leu Glu Val Thr Thr Pro Met Val Gly Ala Gin Thr 50 55 60
Gin Ala Asp lie Arg Arg Leu Thr Ser Leu Gly Leu Asn Ala Lys lie 65 70 75 80
Leu Thr His Val Arg Cys His Leu Glu Asp Val Gin Arg Ala Val Asp 85 90 95 -126 146966-序列表.doc 201037078
Leu Gly Val Asp Gly Leu Asp Leu Leu Phe Gly Thr Ser Ser Phe Leu 100 105 110
Arg Glu Phe Ser His Gly Lys Ser lie Ala Gin lie lie Asp Thr Ala 115 120 125
Ser Glu Val lie Gly Trp lie Lys Gin Asn His Pro Asp Leu Glu lie 130 135 140
Arg Phe Ser Ala Glu Asp Thr Phe Arg Ser Glu Glu Ala Asp Leu Met 145 150 155 160
Ala Val Tyr Arg Ala Val Ser Asp Leu Gly Val His Arg Val Gly Leu 165 170 175
Ala Asp Thr Val Gly Val Ala Thr Pro Arg Gin Val Tyr Thr Leu Val 180 185 190
Arg Glu Val Arg Lys Val lie His Ala Glu Cys Gly lie Glu Phe His 195 200 205
Gly His Asn Asp Thr Gly Cys Ala Val Ser Asn Ala Tyr Glu Ala lie 210 215 220
Glu Ala Gly Ala Thr His lie Asp Thr Thr lie Leu Gly lie Gly Glu 225 230 235 240
Leu Ala Arg Met Phe Thr 255
Arg Asn Gly lie Thr Pro Leu Gly Gly Phe 245 250
Phe Asp Pro Gin Gly Leu lie Asp Lys Tyr Asn Leu Glu Leu Leu Pro 260 265 270
Glu Leu Asp Arg Leu lie Ala Arg Leu Val Asp Leu Pro lie Pro Trp 275 280 285
Asn Asn Tyr Leu Thr Gly Glu Phe Ala Tyr Asn His Lys Ala Gly Met 290 295 300
His Leu Lys Ala 工le Tyr Leu Asn Pro Gly Ala Tyr Glu Ala lie Pro 305 310 315 320
Pro Ser Val Phe Gly Val Gly Arg Arg lie Gin Ala Ala Ser Lys Val 325 330 335 127 146966-序列表.doc 201037078
Thr Gly Lys His Ala 工le Ala His Lys Ala Arg Glu Leu Gly Leu His 340 345 350
Tyr Gly Glu Asp Ala Leu Arg Arg Val Thr Asp His lie Lys Ala Leu 355 360 365
Ala Glu Glu Gly Glu Leu Asp Asp Ala His Leu Glu Gin Val Leu Arg 370 375 380
Glu Trp Val Arg Ala 385 <210> 72 <211> 553
<212> PRT <213>硫磺礦硫化葉菌 <400〉 72
Met Ala Leu Lys Met Lys Tyr Asp Phe Leu Leu Leu Ser Leu Lys Leu 15 10 15
Leu Asn Leu Pro lie lie Phe His Leu Cys Ser Val Ser Lys Lys Ser 20 25 30
Val Glu Val Leu Asp Thr Thr Leu Arg Asp Gly Ser Gin Gly Ala Asn 35 40 45 lie Ser Phe Thr Leu Asn Asp Lys lie Lys lie Ala Leu Leu Leu Asp 50 55 60
Glu Leu Gly Val Asp Tyr lie Glu Gly Gly Trp Pro Gly Ser Asn Pro 65 70 75 80
Lys Asp Glu Glu Phe Phe Arg Glu lie Lys Lys Tyr Arg Leu Ser Lys 85 90 95
Ala Lys lie Ala Ala Phe Gly Ser Thr Lys Arg Lys Asp Val Ser Val 100 105 110
Lys Glu Asp lie Ser Leu Asn Ser lie Val Lys Ala Asp Val Asp Val 115 120 125
Ala Val lie Phe Gly Lys Ser Trp Ser Leu His Ala Thr Glu Val Leu 130 135 140
Lys Val Thr Lys Gin Asp Asn Leu Asp lie Val Tyr Asp Ser lie Asn 128- 146966-序列表.doc 201037078 145 150 155 160
Tyr Leu Lys Ser His Gly Leu Lys Val lie Phe Asp Ala Glu His Phe 165 170 175
Tyr Gin Gly Phe Lys Glu Asp Pro Glu Tyr Ala Leu Glu Val Val Lys 180 185 190
Thr Ala Glu Ser Ala Gly Ala Asp Val lie Ala Leu Ala Asp Thr Asn 195 200 205
Gly Gly Thr Pro Pro Phe Glu Val Tyr Glu lie Thr Lys Lys Val Arg 210 215 220
Glu Val Leu Gin Val Lys Leu Gly lie His Ala His Asn Asp lie Gly 225 230 235 240
Cys Ala Val Ala Asn Ser Leu Met Ala lie Lys Ala Gly Ala Arg His 245 250 255
Val Gin Gly Thr lie Asn Gly lie Gly Glu Arg Thr Gly Asn Ala Asp 260 265 270
Leu lie Gin lie lie Pro Thr Leu lie Leu Lys Met Gly Leu Asn Ala 275 280 285
Leu Asn Gly Gin Glu Ser Leu Arg Lys Leu Arg Glu Val Ser Arg lie 290 295 300
Val Tyr Glu lie Leu Gly Leu Pro Pro Asn Pro Tyr Gin Pro Tyr Val 305 310 315 320
Gly Asp Asn Ala Phe Ala His 325
Lys Ala Gly Val His Val Asp Ala Val 330 335
Met Lys Val Pro Arg Ala Tyr Glu His Val Asp Pro Ser Leu Val Gly 340 345 350
Asn Asp Arg Lys Phe Val lie Ser Glu Leu Ser Gly Thr Ala Asn Leu 355 360 365
Val Ser Tyr Leu Gin Gly Leu Gly lie Ala Val Asp Lys Lys Asp Glu 370 375 380
Arg Leu Lys Lys Ala Leu Asn Lys lie Lys Glu Leu Glu Ala Arg Gly 385 390 395 400 129- 146966·序列表.doc 201037078
Tyr Ser Phe Asp Val Gly Pro Ala Ser Ala lie Leu lie Thr Leu Lys 405 410 415
Glu Leu Asn lie Tyr Lys Asn Tyr lie Asn Leu Glu Tyr Trp Lys Val 420 425 430 lie Asn Glu Asn Asn Gly Leu Ser lie Gly lie Val Lys Val Asn Ser 435 440 445
Gin Leu Glu Val Ala Glu Gly Val Gly Pro Val Asn Ala 工le Asp Arg 450 455 460
Ala Leu Arg Met Ala Leu Gin Arg Val Tyr Pro Glu lie Gly Glu Val 465 470 475 480
Lys Leu lie Asp Tyr Arg Val lie Leu Pro Ser Glu lie Lys Asn Thr 485 490 495
Glu Ser Val Val Arg Val Thr lie 500 505
Glu Phe Thr Asp Asn Lys Met Asn 510
Trp Arg Thr Glu Gly Val Ser Lys 515 520
Ser Val Val Glu Ala Ser Val Met 525
Ala Leu Val Asp Gly Leu Asp Tyr Tyr Leu Gin Leu Lys Lys Thr Leu 530 535 540
Lys Thr Ala Val Asp Asn Tyr lie Val 545 550 <210> 73 <211> 361 <212> PRT <213>柯氏熱球菌 <400> 73
Met Val Leu Asp Ser Thr Leu Arg Glu Gly Glu Gin Thr Pro Gly Val 15 10 15
Asn Phe Ser Pro Glu Asp Arg Leu Arg lie Gly lie Ala Leu Asp Glu 20 25 30
Val Gly Val Asp Phe lie Glu Ala Gly His Pro Ala Val Ser Gly Glu 35 40 45 -130- 146966-序列表.doc 201037078 lie Leu Glu Gly lie Arg Leu Leu Ala Ser His Gly Leu Asn Ala Asn 50 55 60 lie Leu Ala His Ser Arg Ala Leu Arg Ser Asp lie Asp Leu Val Leu 65 70 75 80
Lys Ala Glu Ala Glu Trp lie Gly lie Phe Met Cys Leu Ser Gin Arg 85 90 95
Cys Leu Glu Arg Arg Phe Arg Thr Asp Leu Ser Gly Ala Leu Thr Arg 100 105 110
Val Glu Asp Ala lie Leu Tyr Ala Lys Asp His Gly Leu Lys lie Arg 115 120 125
〇
Phe Thr Pro Glu Asp Thr Thr Arg Thr Glu Trp Lys Asn Leu Thr Ala 130 135 140
Ala Leu Asn Leu Ala Arg Glu Leu Lys Val Asp Arg Val Ser lie Ala 145 150 155 160
Asp Thr Thr Gly Ala Ala His Pro Leu Glu Phe Tyr Asp Leu Val Lys 165 170 175
Arg Val Val Glu Phe Gly lie Pro Val Asn Val His Cys His Asn Asp 180 185 190
Leu Gly Leu Ala Leu Ala Asn Ala lie Met Gly lie Glu Ala Gly Ala 195 200 205
Thr Leu Val Asp Ala Thr Val Asn Gly lie Gly Glu Arg Ala Gly lie 210 215 220
Val Asp Leu Ser His Leu Leu Ala Ala Leu Tyr Tyr His Tyr Gly Val 225 230 235 240
Lys Lys Tyr Arg Leu Glu Lys Leu Tyr Ser Leu Ser Arg Leu Val Ser 245 250 255
Glu lie Thr Gly Leu Gin Val Gin Val Asn Tyr Pro lie Val Gly Gin 260 265 270
Asn Ala Phe Thr His Lys Ala Gly Leu His Val Ser Ala Val Val Arg 275 280 285
Asp Pro Ser Phe Tyr Glu Phe Leu Pro Ala Glu Thr Phe Gly Arg Glu • 131· 146966-序列表.doc 201037078 290 295 300
Arg Thr lie Tyr Val Asp Arg Phe Ala Gly Arg Glu Thr lie Arg Phe 305 310 315 320
His Leu Ser Arg Phe Gly lie His Asp Glu Glu lie lie Glu Glu Leu 325 330 335
Leu Arg Arg Val Lys Ala Ser Arg Arg Pro Phe Thr Pro Glu Met Leu 340 345 350
Ala Glu Glu Ala Arg Arg Met Met Thr 355 360 <210> 74 <211> 361 <212> PRT <213>超嗜熱火球菌 <400> 74
Met lie Leu Asp Ser Thr Leu Arg Glu Gly Glu Gin Thr Pro Gly Val 15 10 15
Asn Tyr Ser Pro Glu Gin Arg Leu Arg lie Ala Leu Ala Leu Asp Glu 20 25 30 lie Gly Val Asp Phe lie Glu Val Gly His Pro Ala Val Ser Lys Asp 35 40 45
Val Phe lie Gly lie Lys Leu lie Ala Ser Gin Asp Leu Asn Ala Asn 50 55 60
Leu Leu Ala His Ser Arg Ala Leu Leu Glu Asp lie Asp Tyr Val lie 65 70 75 80
Gin Ala Asp Val Glu Trp Val Gly lie Phe Phe Cys Leu Ser Asn Ala 85 90 95
Cys Leu Arg Lys Arg Phe Arg Met Ser Leu Ser Gin Ala Leu Glu Arg 100 105 110 lie Ser Lys Ala lie Glu Tyr Ala Lys Asp His Gly Leu Lys Val Arg 115 120 125
Phe Thr Pro Glu Asp Thr Thr Arg Thr Glu Trp Glu Asn Leu Arg Arg 130 135 140 -132- 146966-序列表.doc 201037078
Ala lie Glu Leu Ala Lys Glu Leu Lys Val Asp Arg lie Ser Val Ala 145 150 155 160
Asp Thr Thr Gly Gly Thr His Pro Leu Arg Phe Tyr Thr Leu Val Lys 165 170 175
Lys Val Val Asn Phe Gly lie Pro Val Asn Val His Cys His Asn Asp 180 185 190
Leu Gly Leu Ala Leu Ala Asn Ala 工le Met Gly lie Glu Gly Gly Ala 195 200 205
Thr Val Val Asp Ala Thr Val Asn Gly Leu Gly Glu Arg Ala Gly lie 210 215 220
Val Asp Leu Ala Gin lie Val Thr Val Leu Tyr Tyr His Tyr Gly Val 225 230 235 240
Lys Lys Tyr Arg Leu Asp Lys Leu Tyr Glu lie Ser Arg Met Val Ser 245 250 255
Glu lie Thr Gly lie Ala Leu Gin Pro Asn Tyr Pro 工le Val Gly Glu 260 265 270
Asn Ala Phe Thr His Lys Ala Gly Leu His Val Ser Ala Val Leu Lys 275 280 285
Asp Pro Arg Phe Tyr Glu Phe Leu Pro Ala Glu Val Phe Gly Arg Glu 290 295 300
Arg Thr lie Tyr Val Asp Arg Phe Ala Gly Lys Asp Thr lie Arg Tyr 305 310 315 320
Tyr Leu Gin Lys Leu Gly lie Asn Asp Glu Glu Phe Val Lys Val Leu 325 330 335
Leu Lys Arg Val Lys Ser Ser Arg Glu Pro Phe Thr Trp Asp Lys Phe 340 345 350 lie Glu Glu Val Arg Arg Leu Lys Thr 355 360 <210> 75 <211> 385 <212> PRT <213>棕色固氮菌 -133 146966-序列表.doc 201037078 <400> 75
Met Ala Ser Val lie lie Asp Asp Thr Thr Leu Arg Asp Gly Glu Gin 15 10 15
Ser Ala Gly Val Ala Phe Asn Ala Asp Glu Lys lie Ala lie Ala Arg 20 25 30
Ala Leu Ala Glu Leu Gly Val Pro Glu Leu Glu lie Gly lie Pro Ser 35 40 45
Met Gly Glu Glu Glu Arg Glu Val Met His Ala lie Ala Gly Leu Gly 50 55 60
Leu Ser Ser Arg Leu Leu Ala Trp Cys Arg Leu Cys Asp Val Asp Leu 65 70 75 80
Ala Ala Ala Arg Ser Thr Gly Val Thr Met Val Asp Leu Ser Leu Pro 85 90 95
Val Ser Asp Leu Met Leu His His Lys Leu Asn Arg Asp Arg Asp Trp 100 105 110
Ala Leu Arg Glu Val Ala Arg Leu Val Gly Glu Ala Arg Met Ala Gly 115 120 125
Leu Glu Val Cys Leu Gly Cys Glu Asp Ala Ser Arg Ala Asp Leu Glu 130 135 140
Phe Val Val Gin Val Gly Glu Val Ala Gin Ala Ala Gly Ala Arg Arg 145 150 155 160
Leu Arg Phe Ala Asp Thr Val Gly Val Met Glu Pro Phe Gly Met Leu 165 170 175
Asp Arg Phe Arg Phe Leu Ser Arg Arg Leu Asp Met Glu Leu Glu Val 180 185 190
His Ala His Asp Asp Phe Gly Leu Ala Thr Ala Asn Thr Leu Ala Ala 195 200 205
Val Met Gly Gly Ala Thr His lie Asn Thr Thr Val Asn Gly Leu Gly 210 215 220
Glu Arg Ala Gly Asn Ala Ala Leu Glu Glu Cys Val Leu Ala Leu Lys 225 230 235 240 134- 146966-序列表.doc 201037078
Asn Leu His Gly lie Asp Thr Gly lie Asp Thr Arg Gly lie Pro Ala 245 250 255 lie Ser Ala Leu Val Glu Arg Ala Ser Gly Arg Gin Val Ala Trp Gin 260 265 270
Lys Ser Val Val Gly Ala Gly Val Phe Thr His Glu Ala Gly lie His 275 280 285
Val Asp Gly Leu Leu Lys His Arg Arg Asn Tyr Glu Gly Leu Asn Pro 290 295 300
Asp Glu Leu Gly Arg Ser His Ser Leu Val Leu Gly Lys His Ser Gly 305 310 315 320
G
Ala His Met Val Arg Asn Thr Tyr Arg Asp Leu Gly lie Glu Leu Ala 325 330 335
Asp Trp Gin Ser Gin Ala Leu Leu Gly Arg 工le Arg Ala Phe Ser Thr 340 345 350
Arg Thr Lys Arg Arg Ser Pro Gin Pro Ala Glu Leu Gin Asp Phe Tyr 355 360 365
Arg Gin Leu Cys Glu Gin Gly Asn Pro Glu Leu Ala Ala Gly Gly Met 370 375 380
Ala 385 <210> 76 <211> 381
<212> PRT <213>肺炎克雷伯氏桿菌 <400> Ί6
Met Glu Arg Val Leu lie Asn Asp Thr Thr Leu Arg Asp Gly Glu Gin 1 5 10 15
Ser Pro Gly Val Ala Phe Arg Thr Ser Glu Lys Val Ala He Ala Glu 20 25 30
Ala Leu Tyr Ala Ala Gly lie Thr Ala Met Glu Val Gly Thr Pro Ala 35 40 45 135· 146966-序列表.doc 201037078
Met Gly Asp Glu Glu lie Ala Arg 50 55 lie Gin Leu Val Arg Arg Gin Leu 60
Pro Asp Ala Thr Leu Met Thr Trp Cys Arg Met Asn Ala Leu Glu lie 65 70 75 80
Arg Gin Ser Ala Asp Leu Gly lie Asp Trp Val Asp lie Ser lie Pro 85 90 95
Ala Ser Asp Lys Leu Arg Gin Tyr Lys Leu Arg Glu Pro Leu Ala Val 100 105 110
Leu Leu Glu Arg Leu Ala Met Phe lie His Leu Ala His Thr Leu Gly 115 120 125
Leu Lys Val Cys lie Gly Cys Glu Asp Ala Ser Arg Ala Ser Gly Gin 130 135 140
Thr Leu Arg Ala lie Ala Glu Val Ala Gin Asn Ala Pro Ala Ala Arg 145 150 155 160
Leu Arg Tyr Ala Asp Thr Val Gly Leu Leu Asp Pro Phe Thr Thr Ala 165 170 175
Ala Gin lie Ser Ala Leu Arg Asp Val Trp Ser Gly Glu lie Glu Met 180 185 190
His Ala His Asn Asp Leu Gly Met Ala Thr Ala Asn Thr Leu Ala Ala 195 200 205
Val Ser Ala Gly Ala Thr Ser Val Asn Thr Thr Val Leu Gly Leu Gly 210 215 220
Glu Arg Ala Gly Asn Ala Ala Ala Trp Lys Pro Ser Ala Leu Gly Leu 225 230 235 240
Glu Arg Cys Leu Gly Val Glu Thr Gly Val His Phe Ser Ala Leu Pro 245 250 255
Ala Leu Cys Gin Arg Val Ala Glu Ala Ala Gin Arg Ala lie Asp Pro 260 265 270
Gin Gin Pro Leu Val Gly Glu Leu Val Phe Thr His Glu Ser Gly Val 275 280 285
His Val Ala Ala Leu Leu Arg Asp Ser Glu Ser Tyr Gin Ser 工le Ala 136- 146966·序列表.doc 201037078 290 295 300
Pro Ser Leu Met Gly Arg Ser Tyr Arg Leu Val Leu Gly Lys His Ser 305 310 315 320
Gly Arg Gin Ala Val Asn Gly Val Phe Asp Gin Met Gly Tyr His Leu 325 330 335
Asn Ala Ala Gin lie Asn Gin Leu Leu Pro Ala lie Arg Arg Phe Ala 340 345 350
Glu Asn Trp Lys Arg Ser Pro Lys Asp Tyr Glu Leu Val Ala lie Tyr 355 360 365
Asp Glu Leu Cys Gly Glu Ser Ala Leu Arg Ala Arg Gly 370 375 380
<210> 77 <211> 381 <212> PRT <213>施氏假單胞菌 <400> 77
Met Ser lie Val lie Asp Asp Thr Thr Leu Arg Asp Gly Glu Gin Ser 15 10 15
Ala Gly Val Ala Phe Ser Ala Glu Glu Lys Leu Ala lie Ala Arg Ala 20 25 30
Leu Ala Gin Leu Gly Val Pro Glu Leu Glu lie Gly lie Pro Ser Met 35 40 45
Gly Glu Glu Glu Cys Glu Val Met Arg Ala lie Ala Gly Leu Ala Leu 50 55 60
Pro Val Arg Leu Leu Ala Trp Cys Arg Leu Cys Asp Ala Asp Leu Leu 65 70 75 80
Ala Ala Gly Gly Thr Gly Val Gly Met Val Asp Leu Ser Leu Pro Val 85 90 95
Ser Asp Leu Met Leu Gin His Lys Leu Gly Arg Asp Arg Asp Trp Ala 100 105 110
Leu Arg Glu Ala Ala Arg Leu Val Gly Ala Ala Arg Asp Ala Gly Leu 115 120 125 -137- 146966-序列表.doc 201037078
Glu Val Cys Leu Gly Cys Glu Asp Ala Ser Arg Ala Asp Pro Glu Phe 130 135 140 lie Val Arg Val Ala Glu Val Ala Gin Ala Ala Gly Ala Arg Arg Leu 145 150 155 160
Arg Phe Ala Asp Thr Val Gly Val Met Glu Pro Phe Ala Met His Ala 165 170 175
Arg Phe Arg Phe Leu Ala Glu Arg Leu Asp Leu Glu Leu Glu Val His 180 185 190
Ala His Asp Asp Phe Gly Leu Ala Thr Ala Asn Thr Leu Ala Ala Val 195 200 205
Arg Gly Gly Ala Thr His lie Asn Thr Thr Val Asn Gly Leu Gly Glu 210 215 220
Arg Ala Gly Asn Ala Ala Leu Glu Glu Cys Ala Leu Ala Leu Lys His 225 230 235 240
Leu His Gly lie Asp Cys Gly lie Asp Val Arg Gly lie Pro Ser lie 245 250 255
Ser Ala Leu Val Glu Gin Ala Ser Gly Arg Gin Val Ala Trp Gin Lys 260 265 270
Ser Val Val Gly Ala Gly Val Phe Thr His Glu Ala Gly lie His Val 275 280 285
Asp Gly Leu Leu Lys His Arg Arg Asn Tyr Glu Gly Leu Asn Pro Asp 290 295 300
Glu Leu Gly Arg Ser His Ser Leu Val Leu Gly Lys His Ser Gly Ala 305 310 315 320
His Met Val Glu Leu Ser Tyr Arg Glu Leu Gly lie Glu Leu Gin Gin 325 330 335
Trp Gin Ser Arg Ala Leu Leu Gly Cys lie Arg Arg Phe Ser Thr Gin 340 345 350
Thr Lys Arg Ser Pro Gin Ser Ala Asp Leu Gin Gly Phe Tyr Gin Gin 355 360 365 -138 146966-序列表.doc 201037078
Leu Cys Glu Gin Gly Leu Ala Leu Ala Gly Gly Ala Ala 370 375 380 <210> 78 <211> 477
<212> PRT <213> 不動桿菌 NCIMB9871 <400> 78
Met Asn Tyr Pro Asn lie Pro Leu Tyr lie Asn Gly Glu Phe Leu Asp 1 5 10 15
His Thr Asn Arg Asp Val Lys Glu Val Phe Asn Pro Val Asn His Glu 20 25 30
Cys lie Gly Leu Met Ala Cys Ala Ser Gin Ala Asp Leu Asp Tyr Ala 35 40 45
Leu Glu Ser Ser Gin Gin Ala Phe Leu Arg Trp Lys Lys Thr Ser Pro 50 55 60 lie Thr Arg Ser Glu lie Leu Arg Thr Phe Ala Lys Leu Ala Arg Glu 65 70 75 80
Lys Ala Ala Glu lie Gly Arg Asn lie Thr Leu Asp Gin Gly Lys Pro 85 90 95
Leu Lys Glu Ala lie Ala Glu Val Thr Val Cys Ala Glu His Ala Glu 100 105 110
Trp His Ala Glu Glu Cys Arg Arg lie Tyr Gly Arg Val lie Pro Pro 115 120 125 〇
Arg Asn Pro Asn Val Gin Gin Leu Val Val Arg Glu Pro Leu Gly Val 130 135 140
Cys Leu Ala Phe Ser Pro Trp Asn Phe Pro Phe Asn Gin Ala lie Arg 145 150 155 160
Lys lie Ser Ala Ala lie Ala Ala Gly Cys Thr lie 工le Val Lys Gly 165 170 175
Ser Gly Asp Thr Pro Ser Ala Val Tyr Ala lie Ala Gin Leu Phe His 180 185 190
Glu Ala Gly Leu Pro Asn Gly Val Leu Asn Val lie Trp Gly Asp Ser 195 200 205 -139· 146966-序列表.doc 201037078
Asn Phe lie Ser Asp Tyr Met lie Lys Ser Pro lie lie Gin Lys lie 210 215 220
Ser Phe Thr Gly Ser Thr Pro Val Gly Lys Lys Leu Ala Ser Gin Ala 225 230 235 240
Ser Leu Tyr Met Lys Pro Cys Thr Met Glu Leu Gly Gly His Ala Pro 245 250 255
Val lie Val Cys Asp Asp Ala Asp lie Asp Ala Ala Val Glu His Leu 260 265 270
Val Gly Tyr Lys Phe Arg Asn Ala Gly Gin Val Cys Val Ser Pro Thr 275 280 285
Arg Phe Tyr Val Gin Glu Gly lie Tyr Lys Glu Phe Ser Glu Lys Val 290 295 300
Val Leu Arg Ala Lys Gin lie Lys Val Gly Cys Gly Leu Asp Ala Ser 305 310 315 320
Ser Asp Met Gly Pro Leu Ala Gin Ala Arg Arg Met His Ala Met Gin 325 330 335
Gin lie Val Glu Asp Ala Val His Lys Gly Ser Lys Leu Leu Leu Gly 340 345 350
Gly Asn Lys lie Ser Asp Lys Gly Asn Phe Phe Glu Pro Thr Val Leu 355 360 365
Gly Asp Leu Cys Asn Asp Thr Gin Phe Met Asn Asp Glu Pro Phe Gly 370 375 380
Pro lie lie Gly Leu lie Pro Phe Asp Thr lie Asp His Val Leu Glu 385 390 395 400
Glu Ala Asn Arg Leu Pro Phe Gly Leu Ala Ser Tyr Ala Phe Thr Thr 405 410 415
Ser Ser Lys Asn Ala His Gin lie Ser Tyr Gly Leu Glu Ala Gly Met 420 425 430
Val Ser lie Asn His Met Gly Leu Ala Leu Ala Glu Thr Pro Phe Gly 435 440 445 140- 146966-序列表.doc 201037078
Gly lie Lys Asp Ser Gly Phe Gly Ser Glu Gly Gly He Glu Thr Phe 450 455 460
Asp Gly Tyr Leu Arg Thr Lys Phe lie Thr Gin Leu Asn 465 470 475 <210> 79 <211> 473
<212> PRT
<213>地中海熱布氏桿菌16M <400> 79
Met Arg lie Gly Lys Met Glu Met Gin Thr Arg Tyr Pro Asp Val Lys 1 5 10 15
Leu Phe lie Asp Gly Thr Trp Arg Asp Gly Ser Arg Gly Glu Thr lie 20 25 30
Glu lie Phe Asn Pro Ala Thr Asp Glu Val lie Gly His lie Ala Arg 35 40 45
Ala Thr Thr Ala Asp Leu Asp Asp Ala Leu Ala Ala Val Asp Arg Gly 50 55 60
Phe Glu Ala Trp Ser Lys Val Ser Ala Phe Asp Arg Tyr Lys lie Met 65 70 75 80
Arg Arg Ala Ala Asp lie Phe Arg Ser Arg Gly Glu Glu Val Ala Arg 85 90 95
Leu Leu Thr Met Glu Gin Gly Lys Pro Leu Ala Glu Ala Arg lie Glu 100 105 110
Ala Ala Ala Ala Cys Asp Leu lie Asp Trp Phe Ala Glu Glu Ala Arg 115 120 125
Arg Ser Tyr Gly Arg lie Val Pro Pro Arg Gin Ala Tyr Val Met Gin 130 135 140
Ala Glu Val Lys Glu Pro Val Gly Pro Val Ala Ala Phe Thr Pro Trp 145 150 155 160
Asn Phe Pro lie Asn Gin Ala Val Arg Lys lie Ser Ala Ala Leu Ala 165 170 175
Ala Gly Cys Ser lie Leu Leu Lys Ala Ala Glu Asp Thr Pro Ala Ala -141 - 146966·序列表.doc 201037078 180 185 190
Pro Ala Glu Leu Val Arg Ala Phe Ala Glu Ala Gly Leu Pro Asp Gly 195 200 205
Ala lie Asn Leu Val Tyr Gly Asp Pro Ala Glu lie Ser Ala Tyr Leu 210 215 220 lie Pro His Pro Val lie Arg Lys Val Ser Phe Thr Gly Ser Thr Gin 225 230 235 240
Val Gly Lys Gin Leu Ala Ala Leu Ala Gly Leu His Met Lys Arg Val 245 250 255
Thr Met Glu Leu Gly Gly His Ala Pro Val lie lie Ala Ala Asp Ala 260 265 270
Asp Val Glu Gin Ala lie Lys Val Val Ser Gly Ser Lys Phe Arg Asn 275 280 285
Ala Gly Gin Val Cys lie Ser Pro Thr Arg Phe Leu lie Glu Asn Ser 290 295 300
Val Tyr Asp Gin Val Val Glu Gly Met Ala Ala Tyr Ala Thr Ser Leu 305 310 315 320
Lys Val Gly Asp Gly Leu Glu Ala Gly Thr Thr Met Gly Pro Leu Val 325 330 335
Asn Ala Lys Arg Val Asn Ala Met Glu Arg Leu Val Gin Asp Ala Arg 340 345 350
Glu His Lys Ala Arg Val Val Thr Gly Gly Glu Arg lie Gly Asn Arg 355 360 365
Gly Asn Phe Phe Glu Pro Thr lie Leu Ala Asp Val Pro Arg Asp Ala 370 375 380
Ala lie Met Asn Glu Glu Pro Phe Gly Pro Val Ala Leu Leu Asn Arg 385 390 395 400
Phe Asp Ala Leu Asp Glu Ala Leu Ser Glu Ala Asn Arg Leu Asn Tyr 405 410 415
Gly Leu Ala Ala Tyr Ala Phe Thr Gly Ser Ser Ala Lys Ala Ala Arg 420 425 430 -142- 146966-序列表.doc 201037078 lie Ser Ser Thr Val Arg Ser Gly Met lie Thr lie Asn Gin Leu Arg 435 440 445
Ser Gly Pro Ala Gly Ser Ala Leu Arg Arg Asp Gin Arg Phe Arg Leu 450 455 460
Trp Asn Gly Arg Arg Cys Arg Arg Ala 465 470 <210> 80 <211> 530 <212> PRT <213>鮑曼不動桿菌 <400> 80
Met Arg Leu lie Met Leu Asn lie Thr Gly Gin Asn Phe lie Ala Gly
Gin Arg Ser Ser Ala Gly Ser Lys Phe Val Leu Ser Tyr Asp Ala Ala 20 25 30
Thr Asp Glu Ala Leu Pro Tyr Gin Phe Ala Gin Ala Thr Pro Glu Glu 35 40 45 lie Asp Gin Ala Ala Gin Ala Ala Ala Leu Ala Tyr Pro Ala Phe Arg 50 55 60
Gin Thr Thr Pro Glu Gin Arg Ala Val Phe Leu Glu Thr lie Ala Ser 65 70 75 80
Glu lie Asp Ala Leu Asp Asp Gin Phe lie Ala Thr Val Cys Gin Glu 85 90 95
Thr Ala Leu Pro Glu Ala Arg lie Arg Gly Glu Arg Gly Arg Thr Thr 100 105 110
Gly Gin Leu Arg Leu Phe Ala Gin Val Leu Arg Arg Gly Asp Tyr Leu 115 120 125
Gly Ala Arg lie Asp Leu Ala Leu Pro Glu Arg Gin Pro Leu Pro Arg 130 135 140
Pro Asp Leu Arg Gin Tyr Lys lie Gly Val Gly Pro Val Ala Val Phe 145 150 155 160 -143- 146966-序列表.doc 201037078
Gly Ala Ser Asn Phe Pro Leu Ala Phe Ser Thr Ala Gly Gly Asp Thr 165 170 175
Ala Ser Ala Leu Ala Ala Gly Cys Pro Val He Val Lys Ala His Ser 180 185 19〇
Gly His Met Ala Thr Ala Glu Ser He Ala Asn Ala He Cys Ser Ala 195 200 205 lie Glu Lys Cys Ala Met Pro Lys Gly lie Phe Ser Met He Tyr Gly 210 215 220
Gin Gly Val Gly Glu Pro Leu Val Lys His Pro Ala lie Lys Ala Val 225 230 235 240
Gly Phe Thr Gly Ser Leu Lys Gly Gly Arg Ala Leu Cys Asp Leu Ala 245 250 255
Ala Ala Arg Pro Glu Pro lie Pro Val Phe Ala Glu Met Ser Ser lie 260 265 270
Asn Pro Met lie Leu Leu Pro Glu Ala Leu Lys Val Arg Gly Asp Lys 275 280 285 lie Ala Thr Glu Leu Ser Gly Ser Val Val Leu Gly Cys Gly Gin Phe 290 295 300
Cys Thr Asn Pro Gly Leu lie lie Gly lie Lys Ser Pro Glu Phe Ser 305 310 315 320
Gin Phe Leu Asp His Phe Lys Ala Ala Met Ala Gin Gin Pro Pro Gin 325 330 335
Thr Met Leu Asn Lys Gly Thr Leu Arg Ser Tyr Glu His Gly Leu Lys 340 345 350
Glu Leu Leu Ala His Asp Lys lie Glu His Leu Ala Gly Gin Pro Gin 355 360 365
Gin Gly Pro Gin Ala Tyr Pro Gin Leu Phe Lys Ala Asp Val Ser Leu 370 375 380
Leu Leu Glu His Asp Glu Phe Leu Gin Glu Glu Val Phe Gly Pro Thr 385 390 395 400
Thr lie Val lie Glu Val Glu Ser Ala Glu Gin Leu Ala Leu Ala Leu -144· 146966-序列表.doc 201037078 405 410 415
Asn Gly Leu Arg Gly Gin Leu Thr Ala Ser Leu lie Ala Glu Pro Gin 420 425 430
Asp Phe Glu Asn Phe Ala Thr Leu lie Pro Leu Leu Glu Glu Lys Ala 435 440 445
Gly Arg Leu Leu Leu Asn Gly Tyr Pro Thr Gly Val Glu Val Cys Asp 450 455 460
Ala Met Val His Gly Gly Pro Tyr Pro Ala Thr Ser Asp Ala Arg Gly 465 470 475 480
Thr Ser Val Gly Thr Leu Ala lie Glu Arg Tyr Leu Arg Pro Val Cys 485 490 495 o
Tyr Gin Asn Tyr Pro Asp His Leu Leu Pro Leu Ala Leu Gin Asn Ala 500 505 510
Asn Pro Leu Gly lie Ala Arg Leu Val Asn Gly Glu Met Ser Lys Ala 515 520 525
Ala Leu 530 <210> 81 <211> 481 <212> PRT <213> 巴西固氮螺菌 <400> 81
Met Ala Asn Val Thr Tyr Thr Asp Thr Gin Leu Leu lie Asp Gly Glu 15 10 15
Trp Val Asp Ala Ala Ser Gly Lys Thr lie Asp Val Val Asn Pro Ala 20 25 30
Thr Gly Lys Pro He Gly Arg Val Ala His Ala Gly He Ala Asp Leu 35 40 45
Asp Arg Ala Leu Ala Ala Ala Gin Ser Gly Phe Glu Ala Trp Arg Lys 50 55 60
Val Pro Ala His Glu Arg Ala Ala Thr Met Arg Lys Ala Ala Ala Leu 65 70 75 80 •145· 146966-序列表.doc 201037078
Val Arg Glu Arg Ala Asp Ala lie Ala Gin Leu Met Thr Gin Glu Gin 85 90 95
Gly Lys Pro Leu Thr Glu Ala Arg Val Glu Val Leu Ser Ala Ala Asp 100 105 110 lie lie Glu Trp Phe Ala Asp Glu Gly Arg Arg Val Tyr Gly Arg lie 115 120 125
Val Pro Pro Arg Asn Leu Gly Ala Gin Gin Thr Val Val Lys Glu Pro 130 135 140
Val Gly Pro Val Ala Ala Phe Thr Pro Trp Asn Phe Pro Val Asn Gin 145 150 155 160
Val Val Arg Lys Leu Ser Ala Ala Leu Ala Thr Gly Cys Ser Phe Leu 165 170 175
Val Lys Ala Pro Glu Glu Thr Pro Ala Ser Pro Ala Ala Leu Leu Arg 180 185 190
Ala Phe Val Asp Ala Gly Val Pro Ala Gly Val lie Gly Leu Val Tyr 195 200 205
Gly Asp Pro Ala Glu lie Ser Ser Tyr Leu lie Pro His Pro Val lie 210 215 220
Arg Lys Val Thr Phe Thr Gly Ser Thr Pro Val Gly Lys Gin Leu Ala 225 230 235 240
Ser Leu Ala Gly Leu His Met Lys Arg Ala Thr Met Glu Leu Gly Gly 245 250 255
His Ala Pro Val lie Val Ala Glu Asp Ala Asp Val Ala Leu Ala Val 260 265 270
Lys Ala Ala Gly Gly Ala Lys Phe Arg Asn Ala Gly Gin Val Cys lie 275 280 285
Ser Pro Thr Arg Phe Leu Val His Asn Ser lie Arg Asp Glu Phe Thr 290 295 300
Arg Ala Leu Val Lys His Ala Glu Gly Leu Lys Val Gly Asn Gly Leu 305 310 315 320 146· 146966-序列表.doc 201037078
Glu Glu Gly Thr Thr Leu Gly Ala Leu Ala Asn Pro Arg Arg Leu Thr 325 330 335
Ala Met Ala Ser Val lie Asp Asn Ala Arg Lys Val Gly Ala Ser lie 340 345 350
Glu Thr Gly Gly Glu Arg lie Gly Ser Glu Gly Asn Phe Phe Ala Pro 355 360 365
Thr Val lie Ala Asn Val Pro Leu Asp Ala Asp Val Phe Asn Asn Glu 370 375 380
Pro Phe Gly Pro Val Ala Ala lie Arg Gly Phe Asp Lys Leu Glu Glu 385 390 395 400
Ala lie Ala Glu Ala Asn Arg Leu Pro Phe Gly Leu Ala Gly Tyr Ala 405 410 415
Phe Thr Arg Ser Phe Ala Asn Val His Leu Leu Thr Gin Arg Leu Glu 420 425 430
Val Gly Met Leu Trp lie Asn Gin Pro Ala Thr Pro Trp Pro Glu Met 435 440 445
Pro Phe Gly Gly Val Lys Asp Ser Gly Tyr Gly Ser Glu Gly Gly Pro 450 455 460
Glu Ala Leu Glu Pro Tyr Leu Val Thr Lys Ser Val Thr Val Met Ala 465 470 475 480
Val <210> <211> <212> <213>
82 1350 DNA 韋氏芽胞桿菌 <220> <221> CDS <222> (1)..(1350) <400> 82 gtg caa gcg acg gag caa aca caa agt ttg aaa aaa aca gat gaa aag Val Gin Ala Thr Glu Gin Thr Gin Ser Leu Lys Lys Thr Asp Glu Lys 15 10 15 tac ctt tgg cat gcg atg aga gga gca gcc cct agt cca acg aat tta Tyr Leu Trp His Ala Met Arg Gly Ala Ala Pro Ser Pro Thr Asn Leu 48 146966·序列表.doc •147- 96 201037078 20 25 30 att ate aca aaa gca gaa ggg gca tgg gtg aeg gat att gat gga aac 144 工le lie Thr Lys Ala Glu Gly Ala Trp Val Thr Asp lie Asp Gly Asn 35 40 45 cgt tat tta gac ggt atg tee ggt ett tgg tgc gtg aat gtt ggg tat 192
Arg Tyr Leu Asp Gly Met Ser Gly Leu Trp Cys Val Asn Val Gly Tyr 50 55 60 ggt ega aaa gaa ett gca aga geg geg ttt gaa cag ett gaa gaa atg 240
Gly Arg Lys Glu Leu Ala Arg Ala Ala Phe Glu Gin Leu Glu Glu Met 65 70 75 80 ccg tat ttc ect ctg act caa agt cat gtt cct get att aaa tta gca 288
Pro Tyr Phe Pro Leu Thr Gin Ser His Val Pro Ala lie Lys Leu Ala 85 90 95 gaa aaa ttg aat gaa tgg ett gat gat gaa tac gtc att ttc ttt tet 336
Glu Lys Leu Asn Glu Trp Leu Asp Asp Glu Tyr Val lie Phe Phe Ser 100 105 110 aac agt gga teg gaa geg aat gaa aca gca ttt aaa att get cgt caa 384
Asn Ser Gly Ser Glu Ala Asn Glu Thr Ala Phe Lys lie Ala Arg Gin 115 120 125 tat cat caa caa aaa ggt gat cat gga ege tat aag ttt att tee ege 432
Tyr His Gin Gin Lys Gly Asp His Gly Arg Tyr Lys Phe lie Ser Arg 130 135 140 tac ege get tat cac ggt aac tea atg gga get ett gca gca aca ggt 480
Tyr Arg Ala Tyr His Gly Asn Ser Met Gly Ala Leu Ala Ala Thr Gly 145 150 155 160 caa gca cag ega aag tat aaa tat gaa cca etc ggg caa gga ttc ctg 528
Gin Ala Gin Arg Lys Tyr Lys Tyr Glu Pro Leu Gly Gin Gly Phe Leu 165 170 175 cat gta gca ccg cct gat aeg tat ega aat cca gag gat gtt cat aca 576
His Val Ala Pro Pro Asp Thr Tyr Arg Asn Pro Glu Asp Val His Thr 180 185 190 ctg gca agt get gag gaa ate gat cgt gtc atg aca tgg gag tta age 624
Leu Ala Ser Ala Glu Glu lie Asp Arg Val Met Thr Trp Glu Leu Ser 195 200 205 caa aca gta gee ggt gtg att atg gag cca ate att act ggg ggc gga 672
Gin Thr Val Ala Gly Val lie Met Glu Pro lie lie Thr Gly Gly Gly 210 215 220 att tta atg cct cct gat gga tat atg gga aaa gta aaa gaa att tgc 720 lie Leu Met Pro Pro Asp Gly Tyr Met Gly Lys Val Lys Glu lie Cys 225 230 235 240 gag aag cac ggt geg ttg etc att tgt gat gaa gtt ata tgt gga ttt 768
Glu Lys His Gly Ala Leu Leu lie Cys Asp Glu Val lie Cys Gly Phe 245 250 255 ggc egg aca ggg aag cca ttt gga ttt atg aat tat ggc gtc aaa cca 816
Gly Arg Thr Gly Lys Pro Phe Gly Phe Met Asn Tyr Gly Val Lys Pro 260 265 270 -148 146966-序列表.doc 864 201037078 gat ate att aca atg gca aaa ggt att aca agt geg tat ett cct ttg
Asp lie lie Thr Met Ala Lys Gly lie Thr Ser Ala Tyr Leu Pro Leu 275 280 285 912 tea gca aca gca gtt aga ega gag gtt tat gag gca ttc gta ggt agt
Ser Ala Thr Ala Val Arg Arg Glu Val Tyr Glu Ala Phe Val Gly Ser 290 295 300
gat gat tat gat ege ttc ege cat gta aat aeg ttc gga ggg aat cct Asp Asp Tyr Asp Arg Phe Arg His Val Asn Thr Phe Gly Gly Asn Pro 305 310 315 320 get get tgc get tta get ttg aag aat tta gaa att atg gag aat gag Ala Ala Cys Ala Leu Ala Leu Lys Asn Leu Glu lie Met Glu Asn Glu 325 330 335 aaa etc att gaa cgt tee aaa gaa ttg ggt gaa ega ctg tta tat gag Lys Leu lie Glu Arg Ser Lys Glu Leu Gly Glu Arg Leu Leu Tyr Glu 340 345 350 eta gag gat gta aaa gag cat cca aac gta ggg gat gtt ege gga aag Leu Glu Asp Val Lys Glu His Pro Asn Val Gly Asp Val Arg Gly Lys 355 360 365 ggc ett ett tta ggc att gaa eta gtg gaa gat aag caa aca aaa gaa Gly Leu Leu Leu Gly lie Glu Leu Val Glu Asp Lys Gin Thr Lys Glu 370 375 380 ccg get tee att gaa aag atg aac aaa gtc ate aat get tgt aaa gaa Pro Ala Ser lie Glu Lys Met Asn Lys Val lie Asn Ala Cys Lys Glu 385 390 395 400 aaa ggt eta att att ggt aaa aat ggt gac act gtc gca ggt tac aat Lys Gly Leu lie lie Gly Lys Asn Gly Asp Thr Val Ala Gly Tyr Asn 405 410 415 aat att ttg cag ett gca cct cca tta age ate aca gag gaa gac ttt Asn lie Leu Gin Leu Ala Pro Pro Leu Ser lie Thr Glu Glu Asp Phe 420 425 430 act ttt ate gtt aaa aca atg aaa gaa tgt tta tee ege att aac ggg Thr Phe lie Val Lys Thr Met Lys Glu Cys Leu Ser Arg lie Asn Gly 435 440 445 cag taa 1350 Gin <210> 83 <211> 449 <212> PRT <213>韋氏芽胞桿菌 <400> 83
Val Gin Ala Thr Glu Gin Thr Gin Ser Leu Lys Lys Thr Asp Glu Lys 1 5 10 15 146966-序列表.doc -149- 960 1008 1056 1104 1152 1200 1248 1296 1344 201037078
Tyr Leu Trp His Ala Met Arg Gly Ala Ala Pro Ser Pro Thr Asn Leu 20 25 30 lie lie Thr Lys Ala Glu Gly Ala Trp Val Thr Asp lie Asp Gly Asn 35 40 45
Arg Tyr Leu Asp Gly Met Ser Gly Leu Trp Cys Val Asn Val Gly Tyr 50 55 60
Gly Arg Lys Glu Leu Ala Arg Ala Ala Phe Glu Gin Leu Glu Glu Met 65 70 75 80
Pro Tyr Phe Pro Leu Thr Gin Ser His Val Pro Ala lie Lys Leu Ala 85 90 95
Glu Lys Leu Asn Glu Trp Leu Asp Asp Glu Tyr Val lie Phe Phe Ser 100 105 110
Asn Ser Gly Ser Glu Ala Asn Glu Thr Ala Phe Lys lie Ala Arg Gin 115 120 125
Tyr His Gin Gin Lys Gly Asp His Gly Arg Tyr Lys Phe lie Ser Arg 130 135 140
Tyr Arg Ala Tyr His Gly Asn Ser Met Gly Ala Leu Ala Ala Thr Gly 145 150 155 160
Gin Ala Gin Arg Lys Tyr Lys Tyr Glu Pro Leu Gly Gin Gly Phe Leu 165 170 175
His Val Ala Pro Pro Asp Thr Tyr Arg Asn Pro Glu Asp Val His Thr 180 185 190
Leu Ala Ser Ala Glu Glu lie Asp Arg Val Met Thr Trp Glu Leu Ser 195 200 205
Gin Thr Val Ala Gly Val He Met Glu Pro He lie Thr Gly Gly Gly 210 215 220 lie Leu Met Pro Pro Asp Gly Tyr Met Gly Lys Val Lys Glu lie Cys 225 230 235 240
Glu Lys His Gly Ala Leu Leu lie Cys Asp Glu Val lie Cys Gly Phe 245 250 255
Gly Arg Thr Gly Lys Pro Phe Gly Phe Met Asn Tyr Gly Val Lys Pro •150- 146966-序列表.doc 201037078 260 265 270
Asp 工le lie Thr Met Ala Lys Gly lie Thr Ser Ala Tyr Leu Pro Leu 275 280 285
Ser Ala Thr Ala Val Arg Arg Glu Val Tyr Glu Ala Phe Val Gly Ser 290 295 300
Asp Asp Tyr Asp Arg Phe Arg His Val Asn Thr Phe Gly Gly Asn Pro 305 310 315 320
Ala Ala Cys Ala Leu Ala Leu Lys Asn Leu Glu lie Met Glu Asn Glu 325 330 335
Lys Leu lie Glu Arg Ser Lys Glu Leu Gly Glu Arg Leu Leu Tyr Glu 340 345 350
O
Leu Glu Asp Val Lys Glu His Pro Asn Val Gly Asp Val Arg Gly Lys 355 360 365
Gly Leu Leu Leu Gly lie Glu Leu Val Glu Asp Lys Gin Thr Lys Glu 370 375 380
Pro Ala Ser lie Glu Lys Met Asn Lys Val lie Asn Ala Cys Lys Glu 385 390 395 400
Lys Gly Leu lie lie Gly Lys Asn Gly Asp Thr Val Ala Gly Tyr Asn 405 410 415
Asn lie Leu Gin Leu Ala Pro Pro Leu Ser lie Thr Glu Glu Asp Phe 420 425 430 o
Thr Phe lie Val Lys Thr Met Lys Glu Cys Leu Ser Arg lie Asn Gly 435 440 445
Gin <210> 84 <211> 1350 <212> DNA <213> 人工 <220> <223>韋氏芽胞桿菌KBAB4胺基轉移酶之密碼子優化基因 <400> 84 atgcaggcta ccgaacaaac ccaatctctg aaaaagactg acgaaaaata tctgtggcac • 151 - 146966·序列表.doc 120 201037078 gcgatgcgcg gtgcagctcc gtctccgacc aacctgatta ttaccaaagc tgaaggcgcg tgggtgaccg acattgacgg taaccgttat ctggatggca tgagcggcct gtggtgtgtt aatgtcggtt atggccgtaa ggagctggcg cgcgcggcat ttgaacaact ggaagaaatg ccgtacttcc cgctgactca aagccatgtg ccggctatca aactggcgga aaaactgaac gaatggctgg acgacgaata cgtgattttc ttctctaatt ctggctccga agcaaacgaa accgcattca aaatcgcccg tcaatatcac cagcagaaag gtgaccacgg ccgctataaa ttcatcagcc gttatcgtgc ataccatggt aattctatgg gtgcgctggc tgctaccggt caggctcagc gcaaatacaa gtacgaaccg ctgggtcagg gttttctgca cgttgcacca ccggatacct accgtaaccc ggaagacgtc cacaccctgg cttctgccga agaaatcgat cgtgttatga cctgggagct gtcccagact gttgcgggtg ttatcatgga acctattatt accggtggtg gcattctgat gccgccggac ggttatatgg gtaaagtcaa ggaaatctgc gaaaaacacg gcgcgctgct gatctgcgat gaagttatct gtggcttcgg tcgcaccggc aaaccatttg gcttcatgaa ttatggcgta aaacctgaca ttattaccat ggctaaaggc attacttccg cttatctgcc gctgagcgcg accgcagttc gccgcgaagt ttatgaagcg tttgttggtt ctgatgatta cgaccgtttc cgtcatgtaa acacgtttgg cggtaaccca gcggcatgtg cgctggcgct gaaaaacctg gaaatcatgg aaaacgaaaa gctgatcgaa cgtagcaaag aactgggtga acgtctgctg tacgaactgg aagatgtcaa agaacacccg aacgtgggcg atgttcgcgg taaaggcctg ctgctgggta ttgaactggt tgaagacaaa cagaccaagg aaccggcttc cattgaaaag atgaacaaag tgattaacgc gtgcaaagag aaaggcctga tcattggtaa gaacggtgat accgtggcag gttataacaa cattctgcag ctggcgccgc ctctgagcat cactgaagaa gatttcacct tcatcgtcaa aactatgaag gagtgcctga gccgcatcaa tggtcagtaa 1350 <210> 85 <211> 1371 <212> DNA <213>綠膿桿菌 <220> <221> CDS <222> (1) .. (1371) <400> 85 atg aac age caa ate acc aac gee aag acc cgt gag tgg cag geg ttg Met Asn Ser Gin lie Thr Asn Ala Lys Thr Arg Glu Trp Gin Ala Leu 15 10 15 146966-序列表.doc -152- 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 96 201037078 age ege gac cac cat ctg ccg ccg ttc acc gac tac aag cag ttg aac
Ser Arg Asp His His Leu Pro Pro Phe Thr Asp Tyr Lys Gin Leu Asn 20 25 30 144 gag aag ggc geg egg ate ate acc aag gee gaa ggc gtc tat ate tgg
Glu Lys Gly Ala Arg lie lie Thr Lys Ala Glu Gly Val Tyr 工le Trp 35 40 45
gac age gag ggc aac aag ate etc gat geg atg gee ggc etc tgg tgc Asp Ser Glu Gly Asn Lys lie Leu Asp Ala Met Ala Gly Leu Trp Cys 50 55 60 gtc aac gtc ggc tac ggc ege gag gag ctg gtc cag gee gee acc egg Val Asn Val Gly Tyr Gly Arg Glu Glu Leu Val Gin Ala Ala Thr Arg 65 70 75 80 cag atg ege gag ttg ccg ttc tac aac ctg ttc ttc cag acc gee cac Gin Met Arg Glu Leu Pro Phe Tyr Asn Leu Phe Phe Gin Thr Ala His 85 90 95 ccg ccg gtg gtc gag ctg gee aag geg ate gee gac gtc get ccg gaa Pro Pro Val Val Glu Leu Ala Lys Ala lie Ala Asp Val Ala Pro Glu 100 105 110 ggc atg aac cac gtg ttc ttc acc ggc tee ggc tee gag gee aac gac Gly Met Asn His Val Phe Phe Thr Gly Ser Gly Ser Glu Ala Asn Asp 115 120 125 acc gtg ctg cgt atg gtc ege cac tat tgg geg acc aag ggc cag ccg Thr Val Leu Arg Met Val Arg His Tyr Trp Ala Thr Lys Gly Gin Pro 130 135 140 cag aag aaa gtg gtg ate ggc ege tgg aac ggc tac cac ggc tee acc Gin Lys Lys Val Val lie Gly Arg Trp Asn Gly Tyr His Gly Ser Thr 145 150 155 160 gtc gee ggc gtc age ctg ggc ggc atg aag geg ttg cat gag cag ggt Val Ala Gly Val Ser Leu Gly Gly Met Lys Ala Leu His Glu Gin Gly 165 170 175 gat ttc ccc ate ccg ggc ate gtc cac ate gee cag ccc tac tgg tac Asp Phe Pro lie Pro Gly lie Val His lie Ala Gin Pro Tyr Trp Tyr 180 185 190 ggc gag ggc ggc gac atg teg ccg gac gag ttc ggc gtc tgg gee gee Gly Glu Gly Gly Asp Met Ser Pro Asp Glu Phe Gly Val Trp Ala Ala 195 200 205 gag cag ttg gag aag aag att etc gaa gtg ggc gag gaa aac gtc gee Glu Gin Leu Glu Lys Lys lie Leu Glu Val Gly Glu Glu Asn Val Ala 210 215 220 gee ttc ate gee gag ccg ate cag ggc gee ggc ggc gtg ate gtc ccg Ala Phe lie Ala Glu Pro lie Gin Gly Ala Gly Gly Val lie Val Pro 225 230 235 240 ccg gac acc tac tgg ccg aag ate ege gag ate etc gee aag tac gac Pro Asp Thr Tyr Trp Pro Lys lie Arg Glu lie Leu Ala Lys Tyr Asp 245 250 255 ate ctg ttc ate gee gac gaa gtg ate tgc ggc ttc ggc cgt acc ggc 146966-序列表.doc 153- 192 240 288 336 384 432 480 528 576 624 672 720 816 864201037078 lie Leu Phe lie Ala Asp Glu Val lie Cys Gly Phe Gly Arg Thr Gly 260 265 270 gag tgg ttc ggc age cag tac tac ggc aac gcc ccg gac ctg atg ccg Glu Trp Phe Gly Ser Gin Tyr Tyr Gly Asn Ala Pro Asp Leu Met Pro 275 280 285 ate gcc aag ggc etc acc tcc ggc tac ate ccc atg ggc ggg gtg gtg lie Ala Lys Gly Leu Thr Ser Gly Tyr lie Pro Met Gly Gly Val Val 290 295 300 gtg ege gac gag ate gtc gaa gtg etc aac cag ggc ggc gag ttc tac Val Arg Asp Glu lie Val Glu Val Leu Asn Gin Gly Gly Glu Phe Tyr 305 310 315 320 cac ggc ttc acc tat tcc ggt cac ccg gtg geg gcc gcc gtg gcc ctg His Gly Phe Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala Leu 325 330 335 gag aac ate ege ate ctg ege gaa gag aag ate ate gag aag gtg aag Glu Asn lie Arg lie Leu Arg Glu Glu Lys lie lie Glu Lys Val Lys 340 345 350 geg gaa aeg gca ccg tat ttg cag aaa ege tgg cag gag ctg gcc gac Ala Glu Thr Ala Pro Tyr Leu Gin Lys Arg Trp Gin Glu Leu Ala Asp 355 360 365 cac ccg ttg gtg ggc gaa geg ege ggg gtc ggc atg gtc gcc gcc ctg His Pro Leu Val Gly Glu Ala Arg Gly Val Gly Met Val Ala TVla Leu 370 375 380 gag ctg gtc aag aac aag aag acc ege gag cgt ttc acc gac aag ggc Glu Leu Val Lys Asn Lys Lys Thr Arg Glu Arg Phe Thr Asp Lys Gly 385 390 395 400 gtc ggg atg ctg tgc egg gaa cat tgt ttc ege aac ggt ttg ate atg Val Gly Met Leu Cys Arg Glu His Cys Phe Arg Asn Gly Leu lie Met 405 410 415 ege geg gtg ggc gac act atg att ate teg ccg ccg ctg gtg ate gat Arg Ala Val Gly Asp Thr Met lie lie Ser Pro Pro Leu Val lie Asp 420 425 430 ccg teg cag ate gat gag ttg ate acc ctg geg ege aag tgc etc gat Pro Ser Gin lie Asp Glu Leu lie Thr Leu Ala Arg Lys Cys Leu Asp 435 440 445 cag acc gcc gcc gcc gtc ctg get tga 1371 Gin Thr Ala Ala Ala Val Leu Ala 450 455 912 960 1008 1056 1104 1152 1200 1248 1296 1344 <210> 86 <211> 456 <212> PRT <213>綠腺桿菌 <400> 86 Met Asn Ser Gin lie Thr Asn Ala Lys Thr Arg Glu Trp Gin Ala Leu 1 5 10 15 146966·序列表.doc -154 201037078
Ser Arg Asp His His Leu Pro Pro Phe Thr Asp Tyr Lys Gin Leu Asn 20 25 30
Glu Lys Gly Ala Arg lie lie Thr Lys Ala Glu Gly Val Tyr lie Trp 35 40 45
Asp Ser Glu Gly Asn Lys lie Leu Asp Ala Met Ala Gly Leu Trp Cys 50 55 60
Val Asn Val Gly Tyr Gly Arg Glu Glu Leu Val Gin Ala Ala Thr Arg 65 70 75 80
Gin Met Arg Glu Leu Pro Phe Tyr Asn Leu Phe Phe Gin Thr Ala His 85 90 95
Pro Pro Val Val Glu Leu Ala Lys Ala lie Ala Asp Val Ala Pro Glu 100 105 110
Gly Met Asn His Val Phe Phe Thr Gly Ser Gly Ser Glu Ala Asn Asp 115 120 125
Thr Val Leu Arg Met Val Arg His Tyr Trp Ala Thr Lys Gly Gin Pro 130 135 140
Gin Lys Lys Val Val lie Gly Arg Trp Asn Gly Tyr His Gly Ser Thr 145 150 155 160
Val Ala Gly Val Ser Leu Gly Gly Met Lys Ala Leu His Glu Gin Gly 165 170 175
Asp Phe Pro lie Pro Gly lie Val His lie Ala Gin Pro Tyr Trp Tyr 180 185 190
Gly Glu Gly Gly Asp Met Ser Pro Asp Glu Phe Gly Val Trp Ala Ala 195 200 205
Glu Gin Leu Glu Lys Lys lie Leu Glu Val Gly Glu Glu Asn Val Ala 210 215 220
Ala Phe lie Ala Glu Pro lie Gin Gly Ala Gly Gly Val lie Val Pro 225 230 235 240
Pro Asp Thr Tyr Trp Pro Lys lie Arg Glu lie Leu Ala Lys Tyr Asp 245 250 255 155- 146966-序列表.doc 201037078 lie Leu Phe lie Ala Asp Glu Val lie Cys Gly Phe Gly Arg Thr Gly 260 265 270
Glu Trp Phe Gly Ser Gin Tyr Tyr Gly Asn Ala Pro Asp Leu Met Pro 275 280 285 lie Ala Lys Gly Leu Thr Ser Gly Tyr lie Pro Met Gly Gly Val Val 290 295 300
Val Arg Asp Glu lie Val Glu Val Leu Asn Gin Gly Gly Glu Phe Tyr 305 310 315 320
His Gly Phe Thr Tyr Ser Gly His Pro Val Ala Ala Ala Val Ala Leu 325 330 335
Glu Asn lie Arg lie Leu Arg Glu Glu Lys lie lie Glu Lys Val Lys 340 345 350
Ala Glu Thr Ala Pro Tyr Leu Gin Lys Arg Trp Gin Glu Leu Ala Asp 355 360 365
His Pro Leu Val Gly Glu Ala Arg Gly Val Gly Met Val Ala Ala Leu 370 375 380
Glu Leu Val Lys Asn Lys Lys Thr Arg Glu Arg Phe Thr Asp Lys Gly 385 390 395 400
Val Gly Met Leu Cys Arg Glu His Cys Phe Arg Asn Gly Leu lie Met 405 410 415
Arg Ala Val Gly Asp Thr Met lie lie Ser Pro Pro Leu Val lie Asp 420 425 430
Pro Ser Gin lie Asp Glu Leu lie Thr Leu Ala Arg Lys Cys Leu Asp 435 440 445
Gin Thr Ala Ala Ala Val Leu Ala 450 455 <210> 87 <211> 70 <212> DNA <213> 人工 <220> <223> 引子 156· 146966-序列表.doc 201037078 <400> 87 ggggacaagt ttgtacaaaa aagcaggcta ggaggaatta accatgaaca gccaaatcac 60 caacgccaag 70 <210> 88 <211> 49 <212> DNA <213> 人工 <220> <223> 引子 <400> 88 ggggaccact ttgtacaaga aagctgggtt caagccagga cggcggcgg 49 <210> 89 <211> 849 <212> DNA 0 <213>枯草桿菌 <220> <221> CDS <222> (1)..(849) <400> 89 atg aag gtt tta gtc aat ggc egg ctg att ggg ege agt gaa gca tea Met Lys Val Leu Val Asn Gly Arg Leu lie Gly Arg Ser Glu Ala Ser 15 10 15 48 ate gat ttg gaa gat ege ggt tat cag ttt ggt gac ggc ate tat gaa lie Asp Leu Glu Asp Arg Gly Tyr Gin Phe Gly Asp Gly lie Tyr Glu 20 25 30 96 gtg ate agg gtg tac aaa gga gta ttg ttc ggc tta cgt gag cat gca Val lie Arg Val Tyr Lys Gly Val Leu Phe Gly Leu Arg Glu His Ala 35 40 45 144
gag cgt ttt ttc aga agt get get gaa ate gga att tea ctg cca ttc Glu Arg Phe Phe Arg Ser Ala Ala Glu lie Gly lie Ser Leu Pro Phe 50 55 60 192 agt ata gaa gat etc gag tgg gac ctg caa aag ett gta cag gaa aat Ser lie Glu Asp Leu Glu Trp Asp Leu Gin Lys Leu Val Gin Glu Asn 65 70 75 80 geg gtc agt gag gga geg gta tac att cag aca aca aga ggt gtg gee Ala Val Ser Glu Gly Ala Val Tyr He Gin Thr Thr Arg Gly Val Ala 85 90 95 240 288 ccg ega aaa cac cag tat gaa gee ggc etc gag ccg cag act act gee Pro Arg Lys His Gin Tyr Glu Ala Gly Leu Glu Pro Gin Thr Thr Ala 100 105 110 336 tat aeg ttt aeg gtg aaa aaa ccg gag caa gag cag gca tac gga gtg Tyr Thr Phe Thr Val Lys Lys Pro Glu Gin Glu Gin Ala Tyr Gly Val 115 120 125 146966-序列表.doe 157- 384 432201037078 gcg gcc att aca gat gag gat ctt cgc tgg tta aga tgt gat ate aaa Ala Ala lie Thr Asp Glu Asp Leu Arg Trp Leu Arg Cys Asp lie Lys 130 135 140 agt ctg aat tta ctg tat aat gtc atg aeg aag caa agg gcc tat gaa Ser Leu Asn Leu Leu Tyr Asn Val Met Thr Lys Gin Arg Ala Tyr Glu 145 150 155 160 gcc gga gca ttt gaa gcc att tta ctt agg gac ggc gtt gtt aeg gag Ala Gly Ala Phe Glu Ala lie Leu Leu Arg Asp Gly Val Val Thr Glu 165 170 175 ggt aca tcc tet aac gtt tat gcc gtt ate aac ggc aca gtg ega aca Gly Thr Ser Ser Asn Val Tyr Ala Val lie Asn Gly Thr Val Arg Thr 180 185 190 cat ccg get aat egg etc att etc aat gga att aca egg atg aat att His Pro Ala Asn Arg Leu 工le Leu Asn Gly lie Thr Arg Met Asn lie 195 200 205 tta gga ctg att gag aag aat ggg ate aaa ctg gat gag act cct gtc Leu Gly Leu lie Glu Lys Asn Gly lie Lys Leu Asp Glu Thr Pro Val 210 215 220 agt gaa gaa gag ttg aaa cag gcg gaa gag ate ttt att teg tea aeg Ser Glu Glu Glu Leu Lys Gin Ala Glu Glu lie Phe lie Ser Ser Thr 225 230 235 240 aeg gca gaa att att ccg gtc gtg aeg etc gat gga caa teg ate gga Thr Ala Glu lie lie Pro Val Val Thr Leu Asp Gly Gin Ser lie Gly 245 250 255 age ggg aaa ccc gga ccg gtg acc aaa cag ctt cag get get ttt caa Ser Gly Lys Pro Gly Pro Val Thr Lys Gin Leu Gin Ala Ala Phe Gin 260 265 270 gaa age att caa cag get get age att tea taa Glu Ser 工le Gin Gin Ala Ala Ser lie Ser 275 280 480 528 576 624 672 720 768 816 849 <210> 90 <211> 282 <212> PRT <213>枯草桿菌 <400> 90 Met Lys Val Leu Val Asn Gly Arg Leu lie Gly Arg Ser Glu Ala Ser 15 10 15 lie Asp Leu Glu Asp Arg Gly Tyr Gin Phe Gly Asp Gly lie Tyr Glu 20 25 30
Val lie Arg Val Tyr Lys Gly Val Leu Phe Gly Leu Arg Glu His Ala 35 40 45
Glu Arg Phe Phe Arg Ser Ala Ala Glu lie Gly lie Ser Leu Pro Phe 146966-序列表.doc -158- 201037078 50 55 60
Ser lie Glu Asp Leu Glu Trp Asp Leu Gin Lys Leu Val Gin Glu Asn 65 70 75 80
Ala Val Ser Glu Gly Ala Val Tyr lie Gin Thr Thr Arg Gly Val Ala 85 90 95
Pro Arg Lys His Gin Tyr Glu Ala Gly Leu Glu Pro Gin Thr Thr Ala 100 105 110
Tyr Thr Phe Thr Val Lys Lys Pro Glu Gin Glu Gin Ala Tyr Gly Val 115 120 125
Ala Ala lie Thr Asp Glu Asp Leu Arg Trp Leu Arg Cys Asp lie Lys 130 135 140 o
Ser Leu Asn Leu Leu Tyr Asn Val Met Thr Lys Gin Arg Ala Tyr Glu 145 150 155 160
Ala Gly Ala Phe Glu Ala lie Leu Leu Arg Asp Gly Val Val Thr Glu 165 170 175
Gly Thr Ser Ser Asn Val Tyr Ala Val lie Asn Gly Thr Val Arg Thr 180 185 190
His Pro Ala Asn Arg Leu lie Leu Asn Gly lie Thr Arg Met Asn lie 195 200 205
Leu Gly Leu lie Glu Lys Asn Gly lie Lys Leu Asp Glu Thr Pro Val 210 215 220 o
Ser Glu Glu Glu Leu Lys Gin Ala Glu Glu lie Phe lie Ser Ser Thr 225 230 235 240
Thr Ala Glu lie lie Pro Val Val Thr Leu Asp Gly Gin Ser lie Gly 245 250 255
Ser Gly Lys Pro Gly Pro Val Thr Lys Gin Leu Gin Ala Ala Phe Gin 260 265 270
Glu Ser lie Gin Gin Ala Ala Ser lie Ser 275 280 <210> 91 <211> 1347 -159- 146966-序列表.doc 201037078 <212> DNA <213>枯草桿菌 <220> <221> CDS <222> (1)..(1347) <400> 91 atg act cat gat ttg ata gaa aaa agt aaa aag cac etc tgg ctg cca 48
Met Thr His Asp Leu lie Glu Lys Ser Lys Lys His Leu Trp Leu Pro 15 10 15 ttt acc caa atg aaa gat tat gat gaa aac ccc tta ate ate gaa age 96
Phe Thr Gin Met Lys Asp Tyr Asp Glu Asn Pro Leu lie lie Glu Ser 20 25 30 ggg act gga ate aaa gtc aaa gac ata aac ggc aag gaa tac tat gac 144
Gly Thr Gly lie Lys Val Lys Asp lie Asn Gly Lys Glu Tyr Tyr Asp 35 40 45 ggt ttt tea teg gtt tgg ett aat gtc cac gga cac ege aaa aaa gaa 192
Gly Phe Ser Ser Val Trp Leu Asn Val His Gly His Arg Lys Lys Glu 50 55 60 eta gat gac gee ata aaa aaa cag etc gga aaa att geg cac tee aeg 240
Leu Asp Asp Ala lie Lys Lys Gin Leu Gly Lys lie Ala His Ser Thr 65 70 75 80 tta ttg ggc atg acc aat gtt cca gca acc cag ett gee gaa aca tta 288
Leu Leu Gly Met Thr Asn Val Pro Ala Thr Gin Leu Ala Glu Thr Leu 85 90 95 ate gac ate age cca aaa aag etc aeg egg gtc ttt tat tea gac age 336 lie Asp lie Ser Pro Lys Lys Leu Thr Arg Val Phe Tyr Ser Asp Ser 100 105 110 ggc gca gag geg atg gaa ata gee eta aaa atg geg ttt cag tat tgg 384
Gly Ala Glu Ala Met Glu lie Ala Leu Lys Met Ala Phe Gin Tyr Trp 115 120 125 aag aac ate ggg aag ccc gag aaa caa aaa ttc ate gca atg aaa aac 432
Lys Asn lie Gly Lys Pro Glu Lys Gin Lys Phe lie Ala Met Lys Asn 130 135 140 ggg tat cac ggt gat aeg att ggc gee gtc agt gtc ggt tea att gag 480
Gly Tyr His Gly Asp Thr lie Gly Ala Val Ser Val Gly Ser lie Glu 145 150 155 160 ett ttt cac cac gta tac ggc ccg ttg atg ttc gag agt tac aag gee 528
Leu Phe His His Val Tyr Gly Pro Leu Met Phe Glu Ser Tyr Lys Ala 165 170 175 ccg att cct tat gtg tat cgt tet gaa age ggt gat cct gat gag tgc 576
Pro lie Pro Tyr Val Tyr Arg Ser Glu Ser Gly Asp Pro Asp Glu Cys 180 185 190 cgt gat cag tgc etc ega gag ett gca cag ctg ett gag gaa cat cat 624
Arg Asp Gin Cys Leu Arg Glu Leu Ala Gin Leu Leu Glu Glu His His 195 200 205 -160· 146966-序列表.doc 672 201037078 gag gaa att gcc gcg ctt tcc att gaa tea atg gta caa ggc geg tee Glu Glu lie Ala Ala Leu Ser lie Glu Ser Met Val Gin Gly Ala Ser 210 215 220
❹ ggt atg ate gtg atg ccg gaa gga tat ttg gca ggc gtg ege gag eta Gly Met lie Val Met Pro Glu Gly Tyr Leu Ala Gly Val Arg Glu Leu 225 230 235 240 tgt aca aca tac gat gtc tta atg ate gtt gat gaa gtc get aca ggc Cys Thr Thr Tyr Asp Val Leu Met lie Val Asp Glu Val Ala Thr Gly 245 250 255 ttt ggc cgt aca gga aaa atg ttt gcg tgc gag cac gag aat gtc cag Phe Gly Arg Thr Gly Lys Met Phe Ala Cys Glu His Glu Asn Val Gin 260 265 270 cct gat ctg atg get gcc ggt aaa ggc att aca gga ggc tat ttg cca Pro Asp Leu Met Ala Ala Gly Lys Gly lie Thr Gly Gly Tyr Leu Pro 275 280 285 att gcc gtt aeg ttt gcc act gaa gac ate tat aag gca ttc tat gat lie Ala Val Thr Phe Ala Thr Glu Asp lie Tyr Lys Ala Phe Tyr Asp 290 295 300 gat tat gaa aac eta aaa acc ttt ttc cat ggc cat tec tat aca ggc Asp Tyr Glu Asn Leu Lys Thr Phe Phe His Gly His Ser Tyr Thr Gly 305 310 315 320 aat cag ctt ggc tgt gcg gtt gcg ctt gaa aat ctg gca tta ttt gaa Asn Gin Leu Gly Cys Ala Val Ala Leu Glu Asn Leu Ala Leu Phe Glu 325 330 335 tet gaa aac att gtg gaa caa gta gcg gaa aaa agt aaa aag etc cat Ser Glu Asn lie Val Glu Gin Val Ala Glu Lys Ser Lys Lys Leu His 340 345 350 ttt ctt ctt caa gat ctg cac get ctt cct cat gtt ggg gat att egg Phe Leu Leu Gin Asp Leu His Ala Leu Pro His Val Gly Asp lie Arg 355 360 365 cag ctt ggc ttt atg tgc ggt gca gag ctt gta ega tea aag gaa act Gin Leu Gly Phe Met Cys Gly Ala Glu Leu Val Arg Ser Lys Glu Thr 370 375 380 aaa gaa cct tac ccg get gat egg egg att gga tac aaa gtt tcc tta Lys Glu Pro Tyr Pro Ala Asp Arg Arg lie Gly Tyr Lys Val Ser Leu 385 390 395 400 aaa atg aga gag tta gga atg ctg aca aga ccg ctt ggg gac gtg att Lys Met Arg Glu Leu Gly Met Leu Thr Arg Pro Leu Gly Asp Val lie 405 410 415 gca ttt ctt cct cct ctt gcc age aca get gaa gag etc teg gaa atg Ala Phe Leu Pro Pro Leu Ala Ser Thr Ala Glu Glu Leu Ser Glu Met 420 425 430 gtt gcc att atg aaa caa gcg ate cac gag gtt aeg age ctt gaa gat Val Ala lie Met Lys Gin Ala lie His Glu Val Thr Ser Leu Glu Asp 435 440 445 tga 1347 146966-序列表.doc -161 - 720 768 816 864 912 960 1008 1056 1104 1152 1200 1248 1296 1344 201037078 <210> 92 <211> 448 <212> PRT <213>枯草桿菌 <400〉 92
Met Thr His Asp Leu lie Glu Lys Ser Lys Lys His Leu Trp Leu Pro 1 5 10 15 Phe Thr Gin Met Lys Asp Tyr Asp Glu Asn Pro Leu lie lie Glu Ser 20 25 30
Gly Thr Gly lie Lys Val Lys Asp lie Asn Gly Lys Glu Tyr Tyr Asp 35 40 45
Gly Phe Ser Ser Val Trp Leu Asn Val His Gly His Arg Lys Lys Glu 50 55 60
Leu Asp Asp Ala lie Lys Lys Gin Leu Gly Lys lie Ala His Ser Thr 65 70 75 80
Leu Leu Gly Met Thr Asn Val Pro Ala Thr Gin Leu Ala Glu Thr Leu 85 90 95 工le Asp lie Ser Pro Lys Lys Leu Thr Arg Val Phe Tyr Ser Asp Ser 100 105 110
Gly Ala Glu Ala Met Glu lie Ala Leu Lys Met Ala Phe Gin Tyr Trp 115 120 125
Lys Asn lie Gly Lys Pro Glu Lys Gin Lys Phe lie Ala Met Lys Asn 130 135 140
Gly Tyr His Gly Asp Thr He Gly Ala Val Ser Val Gly Ser He Glu 145 150 155 160
Leu Phe His His Val Tyr Gly Pro Leu Met Phe Glu Ser Tyr Lys Ala 165 170 175
Pro lie Pro Tyr Val Tyr Arg Ser Glu Ser Gly Asp Pro Asp Glu Cys 180 185 190
Arg Asp Gin Cys Leu Arg Glu Leu Ala Gin Leu Leu Glu Glu His His 195 200 205 -162- 146966-序列表.doc 201037078
Glu Glu lie Ala Ala Leu Ser lie Glu Ser Met Val Gin Gly Ala Ser 210 215 220
Gly Met lie Val Met Pro Glu Gly Tyr Leu Ala Gly Val Arg Glu Leu 225 230 235 240
Cys Thr Thr Tyr Asp Val Leu Met lie Val Asp Glu Val Ala Thr Gly 245 250 255
Phe Gly Arg Thr Gly Lys Met Phe Ala Cys Glu His Glu Asn Val Gin 260 265 270
Pro Asp Leu Met Ala Ala Gly Lys Gly lie Thr Gly Gly Tyr Leu Pro 275 280 285
lie Ala Val Thr Phe Ala Thr Glu Asp lie Tyr Lys Ala Phe Tyr Asp 290 295 300
Asp Tyr Glu Asn Leu Lys Thr Phe Phe His Gly His Ser Tyr Thr Gly 305 310 315 320
Asn Gin Leu Gly Cys Ala Val Ala Leu Glu Asn Leu Ala Leu Phe Glu 325 330 335
Ser Glu Asn lie Val Glu Gin Val Ala Glu Lys Ser Lys Lys Leu His 340 345 350
Phe Leu Leu Gin Asp Leu His Ala Leu Pro His Val Gly Asp lie Arg 355 360 365
Gin Leu Gly Phe Met Cys Gly Ala Glu Leu Val Arg Ser Lys Glu Thr 370 375 380
Lys Glu Pro Tyr Pro Ala Asp Arg Arg lie Gly Tyr Lys Val Ser Leu 385 390 395 400
Lys Met Arg Glu Leu Gly Met Leu Thr Arg Pro Leu Gly Asp Val lie 405 410 415
Ala Phe Leu Pro Pro Leu Ala Ser Thr Ala Glu Glu Leu Ser Glu Met 420 425 430
Val Ala lie Met Lys Gin Ala lie His Glu Val Thr Ser Leu Glu Asp 435 440 445 <210> 93 -163- 146966-序列表.doc 48201037078 <211> 1467 <212> DNA <213> 類球紅細菌 <220> <22X> CDS <222> (1)..(1467) <400> 93 atg ccc ggt tgc ggg ggc ttg ccc ggg aat gaa ccg aaa tgc gga cga Met Pro Gly Cys Gly Gly Leu Pro Gly Asn Glu Pro Lys Cys Gly Arg 15 10 15 gag ggg agg teg geg atg aeg egg aat gac geg aeg aat get gee gga Glu Gly Arg Ser Ala Met Thr Arg Asn Asp Ala Thr Asn Ala Ala Gly 20 25 30 geg gtg ggc geg geg atg egg gat cac ate etc ttg cct gca cag gaa Ala Val Gly Ala Ala Met Arg Asp His lie Leu Leu Pro Ala Gin Glu 35 40 45 atg geg aag etc ggc aag tee geg cag ccg gtg ctg act cat gee gag Net Ala Lys Leu Gly Lys Ser Ala Gin Pro Val Leu Thr His Ala Glu 50 55 60 ggc ate tat gtc cat acc gag gac ggc ege ege ctg ate gac ggg ccg Gly lie Tyr Val His Thr Glu Asp Gly Arg Arg Leu lie Asp Gly Pro 65 70 75 80 geg ggc atg tgg tgc geg cag gtg ggc tac ggc ege ege gag ate gtc Ala Gly Met Trp Cys Ala Gin Val Gly Tyr Gly Arg Arg Glu lie Val 85 90 95 gat gee atg geg cat cag geg atg gtg ctg ccc tat gee teg ccc tgg Asp Ala Met Ala His Gin Ala Met Val Leu Pro Tyr Ala Ser Pro Trp 100 105 110 tat atg gee aeg age ccc geg geg egg ctg geg gag aag ate gee aeg Tyr Met Ala Thr Ser Pro Ala Ala Arg Leu Ala Glu Lys lie Ala Thr 115 120 125 ctg aeg ccg ggc gat etc aac egg ate ttt ttc acc aeg ggc ggg teg Leu Thr Pro Gly Asp Leu Asn Arg lie Phe Phe Thr Thr Gly Gly Ser 130 135 140 acc geg gtg gac age geg ctg ege ttc teg gaa ttc tac aac aac gtg Thr Ala Val Asp Ser Ala Leu Arg Phe Ser Glu Phe Tyr Asn Asn Val 145 150 155 160 ctg ggc egg ccg cag aag aag ege ate ate gtg ege tac gac ggc tat Leu Gly Arg Pro Gin Lys Lys Arg lie lie Val Arg Tyr Asp Gly Tyr 165 170 175 cac ggc teg aeg geg etc acc gee gee tgc acc ggc ege acc ggc aac His Gly Ser Thr Ala Leu Thr Ala Ala Cys Thr Gly Arg Thr Gly Asn 180 185 190 tgg ccg aac ttc gac ate geg cag gac egg ate teg ttc etc teg age Trp Pro Asn Phe Asp lie Ala Gin Asp Arg lie Ser Phe Leu Ser Ser 195 200 205 96 144 192 240 288 336 384 432 480 528 576 146966-序列表.doe -164 - 624 672 201037078 ccc aat ccg cgc cac gcc ggc aac cgc age cag gag geg ttc etc gac Pro Asn Pro Arg His Ala Gly Asn Arg Ser Gin Glu Ala Phe Leu Asp 210 215 220
gat ctg gtg cag gaa ttc gag gac egg ate gag age etc ggc ccc gac Asp Leu Val Gin Glu Phe Glu Asp Arg lie Glu Ser Leu Gly Pro Asp 225 230 235 240 aeg ate geg gcc ttc ctg gcc gag ccg ate etc gcc teg ggc ggc gtc Thr lie Ala Ala Phe Leu Ala Glu Pro lie Leu Ala Ser Gly Gly Val 245 250 255 att att ccg ccc gca ggc tat cat geg cgc ttc aag geg ate tgc gag lie lie Pro Pro Ala Gly Tyr His Ala Arg Phe Lys Ala lie Cys Glu 260 265 270 aag cac gac ate etc tat ate teg gac gag gtg gtg aeg ggc ttc ggc Lys His Asp lie Leu Tyr lie Ser Asp Glu Val Val Thr Gly Phe Gly 275 280 285 cgt tgc ggc gag tgg ttc gcc teg gag aag gtg ttc ggg gtg gtg ccg Arg Cys Gly Glu Trp Phe Ala Ser Glu Lys Val Phe Gly Val Val Pro 290 295 300 gac ate ate acc ttc gcc aag ggc gtg acc teg ggc tat gtg ccg etc Asp lie lie Thr Phe Ala Lys Gly Val Thr Ser Gly Tyr Val Pro Leu 305 310 315 320 ggc ggc ett geg ate tee gag geg gtg ctg geg egg ate teg ggc gag Gly Gly Leu Ala lie Ser Glu Ala Val Leu Ala Arg lie Ser Gly Glu 325 330 335 aat gcc aag gga age tgg ttc acc aac ggc tat acc tac age aat cag Asn Ala Lys Gly Ser Trp Phe Thr Asn Gly Tyr Thr Tyr Ser Asn Gin 340 345 350 ccg gtg gcc tgc gcc geg geg ett gcc aac ate gag ctg atg gag cgc Pro Val Ala Cys Ala Ala Ala Leu Ala Asn lie Glu Leu Met Glu Arg 355 360 365 gag ggc ate gtc gat cag geg cgc gag atg geg gac tat ttc gcc geg Glu Gly lie Val Asp Gin Ala Arg Glu Met Ala Asp Tyr Phe Ala Ala 370 375 380 geg ctg get teg ctg cgc gat ctg ccg ggc gtg geg gaa acc egg teg Ala Leu Ala Ser Leu Arg Asp Leu Pro Gly Val Ala Glu Thr Arg Ser 385 390 395 400 gtg ggc etc gtg ggt tgc gtg caa tgc ctg etc gac ccg acc egg geg Val Gly Leu Val Gly Cys Val Gin Cys Leu Leu Asp Pro Thr Arg Ala 405 410 415 gac ggc aeg gcc gag gac aag gcc ttc acc ctg aag ate gac gag cgc Asp Gly Thr Ala Glu Asp Lys Ala Phe Thr Leu Lys lie Asp Glu Arg 420 425 430 tgc ttc gag etc ggg ctg ate gtg cgc ccg ctg ggc gat etc tgc gtg Cys Phe Glu Leu Gly Leu lie Val Arg Pro Leu Gly Asp Leu Cys Val 435 440 445 146966-序列表.doc -165 720 768 816 864 912 960 1008 1056 1104 1152 1200 1248 1296 1344 1392 201037078 ate teg ccg ccg etc ate ate teg ege geg cag ate gac gag atg gtc lie Ser Pro Pro Leu lie lie Ser Arg Ala Gin lie Asp Glu Met Val 450 455 460 geg ate atg egg cag gee ate ace gaa gtg age gee gee cac ggt ctg
Ala lie Met Arg Gin Ala lie Thr Glu Val Ser Ala Ala His Gly Leu 465 470 475 480 acc geg aaa gaa ccg gee gee gtc tga 1467
Thr Ala Lys Glu Pro Ala Ala Val 485 <210> 94 <211> 488 <212> PRT <213> 類球紅細菌 <400> 94
Met Pro Gly Cys Gly Gly Leu Pro Gly Asn Glu Pro Lys Cys Gly Arg 15 10 15
Glu Gly Arg Ser Ala Met Thr Arg Asn Asp Ala Thr Asn Ala Ala Gly 20 25 30
Ala Val Gly Ala Ala Met Arg Asp His lie Leu Leu Pro Ala Gin Glu 35 40 45
Met Ala Lys Leu Gly Lys Ser Ala Gin Pro Val Leu Thr His Ala Glu 50 55 60
Gly lie Tyr Val His Thr Glu Asp Gly Arg Arg Leu lie Asp Gly Pro 65 70 75 80
Ala Gly Met Trp Cys Ala Gin Val Gly Tyr Gly Arg Arg Glu lie Val 85 90 95
Asp Ala Met Ala His Gin Ala Met Val Leu Pro Tyr Ala Ser Pro Trp 100 105 110
Tyr Met Ala Thr Ser Pro Ala Ala Arg Leu Ala Glu Lys lie Ala Thr 115 120 125
Leu Thr Pro Gly Asp Leu Asn Arg lie Phe Phe Thr Thr Gly Gly Ser 130 135 140
Thr Ala Val Asp Ser Ala Leu Arg Phe Ser Glu Phe Tyr Asn Asn Val 145 150 155 160
Leu Gly Arg Pro Gin Lys Lys Arg lie lie Val Arg Tyr Asp Gly Tyr -166- 1440 146966-序列表.doc 201037078 165 170 175
His Gly Ser Thr Ala Leu Thr Ala Ala Cys Thr Gly Arg Thr Gly Asn 180 185 190
Trp Pro Asn Phe Asp lie Ala Gin Asp Arg lie Ser Phe Leu Ser Ser 195 200 205
Pro Asn Pro Arg His Ala Gly Asn Arg Ser Gin Glu Ala Phe Leu Asp 210 215 220
Asp Leu Val Gin Glu Phe Glu Asp Arg lie Glu Ser Leu Gly Pro Asp 225 230 235 240
Thr lie Ala Ala Phe Leu Ala Glu Pro lie Leu Ala Ser Gly Gly Val 245 250 255 o lie lie Pro Pro Ala Gly Tyr His Ala Arg Phe Lys Ala lie Cys Glu 260 265 270
Lys His Asp lie Leu Tyr lie Ser Asp Glu Val Val Thr Gly Phe Gly 275 280 285
Arg Cys Gly Glu Trp Phe Ala Ser Glu Lys Val Phe Gly Val Val Pro 290 295 300
Asp lie lie Thr Phe Ala Lys Gly Val Thr Ser Gly Tyr Val Pro Leu 305 310 315 320
Gly Gly Leu Ala lie Ser Glu Ala Val Leu Ala Arg lie Ser Gly Glu 325 330 335 o
Asn Ala Lys Gly Ser Trp Phe Thr Asn Gly Tyr Thr Tyr Ser Asn Gin 340 345 350
Pro Val Ala Cys Ala Ala Ala Leu Ala Asn lie Glu Leu Met Glu Arg 355 360 365
Glu Gly lie Val Asp Gin Ala Arg Glu Met Ala Asp Tyr Phe Ala Ala 370 375 380
Ala Leu Ala Ser Leu Arg Asp Leu Pro Gly Val Ala Glu Thr Arg Ser 385 390 395 400
Val Gly Leu Val Gly Cys Val Gin Cys Leu Leu Asp Pro Thr Arg Ala 405 410 415 .167- 146966-序列表 _doc 201037078
Asp Gly Thr Ala Glu Asp Lys Ala Phe Thr Leu Lys lie Asp Glu Arg 420 425 430
Cys Phe Glu Leu Gly Leu lie Val Arg Pro Leu Gly Asp Leu Cys Val 435 440 445 lie Ser Pro Pro Leu lie lie Ser Arg Ala Gin lie Asp Glu Met Val 450 455 460
Ala lie Met Arg Gin Ala lie Thr Glu Val Ser Ala Ala His Gly Leu 465 470 475 480
Thr Ala Lys Glu Pro Ala Ala Val 485 <210> 95 <211> 837 <212> DNA <213>嗜肺性退伍軍人桿菌 <220> <221> CDS <222> (1)..(837) <400> 95 atg agt ate gca ttt gtt aac ggc aag tat tgt tgt caa tet gaa gca Met Ser lie Ala Phe Val Asn Gly Lys Tyr Cys Cys Gin Ser Glu Ala 15 10 15 aaa att tea ata ttt gat ega ggg ttt ett ttt ggt gac teg gtt tat Lys lie Ser lie Phe Asp Arg Gly Phe Leu Phe Gly Asp Ser Val Tyr 20 25 30 gaa gtg ctg cct gtt tac cat ggg cag cct tac ttt gta gac caa cat Glu Val Leu Pro Val Tyr His Gly Gin Pro Tyr Phe Val Asp Gin His 35 40 45 ett gac ega tta ttc tea aat atg aaa aaa att aag atg att ata cca Leu Asp Arg Leu Phe Ser Asn Met Lys Lys lie Lys Met lie lie Pro 50 55 60 aat tat gat tgg cat ggt tta att cat aga eta ata tea gaa aat aat Asn Tyr Asp Trp His Gly Leu lie His Arg Leu lie Ser Glu Asn Asn 65 70 75 80 ggc ggt aat tta caa gta tat ate caa gtc aca ega ggg aat caa ggg Gly Gly Asn Leu Gin Val Tyr lie Gin Val Thr Arg Gly Asn Gin Gly 85 90 95 gtg ege aag cat gat ate cct act tcc ate aca cct tet gtt ate gca Val Arg Lys His Asp lie Pro Thr Ser 工le Thr Pro Ser Val lie Ala 100 105 110 48 96 144 192 240 288 146966-序列表.doc -168- 336 384201037078 ttc act atg cat aat cca ttt ccc acc etc gaa gat aag gaa cag gga Phe Thr Met His Asn Pro Phe Pro Thr Leu Glu Asp Lys Glu Gin Gly 115 120 125 atg tea gca aaa ctg gtt gaa gat ttt egg tgg atg aga tgt gat ata Met Ser Ala Lys Leu Val Glu Asp Phe Arg Trp Met Arg Cys Asp lie 130 135 140 aaa act act tet tta att gcc aat ata tta ctg aat gat gag get gta Lys Thr Thr Ser Leu lie Ala Asn lie Leu Leu Asn Asp Glu Ala Val 145 150 155 160 tet gca gga ttc cac act gca att ett gcc egg aac ggt eta att aca Ser Ala Gly Phe His Thr Ala lie Leu Ala Arg Asn Gly Leu 工le Thr 165 170 175 gag gga agt agt acc aac gta ttt att gtc gca cag gat ggt gtt att Glu Gly Ser Ser Thr Asn Val Phe lie Val Ala Gin Asp Gly Val lie 180 185 190 432 480 528 576
aag aca cca ccc atg aat aat ttc tgt tta cca gga att act egg caa Lys Thr Pro Pro Met Asn Asn Phe Cys Leu Pro Gly lie Thr Arg Gin 195 200 205 624 gtt gtt att gaa ata att aaa aaa tta gat tta aag ttc aga gaa ata Val Val lie Glu lie lie Lys Lys Leu Asp Leu Lys Phe Arg Glu lie 210 215 220 672 gaa att age att tea gag ett ttt tet get cag gaa gtt tgg ata aca Glu lie Ser lie Ser Glu Leu Phe Ser Ala Gin Glu Val Trp lie Thr 225 230 235 240 720 agt aeg aca aaa gaa gta ttc cct att aca aag att aat gac tet ttg Ser Thr Thr Lys Glu Val Phe Pro lie Thr Lys lie Asn Asp Ser Leu 245 250 255 768 att aat ggc gga aaa gtt ggc gaa tat tgg egg ata att aat gat tcc lie Asn Gly Gly Lys Val Gly Glu Tyr Trp Arg lie lie Asn Asp Ser 260 265 270 816
tac caa caa eta gta aac taa Tyr Gin Gin Leu Val Asn 275 837 <210> 96 <211> 278 <212> PRT <213>嗜肺性退伍軍人桿菌 <400> 96 Met Ser lie Ala Phe Val Asn Gly Lys Tyr Cys Cys Gin Ser Glu Ala 15 10 15
Lys lie Ser lie Phe Asp Arg Gly Phe Leu Phe Gly Asp Ser Val Tyr 20 25 30
Glu Val Leu Pro Val Tyr His Gly Gin Pro Tyr Phe Val Asp Gin His 146966-序列表.doc -169· 201037078 35 40 45
Leu Asp Arg Leu Phe Ser Asn Met Lys Lys lie Lys Met lie lie Pro 50 55 60
Asn Tyr Asp Trp His Gly Leu lie His Arg Leu lie Ser Glu Asn Asn 65 70 75 80
Gly Gly Asn Leu Gin Val Tyr lie Gin Val Thr Arg Gly Asn Gin Gly 85 90 95
Val Arg Lys His Asp lie Pro Thr Ser lie Thr Pro Ser Val lie Ala 100 105 110
Phe Thr Met His Asn Pro Phe Pro Thr Leu Glu Asp Lys Glu Gin Gly 115 120 125
Met Ser Ala Lys Leu Val Glu Asp Phe Arg Trp Met Arg Cys Asp lie 130 135 140
Lys Thr Thr Ser Leu 工le Ala Asn lie Leu Leu Asn Asp Glu Ala Val 145 150 155 160
Ser Ala Gly Phe His Thr Ala lie Leu Ala Arg Aan Gly Leu lie Thr 165 170 175
Glu Gly Ser Ser Thr Asn Val Phe lie Val Ala Gin Asp Gly Val lie 180 185 190
Lys Thr Pro Pro Met Asn Asn Phe Cys Leu Pro Gly lie Thr Arg Gin 195 200 205
Val Val lie Glu lie lie Lys Lys Leu Asp Leu Lys Phe Arg Glu lie 210 215 220
Glu lie Ser lie Ser Glu Leu Phe Ser Ala Gin Glu Val Trp lie Thr 225 230 235 240
Ser Thr Thr Lys Glu Val Phe Pro lie Thr Lys lie Asn Asp Ser Leu 245 250 255 lie Asn Gly Gly Lys Val Gly Glu Tyr Trp Arg lie lie Asn Asp Ser 260 265 270
Tyr Gin Gin Leu Val Asn 275 •170- 146966-序列表.doc 201037078 <210> 97 <211> 861 <212> DNA <213>歐洲亞硝化單胞菌 <220> <221> CDS <222> (1)..(861) <400> 97 atg att tac etc aat ggc aaa ttt ctg ccg atg gaa cag get acc gtt Met lie Tyr Leu Asn Gly Lys Phe Leu Pro Met Glu Gin Ala Thr Val 1 5 10 15 cca gtg ctg gat aga ggc ttc ate ttc ggt gat ggt gtc tat gaa gtc Pro Val Leu Asp Arg Gly Phe lie Phe Gly Asp Gly Val Tyr Glu Val 20 25 30 ata ccg gtt tat tea cgt aaa ccg ttc egg ctg ggc gaa cat ett tee lie Pro Val Tyr Ser Arg Lys Pro Phe Arg Leu Gly Glu His Leu Ser 35 40 45 egg ctg cag cac agt ctg gat ggc ata cgt etc cag aat ccg cac act Arg Leu Gin His Ser Leu Asp Gly lie Arg Leu Gin Asn Pro His Thr 50 55 60 gaa gaa caa tgg get ggt ctg ate gaa ege ate ate gag ctg aat gaa Glu Glu Gin Trp Ala Gly Leu lie Glu Arg lie lie Glu Leu Asn Glu 65 70 75 80 ggt gat gat cag tac ett tac ctg cac att aca ege ggg gtg gca aaa Gly Asp Asp Gin Tyr Leu Tyr Leu His lie Thr Arg Gly Val Ala Lys 85 90 95 cgt gac cat gee ttt cct ege gaa gta aeg ccc act gtc ttc ate atg Arg Asp His Ala Phe Pro Arg Glu Val Thr Pro Thr Val Phe lie Met 100 105 110 age aac ccg ett ccg get cca cct gca aaa ttg etc gtt tee gga gtt Ser Asn Pro Leu Pro Ala Pro Pro Ala Lys Leu Leu Val Ser Gly Val Q 115 120 125 48 96 144 192 240 288 336 384 tea geg att acc gee agg gat aat ege tgg ggg ege tgt gat ate aaa Ser Ala lie Thr Ala Arg Asp Asn Arg Trp Gly Arg Cys Asp lie Lys 130 135 140 gee att tea ctg ttg cca aat ate tta ttg ege cag ett gee gtg gac Ala lie Ser Leu Leu Pro Asn lie Leu Leu Arg Gin Leu Ala Val Asp 145 150 155 160 gca caa gee atg gaa aeg ate ctg tta ege gat ggt ctg ttg acc gaa Ala Gin Ala Met Glu Thr lie Leu Leu Arg Asp Gly Leu Leu Thr Glu 165 170 175 ggg gee gee age aat att ttc ate gta aaa gac gac ctg ctg ctg acc Gly Ala Ala Ser Asn lie Phe lie Val Lys Asp Asp Leu Leu Leu Thr 180 185 190 432 480 528 146966-序列表.doc -171- 576 624 201037078 624 ccc ccc aaa gat cac cgt ata ttg cct ggc att act tat gat gta gta Pro Pro Lys Asp His Arg lie Leu Pro Gly lie Thr Tyr Asp Val Val 195 200 205 ctg gaa ctg get gaa aca cat ggt gtt cca cat geg aca aga gaa ata Leu Glu Leu Ala Glu Thr His Gly Val Pro His Ala Thr Arg Glu lie 210 215 220 tea gag ett gag tta cgt act gca egg gaa ate atg ctg act tet tee Ser Glu Leu Glu Leu Arg Thr Ala Arg Glu lie Met Leu Thr Ser Ser 225 230 235 240 acc aaa gaa att etc ccg ate aca cag ctg gat gga caa ccg ate ggt Thr Lys Glu lie Leu Pro lie Thr Gin Leu Asp Gly Gin Pro lie Gly 245 250 255 aat ggc acc cca ggg cca gta ttt cag caa ctg gat egg etc tat cag Asn Gly Thr Pro Gly Pro Val Phe Gin Gin Leu Asp Arg Leu Tyr Gin 260 265 270 gca tat aag ctg gaa gtc atg ege ggg cat get cca ege cag taa Ala Tyr Lys Leu Glu Val Met Arg Gly His Ala Pro Arg Gin 275 280 285 672 720 768 816 861 <210> 98
<211> 286 <212> PRT <213〉歐洲亞硝化單胞菌 <400> 98
Met lie Tyr Leu Asn Gly Lys Phe Leu Pro Met Glu Gin Ala Thr Val 1 5 10 15
Pro Val Leu Asp Arg Gly Phe lie Phe Gly Asp Gly Val Tyr Glu Val 20 25 30 lie Pro Val Tyr Ser Arg Lys Pro Phe Arg Leu Gly Glu His Leu Ser 35 40 45
Arg Leu Gin His Ser Leu Asp Gly lie Arg Leu Gin Asn Pro His Thr 50 55 60
Glu Glu Gin Trp Ala Gly Leu lie Glu Arg lie lie Glu Leu Asn Glu 65 70 75 80
Gly Asp Asp Gin Tyr Leu Tyr Leu His lie Thr Arg Gly Val Ala Lys 85 90 95
Arg Asp His Ala Phe Pro Arg Glu Val Thr Pro Thr Val Phe lie Met 100 105 110
Ser Asn Pro Leu Pro Ala Pro Pro Ala Lys Leu Leu Val Ser Gly Val 172· 146966-序列表.doc 201037078 115 120 125
Ser Ala lie Thr Ala Arg Asp Asn Arg Trp Gly Arg Cys Asp He Lys 130 135 140
Ala lie Ser Leu Leu Pro Asn lie Leu Leu Arg Gin Leu Ala Val Asp 145 150 155 160
Ala Gin Ala Met Glu Thr lie Leu Leu Arg Asp Gly Leu Leu Thr Glu 165 170 175
Gly Ala Ala Ser Asn lie Phe lie Val Lys Asp Asp Leu Leu Leu Thr 180 185 190
Pro Pro Lys Asp His Arg lie Leu Pro Gly lie Thr Tyr Asp Val Val ▲ 195 200 205 o
Leu Glu Leu Ala Glu Thr His Gly Val Pro His Ala Thr Arg Glu lie 210 215 220
Ser Glu Leu Glu Leu Arg Thr Ala Arg Glu lie Met Leu Thr Ser Ser 225 230 235 240
Thr Lys Glu lie Leu Pro lie Thr Gin Leu Asp Gly Gin Pro He Gly 245 250 255
Asn Gly Thr Pro Gly Pro Val Phe Gin Gin Leu Asp Arg Leu Tyr Gin 260 265 270
Ala Tyr Lys Leu Glu Val Met Arg Gly His Ala Pro Arg Gin 275 280 285 〇 <210> 99 <211> 1293 <212> DNA <213>淋病奈瑟菌 <220> <221> CDS <222> (1)··(1293) <400> 99 atg agg ata aat atg aac cgt aac gaa att tta ttc gac cgc gcc aag
Met Arg lie Asn Met Asn Arg Asn Glu lie Leu Phe Asp Arg Ala Lys 15 10 15 gcc ate ate ccc ggc ggc gtg aat teg ccc gtg cgc gca ttc ggc age
Ala lie lie Pro Gly Gly Val Asn Ser Pro Val Arg Ala Phe Gly Ser 20 25 30 • 173 48 146966-序列表.doc 96 201037078 gtc ggc ggc gtg ccg cgc ttc ate aaa aaa gee gaa ggc geg tat gtt 144
Val Gly Gly Val Pro Arg Phe lie Lys Lys Ala Glu Gly Ala Tyr Val 35 40 45 tgg gac gaa aac ggc aeg cgc tac acc gat tat gtc ggc tet tgg ggg 192
Trp Asp Glu Asn Gly Thr Arg Tyr Thr Asp Tyr Val Gly Ser Trp Gly 50 55 60 cct geg att gtc gga cac geg cat ccc gaa gtc gtc gaa gee gtg cgc 240
Pro Ala lie Val Gly His Ala His Pro Glu Val Val Glu Ala Val Arg 65 70 75 80 gaa get geg ttg ggc ggt ttg teg ttc ggc geg ccc acc gaa ggc gaa 288
Glu Ala Ala Leu Gly Gly Leu Ser Phe Gly Ala Pro Thr Glu Gly Glu 85 90 95 ate gee att gee gaa caa att gee gaa att atg ccg tet gtc gaa egg 336 lie Ala lie Ala Glu Gin lie Ala Glu lie Met Pro Ser Val Glu Arg 100 105 110 ctg cgc etc gtc age tee ggc aeg gaa geg aeg atg act gee ate cgt 384
Leu Arg Leu Val Ser Ser Gly Thr Glu Ala Thr Met Thr Ala lie Arg 115 120 125 ctg gca cgc ggt ttt acc ggc cgc gac aaa ate ate aaa ttt gaa ggc 432
Leu Ala Arg Gly Phe Thr Gly Arg Asp Lys lie lie Lys Phe Glu Gly 130 135 140 tgc tac cac ggc cat tee gac age ctg ttg gtg aaa gca ggc age ggt 480
Cys Tyr His Gly His Ser Asp Ser Leu Leu Val Lys Ala Gly Ser Gly 145 150 155 160 ctg ett acc ttc ggc aat cct tet tee gee ggt gtg cct gee gac ttt 528
Leu Leu Thr Phe Gly Asn Pro Ser Ser Ala Gly Val Pro Ala Asp Phe 165 170 175 acc aaa cat act ttg gta etc gaa tac aac aac ate gee caa etc gaa 576
Thr Lys His Thr Leu Val Leu Glu Tyr Asn Asn lie Ala Gin Leu Glu 180 185 190 gaa gee ttt gee caa age ggc gac gaa ate gee tgc gtg att gtc gaa 624
Glu Ala Phe Ala Gin Ser Gly Asp Glu lie Ala Cys Val lie Val Glu 195 200 205 ccc ttc gtc ggc aat atg aac etc gtc cgc ccg acc gaa gee ttt gtc 672
Pro Phe Val Gly Asn Met Asn Leu Val Arg Pro Thr Glu Ala Phe Val 210 215 220 aaa gee ttg cgc gga ttg acc gaa aaa cac ggc geg gtg ttg att tac 720
Lys Ala Leu Arg Gly Leu Thr Glu Lys His Gly Ala Val Leu lie Tyr 225 230 235 240 gac gaa gtg atg acc ggt ttc cgc gtc geg etc ggc ggc geg cag teg 768
Asp Glu Val Met Thr Gly Phe Arg Val Ala Leu Gly Gly Ala Gin Ser 245 250 255 ctg cac ggc ate aeg ccc gac ctg acc aeg atg ggc aaa gtc ate ggc 816
Leu His Gly lie Thr Pro Asp Leu Thr Thr Met Gly Lys Val lie Gly 260 265 270 146966-序列表.doc -174 - 864 201037078 ggc ggt atg ccg ctt gcc gcg ttc ggc gga cgc aaa gac ate atg gaa
Gly Gly Met Pro Leu Ala Ala Phe Gly Gly Arg Lys Asp lie Met Glu 275 280 285 tgt att tcc ccg ttg ggc ggc gtg tat cag gca ggt aca tta tea ggc
Cys lie Ser Pro Leu Gly Gly Val Tyr Gin Ala Gly Thr Leu Ser Gly 290 295 300 912 aac ccg att gcc gtc gcc gcc ggc ttg aaa aeg ctg gaa ate ate cag Asn Pro lie Ala Val Ala Ala Gly Leu Lys Thr Leu Glu lie lie Gin 305 310 315 320 cgc gaa ggc ttc tat gaa aac ctg acc gcc ttg aca caa cgc ctt gcc Arg Glu Gly Phe Tyr Glu Asn Leu Thr Ala Leu Thr Gin Arg Leu Ala 325 330 335 aac ggt att gcc gcc gcc aaa gcg cac ggt ate gag ttt gcc gcc gac Asn Gly lie Ala Ala Ala Lys Ala His Gly lie Glu Phe Ala Ala Asp 340 345 350 age gtg ggc ggt atg ttc ggt ctg tat ttc gcc gca cac gtg ccg ega Ser Val Gly Gly Met Phe Gly Leu Tyr Phe Ala Ala His Val Pro Arg 355 360 365 960 1008 1056 1104 aac tat gcc gat atg gcg cgc tcc aat ate gac get ttc aaa cgc ttc Asn Tyr Ala Asp Met Ala Arg Ser Asn lie Asp Ala Phe Lys Arg Phe 370 375 380 ttc cac ggc atg etc gac cgc ggc att gcc ttc ggc ccg tcc get tat Phe His Gly Met Leu Asp Arg Gly lie Ala Phe Gly Pro Ser Ala Tyr 385 390 395 400 gaa gcg ggt ttc gtt tcc gcc gcg cat aeg ccc gag ctg att gac gaa Glu Ala Gly Phe Val Ser Ala Ala His Thr Pro Glu Leu lie Asp Glu 405 410 415 1152 1200 1248 1293 aeg gtt gcg gtt gcg gtt gaa gtg ttc aag gcg atg get gca tga Thr Val Ala Val Ala Val Glu Val Phe Lys Ala Met Ala Ala 420 425 430 <210> 100 <211> 430 <212> PRT <213> 淋病奈瑟菌 <400> 100
Met Arg lie Asn Met Asn Arg Asn Glu lie Leu Phe Asp Arg Ala Lys 15 10 15
Ala lie lie Pro Gly Gly Val Asn Ser Pro Val Arq Ala Phe Gly Ser 20 25 30
Val Gly Gly Val Pro Arg Phe lie Lys Lys Ala Glu Gly Ala Tyr Val 35 40 45
Trp Asp Glu Asn Gly Thr Arg Tyr Thr Asp Tyr Val Gly Ser Trp Gly -175· 146966-序列表.doc 201037078 50 55 60
Pro Ala lie Val Gly His Ala His Pro Glu Val Val Glu Ala Val Arg 65 70 75 80
Glu Ala Ala Leu Gly Gly Leu Ser Phe Gly Ala Pro Thr Glu Gly Glu 85 90 95 lie Ala lie Ala Glu Gin lie Ala Glu lie Met Pro Ser Val Glu Arg 100 105 110
Leu Arg Leu Val Ser Ser Gly Thr Glu Ala Thr Met Thr Ala lie Arg 115 120 125
Leu Ala Arg Gly Phe Thr Gly Arg Asp Lys lie lie Lys Phe Glu Gly 130 135 140
Cys Tyr His Gly His Ser Asp Ser Leu Leu Val Lys Ala Gly Ser Gly 145 150 155 160
Leu Leu Thr Phe Gly Asn Pro Ser Ser Ala Gly Val Pro Ala Asp Phe 165 170 175
Thr Lys His Thr Leu Val Leu Glu Tyr Asn Asn lie Ala Gin Leu Glu 180 185 190
Glu Ala Phe Ala Gin Ser Gly Asp Glu lie Ala Cys Val lie Val Glu 195 200 205
Pro Phe Val Gly Asn Met Asn Leu Val Arg Pro Thr Glu Ala Phe Val 210 215 220
Lys Ala Leu Arg Gly Leu Thr Glu Lys His Gly Ala Val Leu lie Tyr 225 230 235 240
Asp Glu Val Met Thr Gly Phe Arg Val Ala Leu Gly Gly Ala Gin Ser 245 250 255
Leu His Gly lie Thr Pro Asp Leu Thr Thr Met Gly Lys Val lie Gly 260 265 270
Gly Gly Met Pro Leu Ala Ala Phe Gly Gly Arg Lys Asp lie Met Glu 275 280 285
Cys lie Ser Pro Leu Gly Gly Val Tyr Gin Ala Gly Thr Leu Ser Gly 290 295 300 -176- 146966-序列表.doc 201037078
Asn Pro lie Ala Val Ala Ala Gly Leu Lys Thr Leu Glu lie lie Gin 305 310 315 320
Arg Glu Gly Phe Tyr Glu Asn Leu Thr Ala Leu Thr Gin Arg Leu Ala 325 330 335
Asn Gly 工le Ala Ala Ala Lys Ala His Gly lie Glu Phe Ala Ala Asp 340 345 350
Ser Val Gly Gly Met Phe Gly Leu Tyr Phe Ala Ala His Val Pro Arg 355 360 365
Asn Tyr Ala Asp Met Ala Arg Ser Asn lie Asp Ala Phe Lys Arg Phe 370 375 380
Phe His Gly Met Leu Asp Arg Gly lie Ala Phe Gly Pro Ser Ala Tyr 385 390 395 400
Glu Ala Gly Phe Val Ser Ala Ala His Thr Pro Glu Leu 工le Asp Glu 405 410 415
Thr Val Ala Val Ala Val Glu Val Phe Lys Ala Met Ala Ala 420 425 430 <210> 101 <211> 924 <212> DNA <213〉 綠膿桿菌 <220> <221> CDS <222> (1)..(924) <400> 101 atg teg atg gcc gat cgt gat ggc gtg ate tgg tat gac ggt gaa ctg Met Ser Met Ala Asp Arg Asp Gly Val lie Trp Tyr Asp Gly Glu Leu 15 10 15 gtg cag tgg ege gac geg acc aeg cac gtg ctg acc cat acc ctg cac Val Gin Trp Arg Asp Ala Thr Thr His Val Leu Thr His Thr Leu His 20 25 30 tat gga atg ggc gtg ttc gag ggc gtg ege gcc tac gac acc ccg cag Tyr Gly Met Gly Val Phe Glu Gly Val Arg Ala Tyr Asp Thr Pro Gin 35 40 45 ggc aeg geg ate ttc ege ctg cag geg cat acc gac egg ctg ttc gac Gly Thr Ala lie Phe Arg Leu Gin Ala His Thr Asp Arg Leu Phe Asp 50 55 60 48 96 144 146966-序列表.doc -177- 192 201037078 tcc gcg cac ate atg aac atg cag ate ccg tac age ege gac gag ate 240
Ser Ala His lie Met Asn Met Gin lie Pro Tyr Ser Arg Asp Glu lie 65 70 75 80 aac gag gcg acc ege gee gee gtg ege gag aac aac ctg gaa age gee 288
Asn Glu Ala Thr Arg Ala Ala Val Arg Glu Asn Asn Leu Glu Ser Ala 85 90 95 tat ate ege ccg atg gtg ttc tac gga age gaa ggc atg ggc ctg ege 336
Tyr lie Arg Pro Met Val Phe Tyr Gly Ser Glu Gly Met Gly Leu Arg 100 105 110 gee age ggc ctg aag gtc cat gtg ate ate gee gee tgg age tgg ggc 384
Ala Ser Gly Leu Lys Val His Val lie lie Ala Ala Trp Ser Trp Gly 115 120 125 gee tac atg ggc gag gaa gee ctg cag caa ggc ate aag gtg ege acc 432
Ala Tyr Met Gly Glu Glu Ala Leu Gin Gin Gly lie Lys Val Arg Thr 130 135 140 agt tec ttc acc ege cac cac gtc aac ate teg atg acc ege gee aag 480
Ser Ser Phe Thr Arg His His Val Asn lie Ser Met Thr Arg Ala Lys 145 150 155 160 tec aac ggc gee tac ate aac teg atg ctg gee etc cag gaa gcg ate 528
Ser Asn Gly Ala Tyr lie Asn Ser Met Leu Ala Leu Gin Glu Ala lie 165 170 175 tec ggc ggc gee gac gag gee atg atg etc gat ccg gaa ggc tac gtg 576
Ser Gly Gly Ala Asp Glu Ala Met Met Leu Asp Pro Glu Gly Tyr Val 180 185 190 gee gaa ggc tec ggc gag aac ate ttc ate ate aag gat ggc gtg ate 624
Ala Glu Gly Ser Gly Glu Asn lie Phe lie lie Lys Asp Gly Val lie 195 200 205 tac acc ccg gaa gtc acc gee tgc ctg aac ggc ate act cgt aac act 672
Tyr Thr Pro Glu Val Thr Ala Cys Leu Asn Gly lie Thr Arg Asn Thr 210 215 220 ate ctg acc ctg gee gee gaa cac ggt ttt aaa ctg gtc gag aag ege 720 lie Leu Thr Leu Ala Ala Glu His Gly Phe Lys Leu Val Glu Lys Arg 225 230 235 240 ate acc ege gac gag gtg tac ate gee gac gag gee ttc ttc act ggc 768 lie Thr Arg Asp Glu Val Tyr lie Ala Asp Glu Ala Phe Phe Thr Gly 245 250 255 act gee gcg gaa gtc aeg ccg ate ege gaa gtg gac ggt ege aag ate 816
Thr Ala Ala Glu Val Thr Pro lie Arg Glu Val Asp Gly Arg Lys lie 260 265 270 ggc gee ggc ege cgt ggc ccg gtc acc gaa aag ctg cag aaa gee tat 864
Gly Ala Gly Arg Arg Gly Pro Val Thr Glu Lys Leu Gin Lys Ala Tyr 275 280 285 ttc gac ctg gtc age ggc aag acc gag gee cac gee gag tgg cgt acc 912
Phe Asp Leu Val Ser Gly Lys Thr Glu Ala His Ala Glu Trp Arg Thr 290 295 300 ctg gtc aag taa 924 146966-序列表.doc - 178- 201037078
Leu Val Lys 305 <210> 102 <211> 307 <212> PRT <213>綠膿桿菌 <400> 102
Met Ser Met Ala Asp Arg Asp Gly Val lie Trp Tyr Asp Gly Glu Leu 15 10 15
Val Gin Trp Arg Asp Ala Thr Thr His Val Leu Thr His Thr Leu His 20 25 30
Tyr Gly Met Gly Val Phe Glu Gly Val Arg Ala Tyr Asp Thr Pro Gin 35 40 45
Gly Thr Ala lie Phe Arg Leu Gin Ala His Thr Asp Arg Leu Phe Asp 50 55 60
Ser Ala His lie Met Asn Met Gin lie Pro Tyr Ser Arg Asp Glu lie 65 70 75 80
Asn Glu Ala Thr Arg Ala Ala Val Arg Glu Asn Asn Leu Glu Ser Ala 85 90 95
Tyr lie Arg Pro Met Val Phe Tyr Gly Ser Glu Gly Met Gly Leu Arg 100 105 110
Ala Ser Gly Leu Lys Val His Val lie lie Ala Ala Trp Ser Trp Gly 115 120 125 ❹
Ala Tyr Met Gly Glu Glu Ala Leu Gin Gin Gly lie Lys Val Arg Thr 130 135 140
Ser Ser Phe Thr Arg His His Val Asn lie Ser Met Thr Arg Ala Lys 145 150 155 160
Ser Asn Gly Ala Tyr lie Asn Ser Met Leu Ala Leu Gin Glu Ala lie 165 170 175
Ser Gly Gly Ala Asp Glu Ala Met Met Leu Asp Pro Glu Gly Tyr Val 180 185 190
Ala Glu Gly Ser Gly Glu Asn lie Phe lie lie Lys Asp Gly Val lie 195 200 205 •179- 146966-序列表.doc 201037078
Tyr Thr Pro Glu Val Thr Ala Cys Leu Asn Gly lie Thr Arg Asn Thr 210 215 220 lie Leu Thr Leu Ala Ala Glu His Gly Phe Lys Leu Val Glu Lys Arg 225 230 235 240 lie Thr Arg Asp Glu Val Tyr lie Ala Asp Glu Ala Phe Phe Thr Gly 245 250 255
Thr Ala Ala Glu Val Thr Pro lie Arg Glu Val Asp Gly Arg Lys lie 260 265 270
Gly Ala Gly Arg Arg Gly Pro Val Thr Glu Lys Leu Gin Lys Ala Tyr 275 280 285
Phe Asp Leu Val Ser Gly Lys Thr Glu Ala His Ala Glu Trp Arg Thr 290 295 300
Leu Val Lys 305 <210〉 103 <211> 1407 <212> DNA <213>沼澤紅假單胞菌 <220> <221> CDS <222> (1)..(1407) <400> 103 atg aag ctg ata ccg tgc cgc gcc ttt cac ccc ccg gcc gcg cag tgc Met Lys Leu lie Pro Cys Arg Ala Phe His Pro Pro Ala Ala Gin Cys 1 5 10 15 atg agg age gcc atg tta gac aag ate aag ccc aeg tcc gcc gtc aac Met Arg Ser Ala Met Leu Asp Lys lie Lys Pro Thr Ser Ala Val Asn 20 25 30 gcg ccg aac gat etc aac gcg ttc tgg atg ccg ttc acc gcg aac egg Ala Pro Asn Asp Leu Asn Ala Phe Trp Met Pro Phe Thr Ala Asn Arg 35 40 45 gcc ttc aag cgc gcg ccg aag atg gtc gtg ggt gcc gaa ggc atg cac Ala Phe Lys Arg Ala Pro Lys Met Val Val Gly Ala Glu Gly Met His 50 55 60 tac ate acc gcc gat ggt cgc aag ate ate gac gcc gcc teg ggc atg Tyr lie Thr Ala Asp Gly Arg Lys lie lie Asp Ala Ala Ser Gly Met 65 70 75 80 48 96 144 192 146966-序列表.doc 180· 240 288 201037078 tgg tgc acc aat gcg ggc cat ggc cgc aag gaa ate gee gag geg ate
Trp Cys Thr Asn Ala Gly His Gly Arg Lys Glu lie Ala Glu Ala lie 85 90 95 336 384 432 480 528 576 624 672 720 768 816 aag gcg cag gee gat gaa etc gac ttc teg ccg ccg ttc cag ttc ggc
Lys Ala Gin Ala Asp Glu Leu Asp Phe Ser Pro Pro Phe Gin Phe Gly 100 105 110 cag ccg aag gcg ttc gaa etc gee age egg ate gee gat ctg gcg ccg
Gin Pro Lys Ala Phe Glu Leu Ala Ser Arg lie Ala Asp Leu Ala Pro 115 120 125 gaa ggc etc gat cac gtg ttc ttc tgc aat teg ggc teg gaa gee ggc
Glu Gly Leu Asp His Val Phe Phe Cys Asn Ser Gly Ser Glu Ala Gly 130 135 140 gac acc gcg ctg aag ate gcg gtc gee tat cag cag ate aag ggc cag
Asp Thr Ala Leu Lys lie Ala Val Ala Tyr Gin Gin lie Lys Gly Gin 145 150 155 160
ggc tea cgc acc cgc ctg ate ggc cgc gag cgc ggc tat cac ggc gtc Gly Ser Arg Thr Arg Leu lie Gly Arg Glu Arg Gly Tyr His Gly Val 165 170 175 ggc ttc ggc ggc acc gcg gtc ggc ggc ate ggc aac aac cgc aag atg Gly Phe Gly Gly Thr Ala Val Gly Gly lie Gly Asn Asn Arg Lys Met 180 185 190 ttc ggt ccg ctg etc aac ggc gtc gat cat ctg cct gcg act tat gat Phe Gly Pro Leu Leu Asn Gly Val Asp His Leu Pro Ala Thr Tyr Asp 195 200 205 cgc gac aag cag get ttc acc ate ggc gag ccg gaa tac ggc gcg cac
Arg Asp Lys Gin Ala Phe Thr lie Gly Glu Pro Glu Tyr Gly Ala His 210 215 220 ttc gee gaa gcg ett gaa ggc etc gtc aat ctg cac ggc gee aac acc
Phe Ala Glu Ala Leu Glu Gly Leu Val Asn Leu His Gly Ala Asn Thr 225 230 235 240 ate gcg gcg gtg ate gtc gag ccg atg gee ggc tee acc ggc gtg ctg lie Ala Ala Val lie Val Glu Pro Met Ala Gly Ser Thr Gly Val Leu 245 250 255 ccg gcg ccg aag ggc tat etc aag aag ctg cgc gag ate acc aag aag Pro Ala Pro Lys Gly Tyr Leu Lys Lys Leu Arg Glu lie Thr Lys Lys 260 265 270 cac ggc ate ctg ctg ate ttc gac gag gtc ate acc ggc tac ggc cgt 864
His Gly lie Leu Leu lie Phe Asp Glu Val lie Thr Gly Tyr Gly Arg 275 280 285 etc ggc tat gee ttc gcg tee gaa cgt tac ggc gtc acc ccg gac atg 912
Leu Gly Tyr Ala Phe Ala Ser Glu Arg Tyr Gly Val Thr Pro Asp Met 290 295 300 ate acc ttc gee aag ggc gtc acc aat ggt gcg gtg ccg atg ggc ggc 960 lie Thr Phe Ala Lys Gly Val Thr Asn Gly Ala Val Pro Met Gly Gly 305 310 315 320 gtg ate acc teg gcg gag ate cac gat gcg ttc atg acc ggc ccc gag 1008 181 - 146966-序列表.doe 1056201037078
Val lie Thr Ser Ala Glu lie His Asp Ala Phe Met Thr Gly Pro Glu 325 330 335 cac gcg gtc gag ctg gcg cac ggc tac acc tat teg geg cat ccg etc His Ala Val Glu Leu Ala His Gly Tyr Thr Tyr Ser Ala His Pro Leu 340 345 350 gcc tgc gcg gcc ggc ate gcc acc etc gac ate tac ege gac gag aag Ala Cys Ala Ala Gly lie Ala Thr Leu Asp lie Tyr Arg Asp Glu Lys 355 360 365 ctg ttc gag ege gcc aag gcg ctg gag ccg aag ttt gcc gag gcg gtg Leu Phe Glu Arg Ala Lys Ala Leu Glu Pro Lys Phe Ala Glu Ala Val 370 375 380 atg teg ctg aag teg gcc ccg aac gtg gtc gac ate ege acc gtc ggc Met Ser Leu Lys Ser Ala Pro Asn Val Val Asp lie Arg Thr Val Gly 385 390 395 400 ctg aeg gcg ggt ate gac etc get teg ate gcc gat gcg gtc ggc aag Leu Thr Ala Gly lie Asp Leu Ala Ser lie Ala Asp Ala Val Gly Lys 405 410 415 cgt ggc ttc gaa gcg atg aat gcc ggc ttc cac gac cac gag ctg atg Arg Gly Phe Glu Ala Met Asn Ala Gly Phe His Asp His Glu Leu Met 420 425 430 ctg egg ate gcc ggc gac acc ctg gcg ctg acc ccg ccg ctg ate etc Leu Arg lie Ala Gly Asp Thr Leu Ala Leu Thr Pro Pro Leu lie Leu 435 440 445 age gag gac cac ate ggt gag ate gtc gac aag gtc ggc aag gtg ate Ser Glu Asp His lie Gly Glu lie Val Asp Lys Val Gly Lys Val lie 450 455 460 ege gcg gtc gcc tga 1407 Arg Ala Val Ala 465 1104 1152 1200 1248 1296 1344 1392 <210> 104 <211> 468 <212> PRT <213>沼澤紅假單胞菌 <400> 104 Met Lys Leu lie Pro Cys Arg Ala Phe His Pro Pro Ala Ala Gin Cys 15 10 15
Met Arg Ser Ala Met Leu Asp Lys lie Lys Pro Thr Ser Ala Val Asn 20 25 30
Ala Pro Asn Asp Leu Asn Ala Phe Trp Met Pro Phe Thr Ala Asn Arg 35 40 45
Ala Phe Lys Arg Ala Pro Lys Met Val Val Gly Ala Glu Gly Met His 50 55 60 146966-序列表.doc -182- 201037078
Tyr lie Thr Ala Asp Gly Arg Lys lie lie Asp Ala Ala Ser Gly Met 65 70 75 80
Trp Cys Thr Asn Ala Gly His 85
Gly Arg Lys Glu lie Ala Glu Ala lie 90 95
Lys Ala Gin Ala Asp Glu Leu Asp Phe Ser Pro Pro Phe Gin Phe Gly 100 105 110
Gin Pro Lys Ala Phe Glu Leu Ala Ser Arg lie Ala Asp Leu Ala Pro 115 120 125
Glu Gly Leu Asp His Val Phe Phe Cys Asn Ser Gly Ser Glu Ala Gly 130 135 140
Asp Thr Ala Leu Lys lie Ala Val Ala Tyr Gin Gin lie Lys Gly Gin 145 150 155 160
Gly Ser Arg Thr Arg Leu lie Gly Arg Glu Arg Gly Tyr His Gly Val 165 170 175
Gly Phe Gly Gly Thr TVla Val Gly Gly lie Gly Asn Asn Arg Lys Met 180 185 190
Phe Gly Pro Leu Leu Asn Gly Val Asp His Leu Pro Ala Thr Tyr Asp 195 200 205
Arg Asp Lys Gin Ala Phe Thr lie Gly Glu Pro Glu Tyr Gly Ala His 210 215 220
Phe Ala Glu Ala Leu Glu Gly Leu Val Asn Leu His Gly Ala Asn Thr 225 230 235 240 lie Ala Ala Val lie Val Glu Pro Met Ala Gly Ser Thr Gly Val Leu 245 250 255
Pro Ala Pro Lys Gly Tyr Leu Lys Lys Leu Arg Glu lie Thr Lys Lys 260 265 270
His Gly lie Leu Leu lie Phe Asp Glu Val lie Thr Gly Tyr Gly Arg 275 280 285
Leu Gly Tyr Ala Phe Ala Ser Glu Arg Tyr Gly Val Thr Pro Asp Met 290 295 300 •183- 146966-序列表.doc 201037078 lie Thr Phe Ala Lys Gly Val Thr Asn Gly Ala Val Pro Met Gly Gly 305 310 315 320
Val lie Thr Ser Ala Glu lie His Asp Ala Phe Met Thr Gly Pro Glu 325 330 335
His Ala Val Glu Leu Ala His Gly Tyr Thr Tyr Ser Ala His Pro Leu 340 345 350
Ala Cys Ala Ala Gly lie Ala Thr Leu Asp lie Tyr Arg Asp Glu Lys 355 360 365
Leu Phe Glu Arg Ala Lys Ala Leu Glu Pro Lys Phe Ala Glu Ala Val 370 375 380
Met Ser Leu Lys Ser Ala Pro Asn Val Val Asp lie Arg Thr Val Gly 385 390 395 400
Leu Thr Ala Gly lie Asp Leu Ala Ser lie Ala Asp Ala Val Gly Lys 405 410 415
Arg Gly Phe Glu Ala Met Asn Ala Gly Phe His Asp His Glu Leu Met 420 425 430
Leu Arg lie Ala Gly Asp Thr Leu Ala Leu Thr Pro Pro Leu lie Leu 435 440 445
Ser Glu Asp His lie Gly Glu lie Val Asp Lys Val Gly Lys Val lie 450 455 460
Arg Ala Val Ala 465 <210> 105 <211> 1263 <212> DMA <213>大腸桿菌 <220> <221> CDS <222> (1)..(1263) <400> 105 atg cca cat tea ctg ttc age acc gat acc gat etc acc gcc gaa aat Met Pro His Ser Leu Phe Ser Thr Asp Thr Asp Leu Thr Ala Glu Asn 15 10 15 ctg ctg cgt ttg ccc get gaa ttt ggc tgc ccg gtg tgg gtc tac gat 48 96 146966·序列表.doc 184- 144201037078
Leu Leu Arg Leu Pro Ala Glu Phe Gly Cys Pro Val Trp Val Tyr Asp 20 25 30 gcg caa att att cgt egg cag att gca geg ctg aaa cag ttt gat gtg Ala Gin lie lie Arg Arg Gin lie Ala Ala Leu Lys Gin Phe Asp Val 35 40 45 gtg ege ttt gca cag aaa gcc tgt tcc aat att cat att ttg ege tta Val Arg Phe Ala Gin Lys Ala Cys Ser Asn lie His lie Leu Arg Leu 50 55 60 atg cgt gag cag ggc gtg aaa gtg gat tcc gtc teg tta ggc gaa ata Met Arg Glu Gin Gly Val Lys Val Asp Ser Val Ser Leu Gly Glu lie 65 70 75 80 192 240 gag cgt gcg ttg gcg gcg ggt tac aat ccg caa aeg cac ccc gat gat Glu Arg Ala Leu Ala Ala Gly Tyr Asn Pro Gin Thr His Pro Asp Asp 85 90 95 288
att gtt ttt aeg gca gat gtt ate gat cag gcg aeg ett gaa ege gtc lie Val Phe Thr Ala Asp Val lie Asp Gin Ala Thr Leu Glu Arg Val 100 105 110 agt gaa ttg caa att ccg gtg aat gcg ggt tet gtt gat atg etc gac Ser Glu Leu Gin lie Pro Val Asn Ala Gly Ser Val Asp Met Leu Asp 115 120 125 336 384 caa ctg ggc cag gtt teg cca ggg cat egg gta tgg ctg ege gtt aat Gin Leu Gly Gin Val Ser Pro Gly His Arg Val Trp Leu Arg Val Asn 130 135 140 432 ccg ggg ttt ggt cac gga cat age caa aaa acc aat acc ggt ggc gaa Pro Gly Phe Gly His Gly His Ser Gin Lys Thr Asn Thr Gly Gly Glu 145 150 155 160 480 aac age aag cac ggt ate tgg tac acc gat ctg ccc gcc gca ctg gac Asn Ser Lys His Gly lie Trp Tyr Thr Asp Leu Pro Ala Ala Leu Asp 165 170 175 528
gtg ata caa cgt cat cat ctg cag ctg gtc ggc att cac atg cac att Val lie Gin Arg His His Leu Gin Leu Val Gly lie His Met His lie 180 185 190 ggt tet ggc gtt gat tat gcc cat ctg gaa cag gtg tgt ggt get atg Gly Ser Gly Val Asp Tyr Ala His Leu Glu Gin Val Cys Gly Ala Met 195 200 205 576 624 gtg cgt cag gtc ate gaa ttc ggt cag gat tta cag get att tet gcg Val Arg Gin Val lie Glu Phe Gly Gin Asp Leu Gin Ala lie Ser Ala 210 215 220 672 ggc ggt ggg ett tet gtt cct tat caa cag ggt gaa gag gcg gtt gat Gly Gly Gly Leu Ser Val Pro Tyr Gin Gin Gly Glu Glu Ala Val Asp 225 230 235 240 720 acc gaa cat tat tat ggt ctg tgg aat gcc gcg cgt gag caa ate gcc Thr Glu His Tyr Tyr Gly Leu Trp Asn Ala Ala Arg Glu Gin lie Ala 245 250 255 768 ege cat ttg ggc cac cct gtg aaa ctg gaa att gaa ccg ggt ege ttc Arg His Leu Gly His Pro Val Lys Leu Glu lie Glu Pro Gly Arg Phe 146966·序列表.doc -185- 816 864201037078 260 265 270 ctg gta gcg cag tct ggc gta tta att act cag gtg egg age gtc aaa Leu Val Ala Gin Ser Gly Val Leu lie Thr Gin Val Arg Ser Val Lys 275 280 285 caa atg ggg age ege cac ttt gtg ctg gtt gat gee ggg ttc aac gat Gin Met Gly Ser Arg His Phe Val Leu Val Asp Ala Gly Phe Asn Asp 290 295 300 ctg atg ege ccg gca atg tac ggt agt tac cac cat ate agt gee ctg Leu Met Arg Pro Ala Met Tyr Gly Ser Tyr His His lie Ser Ala Leu 305 310 315 320 gca get gat ggt cgt tct ctg gaa cac gcg cca aeg gtg gaa acc gtc Ala Ala Asp Gly Arg Ser Leu Glu His Ala Pro Thr Val Glu Thr Val 325 330 335 gtc gee gga ccg tta tgt gaa teg ggc gat gtc ttt acc cag cag gaa Val Ala Gly Pro Leu Cys Glu Ser Gly Asp Val Phe Thr Gin Gin Glu 340 345 350 ggg gga aat gtt gaa acc ege gee ttg ccg gaa gtg aag gca ggt gat Gly Gly Asn Val Glu Thr Arg Ala Leu Pro Glu Val Lys Ala Gly Asp 355 360 365 tat ctg gta ctg cat gat aca ggg gca tat ggc gca tea atg tea tee Tyr Leu Val Leu His Asp Thr Gly Ala Tyr Gly Ala Ser Met Ser Ser 370 375 380 aac tac aat age cgt ccg ctg tta cca gaa gtt ctg ttt gat aat ggt Asn Tyr Asn Ser Arg Pro Leu Leu Pro Glu Val Leu Phe Asp Asn Gly 385 390 395 400 cag gcg egg ttg att ege cgt ege cag acc ate gaa gaa tta ctg gcg Gin Ala Arg Leu lie Arg Arg Arg Gin Thr lie Glu Glu Leu Leu Ala 405 410 415 ctg gaa ttg ett taa 1263 Leu Glu Leu Leu 420 912 960 1008 1056 1104 1152 1200 1248 <210> 106 <211> 420 <212> PRT <213>大腸桿菌 <400> 106 Met Pro His Ser Leu Phe Ser Thr Asp Thr Asp Leu Thr Ala Glu Asn 15 10 15
Leu Leu Arg Leu Pro Ala Glu Phe Gly Cys Pro Val Trp Val Tyr Asp 20 25 30 Ala Gin lie lie Arg Arg Gin lie Ala Ala Leu Lys Gin Phe Asp Val 35 40 45 146966-序列表.doc - 186- 201037078
Val Arg Phe Ala Gin Lys Ala Cys Ser Asn lie His lie Leu Arg Leu 50 55 60
Met Arg Glu Gin Gly Val Lys Val Asp Ser Val Ser Leu Gly Glu lie 65 70 75 80
Glu Arg Ala Leu Ala Ala Gly Tyr Asn Pro Gin Thr His Pro Asp Asp 85 90 95 lie Val Phe Thr Ala Asp Val lie Asp Gin Ala Thr Leu Glu Arg Val 100 105 110
Ser Glu Leu Gin lie Pro Val Asn Ala Gly Ser Val Asp Met Leu Asp 115 120 125
Gin Leu Gly Gin Val Ser Pro Gly His Arg Val Trp Leu Arg Val Asn 130 135 140
Pro Gly Phe Gly His Gly His Ser Gin Lys Thr Asn Thr Gly Gly Glu 145 150 155 160
Asn Ser Lys His Gly lie Trp Tyr Thr Asp Leu Pro Ala Ala Leu Asp 165 170 175
Val lie Gin Arg His His Leu Gin Leu Val Gly lie His Met His lie 180 185 190
Gly Ser Gly Val Asp Tyr Ala His Leu Glu Gin Val Cys Gly Ala Met 195 200 205
Val Arg Gin Val lie Glu Phe Gly Gin Asp Leu Gin Ala lie Ser Ala 210 215 220
Gly Gly Gly Leu Ser Val Pro Tyr Gin Gin Gly Glu Glu Ala Val Asp 225 230 235 240
Thr Glu His Tyr Tyr Gly Leu Trp Asn Ala Ala Arg Glu Gin lie Ala 245 250 255
Arg His Leu Gly His Pro Val Lys Leu Glu lie Glu Pro Gly Arg Phe 260 265 270
Leu Val Ala Gin Ser Gly Val Leu lie Thr Gin Val Arg Ser Val Lys 275 280 285 •187· 146966-序列表.doc 201037078
Gin Met Gly Ser Arg His Phe Val Leu Val Asp Ala Gly Phe Asn Asp .290 295 300
Leu Met Arg Pro Ala Met Tyr Gly Ser Tyr His His lie Ser Ala Leu 305 310 315 320
Ala Ala Asp Gly Arg Ser Leu Glu His Ala Pro Thr Val Glu Thr Val 325 330 335
Val Ala Gly Pro Leu Cys Glu Ser Gly Asp Val Phe Thr Gin Gin Glu 340 345 350
Gly Gly Asn Val Glu Thr Arg Ala Leu Pro Glu Val Lys Ala Gly Asp 355 360 365
Tyr Leu Val Leu His Asp Thr Gly Ala Tyr Gly Ala Ser Met Ser Ser 370 375 380
Asn Tyr Asn Ser Arg Pro Leu Leu Pro Glu Val Leu Phe Asp Asn Gly 385 390 395 400
Gin Ala Arg Leu lie Arg Arg Arg Gin Thr lie Glu Glu Leu Leu Ala 405 410 415
Leu Glu Leu Leu 420 <210> 107 <211> 1265 <212> DNA <213> 人工 <220> <223>大腸桿菌二胺基庚二酸脫羧酶LysA密碼子優化基因 <400> 107 atatgccaca ctctctgttt tctactgata ctgatctgac tgcggaaaac ctgctgcgtc tgccggctga attcggttgt ccggtatggg tgtacgacgc tcagattatt cgtcgccaga tcgcagcact gaagcagttc gatgtagtgc gttttgcaca gaaggcgtgc tccaacatcc atatcctgcg cctgatgcgt gagcagggcg ttaaagttga ctccgtctct ctgggtgaga ttgagcgcgc cctggcagcc ggctataacc cacagaccca tcctgacgac attgtattta ctgccgacgt gatcgaccag gctactctgg aacgcgtttc tgaactgcag atcccggtta atgctggttc tgtggacatg ctggaccagc tgggccaggt atccccaggt catcgtgtgt ggctgcgtgt caacccaggt ttcggccacg gccactctca gaaaactaac actggtggtg •188- 60 120 180 240 300 360 420 146966-序列表.doc 480 540201037078 agaactccaa gcatggcatt tggtataccg atctgccggc tgcactggac gtaatccagc gtcaccacct gcagctggtg ggcatccaca tgcacattgg ctccggcgta gactacgccc acctggagca agtctgcggt gctatggtac gtcaggtaat cgagttcggc caagatctgc aggcaatcag cgctggtggc ggcctgtctg taccttatca gcagggcgag gaggcggttg acactgagca ctactacggt ctgtggaacg ccgctcgtga gcaaattgca cgtcacctgg gccacccggt gaaactggag atcgagccgg gccgcttcct ggtagcacag tccggcgtac tgattaccca ggtacgctct gttaaacaga tgggctcccg tcactttgtg ctggtagacg caggcttcaa cgacctgatg cgtccggcta tgtatggttc ctatcatcac atctctgcgc tggccgccga cggccgctct ctggaacacg cgccgacggt tgaaacggtg gtggctggtc o cgctgtgcga gtccggcgac gttttcactc agcaggaggg cggcaatgta gagacgcgtg cgctgccgga agtgaaagcc ggtgattatc tggtgctgca tgataccggc gcctatggtg cgagcatgag cagcaactac aactctcgcc cgctgctgcc ggaggtcctg ttcgataacg gccaagcccg cctgatccgt cgtcgtcaga ccatcgagga actgctggca ctggagctgc tgtaa 1265 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 <210> 108 <211> 1692 <212> DNA <213〉釀酒酵母
<220> <221> CDS <222> (1) . . (1692) <400> 108 atg tct gaa att act ttg ggt aaa tat ttg ttc gaa aga tta aag caa Met Ser Glu lie Thr Leu Gly Lys Tyr Leu Phe Glu Arg Leu Lys Gin 15 10 15 gtc aac gtt aac acc gtt ttc ggt ttg cca ggt gac ttc aac ttg tcc Val Asn Val Asn Thr Val Phe Gly Leu Pro Gly Asp Phe Asn Leu Ser 20 25 30 ttg ttg gac aag ate tac gaa gtt gaa ggt atg aga tgg get ggt aac Leu Leu Asp Lys lie Tyr Glu Val Glu Gly Met Arg Trp Ala Gly Asn 35 40 45 gcc aac gaa ttg aac get get tac gcc get gat ggt tac get cgt ate Ala Asn Glu Leu Asn Ala Ala Tyr Ala Ala Asp Gly Tyr Ala Arg lie 50 55 60 aag ggt atg tct tgt ate ate acc acc ttc ggt gtc ggt gaa ttg tct Lys Gly Met Ser Cys lie 工le Thr Thr Phe Gly Val Gly Glu Leu Ser 65 70 75 80 48 96 144 192 146966-序列表.doc -189- 240 201037078 get ttg aac ggt att gee ggt tet tac get gaa cac gtc ggt gtt ttg 288
Ala Leu Asn Gly lie Ala Gly Ser Tyr Ala Glu His Val Gly Val Leu 85 90 95 cac gtt gtt ggt gtc cca tee ate tet get caa get aag caa ttg ttg 336
His Val Val Gly Val Pro Ser lie Ser Ala Gin Ala Lys Gin Leu Leu 100 105 110 ttg cac cac acc ttg ggt aac ggt gac ttc act gtt ttc cac aga atg 384
Leu His His Thr Leu Gly Asn Gly Asp Phe Thr Val Phe His Arg Met 115 120 125 tet gee aac att tet gaa acc act get atg ate act gac att get acc 432
Ser Ala Asn lie Ser Glu Thr Thr Ala Met lie Thr Asp lie Ala Thr 130 135 140 gee cca get gaa att gac aga tgt ate aga acc act tac gtc acc caa 480
Ala Pro Ala Glu lie Asp Arg Cys lie Arg Thr Thr Tyr Val Thr Gin 145 150 155 160 aga cca gtc tac tta ggt ttg cca get aac ttg gtc gac ttg aac gtc 528
Arg Pro Val Tyr Leu Gly Leu Pro Ala Asn Leu Val Asp Leu Asn Val 165 170 175 cca get aag ttg ttg caa act cca att gac atg tet ttg aag cca aac 576
Pro Ala Lys Leu Leu Gin Thr Pro lie Asp Met Ser Leu Lys Pro Asn 180 185 190 gat get gaa tee gaa aag gaa gtc att gac acc ate ttg get ttg gtc 624
Asp Ala Glu Ser Glu Lys Glu Val lie Asp Thr lie Leu Ala Leu Val 195 200 205 aag gat get aag aac cca gtt ate ttg get gat get tgt tgt tee aga 672
Lys Asp Ala Lys Asn Pro Val lie Leu Ala Asp Ala Cys Cys Ser Arg 210 215 220 cac gac gtc aag get gaa act aag aag ttg att gac ttg act caa ttc 720
His Asp Val Lys Ala Glu Thr Lys Lys Leu lie Asp Leu Thr Gin Phe 225 230 235 240 cca get ttc gtc acc cca atg ggt aag ggt tee att gac gaa caa cac 768
Pro Ala Phe Val Thr Pro Met Gly Lys Gly Ser lie Asp Glu Gin His 245 250 255 cca aga tac ggt ggt gtt tac gtc ggt acc ttg tee aag cca gaa gtt 816
Pro Arg Tyr Gly Gly Val Tyr Val Gly Thr Leu Ser Lys Pro Glu Val 260 265 270 aag gaa gee gtt gaa tet get gac ttg att ttg tet gtc ggt get ttg 864
Lys Glu Ala Val Glu Ser Ala Asp Leu lie Leu Ser Val Gly Ala Leu 275 280 285 ttg tet gat ttc aac acc ggt tet ttc tet tac tet tac aag acc aag 912
Leu Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr Ser Tyr Lys Thr Lys 290 295 300 aac att gtc gaa ttc cac tee gac cac atg aag ate aga aac gee act 960
Asn lie Val Glu Phe His Ser Asp His Met Lys lie Arg Asn Ala Thr 305 310 315 320 ttc cca ggt gtc caa atg aaa ttc gtt ttg caa aag ttg ttg acc act 1008 -190· 146966-序列表.doe 1056 201037078
Phe Pro Gly Val Gin Met Lys Phe Val Leu Gin Lys Leu Leu Thr Thr 325 330 335 att get gac gcc get aag ggt tac aag cca gtt get gtc cca get aga lie Ala Asp Ala Ala Lys Gly Tyr Lys Pro Val Ala Val Pro Ala Arg 340 345 350 act cca get aac get get gtc cca get tet acc cca ttg aag caa gaa
Thr Pro Ala Asn Ala Ala Val Pro Ala Ser Thr Pro Leu Lys Gin Glu 355 360 365 tgg atg tgg aac caa ttg ggt aac ttc ttg caa gaa ggt gat gtt gtc
Trp Met Trp Asn Gin Leu Gly Asn Phe Leu Gin Glu Gly Asp Val Val 370 375 380 att get gaa acc ggt acc tee get ttc ggt ate aac caa acc act ttc lie Ala Glu Thr Gly Thr Ser Ala Phe Gly lie Asn Gin Thr Thr Phe 385 390 395 400 cca aac aac acc tac ggt ate tet caa gtc tta tgg ggt tee att ggt
Pro Asn Asn Thr Tyr Gly lie Ser Gin Val Leu Trp Gly Ser lie Gly 405 410 415 ttc acc act ggt get acc ttg ggt get get ttc get get gaa gaa att
Phe Thr Thr Gly Ala Thr Leu Gly Ala Ala Phe Ala Ala Glu Glu lie 420 425 430 gat cca aag aag aga gtt ate tta ttc att ggt gac ggt tet ttg caa
Asp Pro Lys Lys Arg Val lie Leu Phe lie Gly Asp Gly Ser Leu Gin 435 440 445 ttg act gtt caa gaa ate tee acc atg ate aga tgg ggc ttg aag cca
Leu Thr Val Gin Glu lie Ser Thr Met lie Arg Trp Gly Leu Lys Pro 450 455 460 tac ttg ttc gtc ttg aac aac gat ggt tac acc att gaa aag ttg att Tyr Leu Phe Val Leu Asn Asn Asp Gly Tyr Thr lie Glu Lys Leu lie 465 470 475 480 cac ggt cca aag get caa tac aac gaa att caa ggt tgg gac cac eta
His Gly Pro Lys Ala Gin Tyr Asn Glu lie Gin Gly Trp Asp His Leu 485 490 495 tee ttg ttg cca act ttc ggt get aag gac tat gaa acc cac aga gtc
Ser Leu Leu Pro Thr Phe Gly Ala Lys Asp Tyr Glu Thr His Arg Val 500 505 510 get acc acc ggt gaa tgg gac aag ttg acc caa gac aag tet ttc aac Ala Thr Thr Gly Glu Trp Asp Lys Leu Thr Gin Asp Lys Ser Phe Asn 515 520 525 gac aac tet aag ate aga atg att gaa ate atg ttg cca gtc ttc gat Asp Asn Ser Lys 工le Arg Met lie Glu lie Met Leu Pro Val Phe Asp 530 535 540 get cca caa aac ttg gtt gaa caa get aag ttg act get get acc aac Ala Pro Gin Asn Leu Val Glu Gin Ala Lys Leu Thr Ala Ala Thr Asn 545 550 555 560 get aag caa taa 1692
Ala Lys Gin 146966·序列表.doc 1104 1152 1200 1248
1296 1344 1392 1440 1488 1536 1584 1632 1680 -191 - 201037078 <210> 109 <211> 563 <212> PRT <213>釀酒酵母 <400> 109
Met Ser Glu lie Thr Leu Gly Lys Tyr Leu Phe Glu Arg Leu Lys Gin 15 10 15
Val Asn Val Asn Thr Val Phe Gly Leu Pro Gly Asp Phe Asn Leu Ser 20 25 30
Leu Leu Asp Lys lie Tyr Glu Val Glu Gly Met Arg Trp Ala Gly Asn 35 40 45
Ala Asn Glu Leu Asn Ala Ala Tyr Ala Ala Asp Gly Tyr Ala Arg lie 50 55 60
Lys Gly Met Ser Cys lie lie Thr Thr Phe Gly Val Gly Glu Leu Ser 65 70 75 80
Ala Leu Asn Gly lie Ala Gly Ser Tyr Ala Glu His Val Gly Val Leu 85 90 95
His Val Val Gly Val Pro Ser lie Ser Ala Gin Ala Lys Gin Leu Leu 100 105 110
Leu His His Thr Leu Gly Asn Gly Asp Phe Thr Val Phe His Arg Met 115 120 125
Ser Ala Asn lie Ser Glu Thr Thr Ala Met lie Thr Asp lie Ala Thr 130 135 140
Ala Pro Ala Glu lie Asp Arg Cys 工le Arg Thr Thr Tyr Val Thr Gin 145 150 155 160
Arg Pro Val Tyr Leu Gly Leu Pro Ala Asn Leu Val Asp Leu Asn Val 165 170 175
Pro Ala Lys Leu Leu Gin Thr Pro lie Asp Met Ser Leu Lys Pro Asn 180 185 190
Asp Ala Glu Ser Glu Lys Glu Val lie Asp Thr lie Leu Ala Leu Val 195 200 205 192- 146966-序列表.doc 201037078
Lys Asp Ala Lys Asn Pro Val lie Leu Ala Asp Ala Cys Cys Ser Arg 210 215 220
His Asp Val Lys Ala Glu Thr Lys Lys Leu lie Asp Leu Thr Gin Phe 225 230 235 240
Pro Ala Phe Val Thr Pro Met Gly Lys Gly Ser lie Asp Glu Gin His 245 250 255,
Pro Arg Tyr Gly Gly Val Tyr Val Gly Thr Leu Ser Lys Pro Glu Val 260 265 270
Lys Glu Ala Val Glu Ser Ala Asp Leu lie Leu Ser Val Gly Ala Leu 275 280 285
o
Leu Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr Ser Tyr Lys Thr Lys 290 295 300
Asn lie Val Glu Phe His Ser Asp His Net Lys lie Arg Asn Ala Thr 305 310 315 320
Phe Pro Gly Val Gin Met Lys Phe Val Leu Gin Lys Leu Leu Thr Thr 325 330 335 lie Ala Asp Ala Ala Lys Gly Tyr Lys Pro Val Ala Val Pro Ala Arg 340 345 350
Thr Pro Ala Asn Ala Ala Val Pro Ala Ser Thr Pro Leu Lys Gin Glu 355 360 365
Trp Met Trp Asn Gin Leu Gly Asn Phe Leu Gin Glu Gly Asp Val Val 370 375 380 lie Ala Glu Thr Gly Thr Ser Ala Phe Gly lie Asn Gin Thr Thr Phe 385 390 395 400
Pro Asn Asn Thr Tyr Gly lie Ser Gin Val Leu Trp Gly Ser lie Gly 405 410 415
Phe Thr Thr Gly Ala Thr Leu Gly Ala Ala Phe Ala Ala Glu Glu lie 420 425 430
Asp Pro Lys Lys Arg Val lie Leu Phe lie Gly Asp Gly Ser Leu Gin 435 440 445 -193· 146966·序列表.doc 201037078
Leu Thr Val Gin Glu lie Ser Thr Met lie Arg Trp Gly Leu Lys Pro 450 455 460
Tyr Leu Phe Val Leu Asn Asn Asp Gly Tyr Thr lie Glu Lys Leu lie 465 470 475 480
His Gly Pro Lys Ala Gin Tyr Asn Glu lie Gin Gly Trp Asp His Leu 485 490 495
Ser Leu Leu Pro Thr Phe Gly Ala Lys Asp Tyr Glu Thr His Arg Val 500 505 510
Ala Thr Thr Gly Glu Trp Asp Lys Leu Thr Gin Asp Lys Ser Phe Asn 515 520 525
Asp Asn Ser Lys lie Arg Met lie Glu lie Met Leu Pro Val Phe Asp 530 535 540
Ala Pro Gin Asn Leu Val Glu Gin Ala Lys Leu Thr Ala Ala Thr Asn 545 550 555 560
Ala Lys Gin <210> 110 <211> 1692 <212> DNA <213> 人工 <220> <223> 釀酒酵母丙酮酸脫羧酶Pdc密碼子優化基因 <400> 110 atgtccgaga tcactctggg caaatacctg tttgaacgtc tgaaacaggt gaacgttaat accgtattcg gcctgccggg tgatttcaac ctgtccctgc tggacaaaat ctatgaagtt gaaggtatgc gttgggctgg caacgctaac gagctgaacg cagcgtacgc ggcagatggt tacgctcgta tcaaaggtat gtcttgtatc atcaccacct tcggtgttgg tgagctgagc gccctgaacg gcatcgccgg ctcctatgca gagcacgtgg gcgtgctgca cgttgtgggt gtaccgtcca tcagcgccca ggcaaaacag ctgctgctgc accacaccct gggtaacggc gactttaccg ttttccatcg tatgtctgcg aacatcagcg aaactactgc aatgattact gacatcgcta cggcaccggc agaaatcgac cgttgcattc gtaccacgta cgttactcag cgcccggttt atctgggcct gccagccaac ctggtggatc tgaacgtccc ggctaaactg ctgcagactc cgatcgatat gtctctgaaa cctaacgacg cagaatctga gaaagaagtt -194- 60 120 180 240 300 360 420 480 540 146966-序列表.doc 600 660 201037078
atcgatacta ttctggctct tgttgctctc gtcatgatgt ccggcgttcg ttaccccgat ggtgtatacg ttggcacgct ctgatcctgt ctgttggtgc tacaagacca aaaacattgt tttcctggtg tgcagatgaa gcgaaaggtt ataaaccggt gcatccactc cactgaagca ggcgacgttg taatcgcaga ccaaacaaca cctacggtat gcaaccctgg gtgcggcttt ttcatcggtg acggctccct ggcctgaaac cgtacctgtt cacggtccga aagcacagta acctttggcg ctaaagacta ctgacgcagg ataaatcctt ccggtctttg atgctccgca gctaaacagt aa <210> 111 <211> 1707 <212> DNA <213>運動醱酵單胞菌 ggtgaaagat gcaaagaacc aaaggcagaa accaaaaagc gggcaagggt tccatcgatg gtccaaaccg gaggtaaaag actgctgagc gacttcaaca ggagtttcac tccgatcaca attcgtactg cagaaactgc agctgtgccg gcacgtaccc ggaatggatg tggaatcagc aaccggcact agcgcgtttg cagccaagtc ctgtggggct cgctgctgag gagatcgacc gcagctgacc gtccaggaga tgtgctgaac aacgacggct taatgagatc cagggttggg cgagacccac cgcgtggcta caatgacaat agcaagattc gaacctggta gagcaagcaa cagttatcct ggctgacgca tgatcgacct gacgcagttc agcagcaccc gcgttatggt aagcggttga aagcgcagat ccggttcttt ctcctatagc tgaaaatccg caacgcgacc tgaccaccat cgccgacgct cggcgaacgc cgcggttcct tgggtaattt cctgcaagaa gcattaacca gacgaccttc ctatcggctt caccaccggt cgaagaaacg tgttatcctg tttctaccat gatccgctgg acactattga gaaactgatc atcatctgtc tctgctgccg ccaccggcga gtgggataaa gtatgatcga aatcatgctg aactgaccgc ggcaactaac 1692 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 <220> <221> CDS <222> (1)..(1707) <400> 111 atg agt tat act gtc ggt acc tat tta gcg gag egg ett gtc cag att Met Ser Tyr Thr Val Gly Thr Tyr Leu Ala Glu Arg Leu Val Gin lie 15 10 15 48 ggt etc aag cat cac ttc gca gtc gcg ggc gac tac aac etc gtc ett Gly Leu Lys His His Phe Ala Val Ala Gly Asp Tyr Asn Leu Val Leu 20 25 30 96 ett gac aac ctg ett ttg aac aaa aac atg gag cag gtt tat tgc tgt Leu Asp Asn Leu Leu Leu Asn Lys Asn Met Glu Gin Val Tyr Cys Cys 146966-序列表.doc •195- 144 201037078 35 40 45 aac gaa ctg aac tgc ggt ttc agt gca gaa ggt tat get cgt gee aaa 192
Asn Glu Leu Asn Cys Gly Phe Ser Ala Glu Gly Tyr Ala Arg Ala Lys 50 55 60 ggc gca gca gca gee gtc gtt acc tac age gtc ggt geg ett tee gca 240
Gly Ala Ala Ala Ala Val Val Thr Tyr Ser Val Gly Ala Leu Ser Ala 65 70 75 80 ttt gat get ate ggt ggc gee tat gca gaa aac ett ccg gtt ate ctg 288
Phe Asp Ala lie Gly Gly Ala Tyr Ala Glu Asn Leu Pro Val lie Leu 85 90 95 ate tee ggt get ccg aac aac aat gat cac get get ggt cac gtg ttg 336 lie Ser Gly Ala Pro Asn Asn Asn Asp His Ala Ala Gly His Val Leu 100 105 110 cat cac get ett ggc aaa acc gac tat cac tat cag ttg gaa atg gee 384
His His Ala Leu Gly Lys Thr Asp Tyr His Tyr Gin Leu Glu Met Ala 115 120 125 aag aac ate aeg gee gee get gaa geg att tac acc ccg gaa gaa get 432
Lys Asn lie Thr Ala Ala Ala Glu Ala lie Tyr Thr Pro Glu Glu Ala 130 135 140 ccg get aaa ate gat cac gtg att aaa act get ett cgt gag aag aag 480
Pro Ala Lys lie Asp His Val lie Lys Thr Ala Leu Arg Glu Lys Lys 145 150 155 160 ccg gtt tat etc gaa ate get tgc aac att get tee atg ccc tgc gee 528
Pro Val Tyr Leu Glu lie Ala Cys Asn lie Ala Ser Met Pro Cys Ala 165 170 175 get cct gga ccg gca age gca ttg ttc aat gac gaa gee age gac gaa 576
Ala Pro Gly Pro Ala Ser Ala Leu Phe Asn Asp Glu Ala Ser Asp Glu 180 185 190 get tet ttg aat gca geg gtt gaa gaa acc ctg aaa ttc ate gee aac 624
Ala Ser Leu Asn Ala Ala Val Glu Glu Thr Leu Lys Phe lie Ala Asn 195 200 205 ege gac aaa gtt gee gtc etc gtc ggc age aag ctg ege gca get ggt 672
Arg Asp Lys Val Ala Val Leu Val Gly Ser Lys Leu Arg Ala Ala Gly 210 215 220 get gaa gaa get get gtc aaa ttt get gat get etc ggt ggc gca gtt 720
Ala Glu Glu Ala Ala Val Lys Phe Ala Asp Ala Leu Gly Gly Ala Val 225 230 235 240 get acc atg get get gca aaa age ttc ttc cca gaa gaa aac ccg cat 768
Ala Thr Met Ala Ala Ala Lys Ser Phe Phe Pro Glu Glu Asn Pro His 245 250 255 tac ate ggc acc tea tgg ggt gaa gtc age tat ccg ggc gtt gaa aag 816
Tyr lie Gly Thr Ser Trp Gly Glu Val Ser Tyr Pro Gly Val Glu Lys 260 265 270 aeg atg aaa gaa gee gat geg gtt ate get ctg get cct gtc ttc aac 864
Thr Met Lys Glu Ala Asp Ala Val lie Ala Leu Ala Pro Val Phe Asn 275 280 285 •196- 146966-序列表.doc 912 201037078 gac tac tcc acc act ggt tgg acg gat att cct gat cct aag aaa ctg Asp Tyr Ser Thr Thr Gly Trp Thr Asp lie Pro Asp Pro Lys Lys Leu 290 295 300 gtt etc get gaa ccg cgt tet gtc gtc gtt aac ggc att ege ttc ccc Val Leu Ala Glu Pro Arg Ser Val Val Val Asn Gly lie Arg Phe Pro 305 310 315 320 age gtc cat ctg aaa gac tat ctg acc cgt ttg get cag aaa gtt tec Ser Val His Leu Lys Asp Tyr Leu Thr Arg Leu Ala Gin Lys Val Ser 325 330 335 aag aaa acc ggt gca ttg gac ttc ttc aaa tec etc aat gca ggt gaa Lys Lys Thr Gly Ala Leu Asp Phe Phe Lys Ser Leu Asn Ala Gly Glu 340 345 350 960 1008 1056 1104 ctg aag aaa gee get ccg get gat ccg agt get ccg ttg gtc aac gca Leu Lys Lys Ala Ala Pro Ala Asp Pro Ser Ala Pro Leu Val Asn Ala 355 360 365
1152 gaa ate gee cgt cag gtc gaa get ett ctg acc ccg aac acg acg gtt Glu lie Ala Arg Gin Val Glu Ala Leu Leu Thr Pro Asn Thr Thr Val 370 375 380
att get gaa acc ggt gac tet tgg ttc aat get cag ege atg aag etc lie Ala Glu Thr Gly Asp Ser Trp Phe Asn Ala Gin Arg Met Lys Leu 385 390 395 400 ccg aac ggt get ege gtt gaa tat gaa atg cag tgg ggt cac att ggt Pro Asn Gly Ala Arg Val Glu Tyr Glu Met Gin Trp Gly His lie Gly 405 410 415 tgg tec gtt cct gee gee ttc ggt tat gee gtc ggt get ccg gaa cgt Trp Ser Val Pro Ala Ala Phe Gly Tyr Ala Val Gly Ala Pro Glu Arg 420 425 430 ege aac ate etc atg gtt ggt gat ggt tec ttc cag ctg acg get cag Arg Asn lie Leu Met Val Gly Asp Gly Ser Phe Gin Leu Thr Ala Gin 435 440 445 1200 1248 1296 1344 gaa gtc get cag atg gtt ege ctg aaa ctg ccg gtt ate ate ttc ttg Glu Val Ala Gin Met Val Arg Leu Lys Leu Pro Val lie lie Phe Leu 450 455 460 1392 ate aat aac tat ggt tac acc gee gaa gtt atg ate cat gat ggt ccg lie Asn Asn Tyr Gly Tyr Thr Ala Glu Val Met lie His Asp Gly Pro 465 470 475 480 1440 tac aac aac ate aag aac tgg gat tat gee ggt ctg atg gaa gtg ttc Tyr Asn Asn lie Lys Asn Trp Asp Tyr Ala Gly Leu Met Glu Val Phe 485 490 495 aac ggt aac ggt ggt tat gac age ggt get ggt aaa ggc ctg aag get Asn Gly Asn Gly Gly Tyr Asp Ser Gly TVla Gly Lys Gly Leu Lys Ala 500 505 510 1488 1536 aaa acc ggt ggc gaa ctg gca gaa get ate aag gtt get ctg gca aac Lys Thr Gly Gly Glu Leu Ala Glu Ala lie Lys Val Ala Leu Ala Asn 515 520 525 1584 146966-序列表.doc -197- 201037078 acc gac ggc cca acc ctg ate gaa tgc ttc ate ggt cgt gaa gac tgc 1632 Thr Asp Gly Pro Thr Leu lie Glu Cys Phe lie Gly Arg Glu Asp Cys 530 535 540 act gaa gaa ttg gtc aaa tgg ggt aag ege gtt get gcc gcc aac age 1680 Thr Glu Glu Leu Val Lys Trp Gly Lys Arg Val Ala Ala Ala Asn Ser 545 550 555 560 cgt aag cct gtt aac aag etc etc tag 1707 Arg Lys Pro Val Asn Lys Leu Leu 565 <210> 112 <211> 568 <212> PRT <213>運動醱酵單胞菌 <400> 112
Met Ser Tyr Thr Val Gly Thr Tyr Leu Ala Glu Arg Leu Val Gin lie 1 5 10 15
Gly Leu Lys His His Phe Ala Val Ala Gly Asp Tyr Asn Leu Val Leu 20 25 30
Leu Asp Asn Leu Leu Leu Asn Lys Asn Met Glu Gin Val Tyr Cys Cys 35 40 45
Asn Glu Leu Asn Cys Gly Phe Ser Ala Glu Gly Tyr Ala Arg Ala Lys 50 55 6 0
Gly Ala Ala Ala Ala Val Val Thr Tyr Ser Val Gly Ala Leu Ser Ala 65 70 75 80
Phe Asp Ala lie Gly Gly Ala Tyr Ala Glu Asn Leu Pro Val lie Leu 85 90 95 lie Ser Gly Ala Pro Asn Asn Asn Asp His Ala Ala Gly His Val Leu 100 105 110
His His Ala Leu Gly Lys Thr Asp Tyr His Tyr Gin Leu Glu Met Ala 115 120 125
Lys Asn lie Thr Ala Ala Ala Glu Ala lie Tyr Thr Pro Glu Glu Ala 130 135 140
Pro Ala Lys 工le Asp His Val lie Lys Thr Ala Leu Arg Glu Lys Lys 145 150 155 160
Pro Val Tyr Leu Glu lie Ala Cys Asn lie Ala Ser Met Pro Cys Ala •198· 146966-序列表.doc 201037078 165 170 175 Ala Pro Gly Pro Ala Ser Ala Leu Phe Asn Asp Glu Ala Ser Asp Glu 180 185 190
Ala Ser Leu Asn Ala Ala Val Glu Glu Thr Leu Lys Phe lie Ala Asn 195 200 205
Arg Asp Lys Val Ala Val Leu Val Gly Ser Lys Leu Arg Ala Ala Gly 210 215 220
Ala Glu Glu Ala Ala Val Lys Phe Ala Asp Ala Leu Gly Gly Ala Val 225 230 235 240
Ala Thr Met Ala Ala Ala Lys Ser Phe Phe Pro Glu Glu Asn Pro His 245 250 255 ❹
Tyr lie Gly Thr Ser Trp Gly Glu Val Ser Tyr Pro Gly Val Glu Lys 260 265 270
Thr Met Lys Glu Ala Asp Ala Val lie Ala Leu Ala Pro Val Phe Asn 275 280 285
Asp Tyr Ser Thr Thr Gly Trp Thr Asp lie Pro Asp Pro Lys Lys Leu 290 295 300
Val Leu Ala Glu Pro Arg Ser Val Val Val Asn Gly lie Arg Phe Pro 305 310 315 320
Ser Val His Leu Lys Asp Tyr Leu Thr Arg Leu Ala Gin Lys Val Ser 325 330 335 o
Lys Lys Thr Gly Ala Leu Asp Phe Phe Lys Ser Leu Asn Ala Gly Glu 340 345 350
Leu Lys Lys Ala Ala Pro Ala Asp Pro Ser Ala Pro Leu Val Asn Ala 355 360 365
Glu lie Ala Arg Gin Val Glu Ala Leu Leu Thr Pro Asn Thr Thr Val 370 375 380 lie Ala Glu Thr Gly Asp Ser Trp Phe Asn Ala Gin Arg Met Lys Leu 385 390 395 400
Pro Asn Gly Ala Arg Val Glu Tyr Glu Met Gin Trp Gly His lie Gly 405 410 415 •199 146966-序列表.doc 201037078
Trp Ser Val Pro Ala Ala Phe Gly Tyr Ala Val Gly Ala Pro Glu Arg 420 425 430
Arg Asn lie Leu Met Val Gly Asp Gly Ser Phe Gin Leu Thr Ala Gin 435 440 445
Glu Val Ala Gin Met Val Arg Leu Lys Leu Pro Val lie lie Phe Leu 450 455 460 lie Asn Asn Tyr Gly Tyr Thr Ala Glu Val Met lie His Asp Gly Pro 465 470 475 480
Tyr Asn Asn lie Lys Asn Trp Asp Tyr Ala Gly Leu Met Glu Val Phe 485 490 495
Asn Gly Asn Gly Gly Tyr Asp Ser Gly Ala Gly Lys Gly Leu Lys Ala 500 505 510
Lys Thr Gly Gly Glu Leu Ala Glu Ala lie Lys Val Ala Leu Ala Asn 515 520 525
Thr Asp Gly Pro Thr Leu lie Glu Cys Phe lie Gly Arg Glu Asp Cys 530 535 540
Thr Glu Glu Leu Val Lys Trp Gly Lys Arg Val Ala Ala Ala Asn Ser 545 550 555 560
Arg Lys Pro Val Asn Lys Leu Leu 565 <210> 113 <211> 1707 <212> DNA <213> 人工 <220> <223>運動醱酵單胞菌丙酮酸脫羧酶PdcI472A密碼子優化基因 <400> 113 atgtcttata ctgttggtac ttatctggct gagcgtctgg tgcaaatcgg cctgaaacac cactttgcag ttgctggcga ctacaacctg gttctgctgg ataacctgct gctgaacaaa aacatggagc aagtttattg ctgtaacgag ctgaactgcg gcttctctgc ggagggttat gcgcgtgcga aaggtgccgc tgcagcagtc gtaacctact ctgtgggcgc tctgtccgcg ttcgacgcaa tcggtggcgc ttacgctgaa aacctgccgg tgatcctgat tagcggtgcg 200- 60 120 180 240 146966-序列表.doc 300 201037078
ccgaataata acgaccatgc tgctggccac gttctgcacc acgccctggg taaaactgat 360 taccattacc agctggagat ggctaaaaac atcactgcag cagcagaagc gatctacacc 420 ccggaagagg ctccggcaaa aatcgaccac gtgattaaaa ccgctctgcg tgagaaaaag 480 ccggtatacc tggaaatcgc gtgcaacatc gcgtctatgc cgtgcgccgc accgggtccg 540 gcttctgccc tgttcaacga tgaggcgagc gatgaggcat ctctgaacgc agcagtagaa 600 gaaaccctga aatttatcgc aaaccgtgac aaagtagcag tcctggtagg ttctaaactg 660 cgtgcggctg gtgcggaaga ggctgcggta aagttcgcgg atgctctggg cggtgcagtg 720 gcgaccatgg cagcggctaa atccttcttc ccagaggaga acccgcatta cattggtacc 780 tcctggggcg aagtttccta ccctggtgtg gagaaaacca tgaaagaagc cgatgctgtg 840 attgccctgg cgcctgtatt caacgattat tccaccaccg gttggaccga tatcccggac 900 ccgaagaaac tggtcctggc tgaaccgcgc tccgtagtag tgaatggcat tcgtttcccg 960 tccgtacacc tgaaggatta cctgacgcgt ctggcacaga aagtatccaa gaaaactggc 1020 gcgctggact tctttaaatc cctgaacgct ggtgagctga aaaaggcggc tccggccgat 1080 ccgtccgcac cgctggtgaa cgcagagatt gcacgtcagg ttgaggcact gctgacgccg 1140 aacaccaccg taatcgcgga aacgggcgac tcttggttca acgcacagcg catgaaactg 1200 ccgaacggtg cccgcgttga atatgaaatg cagtggggtc acatcggctg gtctgtccca 1260 gcagcgtttg gttacgcggt tggtgcaccg gagcgtcgca acatcctgat ggtgggtgac 1320 ggctccttcc agctgactgc tcaggaggtg gcgcagatgg tgcgcctgaa gctgccggtt 1380 atcattttcc tgatcaacaa ctacggctac accgccgagg taatgatcca cgatggtccg 1440 tacaacaaca tcaaaaactg ggactacgcc ggtctgatgg aggtttttaa cggtaacggc 1500 ggttacgaca gcggtgctgg taagggtctg aaagccaaaa ccggtggcga actggcagag 1560 gcgattaaag ttgcgctggc aaacaccgat ggcccgaccc tgatcgagtg cttcatcggc 1620 cgtgaggact gcaccgagga gctggtcaaa tggggcaaac gtgtggcggc tgctaactct 1680 cgcaagccgg taaacaaact gctgtaa 1707 <210> 114 <211> 1644 <212> DNA <213> 乳酸乳球菌 <220> <221> CDS <222> (1) _·(1644) <400> 114 -201 - 146966-序列表.doc 201037078 atg tat aca gta gga gat tac ctg tta gac cga tta cac gag ttg gga 48
Met Tyr Thr Val Gly Asp Tyr Leu Leu Asp Arg Leu His Glu Leu Gly 15 10 15 att gaa gaa att ttt gga gtt cct ggt gac tat aac tta caa ttt tta 96 lie Glu Glu lie Phe Gly Val Pro Gly Asp Tyr Asn Leu Gin Phe Leu 20 25 30 gat caa att att tea ege gaa gat atg aaa tgg att gga aat get aat 144
Asp Gin lie lie Ser Arg Glu Asp Met Lys Trp lie Gly Asn Ala Asn 35 40 45 gaa tta aat get tet tat atg get gat ggt tat get cgt act aaa aaa 192
Glu Leu Asn Ala Ser Tyr Met Ala Asp Gly Tyr Ala Arg Thr Lys Lys 50 55 60 get gcc gca ttt etc acc aca ttt gga gtc ggc gaa ttg agt geg ate 240
Ala Ala Ala Phe Leu Thr Thr Phe Gly Val Gly Glu Leu Ser Ala lie 65 70 75 80 aat gga ctg gca gga agt tat gcc gaa aat tta cca gta gta gaa att 288
Asn Gly Leu Ala Gly Ser Tyr Ala Glu Asn Leu Pro Val Val Glu lie 85 90 95 gtt ggt tea cca act tea aaa gta caa aat gac gga aaa ttt gtc cat 336
Val Gly Ser Pro Thr Ser Lys Val Gin Asn Asp Gly Lys Phe Val His 100 105 110 cat aca eta gca gat ggt gat ttt aaa cac ttt atg aag atg cat gaa 384
His Thr Leu Ala Asp Gly Asp Phe Lys His Phe Met Lys Met His Glu 115 120 125 cct gtt aca gca geg egg act tta ctg aca gca gaa aat gcc aca tat 432
Pro Val Thr Ala Ala Arg Thr Leu Leu Thr Ala Glu Asn Ala Thr Tyr 130 135 140 gaa att gac cga gta ett tet caa tta eta aaa gaa aga aaa cca gtc 480
Glu lie Asp Arg Val Leu Ser Gin Leu Leu Lys Glu Arg Lys Pro Val 145 150 155 160 tat att aac tta cca gtc gat gtt get gca gca aaa gca gag aag cct 528
Tyr lie Asn Leu Pro Val Asp Val Ala Ala Ala Lys Ala Glu Lys Pro 165 170 175 gca tta tet tta gaa aaa gaa age tet aca aca aat aca act gaa caa 576
Ala Leu Ser Leu Glu Lys Glu Ser Ser Thr Thr Asn Thr Thr Glu Gin 180 185 190 gtg att ttg agt aag att gaa gaa agt ttg aaa aat gcc caa aaa cca 624
Val lie Leu Ser Lys lie Glu Glu Ser Leu Lys Asn Ala Gin Lys Pro 195 200 205 gta gtg att gca gga cac gaa gta att agt ttt ggt tta gaa aaa aeg 672
Val Val lie Ala Gly His Glu Val lie Ser Phe Gly Leu Glu Lys Thr 210 215 220 gta act cag ttt gtt tea gaa aca aaa eta ccg att aeg aca eta aat 720
Val Thr Gin Phe Val Ser Glu Thr Lys Leu Pro lie Thr Thr Leu Asn 225 230 235 240 ttt ggt aaa agt get gtt gat gaa tet ttg ccc tea ttt tta gga ata 768 -202· 146966·序列表.doc 816201037078
Phe Gly Lys Ser Ala Val Asp Glu Ser Leu Pro Ser Phe Leu Gly lie 245 250 255 tat aac ggg aaa ctt tea gaa ate agt ett aaa aat ttt gtg gag tcc Tyr Asn Gly Lys Leu Ser Glu lie Ser Leu Lys Asn Phe Val Glu Ser 260 265 270 gca gac ttt ate eta atg ctt gga gtg aag ctt aeg gac tcc tea aca Ala Asp Phe lie Leu Met Leu Gly Val Lys Leu Thr Asp Ser Ser Thr 275 280 285 ggt gca ttc aca cat cat tta gat gaa aat aaa atg att tea eta aac Gly Ala Phe Thr His His Leu Asp Glu Asn Lys Met lie Ser Leu Asn 290 295 300 ata gat gaa gga ata att ttc aat aaa gtg gta gaa gat ttt gat ttt 工le Asp Glu Gly lie lie Phe Asn Lys Val Val Glu Asp Phe Asp Phe 305 310 315 320 864 912 960
aga gca gtg gtt tet tet tta tea gaa tta aaa gga ata gaa tat gaa Arg Ala Val Val Ser Ser Leu Ser Glu Leu Lys Gly lie Glu Tyr Glu 325 330 335 1008 gga caa tat att gat aag caa tat gaa gaa ttt att cca tea agt get Gly Gin Tyr 工le Asp Lys Gin Tyr Glu Glu Phe lie Pro Ser Ser Ala 340 345 350 ccc tta tea caa gac cgt eta tgg cag gca gtt gaa agt ttg act caa Pro Leu Ser Gin Asp Arg Leu Trp Gin Ala Val Glu Ser Leu Thr Gin 355 360 365 age aat gaa aca ate gtt get gaa caa gga acc tea ttt ttt gga get Ser Asn Glu Thr 工le Val Ala Glu Gin Gly Thr Ser Phe Phe Gly Ala 370 375 380 1056 1104 1152 tea aca att ttc tta aaa tea aat agt cgt ttt att gga caa cct tta Ser Thr lie Phe Leu Lys Ser Asn Ser Arg Phe lie Gly Gin Pro Leu 385 390 395 400 1200
tgg ggt tet att gga tat act ttt cca geg get tta gga age caa att Trp Gly Ser lie Gly Tyr Thr Phe Pro Ala Ala Leu Gly Ser Gin lie 405 410 415 geg gat aaa gag age aga cac ctt tta ttt att ggt gat ggt tea ctt Ala Asp Lys Glu Ser Arg His Leu Leu Phe lie Gly Asp Gly Ser Leu 420 425 430 1248 1296 caa ctt acc gta caa gaa tta gga eta tea ate aga gaa aaa etc aat Gin Leu Thr Val Gin Glu Leu Gly Leu Ser lie Arg Glu Lys Leu Asn 435 440 445 1344 cca att tgt ttt ate ata aat aat gat ggt tat aca gtt gaa aga gaa Pro lie Cys Phe lie lie Asn Asn Asp Gly Tyr Thr Val Glu Arg Glu 450 455 460 1392 ate cac gga cct act caa agt tat aac gac att cca atg tgg aat tac 工le His Gly Pro Thr Gin Ser Tyr Asn Asp lie Pro Met Trp Asn Tyr 465 470 475 480 1440 teg aaa tta cca gaa aca ttt gga gca aca gaa gat cgt gta gta tea Ser Lys Leu Pro Glu Thr Phe Gly Ala Thr Glu Asp Arg Val Val Ser 146966-序列表.doc -203- 1488 201037078 485 490 495 aaa att gtt aga aca gag aat gaa ttt gtg tct gtc atg aaa gaa gcc Lys lie Val Arg Thr Glu Asn Glu Phe Val Ser Val Met Lys Glu Ala 500 505 510 caa gca gat gtc aat aga atg tat tgg ata gaa eta gtt ttg gaa aaa Gin Ala Asp Val Asn Arg Met Tyr Trp lie Glu Leu Val Leu Glu Lys 515 520 525 gaa gat geg cca aaa tta ctg aaa aaa atg ggt aaa tta ttt get gag Glu Asp Ala Pro Lys Leu Leu Lys Lys Met Gly Lys Leu Phe Ala Glu 530 535 540 caa aat aaa tag 1644 Gin Asn Lys 545 1536 1584 1632 <210> 115 <211> 547 <212> PRT <213> 乳酸乳球菌 <400> 115
Met Tyr Thr Val Gly Asp Tyr Leu Leu Asp Arg Leu His Glu Leu Gly 15 10 15 lie Glu Glu lie Phe Gly Val Pro Gly Asp Tyr Asn Leu Gin Phe Leu 20 25 30
Asp Gin lie lie Ser Arg Glu Asp Met Lys Trp lie Gly Asn Ala Asn 35 40 45
Glu Leu Asn Ala Ser Tyr Met Ala Asp Gly Tyr Ala Arg Thr Lys Lys 50 55 60
Ala Ala Ala Phe Leu Thr Thr Phe Gly Val Gly Glu Leu Ser Ala lie 65 70 75 80
Asn Gly Leu Ala Gly Ser Tyr Ala Glu Asn Leu Pro Val Val Glu lie 85 90 95
Val Gly Ser Pro Thr Ser Lys Val Gin Asn Asp Gly Lys Phe Val His 100 105 110
His Thr Leu Ala Asp Gly Asp Phe Lys His Phe Met Lys Met His Glu 115 120 125
Pro Val Thr Ala Ala Arg Thr Leu Leu Thr Ala Glu Asn Ala Thr Tyr 130 135 140 • 204· 146966-序列表.doc 201037078
Glu 工le Asp Arg Val Leu Ser Gin Leu Leu Lys Glu Arg Lys Pro Val 145 150 155 160
Tyr lie Asn Leu Pro Val Asp Val Ala Ala Ala Lys Ala Glu Lys Pro 165 170 175
Ala Leu Ser Leu Glu Lys Glu Ser Ser Thr Thr Asn Thr Thr Glu Gin 180 185 190
Val 工le Leu Ser Lys lie Glu Glu Ser Leu Lys Asn Ala Gin Lys Pro 195 200 205
Val Val lie Ala Gly His Glu Val lie Ser Phe Gly Leu Glu Lys Thr 210 215 220
Val Thr Gin Phe Val Ser Glu Thr Lys Leu Pro lie Thr Thr Leu Asn 225 230 235 240
Phe Gly Lys Ser Ala Val Asp Glu Ser Leu Pro Ser Phe Leu Gly lie 245 250 255
Tyr Asn Gly Lys Leu Ser Glu lie Ser Leu Lys Asn Phe Val Glu Ser 260 265 270
Ala Asp Phe lie Leu Met Leu Gly Val Lys Leu Thr Asp Ser Ser Thr 275 280 285
Gly Ala Phe Thr His His Leu Asp Glu Asn Lys Met lie Ser Leu Asn 290 295 300 lie Asp Glu Gly lie lie Phe Asn Lys Val Val Glu Asp Phe Asp Phe 305 310 315 320
Arg Ala Val Val Ser Ser Leu Ser Glu Leu Lys Gly lie Glu Tyr Glu 325 330 335
Gly Gin Tyr lie Asp Lys Gin Tyr Glu Glu Phe lie Pro Ser Ser Ala 340 345 350
Pro Leu Ser Gin Asp Arg Leu Trp Gin Ala Val Glu Ser Leu Thr Gin 355 360 365
Ser Asn Glu Thr lie Val Ala Glu Gin Gly Thr Ser Phe Phe Gly Ala 370 375 380 -205- 146966-序列表 _doc 201037078
Ser Thr lie Phe Leu Lys Ser Asn Ser Arg Phe lie Gly Gin Pro Leu 385 390 395 400
Trp Gly Ser lie Gly Tyr Thr Phe Pro Ala Ala Leu Gly Ser Gin lie 405 410 415
Ala Asp Lys Glu Ser Arg His Leu Leu Phe lie Gly Asp Gly Ser Leu 420 425 430
Gin Leu Thr Val Gin Glu Leu Gly Leu Ser lie Arg Glu Lys Leu Asn 435 440 445
Pro lie Cys Phe lie lie Asn Asn Asp Gly Tyr Thr Val Glu Arg Glu 450 455 460 工le His Gly Pro Thr Gin Ser Tyr Asn Asp lie Pro Met Trp Asn Tyr 465 470 475 480
Ser Lys Leu Pro Glu Thr Phe Gly Ala Thr Glu Asp Arg Val Val Ser 485 490 495
Lys lie Val Arg Thr Glu Asn Glu Phe Val Ser Val Met Lys Glu Ala 500 505 510
Gin Ala Asp Val Asn Arg Met Tyr Trp lie Glu Leu Val Leu Glu Lys 515 520 525
Glu Asp Ala Pro Lys Leu Leu Lys Lys Met Gly Lys Leu Phe Ala Glu 530 535 540
Gin Asn Lys 545 <210> 116 <211> 1644 <212> DNA <213> 人工 <220> <223〉乳酸乳球菌支鏈a_酮酸脫羧酶KdcA密碼子優化基因 <400> 116 atgtatactg ttggtgatta tctgctggac cgtctgcatg aactgggcat tgaagaaatc ttcggtgtcc caggcgacta caacctgcag ttcctggacc agatcatctc ccgcgaagat atgaaatgga tcggtaacgc aaacgagctg aacgcgtctt atatggctga tggttatgct cgcaccaaaa aggctgcggc ctttctgacc acctttggtg tgggcgagct gagcgcgatc -206· 60 120 180 146966-序列表.doc 240 201037078 Ο Ο aacggcctgg acctctaagg aagcacttta aacgcaacgt tatatcaatc gagaaggaga tctctgaaaa ctggagaaaa tttggcaagt ctgtccgaga gtgaaactga atcagcctga cgtgctgttg gataaacagt caggcagttg ttcttcggtg tggggttcta tctcgtcatc ctgtctatcc gttgagcgtg tctaaactgc actgagaacg tggatcgaac ctgtttgcgg caggttccta ttcagaacga tgaaaatgca acgagatcga tgccggttga gctccactac acgcacagaa ctgttactca ccgcagttga tctccctgaa ccgactcctc acatcgacga tcagcagcct acgaagagtt agtccctgac cttctaccat tcggttacac tgctgttcat gtgaaaaact aaattcatgg cggaaacctt agttcgtatc tggttctgga agcagaacaa cgctgagaac cggcaaattc cgaaccggtc tcgtgtactg tgtcgctgcg taacaccacc accggtggtt attcgtctct cgaatctctg gaacttcgta taccggtgcg gggtatcatc gtccgagctg tattccgtct gcagtccaac ctttctgaag gttcccggct cggtgatggt gaacccgatt tccgactcag cggtgcaact tgttatgaaa aaaagaggat gtaa ctgccggtag gtacatcaca accgccgctc tcccagctgc gccaaagcag gaacaggtta atcgcaggtc gaaacgaaac ccttctttcc gaatccgctg ttcacgcacc ttcaacaagg aaaggcattg tctgcaccgc gaaactatcg tccaactctc gcgctgggta tccctgcagc tgttttatca tcttacaacg gaggatcgcg gaggcgcagg gcaccgaaac tagaaatcgt ccctggcgga gcactctgct tgaaagaacg agaaaccggc tcctgtccaa acgaggttat tgccgatcac tgggcattta actttatcct atctggatga tagttgaaga agtacgaggg tgagccagga tagcggaaca gctttatcgg gccagatcgc tgactgtaca tcaataacga atattccgat tcgtgagcaa cagatgtgaa tgctgaagaa tggttccccg cggcgatttt gaccgcggaa taaaccggtg actgtccctg aattgaagaa ctccttcggc gaccctgaac caacggcaaa gatgctgggt aaacaaaatg tttcgacttc tcaatacatc ccgcctgtgg aggtacctct tcagccgctg tgataaagag ggaactgggt tggctacact gtggaactac gattgtgcgt ccgcatgtac aatgggtaaa 1644 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 <210> 117 <211> 1647 <212> DNA <213>乳酸乳球菌 <220> <221> CDS <222> (1)..(1647) <400> 117 -207- 146966·序列表.doc 201037078 atg tat aca gta gga gat tac eta tta gac ega tta cac gag tta gga 48
Met Tyr Thr Val Gly Asp Tyr Leu Leu Asp Arg Leu His Glu Leu Gly 1 5 10 15 att gaa gaa att ttt gga gtc cct gga gac tat aac tta caa ttt tta 96 lie Glu Glu lie Phe Gly Val Pro Gly Asp Tyr Asn Leu Gin Phe Leu 20 25 30 gat caa att att tcc cac aag gat atg aaa tgg gtc gga aat get aat 144
Asp Gin lie lie Ser His Lys Asp Met Lys Trp Val Gly Asn Ala Asn 35 40 45 gaa tta aat get tea tat atg get gat ggc tat get cgt act aaa aaa 192
Glu Leu Asn Ala Ser Tyr Met Ala Asp Gly Tyr Ala Arg Thr Lys Lys 50 55 60 get gcc gca ttt ett aca acc ttt gga gta ggt gaa ttg agt gca gtt 240
Ala Ala Ala Phe Leu Thr Thr Phe Gly Val Gly Glu Leu Ser Ala Val 65 70 75 80 aat gga tta gca gga agt tac gcc gaa aat tta cca gta gta gaa ata 288
Asn Gly Leu Ala Gly Ser Tyr Ala Glu Asn Leu Pro Val Val Glu lie 85 90 95 gtg gga tea cct aca tea aaa gtt caa aat gaa gga aaa ttt gtt cat 336
Val Gly Ser Pro Thr Ser Lys Val Gin Asn Glu Gly Lys Phe Val His 100 105 110 cat aeg ctg get gac ggt gat ttt aaa cac ttt atg aaa atg cac gaa 384
His Thr Leu Ala Asp Gly Asp Phe Lys His Phe Met Lys Met His Glu 115 120 125 cct gtt aca gca get ega act tta ctg aca gca gaa aat gca acc gtt 432
Pro Val Thr Ala Ala Arg Thr Leu Leu Thr Ala Glu Asn Ala Thr Val 130 135 140 gaa att gac ega gta ett tet gca eta tta aaa gaa aga aaa cct gtc 480
Glu lie Asp Arg Val Leu Ser Ala Leu Leu Lys Glu Arg Lys Pro Val 145 150 155 160 tat ate aac tta cca gtt gat gtt get get gca aaa gca gag aaa ccc 528
Tyr lie Asn Leu Pro Val Asp Val Ala Ala Ala Lys Ala Glu Lys Pro 165 170 175 tea etc cct ttg aaa aag gaa aac tea act tea aat aca agt gac caa 576
Ser Leu Pro Leu Lys Lys Glu Asn Ser Thr Ser Asn Thr Ser Asp Gin 180 185 190 gaa att ttg aac aaa att caa gaa age ttg aaa aat gcc aaa aaa cca 624
Glu lie Leu Asn Lys lie Gin Glu Ser Leu Lys Asn Ala Lys Lys Pro 195 200 205 ate gtg att aca gga cat gaa ata att agt ttt ggc tta gaa aaa aca 672 lie Val lie Thr Gly His Glu lie lie Ser Phe Gly Leu Glu Lys Thr 210 215 220 gtc act caa ttt att tea aag aca aaa eta cct att aeg aca tta aac 720
Val Thr Gin Phe lie Ser Lys Thr Lys Leu Pro lie Thr Thr Leu Asn 225 230 235 240 ttt ggt aaa agt tea gtt gat gaa gcc etc cct tea ttt tta gga ate 768 • 208 - 146966-序列表.doc 816201037078
Phe Gly Lys Ser Ser Val Asp Glu Ala Leu Pro Ser Phe Leu Gly lie 245 250 255 tat aat ggt aca etc tea gag cct aat ett aaa gaa ttc gtg gaa tea Tyr Asn Gly Thr Leu Ser Glu Pro Asn Leu Lys Glu Phe Val Glu Ser 260 265 270 gcc gac ttc ate ttg atg ett gga gtt aaa etc aca gac tet tea aca Ala Asp Phe lie Leu Met Leu Gly Val Lys Leu Thr Asp Ser Ser Thr 275 280 285 864 gga gcc ttc act cat cat tta aat gaa aat aaa atg att tea ctg aat Gly Ala Phe Thr His His Leu Asn Glu Asn Lys Met lie Ser Leu Asn 290 295 300 ata gat gaa gga aaa ata ttt aac gaa aga ate caa aat ttt gat ttt lie Asp Glu Gly Lys lie Phe Asn Glu Arg lie Gin Asn Phe Asp Phe 305 310 315 320 912 960
gaa tcc etc ate tcc tet etc tta gac eta age gaa ata gaa tac aaa Glu Ser Leu lie Ser Ser Leu Leu Asp Leu Ser Glu lie Glu Tyr Lys 325 330 335 gga aaa tat ate gat aaa aag caa gaa gac ttt gtt cca tea aat geg Gly Lys Tyr lie Asp Lys Lys Gin Glu Asp Phe Val Pro Ser Asn Ala 340 345 350 ett tta tea caa gac ege eta tgg caa gca gtt gaa aac eta act caa Leu Leu Ser Gin Asp Arg Leu Trp Gin Ala Val Glu Asn Leu Thr Gin 355 360 365 age aat gaa aca ate gtt get gaa caa ggg aca tea ttc ttt ggc get Ser Asn Glu Thr lie Val Ala Glu Gin Gly Thr Ser Phe Phe Gly Ala 370 375 380 1008 1056 1104 1152 tea tea att ttc tta aaa tea aag agt cat ttt att ggt caa ccc tta Ser Ser lie Phe Leu Lys Ser Lys Ser His Phe lie Gly Gin Pro Leu 385 390 395 400 tgg gga tea att gga tat aca ttc cca gca gca tta gga age caa att Trp Gly Ser lie Gly Tyr Thr Phe Pro Ala Ala Leu Gly Ser Gin lie \J 405 410 415 gca gat aaa gaa age aga cac ett tta ttt att ggt gat ggt tea ett Ala Asp Lys Glu Ser Arg His Leu Leu Phe lie Gly Asp Gly Ser Leu 420 425 430 caa ett aca gtg caa gaa tta gga tta gca ate aga gaa aaa att aat Gin Leu Thr Val Gin Glu Leu Gly Leu Ala lie Arg Glu Lys lie Asn 435 440 445 cca att tgc ttt att ate aat aat gat ggt tat aca gtc gaa aga gaa Pro lie Cys Phe lie lie Asn Asn Asp Gly Tyr Thr Val Glu Arg Glu 450 455 460 att cat gga cca aat caa age tac aat gat att cca atg tgg aat tac lie His Gly Pro Asn Gin Ser Tyr Asn Asp lie Pro Met Trp Asn Tyr 465 470 475 480 1200 1248 1296 1344 1392 1440 tea aaa tta cca gaa teg ttt gga gca aca gaa gat ega gta gtc tea Ser Lys Leu Pro Glu Ser Phe Gly Ala Thr Glu Asp Arg Val Val Ser 146966-序列表.doc -209- 1488 201037078 485 490 495 aaa ate gtt aga act gaa aat gaa ttt gtg tet gtc atg aaa gaa get Lys lie Val Arg Thr Glu Asn Glu Phe Val Ser Val Met Lys Glu Ala 500 505 510 caa gca gat cca aat aga atg tac tgg att gag tta att ttg gca aaa Gin Ala Asp Pro Asn Arg Met Tyr Trp lie Glu Leu lie Leu Ala Lys 515 520 525 gaa ggt gca cca aaa gta ctg aaa aaa atg ggc aaa eta ttt get gaa Glu Gly Ala Pro Lys Val Leu Lys Lys Met Gly Lys Leu Phe Ala Glu 530 535 540 caa aat aaa tea taa 1647 Gin Asn Lys Ser 545 1536 1584 1632 <210> 118 <211> 548 <212> PRT <213> 乳酸乳球菌 <400> 118
Met Tyr Thr Val Gly Asp Tyr Leu Leu Asp Arg Leu His Glu Leu Gly 15 10 15 lie Glu Glu lie Phe Gly Val Pro Gly Asp Tyr Asn Leu Gin Phe Leu 20 25 30
Asp Gin lie lie Ser His Lys Asp Met Lys Trp Val Gly Asn Ala Asn 35 40 45
Glu Leu Asn Ala Ser Tyr Met Ala Asp Gly Tyr Ala Arg Thr Lys Lys 50 55 60
Ala Ala Ala Phe Leu Thr Thr Phe Gly Val Gly Glu Leu Ser Ala Val 65 70 75 80
Asn Gly Leu Ala Gly Ser Tyr Ala Glu Asn Leu Pro Val Val Glu lie 85 90 95
Val Gly Ser Pro Thr Ser Lys Val Gin Asn Glu Gly Lys Phe Val His 100 105 110
His Thr Leu Ala Asp Gly Asp Phe Lys His Phe Met Lys Met His Glu 115 120 125
Pro Val Thr Ala Ala Arg Thr Leu Leu Thr Ala Glu Asn Ala Thr Val 130 135 140 •210- 146966·序列表.doc 201037078
Glu lie Asp Arg Val Leu Ser Ala Leu Leu Lys Glu Arg Lys Pro Val 145 150 155 160
Tyr lie Asn Leu Pro Val Asp Val Ala Ala Ala Lys Ala Glu Lys Pro 165 170 175
Ser Leu Pro Leu Lys Lys Glu Asn Ser Thr Ser Asn Thr Ser Asp Gin 180 185 190
Glu lie Leu Asn Lys lie Gin Glu Ser Leu Lys Asn Ala Lys Lys Pro 195 200 205 lie Val lie Thr Gly His Glu lie lie Ser Phe Gly Leu Glu Lys Thr 210 215 220
o
Val Thr Gin Phe lie Ser Lys Thr Lys Leu Pro lie Thr Thr Leu Asn 225 230 235 240
Phe Gly Lys Ser Ser Val Asp Glu Ala Leu Pro Ser Phe Leu Gly lie 245 250 255
Tyr Asn Gly Thr Leu Ser Glu Pro Asn Leu Lys Glu Phe Val Glu Ser 260 265 270
Ala Asp Phe lie Leu Met Leu Gly Val Lys Leu Thr Asp Ser Ser Thr 275 280 285
Gly Ala Phe Thr His His Leu Asn Glu Asn Lys Met lie Ser Leu Asn 290 295 300 lie Asp Glu Gly Lys lie Phe Asn Glu Arg lie Gin Asn Phe Asp Phe 305 310 315 320
Glu Ser Leu lie Ser Ser Leu Leu Asp Leu Ser Glu lie Glu Tyr Lys 325 330 335
Gly Lys Tyr lie Asp Lys Lys Gin Glu Asp Phe Val Pro Ser Asn Ala 340 345 350
Leu Leu Ser Gin Asp Arg Leu Trp Gin Ala Val Glu Asn Leu Thr Gin 355 360 365
Ser Asn Glu Thr lie Val Ala Glu Gin Gly Thr Ser Phe Phe Gly Ala 370 375 380 •211 - 146966-序列表.doc 201037078
Ser Ser lie Phe Leu Lys Ser Lys Ser His Phe lie Gly Gin Pro Leu 385 390 395 400
Trp Gly Ser lie Gly Tyr Thr Phe Pro Ala Ala Leu Gly Ser Gin lie 405 410 415
Ala Asp Lys Glu Ser Arg His Leu Leu Phe lie Gly Asp Gly Ser Leu 420 425 430
Gin Leu Thr Val Gin Glu Leu Gly Leu Ala lie Arg Glu Lys lie Asn 435 440 445
Pro lie Cys Phe lie lie Asn Asn Asp Gly Tyr Thr Val Glu Arg Glu 450 455 460 lie His Gly Pro Asn Gin Ser Tyr Asn Asp lie Pro Met Trp Asn Tyr 465 470 475 480
Ser Lys Leu Pro Glu Ser Phe Gly Ala Thr Glu Asp Arg Val Val Ser 485 490 495
Lys lie Val Arg Thr Glu Asn Glu Phe Val Ser Val Met Lys Glu Ala 500 505 510
Gin Ala Asp Pro Asn Arg Net Tyr Trp lie Glu Leu lie Leu Ala Lys 515 520 525
Glu Gly Ala Pro Lys Val Leu Lys Lys Met Gly Lys Leu Phe Ala Glu 530 535 540
Gin Asn Lys Ser 545 <210> 119 <211> 1647 <212> DNA <213> 人工 <220> <223>乳酸乳球菌a-酮異戊酸脫羧酶KivD密碼子優化基因 <400> 119 atgtatactg ttggtgatta cctgctggat cgtctgcatg aactgggcat cgaggaaatt ttcggcgtac ctggtgacta taacctgcag ttcctggatc agatcatttc ccacaaagat atgaaatggg ttggtaacgc gaacgagctg aatgcaagct acatggctga cggttatgca cgcaccaaga aagctgcggc gttcctgact acttttggcg tcggcgagct gtctgcggta 212- 60 120 180 146966·序列表.doc 240 201037078
aacggtctgg ccggctccta cgcggaaaac ctgccggtag tagaaatcgt cggttccccg 300 acctctaaag ttcagaacga gggtaaattc gtgcaccata ctctggccga tggtgacttc 360 aaacacttca tgaagatgca cgaaccggtc actgctgctc gtacgctgct gaccgcggaa 420 aatgcgactg tcgagattga tcgtgtactg agcgcactgc tgaaagaacg caagcctgta 480 tacatcaacc tgccggttga tgtcgcggcc gccaaagcgg aaaaaccatc tctgccgctg 540 aaaaaggaga acagcacctc taacaccagc gaccaggaaa tcctgaacaa gatccaggag 600 tctctgaaga acgctaaaaa gccgatcgta atcaccggcc atgagattat ctctttcggt 660 ctggagaaaa ctgtcaccca gttcatcagc aaaaccaaac tgccgatcac caccctgaac 720 ttcggtaaat cctccgttga cgaagcgctg ccgtcctttc tgggtattta caacggcact 780 ctgtctgagc cgaacctgaa agagttcgtg gagtctgcgg attttatcct gatgctgggc 840 gtgaaactga cggattcctc caccggtgca ttcacccacc acctgaatga gaataaaatg 900 atctctctga acattgatga gggcaaaatc ttcaacgagc gtattcagaa cttcgatttc 960 gaatccctga tctcctccct gctggatctg tccgagattg aatataaagg caaatacatt 1020 gataagaagc aagaggactt cgtaccgtct aacgcgctgc tgagccagga ccgtctgtgg 1080 caagctgtgg aaaacctgac ccagtccaac gaaaccatcg tggcggaaca gggtacctcc 1140 ttcttcggtg ctagctctat cttcctgaaa tctaaaagcc acttcatcgg tcagccactg 1200 tggggctcta ttggctacac cttcccggca gcgctgggtt cccaaatcgc agacaaagaa 1260 tcccgccacc tgctgttcat tggtgacggc tctctgcaac tgaccgtaca ggagctgggt 1320 ctggcgattc gtgagaaaat caacccgatt tgtttcatca tcaacaacga tggctacact 1380 gttgagcgtg agatccacgg cccgaaccag tcctacaacg acattccgat gtggaactac 1440 tctaaactgc cggaatcctt cggtgcgact gaagaccgtg tcgtaagcaa gatcgtccgt 1500 accgaaaacg aattcgtgtc tgtcatgaaa gaagcacagg cggacccgaa ccgcatgtac 1560 tggatcgagc tgattctggc taaagagggc gcgccaaaag tactgaaaaa gatgggtaaa 1620 ctgttcgcag aacagaacaa atcctaa 1647 <210> 120 <211> 3696 <212> DNA <213>結核分枝桿菌 <220> <221> CDS <222> (1) .. (3696) <400> 120 -213- 146966·序列表.doc 201037078 gtg gcc aac ata agt tea cca ttc ggg caa aac gaa tgg ctg gtc gaa 48
Val Ala Asn lie Ser Ser Pro Phe Gly Gin Asn Glu Trp Leu Val Glu 1 5 10 15 gag atg tac ege aag ttc ege gac gac ccc tee teg gtc gat ccc age 96
Glu Met Tyr Arg Lys Phe Arg Asp Asp Pro Ser Ser Val Asp Pro Ser 20 25 30 tgg cac gag ttc ctg gtt gac tac age ccc gaa ccc acc tee caa cca 144
Trp His Glu Phe Leu Val Asp Tyr Ser Pro Glu Pro Thr Ser Gin Pro 35 40 45 get gcc gaa cca acc egg gtt acc teg cca etc gtt gcc gag egg gcc 192
Ala Ala Glu Pro Thr Arg Val Thr Ser Pro Leu Val Ala Glu Arg Ala 50 55 60 get geg gcc gcc ccg cag gca ccc ccc aag ccg gcc gac acc geg gcc 240
Ala Ala Ala Ala Pro Gin Ala Pro Pro Lys Pro Ala Asp Thr Ala Ala 65 70 75 80 geg ggc aac ggc gtg gtc gcc gca ctg gcc gcc aaa act gcc gtt ccc 288
Ala Gly Asn Gly Val Val Ala Ala Leu Ala Ala Lys Thr Ala Val Pro 85 90 95 ccg cca gcc gaa ggt gac gag gta geg gtg ctg ege ggc gcc gcc geg 336
Pro Pro Ala Glu Gly Asp Glu Val Ala Val Leu Arg Gly Ala Ala Ala 100 105 110 gee gtc gtc aag aac atg tee geg teg ttg gag gtg ccg aeg geg acc 384
Ala Val Val Lys Asn Met Ser Ala Ser Leu Glu Val Pro Thr Ala Thr 115 120 125 age gtc egg geg gtc ccg gcc aag eta ctg ate gac aac egg ate gtc 432
Ser Val Arg Ala Val Pro Ala Lys Leu Leu lie Asp Asn Arg 工le Val 130 135 140 ate aac aac cag ttg aag egg acc ege ggc ggc aag ate teg ttc aeg 480 lie Asn Asn Gin Leu Lys Arg Thr Arg Gly Gly Lys lie Ser Phe Thr 145 150 155 160 cat ttg ctg ggc tac gcc ctg gtg cag geg gtg aag aaa ttc ccg aac 528
His Leu Leu Gly Tyr Ala Leu Val Gin Ala Val Lys Lys Phe Pro Asn 165 170 175 atg aac egg cac tac acc gaa gtc gac ggc aag ccc acc geg gtc aeg 576
Met Asn Arg His Tyr Thr Glu Val Asp Gly Lys Pro Thr Ala Val Thr 180 185 190 ccg geg cac acc aat etc ggc ctg geg ate gac ctg caa ggc aag gac 624
Pro Ala His Thr Asn Leu Gly Leu Ala lie Asp Leu Gin Gly Lys Asp 195 200 205 ggg aag cgt tee ctg gtg gtg gcc ggc ate aag egg tgc gag acc atg 672
Gly Lys Arg Ser Leu Val Val Ala Gly lie Lys Arg Cys Glu Thr Met 210 215 220 ega ttc geg cag ttc gtc aeg gcc tac gaa gac ate gta ege egg gcc 720
Arg Phe Ala Gin Phe Val Thr Ala Tyr Glu Asp lie Val Arg Arg Ala 225 230 235 240 ege gac ggc aag ctg acc act gaa gac ttt gcc ggc gtg aeg att teg 768 -214- 146966·序列表.doc 816 201037078
Arg Asp Gly Lys Leu Thr Thr Glu Asp Phe Ala Gly Val Thr lie Ser 245 250 255 ctg acc aat ccc gga acc ate ggc acc gtg cat teg gtg ccg egg ctg
Leu Thr Asn Pro Gly Thr lie Gly Thr Val His Ser Val Pro Arg Leu 260 265 270 864 atg ccc ggc cag ggc gcc ate ate ggc gtg ggc gee atg gaa tac ccc
Met Pro Gly Gin Gly Ala lie lie Gly Val Gly Ala Met Glu Tyr Pro 275 280 285 912 gcc gag ttt caa ggc gcc age gag gaa ege ate gcc gag ctg ggc ate
Ala Glu Phe Gin Gly Ala Ser Glu Glu Arg lie Ala Glu Leu Gly lie 290 295 300 ggc aaa ttg ate act ttg acc tee acc tac gac cac ege ate ate cag Gly Lys Leu lie Thr Leu Thr Ser Thr Tyr Asp His Arg lie lie Gin 305 310 315 320 ggc geg gaa teg ggc gac ttc ctg ege acc ate cac gag ttg ctg etc Gly Ala Glu Ser Gly Asp Phe Leu Arg Thr lie His Glu Leu Leu Leu 325 330 335 teg gat ggc ttc tgg gac gag gtc ttc ege gaa ctg age ate cca tat Ser Asp Gly Phe Trp Asp Glu Val Phe Arg Glu Leu Ser lie Pro Tyr 340 345 350 ctg ccg gtg ege tgg age acc gac aac ccc gac teg ate gtc gac aag Leu Pro Val Arg Trp Ser Thr Asp Asn Pro Asp Ser lie Val Asp Lys 355 360 365 aac get ege gtc atg aac ttg ate geg gcc tac ege aac ege ggc cat Asn Ala Arg Val Met Asn Leu lie Ala Ala Tyr Arg Asn Arg Gly His 370 375 380 ctg atg gcc gat acc gac ccg ctg egg ttg gac aaa get egg ttc ege Leu Met Ala Asp Thr Asp Pro Leu Arg Leu Asp Lys Ala Arg Phe Arg 385 390 395 400 960 1008 1056 1104 1152 1200
agt cac ccc gac etc gaa gtg ctg acc cac ggc ctg aeg ctg tgg gat Ser His Pro Asp Leu Glu Val Leu Thr His Gly Leu Thr Leu Trp Asp 405 410 415 etc gat egg gtg ttc aag gtc gac ggc ttt gcc ggt geg cag tac aag Leu Asp Arg Val Phe Lys Val Asp Gly Phe Ala Gly Ala Gin Tyr Lys 420 425 430 1248 1296 aaa ctg ege gac gtg ctg ggc ttg ctg ege gat gcc tac tgc ege cac Lys Leu Arg Asp Val Leu Gly Leu Leu Arg Asp Ala Tyr Cys Arg His 435 440 445 1344 ate ggc gtg gag tac gcc cat ate etc gac ccc gaa caa aag gag tgg lie Gly Val Glu Tyr Ala His lie Leu Asp Pro Glu Gin Lys Glu Trp 450 455 460 1392 etc gaa caa egg gtc gag acc aag cac gtc aaa ccc act gtg gcc caa Leu Glu Gin Arg Val Glu Thr Lys His Val Lys Pro Thr Val Ala Gin 465 470 475 480 1440 cag aaa tac ate etc age aag etc aac gcc gcc gag gcc ttt gaa aeg Gin Lys Tyr lie Leu Ser Lys Leu Asn Ala Ala Glu Ala Phe Glu Thr 146966-序列表.doc -215- 1488 201037078 485 490 495 ttc eta cag acc aag tac gtc ggc cag aag egg ttc teg ctg gaa ggc Phe Leu Gin Thr Lys Tyr Val Gly Gin Lys Arg Phe Ser Leu Glu Gly 500 505 510 gee gaa age gtg ate ccg atg atg gac geg geg ate gac cag tgc get Ala Glu Ser Val lie Pro Met Met Asp Ala Ala lie Asp Gin Cys Ala 515 520 525 gag cac ggc etc gac gag gtg gtc ate ggg atg ccg cac egg ggc egg Glu His Gly Leu Asp Glu Val Val lie Gly Met Pro His Arg Gly Arg 530 535 540 etc aac gtg ctg gee aac ate gtc ggc aag ccg tac teg cag ate ttc Leu Asn Val Leu Ala Asn lie Val Gly Lys Pro Tyr Ser Gin lie Phe 545 550 555 560 acc gag ttc gag ggc aac ctg aat ccg teg cag geg cac ggc tee ggt Thr Glu Phe Glu Gly Asn Leu Asn Pro Ser Gin Ala His Gly Ser Gly 565 570 575 gac gtc aag tac cac ctg ggc gee acc ggg ctg tac ctg cag atg ttc Asp Val Lys Tyr His Leu Gly Ala Thr Gly Leu Tyr Leu Gin Met Phe 580 585 590 ggc gac aac gac att cag gtg teg ctg acc gee aac ccg teg cat ctg Gly Asp Asn Asp lie Gin Val Ser Leu Thr Ala Asn Pro Ser His Leu 595 600 605 gag gee gtc gac ccg gtg ctg gag gga ttg gtg egg gee aag cag gat Glu Ala Val Asp Pro Val Leu Glu Gly Leu Val Arg Ala Lys Gin Asp 610 615 620 ctg etc gac cac gga age ate gac age gac ggc caa egg geg ttc teg Leu Leu Asp His Gly Ser lie Asp Ser Asp Gly Gin Arg Ala Phe Ser 625 630 635 640 gtg gtg ccg ctg atg ttg cat ggc gat gee geg ttc gee ggt cag ggt Val Val Pro Leu Met Leu His Gly Asp Ala Ala Phe Ala Gly Gin Gly 645 650 655 gtg gtc gee gag aeg ctg aac ctg geg aat ctg ccg ggc tac ege gtc Val Val Ala Glu Thr Leu Asn Leu Ala Asn Leu Pro Gly Tyr Arg Val 660 665 670 ggc ggc acc ate cac ate ate gtc aac aac cag ate ggc ttc acc acc Gly Gly Thr lie His lie lie Val Asn Asn Gin lie Gly Phe Thr Thr 675 680 685 geg ccc gag tat tee agg tee age gag tac tgc acc gac gtc gca aag Ala Pro Glu Tyr Ser Arg Ser Ser Glu Tyr Cys Thr Asp Val Ala Lys 690 695 700 atg ate ggg gca ccg ate ttt cac gtc aac ggc gac gac ccg gag geg Met lie Gly Ala Pro lie Phe His Val Asn Gly Asp Asp Pro Glu Ala 705 710 715 720 tgt gtc tgg gtg geg egg ttg geg gtg gac ttc ega caa egg ttc aag Cys Val Trp Val Ala Arg Leu Ala Val Asp Phe Arg Gin Arg Phe Lys 725 730 735 1536 1584 1632 1690 1728 1776 1824 1872 1920 1968 2016 2064 2112 2160 2208 146966-序列表.doc -216 201037078 aag gac gtc gtc ate gac atg ctg tgc tac ege ege ege ggg cac aac Lys Asp Val Val lie Asp Met Leu Cys Tyr Arg Arg Arg Gly His Asn 740 745 750 gag ggt gac gac ccg teg atg acc aac ccc tac gtg tac gac gtc gtc Glu Gly Asp Asp Pro Ser Met Thr Asn Pro Tyr Val Tyr Asp Val Val 755 760 765 2256 2304 gac acc aag ege ggg gee ege aaa age tac acc gaa gee ctg ate gga Asp Thr Lys Arg Gly Ala Arg Lys Ser Tyr Thr Glu Ala Leu He Gly 770 775 780 2352 cgt ggc gac ate teg atg aag gag gee gag gac geg ctg ege gac tac Arg Gly Asp lie Ser Met Lys Glu Ala Glu Asp Ala Leu Arg Asp Tyr 785 790 795 800 2400 cag ggc cag ctg gaa egg gtg ttc aac gaa gtg ege gag ctg gag aag Gin Gly Gin Leu Glu Arg Val Phe Asn Glu Val Arg Glu Leu Glu Lys 805 810 815 2448
cac ggt gtg cag ccg age gag teg gtc gag tee gac cag atg att ccc His Gly Val Gin Pro Ser Glu Ser Val Glu Ser Asp Gin Met lie Pro 820 825 830 2496 geg ggg ctg gee act geg gtg gac aag teg ctg ctg gee egg ate ggc Ala Gly Leu Ala Thr Ala Val Asp Lys Ser Leu Leu Ala Arg lie Gly 835 840 845 2544 gat geg ttc etc gee ttg ccg aac ggc ttc acc geg cac ccg ega gtc Asp Ala Phe Leu Ala Leu Pro Asn Gly Phe Thr Ala His Pro Arg Val 850 855 860 2592 caa ccg gtg ctg gag aag ege egg gag atg gee tat gaa ggc aag ate Gin Pro Val Leu Glu Lys Arg Arg Glu Met Ala Tyr Glu Gly Lys lie 865 870 875 880 2640 gac tgg gee ttt ggc gag ctg ctg geg ctg ggc teg ctg gtg gee gaa Asp Trp Ala Phe Gly Glu Leu Leu Ala Leu Gly Ser Leu Val Ala Glu 885 890 895 2688
ggc aag ctg gtg ege ttg teg ggg cag gac age ege ege ggc acc ttc Gly Lys Leu Val Arg Leu Ser Gly Gin Asp Ser Arg Arg Gly Thr Phe 900 905 910 2736 tee cag egg cat teg gtt etc ate gac ege cac act ggc gag gag ttc Ser Gin Arg His Ser Val Leu lie Asp Arg His Thr Gly Glu Glu Phe 915 920 925 2784 aca cca ctg cag ctg ctg geg acc aac tee gac ggc age ccg acc ggc Thr Pro Leu Gin Leu Leu Ala Thr Asn Ser Asp Gly Ser Pro Thr Gly 930 935 940 2832 gga aag ttc ctg gtc tac gac teg cca ctg teg gag tac gee gee gtc Gly Lys Phe Leu Val Tyr Asp Ser Pro Leu Ser Glu Tyr Ala Ala Val 945 950 955 960 • 2880 ggc ttc gag tac ggc tac act gtg ggc aat ccg gac gee gtg gtg etc Gly Phe Glu Tyr Gly Tyr Thr Val Gly Asn Pro Asp Ala Val Val Leu 965 970 975 2928 146966-序列表.doe -217- 201037078 tgg gag gcg cag ttc ggc gac ttc gtc aac ggc gcg cag teg ate ate Trp Glu Ala Gin Phe Gly Asp Phe Val Asn Gly Ala Gin Ser lie lie 980 985 990 gac gag ttc ate age tee ggt gag gee aag tgg ggc caa ttg tee aac Asp Glu Phe lie Ser Ser Gly Glu Ala Lys Trp Gly Gin Leu Ser Asn 995 1000 1005 gtc gtg ctg ctg tta ccg cac ggg cac gag ggg cag gga ccc gac Val Val Leu Leu Leu Pro His Gly His Glu Gly Gin Gly Pro Asp 1010 1015 1020 cac act tet gee egg ate gaa ege ttc ttg cag ttg tgg gcg gaa His Thr Ser Ala Arg lie Glu Arg Phe Leu Gin Leu Trp Ala Glu 1025 1030 1035 ggt teg atg acc ate gcg atg ccg teg act ccg teg aac tac ttc Gly Ser Met Thr lie Ala Met Pro Ser Thr Pro Ser Asn Tyr Phe 1040 1045 1050 cac ctg eta ege egg cat gee ctg gac ggc ate caa ege ccg ctg His Leu Leu Arg Arg His Ala Leu Asp Gly lie Gin Arg Pro Leu 1055 1060 1065 ate gtg ttc aeg ccc aag teg atg ttg cgt cac aag gee gee gtc lie Val Phe Thr Pro Lys Ser Met Leu Arg His Lys Ala Ala Val 1070 1075 1080 age gaa ate aag gac ttc acc gag ate aag ttc ege tea gtg ctg Ser Glu lie Lys Asp Phe Thr Glu lie Lys Phe Arg Ser Val Leu 1085 1090 1095 gag gaa ccc acc tat gag gac ggc ate gga gac ege aac aag gtc Glu Glu Pro Thr Tyr Glu Asp Gly lie Gly Asp Arg Asn Lys Val 1100 1105 1110 age egg ate ctg ctg acc agt ggc aag ctg tat tac gag ctg gee Ser Arg lie Leu Leu Thr Ser Gly Lys Leu Tyr Tyr Glu Leu Ala 1115 1120 1125 gee ege aag gee aag gac aac ege aat gac etc gcg ate gtg egg Ala Arg Lys Ala Lys Asp Asn Arg Asn Asp Leu Ala lie Val Arg 1130 1135 1140 ett gaa cag etc gee ccg ctg ccc agg cgt ega ctg cgt gaa aeg Leu Glu Gin Leu Ala Pro Leu Pro Arg Arg Arg Leu Arg Glu Thr 1145 1150 1155 ctg gac ege tac gag aac gtc aag gag ttc ttc tgg gtc caa gag Leu Asp Arg Tyr Glu Asn Val Lys Glu Phe Phe Trp Val Gin Glu 1160 1165 1170 gaa ccg gee aac cag ggt gcg tgg ccg ega ttc ggg etc gaa eta Glu Pro Ala Asn Gin Gly Ala Trp Pro Arg Phe Gly Leu Glu Leu 1175 1180 1185 ccc gag ctg ctg cct gac aag ttg gee ggg ate aag ega ate teg Pro Glu Leu Leu Pro Asp Lys Leu Ala Gly lie Lys Arg lie Ser 1190 1195 1200 ege egg gcg atg tea gee ccg teg tea ggc teg teg aag gtg cac 2976 3024 3069 3114 3159 3204 3249 3294 3339 3384 3429 3474 3519 3564 3609 3654 146966·序列表.doc •218- 201037078
Arg Arg Ala Met Ser Ala Pro Ser Ser Gly Ser Ser Lys Val His 1205 1210 1215 gcc gtc gaa cag cag gag ate etc gac gag geg ttc ggc tga 3696 Ala Val Glu Gin Gin Glu He Leu Asp Glu Ala Phe Gly 1220 1225 1230 <210> 121 <211> 1231 <212> PRT <213>結核分枝桿菌 <400> 121
Val Ala Asn lie Ser Ser Pro Phe Gly Gin Asn Glu Trp Leu Val Glu 15 10 15
Glu Met Tyr Arg Lys Phe Arg Asp Asp Pro Ser Ser Val Asp Pro Ser 20 25 30
Trp His Glu Phe Leu Val Asp Tyr Ser Pro Glu Pro Thr Ser Gin Pro 35 40 45
Ala Ala Glu Pro Thr Arg Val Thr Ser Pro Leu Val Ala Glu Arg Ala 50 55 60
Ala Ala Ala Ala Pro Gin Ala Pro Pro Lys Pro Ala Asp Thr Ala Ala 65 70 75 80
Ala Gly Asn Gly Val Val Ala Ala Leu Ala Ala Lys Thr Ala Val Pro 85 90 95
Pro Pro Ala Glu Gly Asp Glu Val Ala Val Leu Arg Gly Ala Ala Ala 100 105 110
Ala Val Val Lys Asn Met Ser Ala Ser Leu Glu Val Pro Thr Ala Thr 115 120 125
Ser Val Arg Ala Val Pro Ala Lys Leu Leu lie Asp Asn Arg lie Val 130 135 140 lie Asn Asn Gin Leu Lys Arg Thr Arg Gly Gly Lys lie Ser Phe Thr 145 150 155 160
His Leu Leu Gly Tyr Ala Leu Val Gin Ala Val Lys Lys Phe Pro Asn 165 170 175
Met Asn Arg His Tyr Thr Glu Val Asp Gly Lys Pro Thr Ala Val Thr 180 185 190 -219 146966-序列表.doc 201037078
Pro Ala His Thr Asn Leu Gly Leu Ala lie Asp Leu Gin Gly Lys Asp 195 200 205
Gly Lys Arg Ser Leu Val Val Ala Gly lie Lys Arg Cys Glu Thr Met 210 215 220
Arg Phe Ala Gin Phe Val Thr Ala Tyr Glu Asp lie Val Arg Arg Ala 225 230 235 240
Arg Asp Gly Lys Leu Thr Thr Glu Asp Phe Ala Gly Val Thr lie Ser 245 250 255
Leu Thr Asn Pro Gly Thr lie Gly Thr Val His Ser Val Pro Arg Leu 260 265 270
Met Pro Gly Gin Gly Ala lie lie Gly Val Gly Ala Met Glu Tyr Pro 275 280 285
Ala Glu Phe Gin Gly Ala Ser Glu Glu Arg lie Ala Glu Leu Gly lie 290 295 300
Gly Lys Leu 工le Thr Leu Thr Ser Thr Tyr Asp His Arg lie lie Gin 305 310 315 320
Gly Ala Glu Ser Gly Asp Phe Leu Arg Thr lie His Glu Leu Leu Leu 325 330 335
Ser Asp Gly Phe Trp Asp Glu Val Phe Arg Glu Leu Ser lie Pro Tyr 340 345 350
Leu Pro Val Arg Trp Ser Thr Asp Asn Pro Asp Ser lie Val Asp Lys 355 360 365
Asn Ala Arg Val Met Asn Leu lie Ala Ala Tyr Arg Asn Arg Gly His 370 375 380
Leu Met Ala Asp Thr Asp Pro Leu Arg Leu Asp Lys Ala Arg Phe Arg 385 390 395 400
Ser His Pro Asp Leu.Glu Val Leu Thr His Gly Leu Thr Leu Trp Asp 405 410 415
Leu Asp Arg Val Phe Lys Val Asp Gly Phe Ala Gly Ala Gin Tyr Lys 420 425 430 •220 146966-序列表.doc 201037078
Lys Leu Arg Asp Val Leu Gly Leu Leu Arg Asp Ala Tyr Cys Arg His 435 440 445 lie Gly Val Glu Tyr Ala His lie Leu Asp Pro Glu Gin Lys Glu Trp 450 455 460
Leu Glu Gin Arg Val Glu Thr Lys His Val Lys Pro Thr Val Ala Gin 465 470 475 480
Gin Lys Tyr lie Leu Ser Lys Leu Asn Ala Ala Glu Ala Phe Glu Thr 485 490 495
Phe Leu Gin Thr Lys Tyr Val Gly Gin Lys Arg Phe Ser Leu Glu Gly 500 505 510 o
Ala Glu Ser Val lie Pro Met Met Asp Ala Ala lie Asp Gin Cys Ala 515 520 525
Glu His Gly Leu Asp Glu Val Val lie Gly Met Pro His Arg Gly Arg 530 535 540
Leu Asn Val Leu Ala Asn lie Val Gly Lys Pro Tyr Ser Gin lie Phe 545 550 555 560
Thr Glu Phe Glu Gly Asn Leu Asn Pro Ser Gin Ala His Gly Ser Gly 565 570 575
Asp Val Lys Tyr His Leu Gly Ala Thr Gly Leu Tyr Leu Gin Met Phe 580 585 590
Gly Asp Asn Asp lie Gin Val Ser Leu Thr Ala Asn Pro Ser His Leu 595 600 605
Glu Ala Val Asp Pro Val Leu Glu Gly Leu Val Arg Ala Lys Gin Asp 610 615 620
Leu Leu Asp His Gly Ser lie Asp Ser Asp Gly Gin Arg Ala Phe Ser 625 630 635 640
Val Val Pro Leu Met Leu His Gly Asp Ala Ala Phe Ala Gly Gin Gly 645 650 655
Val Val Ala Glu Thr Leu Asn Leu Ala Asn Leu Pro Gly Tyr Arg Val 660 665 670 -221 - 146966-序列表.doc 201037078
Gly Gly Thr lie His lie lie Val Asn Asn Gin lie Gly Phe Thr Thr 675 680 685
Ala Pro Glu Tyr Ser Arg Ser Ser Glu Tyr Cys Thr Asp Val Ala Lys 690 695 700
Met lie Gly Ala Pro lie Phe His Val Asn Gly Asp Asp Pro Glu Ala 705 710 715 720
Cys Val Trp Val Ala Arg Leu Ala Val Asp Phe Arg Gin Arg Phe Lys 725 730 735
Lys Asp Val Val lie Asp Met Leu Cys Tyr Arg Arg Arg Gly His Asn 740 745 750
Glu Gly Asp Asp Pro Ser Met Thr Asn Pro Tyr Val Tyr Asp Val Val 755 760 765
Asp Thr Lys Arg Gly Ala Arg Lys Ser Tyr Thr Glu Ala Leu lie Gly 770 775 780
Arg Gly Asp lie Ser Met Lys Glu Ala Glu Asp Ala Leu Arg Asp Tyr 785 790 795 800
Gin Gly Gin Leu Glu Arg Val Phe Asn Glu Val Arg Glu Leu Glu Lys 805 810 815
His Gly Val Gin Pro Ser Glu Ser Val Glu Ser Asp Gin Met lie Pro 820 825 830
Ala Gly Leu Ala Thr Ala Val Asp Lys Ser Leu Leu Ala Arg lie Gly 835 840 845
Asp Ala Phe Leu Ala Leu Pro Asn Gly Phe Thr Ala His Pro Arg Val 850 855 860
Gin Pro Val Leu Glu Lys Arg Arg Glu Met Ala Tyr Glu Gly Lys lie 865 870 875 880
Asp Trp Ala Phe Gly Glu Leu Leu Ala Leu Gly Ser Leu Val Ala Glu 885 890 895
Gly Lys Leu Val Arg Leu Ser Gly Gin Asp Ser Arg Arg Gly Thr Phe 900 905 910
Ser Gin Arg His Ser Val Leu lie Asp Arg His Thr Gly Glu Glu Phe 222 - 146966-序列表,doc 201037078 915 920 925
Thr Pro Leu Gin Leu Leu Ala Thr Asn Ser Asp Gly Ser Pro Thr Gly 930 935 940
Gly Lys Phe Leu Val Tyr Asp Ser Pro Leu Ser Glu Tyr Ala Ala Val 945 950 955 960
Gly Phe Glu Tyr Gly Tyr Thr Val Gly Asn Pro Asp Ala Val Val Leu 965 970 975
Trp Glu Ala Gin Phe Gly Asp Phe Val Asn Gly Ala Gin Ser lie lie 980 985 990 ❹
Asp Glu Phe lie Ser Ser Gly Glu Ala Lys Trp Gly Gin Leu Ser Asn 995 1000 1005
Val Val Leu Leu Leu Pro His Gly His Glu Gly Gin Gly Pro Asp 1010 1015 1020
His Thr Ser Ala Arg lie Glu Arg Phe Leu Gin Leu Trp Ala Glu 1025 1030 1035
Gly Ser Met Thr lie Ala Met Pro Ser Thr Pro Ser Asn Tyr Phe 1040 1045 1050
His Leu Leu Arg Arg His Ala Leu Asp Gly lie Gin Arg Pro Leu 1055 1060 1065 〇 lie Val Phe Thr Pro Lys Ser Met Leu Arg His Lys Ala Ala Val 1070 1075 1080
Ser Glu lie Lys Asp Phe Thr Glu lie Lys Phe Arg Ser Val Leu 1085 1090 1095
Glu Glu Pro Thr Tyr Glu Asp Gly lie Gly Asp Arg Asn Lys Val 1100 1105 1110
Ser Arg lie Leu Leu Thr Ser Gly Lys Leu Tyr Tyr Glu Leu Ala 1115 1120 1125
Ala Arg Lys Ala Lys Asp Asn Arg Asn Asp Leu Ala lie Val Arg 1130 1135 1140
Leu Glu Gin Leu Ala Pro Leu Pro Arg Arg Arg Leu Arg Glu Thr 1145 1150 1155 146966-序列表.doc -223- 201037078
Leu Asp 1160
Arg Tyr Glu Asn Val 1165
Lys Glu Phe Phe Trp Val Gin Glu 1170
Glu Pro 1175
Ala Asn Gin Gly Ala 1180
Trp Pro Arg Phe Gly Leu Glu Leu 1185
Pro Glu 1190
Leu Leu Pro Asp Lys 1195
Leu Ala Gly lie Lys Arg lie Ser 1200
Arg Arg Ala Met 1205
Ser Ala Pro 1210
Ser Ser Gly Ser Ser Lys Val His 1215
Ala Val 1220
Glu Gin Gin Glu lie 1225
Leu Asp Glu Ala Phe Gly 1230 <210〉122 <211> 3696 <212> DNA <213> 人工 <220> <223>結核分枝桿菌a-網戊二酸脫叛酶Kgd密碼子優化基因 <400> 122 atggctaata tctcctctcc gtttggtcag aatgaatggc tggtagaaga aatgtaccgt 60 aaattccgcg atgacccgtc ctctgtggac ccgtcctggc atgaattcct ggtagactac 120 agcccggagc cgaccagcca accggcagcg gaaccaaccc gcgttacttc tccgctggta 180 gcggaacgtg cagctgctgc cgcgcctcag gcgccgccta aaccggcgga tactgccgca 240 gccggtaacg gtgtggtggc cgcactggct gctaagactg cggttccgcc gccagcagaa 300 ggcgatgaag ttgcagtcct gcgcggtgcg gcggctgcag tggtgaaaaa catgagcgcg 360 tccctggagg taccgaccgc cacgagcgtg cgcgcggtcc ctgctaaact gctgattgat 420 aaccgtattg tgatcaacaa ccagctgaaa cgtacccgtg gtggcaagat ctccttcact 480 catctgctgg gttatgcact ggtacaagcg gttaagaaat tccctaacat gaaccgtcat 540 tacactgagg tcgacggtaa accgacggct gttactccgg cacacacgaa cctgggcctg 600 gcgatcgacc tgcaaggtaa agatggtaag cgctccctgg tagttgcggg tattaaacgt 660 tgcgaaacca tgcgtttcgc acaattcgta accgcctacg aggacattgt ccgccgtgct 720 cgtgatggca aactgaccac cgaagatttt gcgggcgtta ctattagcct gaccaaccca 780 ggcaccatcg gcaccgtgca cagcgtacct cgtctgatgc cgggccaagg tgcgattatc 840 ggtgtgggtg ccatggagta cccggcagaa tttcagggtg cttctgaaga gcgcatcgcc 900 -224- 146966·序列表.doc 960 960
Ο 201037078 gagctgggta ttggtaaact gatcaccctg acttctacct atgaccaccg catcattcag ggcgcagaat ccggtgactt cctgcgcact attcacgaac tgctgctgtc cgacggtttc tgggatgaag tttttcgtga actgagcatc ccatatctgc cagttcgctg gtccaccgac aatccggact ctatcgttga caaaaacgct cgcgtaatga acctgatcgc tgcttatcgt aatcgtggtc acctgatggc tgatacggat ccgctgcgcc tggataaagc tcgtttccgt tcccacccgg acctggaagt gctgacccat ggtctgactc tgtgggatct ggaccgcgtg ttcaaagtag atggtttcgc gggtgctcag tacaagaagc tgcgtgacgt gctgggtctg ctgcgtgatg cgtactgtcg tcacattggt gtggagtacg cccacattct ggatccggaa cagaaagaat ggctggagca gcgtgtcgag accaaacacg taaaaccgac cgtagcgcag cagaaatata tcctgtccaa actgaacgcc gccgaggctt tcgaaacttt cctgcagacc aagtacgtgg gccagaaacg cttcagcctg gagggtgcgg aaagcgttat tccgatgatg gatgcagcta tcgatcagtg cgcggaacat ggtctggatg aagtcgttat cggtatgccg caccgtggtc gcctgaacgt actggcaaac atcgtcggta aaccatattc tcagatcttc acggaattcg agggcaacct gaacccgtcc caagcccacg gctccggcga cgtaaaatat catctgggtg ctaccggcct gtatctgcag atgttcggtg ataacgacat ccaggtatct ctgactgcta acccgagcca cctggaggcg gttgatcctg ttctggaagg tctggttcgc gccaaacagg atctgctgga ccacggctct atcgacagcg atggccagcg tgcattcagc gttgtaccgc tgatgctgca tggcgacgcg gcgttcgccg gtcagggtgt cgtagcagaa actctgaacc tggcgaacct gcctggctat cgcgtgggtg gcaccattca catcatcgtt aacaaccaaa tcggtttcac cacggcaccg gagtatagcc gttctagcga atattgcacc gacgtagcca aaatgatcgg tgcgccgatc ttccatgtaa acggtgacga tccagaggcc tgcgtgtggg tggctcgtct ggccgtagac ttccgccagc gttttaagaa agatgtggtt atcgacatgc tgtgctaccg ccgtcgtggt cacaacgaag gtgatgatcc gtctatgact aacccgtatg tctatgacgt ggtggacacc aagcgtggtg cacgcaaatc ttacacggag gccctgatcg gtcgtggcga catctctatg aaagaagcgg aagacgctct gcgtgattac cagggtcagc tggaacgtgt gttcaatgag gtgcgtgagc tggaaaagca cggcgtacaa ccgtccgaat ccgtagagtc cgatcagatg atccctgctg gtctggcaac tgctgttgat aaaagcctgc tggcgcgtat cggcgacgca ttcctggcgc tgccgaatgg ctttaccgcg cacccgcgcg tacagccggt actggaaaaa cgtcgtgaaa tggcctacga aggtaaaatc gattgggcct tcggtgagct gctggccctg ggctctctgg tggctgaggg caagctggta 146966-序列表.doc •225 - 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 201037078 cgcctgagcg gccaggactc ccgtcgcggc actttttctc agcgtcacag cgtcctgatc 2760 gatcgtcaca ccggcgaaga attcacgccg ctgcaactgc tggctactaa ctccgatggt 2820 agcccgaccg gtggtaagtt cctggtgtac gattccccgc tgtccgaata tgctgcagtt 2880 ggtttcgagt atggttacac cgttggcaac ccggacgcag tggttctgtg ggaagcgcag 2940 ttcggcgatt tcgttaacgg tgcccagtcc attatcgatg agtttattag cagcggcgag 3000 gccaaatggg gccagctgtc taacgttgtg ctgctgctgc ctcacggcca cgagggtcaa 3060 ggcccggacc acacctccgc ccgtatcgaa cgcttcctgc agctgtgggc tgaaggctct 3120 atgaccatcg cgatgccgtc taccccaagc aactacttcc acctgctgcg tcgccacgca 3180 ctggacggca ttcagcgccc gctgatcgtt ttcaccccaa aatccatgct gcgccacaaa 3240 gcagctgttt ctgaaatcaa agattttacg gaaattaaat tccgttctgt gctggaagaa 3300 ccaacctacg aagacggtat tggcgaccgc aacaaggtaa gccgtatcct gctgacctcc 3360 ggcaaactgt actacgagct ggcagcacgt aaggcaaaag ataaccgcaa cgacctggcc 3420 atcgtccgcc tggaacagct ggcgccactg ccacgccgtc gcctgcgtga aaccctggat 3480 cgctacgaaa acgtaaaaga attcttctgg gtgcaggaag aaccggcaaa ccagggtgcg 3540 tggccgcgct ttggtctgga actgccggaa ctgctgccgg ataaactggc aggtatcaag 3600 cgcatcagcc gtcgcgctat gagcgccccg tcttctggta gctctaaagt acacgctgta 3660 gaacagcaag agatcctgga tgaggccttc ggctaa 3696 <210> 123 <211> 74 <212> DNA <213> 人工序列 <220> <223> 用於擴增枯草桿菌胺基轉移酶x之正向引子 <400> 123 ggggacaagt ttgtacaaaa aagcaggcta ggaggaatta accatgaagg ttttagtcaa 60 tggccggctg attg 74 <210> 124 <211> 62 <212> DNA <213> 人工 <220> <223> 用於擴增枯草桿菌胺基轉移酶x之反向引子 <400> 124 ggggaccact ttgtacaaga aagctgggtt tatgaaatgc tagcagcctg ttgaatgctt 60 •226- 146966·序列表.doc 62 201037078 tc <210> <211> <212> <213> <220> <223> 125 82
DNA 人工 用於擴增枯草桿菌胺基轉移酶y之正向引子 <400> 125 ggggacaagt ttgtacaaaa aagcaggcta ggaggaatta accatgactc atgatttgat 82 agaaaaaagt aaaaagcacc tc
O 〇 <210> 126 <211> 57 <212> DNA <213> 人工 <220> <223> 用於擴增枯草桿菌胺基轉移酶y之反向引子 <400> 126 ggggaccact ttgtacaaga aagctgggtt caatcttcaa <210> 127 <211> 64 <212> DNA <213> 人工 <220> <223> 用於擴增類球紅細菌胺基轉移酶之正向引子 <400> 127 ggggacaagt ttgtacaaaa aagcaggcta ggaggaatta cttg <210> 128 <211> 51 <212> DNA <213> 人工 64 <220> <223> 用於擴增類球紅細菌胺基轉移酶之反向引子 <400> 128 ggggaccact ttgtacaaga aagctgggtt cagacggcgg ccggttcttt c <210> 129 146966-序列表.doc - 227 - 201037078 <211> 78 <212> DNA <213> 人工 <220> <223>肖於擴增嗜肺性退伍軍人桿菌胺基轉移酶之正向引子 <400> 129 ggggacaagt y acaaaa aagcaggcta ggaggaatta accatgagta taacggcaag tattgttg tcgcatttgt 78 <210> 130 <211> 67 <212> DNA <213> 人工 <220> <223>用於擴增嗜肺性退伍軍人桿菌胺基轉移酶之反向引子 <400> 130 ggggaccact ttgtacaaga aagctgggtt tagtttacta gttgttggta attatcc ggaatcatta 67 <210> 131 <211> 76 <212> DNA <213> 人工 <220> <223> 用於擴增歐洲亞硝化單胞菌胺基轉移酶之正向引子 <400> 131 ggggacaagt ttgtacaaaa aagcaggcta ggaggaatta accatgattt caaatttctg ccgatg acctcaatgg 76 <210> 132 <211> 50 <212> DNA <213> 人工 <220> <223> 用於擴增歐洲亞硝化單胞菌胺基轉移酶之反向引子 <400〉 132 ggggaccact ttgtacaaga aagctgggtt tactggcgtg gagcatgccc <210> 133 <211> 79 <212> DNA <213> 人工 146966·序列表.doc 228- 201037078 Ο
<220> <223> 用於擴增淋病奈瑟菌胺基轉移酶之正向引子 <400> 133 ggggacaagt ttgtacaaaa aagcaggcta ggaggaatta accatgagga taaatatgaa ccgtaacgaa attttattc 79 <210> 134 <211> 56 <212> DNA <213> 人工 <220> <223> 用於擴增淋病奈瑟菌胺基轉移酶之反向引子 <400> 134 ggggaccact ttgtacaaga aagctgggtt catgcagcca tcgccttgaa cacttc <210> 135 <211> 66 <212> DNA <213> 人工 <220> <223> 用於擴增綠膿桿菌胺基轉移酶之正向引子 <400> 135 ggggacaagt ttgtacaaaa aagcaggcta ggaggaatta accatgtcga tggccgatcg tgatgg 66 <210> 136 <211> 53 <212> DNA <213> 人工 <220> <223〉 用於擴增綠膿桿菌胺基轉移酶之反向引子 <400> 136 ggggaccact ttgtacaaga aagctgggtt tacttgacca gggtacgcca etc <210> 137 <211> 67 <212> DNA <213> 人工 <220> <223>用於擴增沼澤紅假單胞菌胺基轉移酶之正向引子 229- 146966·序列表.doc 201037078 <400> 137 ggggacaagt ttgtacaaaa aagcaggcta ggaggaatta accatgaagc tgataccgtg 60 ccgcgcc 67 <210> 138 <211> 51 <212> DNA <213〉 人工 <220> <223> 用於擴增沼澤紅假單胞菌胺基轉移酶之反向引子 <400> 138 ggggaccact ttgtacaaga aagctgggtt caggcgaccg cgcggatcac C <210> 139 <211> 71 <212> DNA <213> 人工 <220> <223> 用於擴增枯草桿菌胺基轉移酶（gil6077991)之正向引子 <400> 139 ggggacaagt ttgtacaaaa aagcaggcta ggaggaatta accatggaga tgatggggat ggaaaacatt c 71 <210> 140 <211> 65 <212> DNA <213> 人工 <220> <223> 用於擴增枯草桿菌胺基轉移酶（gil6077991)之反向引子 <400> 140 ggggaccact ttgtacaaga aagctgggtt tatatcgttt gaaagctttc tttcaccgtt ttcac 65 <210> 141 <211> 66 <212> DNA <213> 人工 <220> <223> 用於擴增綠腺桿菌胺基轉移酶（gi9951072)之正向引子 <400> 141 51 ggggacaagt ttgtacaaaa aagcaggcta ggaggaatta accatgaacg caagactgca 60 60 60 M6966-序列表.doc -230· 66 201037078 cgccac <210> 142 <211> 48 <212> DNA <213> 人工 <220> <223>用於擴增綠膿桿菌胺基轉移酶（#9951072)之反向引子 <400> 142 ggggaccact ttgtacaaga aagctgggtt taccggtgac cggcgcgg <210> 143 <211> 69 <212> DNA <213> 人工
<220> <223>用於擴增綠膿桿菌胺基轉移酶（gi9951630)之正向引子 <400> 143 ggggacaagt ttgtacaaaa aagcaggcta ggaggaatta accatgacaa tgaatgacga 69 gccgcagtc <210> 144 <211> 49 <212> DNA <213> 人工 <220> <223>用於擴增綠膿桿菌胺基轉移酶（gi9951630)之反向引子 <400> 144 ggggaccact ttgtacaaga aagctgggtt cagacgctgg cgcggatgg <210> 145 <211> 1221 <212> DNA <213>詹氏曱烷球菌 <400> 145 atgacaaaag tgctggtgat gtttatggat ttcttatttg agaacagctg gaaagcagtt tgtccctaca atccaaagtt ggatttaaag gacatttata tttatgacac aaccctaaga gatggagagc aaaccccagg agtttgcttt accaaagaac aaaaattgga gattgcaagg aagttggatg aacttggatt aaagcagatt gaagctggct tcccaatagt atctgaaaga gaagcagata tagttaaaac aattgctaat gaagggctaa atgctgatat cttagcttta tgcagggctt taaagaaaga tatagataaa gcaatagagt gcgatgtaga tgggattatt -231 - 146966-序列表.doc 201037078 accttcatag caacatctcc tctccactta aaatataaat tcaacaacaa aagcttagat 420 gaaatattag agatgggagt tgaggcagtt gagtatgcaa aggaacatgg cttatttgtt 480 gctttctctg cagaggatgc gacaagaaca ccaatagagg acttgattaa agtgcataaa 540 gccgctgaag aggctggagc agatagggtt catatagcag acacaactgg ctgtgctacc 600 ccccaaagta tggagtttat atgtaaaaca ttgaaggaga acttaaaaaa ggcacatatt 660 ggagtgcatt gtcacaacga ctttggattt gcagttataa attcaatata tggtttaatt 720 ggaggagcta aggcagtttc aacaacagtt aatggtattg gagagagggc agggaatgca 780 gctttagaag agctaattat ggctttaact gtcttgtatg atgttgattt gggattaaac 840 ttggaggttc ttccagagtt atgcagaatg gttgaggaat actctggaat aaagatgcca 900 aagaacaaac caatagttgg agagcttgta tttgctcatg aaagtggaat tcacgttgat 960 gctgtcatag agaatccatt aacctatgaa cccttccttc cagagaaaat agggcttaag 1020 agaaatattt tgttagggaa gcattctgga tgcagagccg ttgcctataa gctaaaactt 1080 atgggaattg attacgatag agagatgttg tgcgagattg ttaaaaaggt taaagagatt 1140 agagaggaag gtaaatttat aactgatgaa gtctttaagg agattgttga agaagtttta 1200 aggaagagaa ataaaaatta a 1221 <210> 146 <211> 513 <212> DNA <213>詹氏甲烷球菌 <400> 146 atgattatta agggaagagc tcacaaattt ggggatgatg tagatacaga cgcaataatt 60 ccaggacctt acttaaggac tacagaccct tacgagttag cttcacactg catggcaggg 120 atagatgaaa acttcccgaa aaaggttaag gagggggatg tgatagttgc tggagagaat 180 tttggttgtg gttcaagtag ggagcaggct gtaatagcaa taaaatactg tggtattaag 240 gctgtgatag caaaaagctt tgcaagaata ttctatagaa atgcaataaa cgttggatta 300 ataccaataa tagcaaatac agatgaaatt aaagacggag acatagtaga gattgattta 360 gataaagaag agattgtaat aaccaataaa aacaaaacaa taaagtgtga aacaccaaaa 420 ggtttagaaa gagaaatatt ggctgctggt ggcttagtca attatttaaa aaagagaaaa 480 ctaatacaat caaaaaaagg tgtaaaaaca tga 513 <210> 147 <211> 1263 <212> DNA <213>詹氏曱烷球菌 146966-序列表.doc - 232 - 60 60
201037078 <400> 147 ttgacattgg tagagaagat actatcaaaa aaagttggtt atgaagtttg tgcaggagat agcatagagg ttgaagttga tttggcaatg acacacgatg gaacaacacc tttagcatac aaagctttaa aggaaatgag tgatagtgtt tggaatccag ataaaatagt cgttgccttt gaccacaatg ttccaccaaa cacagttaaa gctgctgaaa tgcaaaaatt agctttggag tttgttaaaa gatttggcat taaaaatttc cataaaggtg gagaaggcat ctgtcatcaa atcttagctg aaaattatgt tttgccaaac atgtttgtag ctggtggaga cagccataca tgcacacatg gagcttttgg agcttttgct actggctttg gagctactga tatggcttac atctatgcaa caggagaaac atggattaaa gtgccaaaaa caattagggt agatatagtt ggaaaaaatg aaaatgtttc tgccaaagat attgttttaa gggtttgtaa ggaaattggg agaagaggag caacatacat ggctattgag tatggtggag aggttgttaa aaacatggac atggatggaa ggctaacttt atgcaacatg gcaatagaga tgggaggaaa aacaggagtg atagaggctg atgaaattac ttatgattat ttaaagaaag agagaggact ttctgatgag gatatagcta aattaaaaaa agagagaata acagtaaata gagatgaagc aaactactat aaggagatag aaattgacat aacagatatg gaagaacaag ttgctgttcc acaccaccca gataacgtaa agccaattag tgatgttgaa gggactgaga taaatcaagt ttttattggg agttgcacaa atggaaggtt gagtgattta agagaagcag ctaaatattt aaaaggtagg gaggttcata aagatgttaa gctaattgtt atcccggcat caaaaaaggt atttttgcaa gcgttaaaag agggtattat agatatcttt gttaaagctg gggcgatgat ttgcactccg ggatgcggac cttgcttagg agctcatcaa ggggttttgg ctgagggaga aatttgttta tcaacaacaa acagaaactt taaaggaagg atggggcata taaatagcta tatttacttg gcatctccaa agattgccgc aataagtgca gttaagggat atataaccaa caaattggat taa 1263 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 <210> 148 <211> 1044 <212> DNA <213>詹氏甲炫球菌 <400> 148 atgatgaagg tgtgtgttat ataaaaatat taaatgagtt tgtttaaaaa aatatggtaa gatattattt tgtttggggc agaaggggat ggaataggaa gggagagttt gaaataataa tgcacttcca gaggatacaa tataacctca ccaaagccag aagaagtgat tccagaggcc aaggagaggc aggattagaa tagaaaaagc taaagaggca gggaagttca aaattataaa 60 120 180 • 233 - 146966-序列表.doc 240 201037078 agccctataa taacgttgag gaagatgttt catttatatg caaatgtaag accaataaac 300 aactttggaa ttggacaatt aattgggaaa attgcagatt atgaattctt aaatgctaag 360 aatattgata tagttattat aagagagaat acggaagatt tatatgttgg tagagagaga 420 ttagaaaatg atacagcaat agctgagagg gttataacaa gaaagggtag cgagagaata 480 ataagatttg catttgaata tgctataaaa aataatagga aaaaggtatc ttgcatccat 540 aaagctaatg ttttaagaat aactgatggt ttattcttag aggtttttaa tgaaataaaa 600 aaacattata atatagaggc agatgattat ttagttgatt caacagctat gaacttaata 660 aaacatcctg aaaaatttga tgttattgtt acaacaaaca tgtttgggga tattttatca 720 gatgaggcat ctgcattaat tggaggactt ggtttagctc cttcagcaaa tataggagat 780 gataaagcat tatttgagcc agttcatggt tcagctccag atatagctgg gaaaggtata 840 gcaaatccaa tggcatctat attaagtatt gctatgcttt ttgattatat tggagagaaa 900 gaaaagggag atttgattag agaggcagtg aaatactgct taataaacaa aaaagttact 960 cctgacttgg gaggggattt aaagacaaaa gatgttggag acgaaattct aaattacatt 1020 agaaagaagt taaagggata ttga 1044 <210> 149 <211> 1155
<212> DNA <213>棕色固氮菌高檸檬酸合成酶 <400> 149 atggctagcg tgatcatcga cgacactacc ctgcgtgacg gtgaacagag tgccggggtc 60 gccttcaatg ccgacgagaa gatcgctatc gcccgcgcgc tcgccgaact gggcgtgccg 120 gagttggaga tcggcattcc cagcatgggc gaggaagagc gcgaggtgat gcacgccatc 180 gccggtctcg gcctgtcgtc tcgcctgctg gcctggtgcc ggctatgcga cgtcgatctc 240 gcggcggcgc gctccaccgg ggtgaccatg gtcgaccttt cgctgccggt ctccgacctg 300 atgctgcacc acaagctcaa tcgcgatcgc gactgggcct tgcgcgaagt ggccaggctg 360 gtcggcgaag cgcgcatggc cgggctcgag gtgtgcctgg gctgcgagga cgcctcgcgg 420 gcggatctgg agttcgtcgt gcaggtgggc gaagtggcgc aggccgccgg cgcccgtcgg 480 ctgcgcttcg ccgacaccgt cggggtcatg gagcccttcg gcatgctcga ccgcttccgt 540 ttcctcagcc ggcgcctgga catggagctg gaagtgcacg cccacgatga tttcgggctg 600 gccacggcca acaccctggc cgcggtgatg ggcggggcga ctcatatcaa caccacggtc 660 aacgggctcg gcgagcgtgc cggcaacgcc gcgctggaag agtgcgtgct ggcgctcaag 720 aacctccacg gtatcgacac cggtatcgat acccgcggca tcccggccat ctccgcgctg 780 -234- 146966·序列表.doc 201037078 gtcgagcggg cctcggggcg ccaggtggcc tggcagaaga gcgtggtcgg cgccggggtg 840 ttcactcacg aggccggtat ccacgtcgac ggactgctca agcatcggcg caactacgag 900 gggctgaatc ccgacgaact cggtcgcagc cacagtctgg tgctgggcaa gcattccggg 960 gcgcacatgg tgcgcaacac gtaccgcgat ctgggtatcg agctggcgga ctggcagagc 1020 caagcgctgc tcggccgcat ccgtgccttc tccaccagga ccaagcgcag cccgcagcct 1080 gccgagctgc aggatttcta tcggcagttg tgcgagcaag gcaatcccga actggccgca 1140 ggaggaatgg catga 1155 <210> 150 <211> 30 <212> DNA <213> 人工
<220> <223> Avine-WT-R-BamHI <400> 150 aaattggatc ctcatgccat tcctcctgcg 30 <210> 151 <211> 75 <212> DNA <213> 人工 <220> <223> Avine-WT-F-SacI <400> 151 aaattgagct ctttctccat acccgttttt ttgggctaac aggaggaatt aaccatggct 60 agcgtgatca tcgac 75
<210> 152 <211> 31 <212> DNA <213> 人工 <220> <223> Avine-WT-R-Hindlll <400> 152 aaattaaagc tttcatgcca ttcctcctgc g 31 <210> 153 <211> 76 <212〉 DNA <213> 人工 <220> <223> Avine-WT-F-Hindll 工 146966·序列表.doc -235· 201037078 <400> 153 aaattaaagc tttttctcca tacccgtttt tttgggctaa caggaggaat taaccatggc tagcgtgatc atcgac Ί6 <210> <211> <212> <213> 154 21 DNA 人工 <220> <223> AksA-Avine-F <400> 154 atggctagcg tgatcatcga c 21 <210> <211> <212> <213> 155 32 DNA 人工 <220> <223> AksA-Avine~Rl <400> 155 aaattggcgc gcctcatgcc attcctcctg eg 32 <210> <211> <212> <213> 156 32 DNA 人工 <220> <223> Pgal2-F2 <400> 156 aaattgttaa ctccagaagg cacatctatt ac 32 <210> <211> <212> <213> 157 49 DNA 人工 <220> <223> Pgal2-R <400> 157 egtegatgat cacgctagcc attatgaaag cctccttttt tttattatg
<210> <211> <212> <213> 158 207 DNA 人工 <220> <223> mtSP 236- 146966-序列表.doc 60201037078 <400> 158 atggcctcca ctcgtgtcct cgcctctcgc ctggcctccc agatggctgc ttccgccaag gttgcccgcc ctgctgtccg cgttgctcag gtcagcaagc gcaccatcca gactggctcc cccctccaga ccctcaagcg cacccagatg acctccatcg tcaacgccac cacccgccag gctttccaga agcgcgccta ctcttcc 207 120 180 <210> 159 <211> 29 <212> DNA <213> 人工
<220> <223> pF113-F-NsiI
<400> 159 aaattatgca tacagcatgg cctgcaacg <210> 160 <211> 31 <212> DNA <213> 人工 29 <220> <223> pF113-R-AgeI <400> 160 aaattaccgg tcagggttat tgtctcatga g 31 <210> 161 <211> 32 <212> DNA <213> 人工 o <220> <223> AT-Vfl_for_Ec <400> 161 aaatttggta ccgctaggag gaattaacca tg 32 <210> 162 <211> 33 <212> DNA <213> 人工 <220> <22 3> Kdc—for—Ec <400> 162 aaatttacta gtggctagga ggaattacat atg 33 <210> 163 <211> 35 146966·序列表.doc 237- 201037078 <212> DNA <213> 人工 <220> <223> Kdc rev Ec <400> 163 aaatttaagc ttattacttg ttctgctccg caaac 35 <210> 164 <211> <212> <213> 57 DNA 人工 <220> <223> AT-Vfl-F <400> 164 aaatttacta gtaagaattt ttgaggaggc aatataaatg aataaaccac agtcttg 57 <210> 165 <211> <212> <213> 32 DNA 人工 <220> <223> AT-Vfl-R <400> 165 aaatttggat cctacaagaa agctgggttt ac 32 <210> 166 <211> <212> <213> 33 DNA 人工 <220> <223> AT-Vfl rev Ec <400> 166 aaatttacta gtaagctggg tttacgcgac ttc 33 <210> 167 <211> <212> <213> 1221 DNA 腸桿菌 <400> 167 atgaccaaag ttctggtaat gttcatggac ttcctgttcg aaaactcctg gaaagcggtt 60 tgcccgtaca acccgaaact ggatctgaaa gacatctaca tctacgacac cactctgcgt 120 gacggtgaac agactccggg cgtttgcttc accaaagagc agaagctgga aatcgctcgt 180 aagctggacg aactgggtct gaagcagatc gaagctggct tcccgatcgt ttctgaacgt 240 gaagctgaca tcgttaaaac tatcgctaac gaaggtctga acgctgacat cctggcactg 300 •238 · 146966-序列表.doc 360 360
201037078 tgccgtgcgc tgaagaaaga catcgacaaa,gcaatcgaat gcgacgttga cggtatcatc actttcatcg caacttctcc gctgcacctg aaatacaaat tcaacaacaa atctctggat gaaatcctgg aaatgggcgt tgaagcggta gaatacgcta aagagcacgg tctgttcgtt gcattctctg cagaagatgc aactcgtact ccgatcgaag atctgatcaa agttcacaaa gcagctgaag aagcgggtgc tgaccgcgtt cacatcgctg acaccactgg ctgcgcaact ccgcagtcta tggaattcat ctgcaaaact ctgaaagaaa acctgaagaa agcacacatc ggcgtacact gccacaacga cttcggtttc gctgttatca actccatcta cggtctgatc ggtggtgcga aagcggtatc tactaccgtt aacggtatcg gtgaacgtgc tggtaacgct gcactggaag agctgatcat ggcgctgacc gtactgtacg acgttgacct gggtctgaac ctggaagttc tgccggaact gtgccgtatg gttgaagaat actccggtat caagatgccg aaaaacaagc caatcgttgg tgaactggta ttcgctcacg aatccggtat ccacgttgac gctgttatcg aaaacccgct gacttacgaa ccgttcctgc cggaaaaaat cggtctgaaa cgtaacatcc tgctgggtaa gcactctggt tgccgtgctg ttgcttacaa gctgaaactg atgggtatcg actacgaccg tgaaatgctg tgcgaaatcg ttaagaaagt taaagaaatc cgtgaagaag gtaaattcat cactgacgaa gttttcaaag agatcgttga agaagttctg cgtaagcgta acaaaaacta a 1221 <210> 168 <211> 1044 <212> DNA <213> AksF_^勝桿菌 <400> 168 atgatgaaag tttgcgttat cgaaggtgac ggtatcggta aagaagttat cccggaagct atcaagatcc tgaacgaact gggtgaattc gaaatcatca aaggtgaagc gggtctggaa tgcctgaaga aatacggtaa cgcactgcca gaagatacca tcgaaaaagc gaaagaagct gacatcatcc tgttcggtgc aatcacttct ccgaagccgg gtgaagttca gaactacaaa tctccgatca tcactctgcg taagatgttc cacctgtacg ctaacgtacg tccgatcaac aacttcggta tcggtcagct gatcggtaag atcgctgact acgagttcct gaacgctaaa aacatcgaca tcgttatcat ccgtgaaaac actgaagatc tgtacgttgg tcgtgaacgt ctggaaaacg acactgctat cgctgagcgc gttatcactc gtaaaggttc tgaacgtatc atccgcttcg cattcgaata cgcaatcaaa aacaaccgta agaaagtttc ctgcatccac aaagctaacg tactgcgtat cactgacggt ctgttcctgg aagtattcaa cgaaatcaag aaacactaca acatcgaagc tgacgactac ctggttgact ccactgcaat gaacctgatc 146966-序列表.doc -239- 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 60 120 180 240 300 360 420 480 540 600 660 201037078 aagcacccgg aaaaattcga cgttatcgtt accactaaca tgttcggtga catcctgtct 720 gacgaagcgt ctgcactgat cggtggtctg ggtctggcac cgtctgctaa catcggtgac 780 gacaaagcgc tgttcgaacc ggttcacggt tctgcaccgg atatcgctgg taaaggtatc 840 gctaacccga tggcttctat cctgtctatc gcgatgctgt tcgactacat cggtgaaaaa 900 gagaaaggcg acctgatccg tgaagcggta aaatactgcc tgatcaacaa gaaagttact 960 ccggatctgg gtggtgacct gaaaaccaaa gacgttggtg acgaaatcct gaactacatc 1020 cgtaagaaac tgaaaggtta ctaa 1044 <210> 169 <211> 1263 <212> DNA <213> AksD#大腸桿菌 <400> 169 atgactctgg ttgagaagat cctctccaag aaagttggtt acgaagtttg cgcaggcgac 60 tccatcgaag ttgaagttga cctggcgatg actcacgacg gtactactcc gctggcttac 120 aaagcgctga aagagatgtc tgactccgta tggaacccgg acaagatcgt tgttgcattc 180 gaccacaacg taccgccgaa caccgttaaa gcagctgaaa tgcagaagct ggcgctggaa 240 ttcgttaagc gcttcggtat caaaaacttc cacaaaggtg gtgaaggtat ctgccaccag 300 atcctggctg aaaactacgt tctgccgaac atgttcgttg ctggcggcga ctctcacacc 360 tgtactcacg gtgcattcgg tgcattcgca actggcttcg gtgcaactga catggcttac 420 atctacgcaa ctggcgaaac ctggatcaaa gttccgaaaa ctatccgcgt tgatatcgtt 480 ggtaaaaacg aaaacgtatc tgcgaaagac atcgttctgc gcgtttgcaa agaaatcggt 540 cgtcgcggtg caacttacat ggctatcgaa tacggtggtg aagttgttaa aaacatggac 600 atggacggtc gtctgactct gtgcaacatg gctatcgaaa tgggtggtaa aactggcgtt 660 atcgaagctg acgaaatcac ttacgactac ctgaagaaag agcgtggtct gtctgacgaa 720 gatatcgcta aactgaagaa agagcgtatc accgttaacc gtgacgaagc taactactac 780 aaagaaatcg aaatcgacat cactgacatg gaagaacagg ttgctgtacc gcaccacccg 840 gataacgtta agccaatctc tgacgttgaa ggtactgaaa tcaaccaggt attcatcggt 900 tcctgcacca acggtcgtct gtctgatctg cgtgaagctg cgaaatacct gaaaggtcgt 960 gaagttcaca aagacgttaa gctgatcgtt atcccggctt ccaagaaagt attcctgcag 1020 gcgctgaaag aaggtatcat cgacatcttc gttaaagcgg gtgcgatgat ctgtactccg 1080 ggttgcggtc cgtgcctggg tgcacaccag ggcgtactgg cagaaggtga aatctgcctg 1140 tctactacca accgtaactt caaaggtcgt atgggtcaca tcaactctta catctacctg 1200 .240- 146966-序列表.doc 1260 201037078 gcttctccga aaatcgctgc tatctctgct gttaaaggtt acatcactaa caagctggat taa 1263 o <210> 170 <211> 513 <212> DNA <213> AksE_ <400> 170 atgatcatca ccaggtccgt atcgacgaaa ttcggttgcg gcggttatcg atcccgatca gataaagaag ggtctggaac ctgattcagt .大腸桿菌 aaggtcgtgc acctccgtac acttcccgaa gttcttcccg ctaaatcttt tcgctaacac aaatcgttat gtgaaatcct ccaagaaagg gcacaagttc tactgacccg gaaagttaaa tgagcaggct cgcacgtatc cgacgaaatc cactaacaaa ggcagctggc cgtaaaaact ggtgacgacg tacgaactgg gaaggtgacg gttatcgcta ttctaccgta aaagacggtg aacaaaacta ggtctggtta taa ttgacactga catctcactg ttatcgttgc tcaaatactg acgcaatcaa acatcgttga tcaagtgcga actacctgaa cgctatcatc catggcgggt tggcgaaaac cggtatcaaa cgtaggtctg aatcgacctg aactccgaaa gaaacgtaag 513 60 120 180 240 300 360 420 480 o <210> 171 <211> 1221 <212> DNA <213> 从5八_釀酒酵母 <400> 171 atgaccaagg ttttggtcat gttcatggac ttcttgtttg aaaactcctg gaaggccgtt tgtccataca acccaaagtt ggacttgaag gacatctaca tctacgacac cactttaaga gatggtgaac aaaccccagg tgtttgtttc accaaggaac aaaaattgga aattgccaga aagttggacg aattgggttt gaaacaaatc gaagctggtt tcccaatcgt ttctgaaaga gaagctgaca ttgtcaagac cattgccaac gaaggtttga acgctgatat cttagctcta tgtagagctt tgaagaagga cattgacaag gccatcgaat gtgatgtcga tggtatcatc actttcattg ctacttctcc attacatttg aaatacaagt tcaacaacaa atctttggac gaaatcttgg aaatgggtgt tgaagctgtc gaatacgcca aggaacacgg tttattcgtt gctttctctg ctgaagatgc taccagaact ccaattgaag atttgatcaa ggtccacaag gctgctgaag aagctggtgc tgaccgtgtc cacattgctg acaccactgg ttgtgccact ccacaatcca tggaatttat ctgtaagact ttgaaggaaa acttgaagaa ggctcacatt ggtgttcact gtcacaacga tttcggtttc gctgtcatca actccatcta cggtttgatt 146966-序列表.doc 60 120 180 240 300 360 420 480 540 600 660 -241 · 720 201037078 ggtggtgcca aggccgtttc caccaccgtc aacggtatcg gtgaaagagc tggtaacgct 780 gctttggaag aattgatcat ggctttgact gtcttatacg atgtcgattt gggtttgaac 840 ttggaagttt tgccagaatt gtgtagaatg gttgaagaat actctggtat caagatgcca 900 aagaacaagc caattgtcgg tgaattggtt ttcgctcatg aatctggtat tcacgttgac 960 gctgtcattg aaaacccatt gacctacgaa cctttcttgc cagaaaagat cggtttgaag 1020 agaaacatcc tattaggtaa gcactctggt tgtcgtgctg ttgcttacaa attgaaattg 1080 atgggtattg actacgacag agaaatgttg tgtgaaattg tcaagaaggt caaggaaatc 1140 agagaagaag gtaagttcat cactgacgaa gttttcaagg aaatcgttga agaagttttg 1200 agaaagagaa acaaaaatta a 1221 <210> 172 <211> 1263 <212> DMA <213V从8〇_釀酒酵母 <400> 172 atgactttag tcgaaaagat cttatccaag aaggtcggtt acgaagtttg tgccggtgac 60 tctattgaag ttgaagttga cttggccatg acccacgacg gtactacccc attggcttac 120 aaggctttga aggaaatgtc tgactccgtc tggaacccag acaagattgt tgttgctttc 180 gaccacaacg ttccaccaaa caccgtcaag gctgctgaaa tgcaaaaatt ggctttggaa 240 tttgtcaaga gattcggtat caagaacttc cacaagggtg gtgaaggtat ctgtcaccaa 300 atcttggctg aaaactacgt tttgccaaac atgttcgttg ctggtggtga ctcccacact 360 tgtacccacg gtgctttcgg tgcctttgct accggtttcg gtgctactga catggcttac 420 atctacgcta ccggtgaaac ctggatcaag gttccaaaga ctatcagagt tgacattgtc 480 ggtaagaacg aaaacgtttc tgccaaggat atcgtcttga gagtttgtaa ggaaattggt 540 agaagaggtg ctacttacat ggccattgaa tacggtggtg aagttgtcaa gaacatggac 600 atggacggta gattgacttt gtgtaacatg gccattgaaa tgggtggtaa gactggtgtc 660 attgaagctg atgaaatcac ctacgactac ttgaagaagg aaagaggtct atccgatgaa 720 gatatcgcca aattgaagaa ggaaagaatc actgttaaca gagatgaagc taactactac 780 aaggaaattg aaattgatat cactgacatg gaagaacaag ttgctgttcc tcatcaccca 840 gacaatgtca agccaatttc tgacgtcgaa ggtactgaaa tcaaccaagt tttcatcggt 900 tcttgtacca acggtagatt atctgattta cgtgaagctg ctaagtactt gaaaggtcgt 960 gaagttcaca aggatgtcaa attgattgtc attccagctt ccaagaaggt tttcttgcaa 1020 gctttgaagg aaggtatcat cgatatcttc gtcaaggctg gtgccatgat ctgtacccca 1080 -242· 146966-序列表.doc 201037078 ggttgtggtc tccaccacca gcttctccaa taa catgtttggg acagaaactt agattgctgc tgctcatcaa caagggtaga catttctgct ggtgtcttgg atgggtcaca gtcaagggtt ctgaaggtga tcaactctta acatcactaa aatctgtttg catctacttg caaattggat 1263 1140 1200 1260
<210> 173 <211> 513 <212> DNA <213> AksE_ <400> 173 atgatcatca ccaggtccat attgacgaaa tttggttgtg gctgtcattg attccaatca gacaaggaag ggtttggaaa ttgatccaat .釀酒酵母 agggtcgtgc acttgagaac acttcccaaa gttcttccag ccaagtcttt ttgctaacac aaattgttat gagaaatctt ccaagaaggg tcacaagttc cactgaccca gaaggtcaag agaacaagct cgctagaatc tgacgaaatc caccaacaag ggctgctggt tgtcaaaacc ggtgacgatg tacgaattgg gaaggtgatg gttattgcca ttctacagaa aaggatggtg aacaagacca ggtttagtca taa ttgacactga cttctcactg tcattgttgc tcaaatactg acgccatcaa acattgttga tcaagtgtga actacttgaa tgctatcatt tatggctggt tggtgaaaac tggtatcaag cgttggtttg aatcgatttg aactccaaag gaagagaaag 513 60 120 180 240 300 360 420 480 <210> 174 <211> 1044 <212> DNA <213> AksF_釀酒酵母 o <400> 174 atgatgaagg tttgtgtcat tgaaggtgac ggtattggta aggaagtcat tccagaagct 60 atcaagatct tgaatgaatt gggtgaattt gaaatcatca agggtgaagc tggtttggaa 120 tgtttgaaga aatacggtaa cgctttgcca gaagatacca ttgaaaaggc caaggaagct 180 gatatcatct tattcggtgc catcacttct ccaaagccag gtgaagttca aaactacaaa 240 tctccaatca tcactttgag aaagatgttc cacttgtacg ctaacgtcag accaatcaac 300 aacttcggta ttggtcaatt gattggtaag attgctgact acgaattttt gaatgccaag 360 aacattgaca ttgtcatcat cagagaaaac actgaagatt tgtacgttgg tcgtgaaaga 420 ttagaaaacg acactgccat tgctgaacgt gttatcacca gaaagggttc tgaaagaatc 480 atcagattcg ctttcgaata cgccatcaag aacaacagaa agaaggtttc ctgtatccac 540 aaggctaacg ttttgagaat caccgatggt ttattcttgg aagttttcaa cgaaatcaag 600 aagcactaca acattgaagc tgatgactac ttggttgact ccactgctat gaacttgatc 660 146966-序列表.doc -243- 201037078 aagcatccag aaaagttcga tgtcattgtc accaccaaca tgttcggtga catcttatct gacgaagctt ctgctttgat tggtggtcta ggtttggctc catctgccaa cattggtgat gacaaggctt tattcgaacc tgttcacggt tctgctccag acattgctgg taagggtatt gccaacccaa tggcttccat cttgtccatt gctatgttgt tcgactacat cggtgaaaag gaaaagggtg acttgatcag agaagctgtc aaatactgtt tgatcaacaa gaaggttact ccagatttgg gtggtgactt gaaaaccaag gatgtcggtg acgaaatctt gaactacatc agaaagaaat tgaaaggcta ctaa 1044 <210> 175 <211> 52 <212> DNA <213> 人工 <220> <223> DC-KdcA-F <400> 175 aaatttggat ccgttgagga ggcctcaaaa atgtatactg ttggtgatta tc <210> 176 <211> 37 <212> DNA <213> 人工 <220〉 <223> DC-KdcA-R <400> 176 aaatttggcg cgccattact tgttctgctc cgcaaac 37 <210> 177 <211> 1161 <212> DNA <213> 人工 <220> <223〉針對大腸桿菌優化之AksA基因密碼子對 <400> 177 atggactgga aagcggtatc tccgtacaac ccgaaactga acctgaaaga ctgctacctg tacgacacca ctctgcgtga cggcgagcag actccgggcg tttgcttcac tcacgaccag aaactggaaa tcgcgaagaa actggacgaa ctgaaaatca agcagatcga agctggcttc ccgatcgttt ctgaaaacga acgtaaagca atcaagtcta tcaccggtga aggtctgaac gctcagatcc tggcactctc tcgcgtactg aaagaagata tcgacaaagc aatcgaatgc gacgttgacg gtatcatcac tttcatcgct gcttctccga tgcacctgaa atacaaactg •244- 720 780 840 900 960 1020 52 60 120 180 240 300 146966-序列表.doc 360 201037078
cacaaatctc cacggtctgt atccgtatcc ctgggttgcg aagaaagcgc atctacggtc cgtgctggta gacatgggtc aagatccgta ggtatccacg aaaatcggtc caccgtctgc aaaaccaaag gttgacaaaa tggatgaagt tcgttgcatt acaaaaacgc caactccgca acatcggtgt tgatcggtgg acgctgctat tgaacactga tcccggaaaa ttgacgctgt agaagcgtaa aggaactggg aaactcgtga tcattaaata tgaagagatg ctctgctgaa tgaagagcac ggcaatgtac tcactgccac tgcgaaagcg cgaagaaatc aatcctgact caagccgctg tctggaaaac gatcatcctg tctggaagca agaaggtact a ggtatgaaag gatgcaactc ggtgctaacc cacatctgct aacgacttcg gtatctacta gttatggcgc gaaatctcca gttggtgaaa ccgctgactt ggtaagcact tctcgtgaag gaaatctctg cggtagaata gtactccggt gcgttcacat ctgaactgtc gtttcgctgt ccgttaacgg tgaaagttct agctggttga tggcattcta acgaaccgtt ctggttgccg agctgtggga acgaagtatt cgctaaagac tgaagatctg cgctgacact ctccaacctg tatcaactcc tatcggtgaa gtacgaccac aaactactcc ccacgaatcc cctgccagaa tgctgttgct aatcgttaag caaaaacatc 1161 420 480 540 600 660 720 780 840 900 960 1020 1080 1140
<210> 178 <211> 1020 <212> DNA <213> 人工 <220> <223>針對大腸桿菌優化之AksF基因密碼子對 <400> 178 * atgcgtaaca ctccgaaaat ctgcgttatc aacggtgacg gtatcggtaa cgaagttatc ccggaaaccg ttcgcgtact gaacgaaatc ggtgacttcg aattcatcga aactcacgct ggttacgaat gcttcaagcg ctgcggtgac gctatcccgg aaaaaactat cgaaatcgct aaagagtctg actccatcct gttcggttct gtaactactc cgaagccgac tgaactgaaa aacaagccgt accgttctcc gattctgact ctgcgtaaag agctggatct gtacgctaac atccgtccga ctttcaactt caaaaacctg gacttcgtta tcatccgtga aaacactgaa ggtctgtacg ttaagaaaga atactacgac gaaaaaaacg aagttgcaac tgctgaacgt atcatctcca aattcggttc ttcccgtatc gttaagttcg cattcgacta cgcactgcag aacaaccgta agaaagtttc ctgcatccac aaagctaacg ttctgcgtat cactgacggt ctgttcctgg gcgtattcga agaaatctcc aagaaatacg agaagctggg tatcgtttct gacgactacc tgatcgacgc aactgcgatg tacctgatcc gtaacccgca gatgttcgac gtaatggtta ccactaacct gttcggtgac atcctgtctg acgaagctgc tggtctgatc 146966-序列表.doc 60 120 180 240 300 360 420 480 540 600 660 720 -245 - 201037078 ggtggtctgg gtatgtcccc gtctgctaac atcggtgaca aaaacggtct gttcgaaccg 780 gttcacggtt ctgcaccgga tatcgctggt aaaggtatct ccaacccaat cgcgactatc 840 ctgtctgctg caatgatgct ggatcacctg aaaatcaaca aagaagctga atacatccgt 900 aacgctgtta agaaaaccgt tgaatgtaaa tacctgactc cggacctggg tggtcacctg 960 aaaacttctg aagttactga aaaaatcatc gaatccatca aatctcagat gattcagtaa 1020 <210> 179 <211> 1257 <212> DNA <213〉 人工 <220> <223>針對大腸桿菌優化之AksD基因密碼子對 <400> 179 atgactctgg ctgaaaaaat catctccaaa aacgttggta aaaacgttta cgctggcgac 60 tccgttgaaa tcgacgttga cgttgcgatg actcacgacg gtactactcc gctgaccgtt 120 aaagcattcg agcagatctc tgacaaagta tgggataacg aaaaaatcgt tatcatcttc 180 gaccacaaca tcccggctaa cacctctaaa gctgctaaca tgcaagttat cactcgtgaa 240 ttcatcaaga agcagggtat caaaaactac tacctggacg gtgaaggtat ctgccaccag 300 gttctgccgg aaaaaggtca cgttaagccg aacatgatca tcgctggtgc tgactctcac 360 acctgtactc acggtgcatt cggtgcattc gcaactggct tcggtgcaac tgacatgggt 420 tacgtttacg caactggtaa aacctggctg cgcgtaccag aaaccattca ggttaacgta 480 actggcgaaa acgaaaacat ctccggtaaa gacatcatcc tgaaaacctg taaagaagtt 540 ggtcgtcgcg gtgcaactta cctctctctg gaatacggtg gtaacgcggt acagaacctg 600 gatatggacg aacgtatggt tctgtctaac atggctatcg aaatgggtgg taaagcgggt 660 atcatcgaag ctgacgacac cacttacaaa tacctggaaa acgctggcgt ttcccgtgaa 720 gaaatcctga acctgaagaa aaacaagatc aaagttaacg aatctgaaga aaactactac 780 aaaactttcg agttcgacat cactgacatg gaagagcaga tcgcttgccc gcaccacccg 840 gacaacgtta aaggcgtttc tgaagtttct ggtatcgaac tggatcaggt attcatcggt 900 tcctgcacca acggtcgtct gaacgatctg cgtatcgctg cgaagcacct gaaaggtaag 960 aaagttaacg aatccactcg tctgatcgtt atcccggctt ccaagtctat cttcaaagaa 1020 gcgctgaaag aaggtctgat cgacaccttc gttgactccg gtgcgctgat ctgtactccg 1080 ggttgcggtc cgtgcctggg tgcacaccag ggcgtactgg gtgacggtga agtttgcctg 1140 gcaactacca accgtaactt caaaggtcgt atgggtaaca ccaagtctga agtttacctc 1200 -246· 146966-序列表.doc 201037078 1257 tcttctccgg caatcgctgc gaagtctgct gttaaaggtt acatcactaa cgagtaa <210> 180 <211> 486 <212> DNA <213> 人工 <220> <223>針對大腸桿菌優化之AksE基因密碼子對 <400> 180
atgaagatca ccggtaaagt tcacgtattc ccgggtgctt acctgaaaac cactgacgaa atcgacgaag atttcccgga aatggttaaa ttcggttgcg gttcttcccg tgagcaggca gcaatcatcg ttgaatcctt cgcacgtatc ttcccgatcg aatgtaaagg tatctccaag gatctggaaa acaagaaagt tatcctgaaa ggtactgcga aagacatcat ggacgaaggt aactaa <210> 181 <211> 1161 <212> DNA <213> 人工 <220> <223> AksA海沼甲烷球菌 <400> 181 atggattgga aagctgtatc tccgtacaac tatgatacga cattgagaga tggtgaacag aaacttgaga tcgccaaaaa actggatgaa ccaattgttt ctgaaaacga gagaaaagcc gcacaaattt tggcgttatc aagagtttta gatgttgatg gaataattac attcattgca cacaaaagcc tcgatgaagt cgaagaaatg cacggacttt tcgtagcatt ctctgcagaa atcagaatcc acaaaaatgc agaagaacac ctcgggtgtg caacaccaca ggcaatgtat aaaaaagcac atatcggggt acactgtcac ggtgacgaca tcgacactga cgctatcatt tacgaactgg cttctcactg catggcgggt gaaggtgact tcctggttgc tggcgaaaac ccgatcgcta tcaaatactg cggtatcaaa ttctaccgta actgcatcaa cctgggcgta cacgttaaag acggtgacct gatcgaactg gacaaagttc tggactgcca catcccgact ggtctgatca actacgctaa gaagcagaaa 486 cctaaattaa atttgaaaga ctgttatttg actcccggag tttgttttac acatgatcaa cttaaaatta aacagatcga agcgggtttt atcaaatcaa ttactggcga aggattaaat aaagaggata ttgataaagc cattgaatgt gcttcaccaa tgcatttgaa atacaaattg ggtatgaaag ccgttgaata cgcaaaagat gatgcgacaa gaactcctgt tgaagacctc ggtgccaata gggtgcatat tgcagatacc catatctgct ctgaattaag cagtaacttg aacgactttg ggttcgcagt tataaactcg 60 120 180 240 300 360 420 480 60 120 180 240 300 360 420 480 540 600 •247· 146966-序列表.doc 660 201037078 atatacggat taattggtgg agcaaaagcg gtatctacaa cagttaacgg aataggcgaa agagcaggaa atgctgcaat tgaagaaatt gtaatggcat tgaaagtact ttacgaccac gatatgggat taaatactga aatactaact gaaatatcga aactcgttga aaactattca aaaattagga ttcccgaaaa taaacctctt gttggggaaa tggcatttta ccatgaaagc ggaatacatg ttgatgcggt tttagagaat cctttaacgt atgaaccgtt tttacctgaa aaaataggtc aaaaaagaaa aattatactt ggaaaacatt ccggatgcag agcagttgca cacagactgc aagaacttgg gcttgaagct tcaagagaag aactttggga aattgtgaaa aaaactaaag aaaccagaga agaaggtact gaaataagcg acgaagtgtt taaaaacatt gtcgataaga ttataaaata a 1161 <210> 182 <211> 1020 <212> DNA <213> 人工 <220> <223> AksF海沼甲烷球菌 <400> 182 atgagaaaca ctcccaaaat ttgtgttatt aatggagatg gcattggaaa cgaagtgatt cctgaaacag tgcgcgtctt gaatgaaatt ggggattttg aatttataga aacacatgcg ggctacgaat gttttaaaag atgtggcgat gcgatacctg aaaagaccat agaaattgca aaagaatctg attctattct ttttggatct gttactaccc caaaaccaac tgaattaaaa aataaaccct atagaagtcc aatattaact ttaagaaaag aactcgacct ttatgcaaat ataagaccga ctttcaactt caaaaacctt gattttgtga taattcgcga aaataccgaa ggtctttatg tgaaaaaaga atattacgac gaaaaaaatg aagttgcgac tgctgaacga attatttcta aatttggaag ctcgagaatt gtaaaatttg cttttgatta tgcacttcaa aacaatagaa aaaaagtatc ctgtattcac aaagcaaatg ttttgaggat cacagatggg ttattcctag gggtatttga agaaatatcg aaaaaatatg aaaaattggg aatagtgtct gatgactatt tgattgatgc aacagcgatg tatttaatta gaaatccgca aatgtttgat gtcatggtta caacaaattt atttggagat attttatcgg atgaagctgc tggacttatc ggaggacttg gaatgtctcc ttcagcaaat attggtgaca aaaacggatt attcgaacca gtgcatggat ccgcaccaga tattgctgga aaaggaattt caaacccgat tgcaacaatt ttaagtgctg caatgatgct tgatcattta aaaataaata aagaagcgga atacataaga aatgcagtta aaaaaactgt tgaatgtaaa tacctaactc cggatcttgg gggacactta 146966-序列表.doc -248- 720 780 840 900 960 1020 1080 1140 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 201037078 aaaacttctg aagttacaga aaaaatcatt gaatcaataa aatctcaaat gattcaatga <210> 183 <211> 1257 <212> DNA <213> 人工 <220> <223> AksD海沼曱娱;球菌 <400> 183 atgacacttg ctgaaaaaat catttctaaa aatgttggaa aaaatgttta cgcgggcgat agcgttgaaa tagacgtgga tgtcgcaatg acgcatgacg ggactacccc tcttacagta aaagcttttg agcagatttc agacaaagtt tgggataatg aaaagatagt tattattttt gaccacaaca tccctgcaaa cacgtcaaaa gctgcgaata tgcaggttat aacgagagaa tttatcaaaa aacagggaat taaaaattat taccttgatg gcgaaggaat atgtcatcag gtacttcctg aaaaaggcca cgtgaagcca aacatgataa ttgcaggagc tgacagtcac acctgtactc atggggcatt cggtgctttt gcgacaggtt ttggtgcaac tgacatgggt tacgtctatg caaccggaaa aacatggctt agagttcctg aaaccattca agtaaatgta accggagaaa atgaaaatat ttctggaaag gacattatct taaaaacttg taaggaagtt ggaagacgtg gagcgacata cctgtcttta gaatacggcg gaaatgcagt ccaaaatctt gacatggacg aaagaatggt tttatcgaac atggccattg aaatgggcgg aaaagctgga attatcgaag ctgacgatac tacttacaaa taccttgaaa atgcaggagt ttcaagagaa gaaattctta acttgaaaaa aaataaaata aaagttaatg aatccgaaga aaattactac aaaacatttg aatttgatat aaccgatatg gaagaacaga ttgcttgccc gcaccaccct gacaatgtaa aaggagtttc tgaagtatca ggaattgaat tagatcaggt attcatcgga 〇 tcttgtacaa acggaagatt aaacgattta agaattgctg caaaacattt gaaaggaaaa aaagttaatg aaagcacccg actaattgta attcctgcat caaaatcaat ctttaaagaa gcgttaaaag aaggattaat cgatactttt gtagattctg gagcattaat ctgcactcct ggatgcggac catgccttgg agcccatcag ggtgttttag gtgatgggga agtatgtctt gctacaacca ataggaactt taaaggaaga atgggaaaca caaaatcgga agtttacctc tcatctcctg caatagctgc aaaatccgca gttaaaggat acattaccaa tgaataa <210> 184 <211> 486 <212> DNA <213> 人工 <220> 146966-序列表.doc 1020
60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 249- 1257 201037078 <223> AksE海沼甲烷球菌 <400> 184 atgaaaataa caggcaaggt gcacgtattt ggggatgaca tcgacacaga tgcgataatt cctggcgctt atttaaaaac aactgatgaa tatgagcttg catcacactg tatggctgga atcgatgaag attttccaga aatggtcaaa gaaggcgact ttttggtagc aggtgagaat ttcggatgcg gaagttcgag agagcaagct ccaattgcaa taaaatactg cggaatcaag gcaataattg ttgaaagttt tgcaaggata ttttatagaa attgtattaa tcttggagtt tttccaattg aatgcaaagg aatatcaaaa cacgtgaaag atggagattt aatagaattg gatctcgaaa ataaaaaagt aattttaaag gacaaggttc tagactgcca cattccaacc ggaactgcaa aagacataat ggatgaaggc gggcttataa attacgcaaa gaaacagaaa aactaa 486 <210> 185 <211> 1251
<212> DNA <213> 史氏甲烷短桿菌ATCC35061 <220> <221> CDS <222> (1) .· (1251) <400> 185 atg aac ate acc gag aag ate ctg tet get aaa geg aag aaa gaa gtt
Met Asn lie Thr Glu Lys lie Leu Ser Ala Lys Ala Lys Lys Glu Val 1 5 10 15 act ccg ggt gaa ate ate gaa ate ccg gtt gat ctg geg atg tet cac
Thr Pro Gly Glu lie lie Glu lie Pro Val Asp Leu Ala Met Ser His 20 25 30 gac ggt act tet ccg cca gca ate aaa act ttc gaa aaa gtt geg act 144 —
Asp Gly Thr Ser Pro Pro Ala lie Lys Thr Phe Glu Lys Val Ala Thr 35 40 45 aaa gta tgg gac aac gag aag att get ate gta ttc gac cac aac gta 192
Lys Val Trp Asp Asn Glu Lys lie Ala lie Val Phe Asp His Asn Val 50 55 60 ccg get aac acc ate ggt tet get gaa ttc cag aaa gtt tgc ege gat 240
Pro Ala Asn Thr lie Gly Ser Ala Glu Phe Gin Lys Val Cys Arg Asp 65 70 75 80 ttc ate aag aag cag aag ate acc aaa aac tac ate cac ggt gac ggt 288
Phe lie Lys Lys Gin Lys 工le Thr Lys Asn Tyr lie His Gly Asp Gly 85 90 95 •250- 146966-序列表.doe 201037078 ate tgc cac cag gta ctg ccg gaa aaa ggt ctg gtt gaa ccg ggt aaa 336 lie Cys His Gin Val Leu Pro Glu Lys Gly Leu Val Glu Pro Gly Lys 100 105 110 gtt ate gtt ggt get gac tet cac act tgc act tac ggt get tac ggc 384
Val lie Val Gly Ala Asp Ser His Thr Cys Thr Tyr Gly Ala Tyr Gly 115 120 125 gca ttc tet acc ggt atg ggt geg act gac ctg geg atg gtt tac gca 432
Ala Phe Ser Thr Gly Met Gly Ala Thr Asp Leu Ala Met Val Tyr Ala 130 135 140 act ggt aaa acc tgg ttc atg gtt ccg gaa get ate aag atg gaa gtt 480
Thr Gly Lys Thr Trp Phe Met Val Pro Glu Ala lie Lys Met Glu Val 145 150 155 160 Ο
tet ggt gaa ctg aac tet tac act gca ccg aaa gac ate ate ctg aaa 528
Ser Gly Glu Leu Asn Ser Tyr Thr Ala Pro Lys Asp lie lie Leu Lys 165 170 175 ate ate ggt gaa gtt ggt att get ggc gca act tac aaa act gca gaa 5Ί6 lie lie Gly Glu Val Gly lie Ala Gly Ala Thr Tyr Lys Thr Ala Glu 180 185 190 ttc tgc ggt gaa acc att gag aag atg ggc gta gaa ggt cgt geg act 624
Phe Cys Gly Glu Thr lie Glu Lys Met Gly Val Glu Gly Arg Ala Thr 195 200 205 ate tgc aac atg get ate gaa atg ggt geg aaa aac ggt ate atg gaa 672 lie Cys Asn Met Ala lie Glu Met Gly Ala Lys Asn Gly lie Met Glu 210 215 220 ccg aac aaa gaa gtt ate cag tac gtt tet cag cgt act ggt aag aaa 720
Pro Asn Lys Glu Val lie Gin Tyr Val Ser Gin Arg Thr Gly Lys Lys 225 230 235 240 gag tet gaa ctg aac ate gtt aag tet gac gaa gat get cag tac tet 768
Glu Ser Glu Leu Asn lie Val Lys Ser Asp Glu Asp Ala Gin Tyr Ser 245 250 255 gaa gaa atg cac ttc gac ate act gac atg gaa ccg cag ate get tgc 816
Glu Glu Met His Phe Asp lie Thr Asp Met Glu Pro Gin lie Ala Cys 260 265 270 ccg aac gac gtt gat aac gtt aaa gac ate tee aaa gtt gaa ggt act 864
Pro Asn Asp Val Asp Asn Val Lys Asp lie Ser Lys Val Glu Gly Thr 275 280 285 •251 · 146966-序列表.doe 201037078 gcg gtt gat cag tgc ctg ate ggt tee tgc acc aac ggt cgt ctg tet 912
Ala Val Asp Gin Cys Leu lie Gly Ser Cys Thr Asn Gly Arg Leu Ser 290 295 300 gac ctg aaa gac get tac gaa ate ctg aaa gac aac gaa ate aac aac 960
Asp Leu Lys Asp Ala Tyr Glu lie Leu Lys Asp Asn Glu lie Asn Asn 305 310 315 320 gac act ege ctg ctg att ctg ccg gca tet gca gaa ate tac aag cag 1008
Asp Thr Arg Leu Leu He Leu Pro Ala Ser Ala Glu He Tyr Lys Gin 325 330 335 get ate cac gaa ggt tac ate gac gca ttc ate gac get ggt get ate 1056
Ala lie His Glu Gly Tyr lie Asp Ala Phe lie Asp Ala Gly Ala lie 340 345 350 ate tgc aac cca ggt tgc ggt ccg tgc ctg ggt ggt cac atg ggc gta 1104 lie Cys Asn Pro Gly Cys Gly Pro Cys Leu Gly Gly His Met Gly Val 355 360 365 ctg tet gaa ggt gaa act tgc ctg tet acc act aac cgt aac ttc aaa 1152
Leu Ser Glu Gly Glu Thr Cys Leu Ser Thr Thr Asn Arg Asn Phe Lys 370 375 380 ggt cgt atg ggc gac ccg aaa tet tee gtt tac ctg get aac tee aaa 1200
Gly Arg Met Gly Asp Pro Lys Ser Ser Val Tyr Leu Ala Asn Ser Lys 385 390 395 400 gtt gtt get gca tet gca ate gaa ggt gtt ate act aac ccg aaa gac 1248
Val Val Ala Ala Ser Ala lie Glu Gly Val lie Thr Asn Pro Lys Asp 405 410 415 ctg 1251
Leu <210> 186 <211> 417
<212> PRT <213〉 史氏曱烷短桿菌ATCC 35061 <400> 186
Met Asn lie Thr Glu Lys lie Leu Ser Ala Lys Ala Lys Lys Glu Val 15 10 15
Thr Pro Gly Glu lie lie Glu lie Pro Val Asp Leu Ala Met Ser His 20 25 30
Asp Gly Thr Ser Pro Pro Ala lie Lys Thr Phe Glu Lys Val Ala Thr 35 40 45
Lys Val Trp Asp Asn Glu Lys lie Ala lie Val Phe Asp His Asn Val 50 55 60
Pro Ala Asn Thr lie Gly Ser Ala Glu Phe Gin Lys Val Cys Arg Asp 65 70 75 80
Phe lie Lys Lys Gin Lys lie Thr Lys Asn Tyr lie His Gly Asp Gly •252 · 146966·序列表.doc 201037078 85 90 95 lie Cys His Gin Val Leu Pro Glu Lys Gly Leu Val Glu Pro Gly Lys 100 105 110
Val lie Val Gly Ala Asp Ser His Thr Cys Thr Tyr Gly Ala Tyr Gly 115 120 125
Ala Phe Ser Thr Gly Met Gly Ala Thr Asp Leu Ala Met Val Tyr Ala 130 135 140
Thr Gly Lys Thr Trp Phe Met Val Pro Glu Ala lie Lys Met Glu Val 145 150 155 160
Ser Gly Glu Leu Asn Ser Tyr Thr Ala Pro Lys Asp lie lie Leu Lys 165 170 175 lie lie Gly Glu Val Gly lie Ala Gly Ala Thr Tyr Lys Thr Ala Glu 180 185 190
Phe Cys Gly Glu Thr lie Glu Lys Met Gly Val Glu Gly Arg Ala Thr 195 200 205 lie Cys Asn Met Ala lie Glu Met Gly Ala Lys Asn Gly lie Met Glu 210 215 220
Pro Asn Lys Glu Val lie Gin Tyr Val Ser Gin Arg Thr Gly Lys Lys 225 230 235 240
Glu Ser Glu Leu Asn lie Val Lys Ser Asp Glu Asp Ala Gin Tyr Ser 245 250 255
Glu Glu Met His Phe Asp lie Thr Asp Met Glu Pro Gin lie Ala Cys 260 265 270
Pro Asn Asp Val Asp Asn Val Lys Asp lie Ser Lys Val Glu Gly Thr 275 280 285
Ala Val Asp Gin Cys Leu lie Gly Ser Cys Thr Asn Gly Arg Leu Ser 290 295 300
Asp Leu Lys Asp Ala Tyr Glu lie Leu Lys Asp Asn Glu lie Asn Asn 305 310 315 320
Asp Thr Arg Leu Leu lie Leu Pro Ala Ser Ala Glu lie Tyr Lys Gin 325 330 335
Ala 工le His Glu Gly Tyr lie Asp Ala Phe lie Asp Ala Gly Ala lie 340 345 350 lie Cys Asn Pro Gly Cys Gly Pro Cys Leu Gly Gly His Met Gly Val 355 360 365
Leu Ser Glu Gly Glu Thr Cys Leu Ser Thr Thr Asn Arg Asn Phe Lys 370 375 380
Gly Arg Met Gly Asp Pro Lys Ser Ser Val Tyr Leu Ala Asn Ser Lys 385 390 395 400
Val Val Ala Ala Ser Ala lie Glu Gly Val lie Thr Asn Pro Lys Asp 405 410 415
Leu <210> 187
<211> 1254 <212> DNA <213> 史氏曱烷短桿菌ATCC35061 <220〉 <223> 野生型 <400> 187 TTGAATATTA CAGAAAAAAT ATTATCAGCT AAAGCAAAAA AAGAGGTTAC ACCCGGAGAA 60
ATTATTGAAA TTCCGGTTGA TTTAGCTATG TCTCATGACG GAACATCACC ACCAGCTATA 120 AAAACCTTTG AAAAAGTAGC TACCAAAGTC TGGGACAATG AAAAAATTGC AATTGTATTT 180 GACCATAATG TTCCTGCAAA TACAATCGGA TCTGCAGAAT TCCAGAAAGT ATGCAGAGAC 240 -253- 146966-序列表.doc 201037078 TTCATAAAAA AACAGAAAAT TACTAAAAAT TACATACATG GTGACGGAAT ATGCCACCAG 300 GTACTTCCTG AAAAAGGACT GGTAGAACCT GGAAAAGTTA TTGTAGGTGC TGACTCACAC 360 ACATGTACTT ATGGTGCATA TGGAGCATTT TCAACAGGAA TGGGTGCTAC TGATTTAGCT 420 ATGGTTTATG CTACAGGTAA AACATGGTTT ATGGTTCCTG AAGCTATTAA. AATGGAAGTC 480 AGCGGAGAGT TAAATTCATA CACTGCTCCA AAAGACATCA TTTTAAAAAT CATCGGTGAA 540 GTGGGAATAG CTGGAGCTAC CTACAAAACT GCAGAATTCT GCGGAGAAAC CATCGAAAAA 600 ATGGGAGTAG AAGGCAGAGC TACAATATGT AACATGGCTA TTGAAATGGG AGCTAAAAAC 660 GGAATAATGG AACCGAATAA AGAAGTTATC CAATATGTTT CCCAAAGAAC CGGAAAAAAA 720 GAAAGTGAAT TGAATATTGT TAAATCCGAT GAAGATGCAC AGTATTCAGA AGAAATGCAC 780
TTTGACAT7VA CTGATATGGA ACCTCAAATA GCCTGCCCTA ATGATGTAGA CAATGTTAAA 840
GACATCTCCA AAGTTGAAGG CACAGCTGTT GACCAATGCT TAATCGGTTC TTGTACAAAC 900
GGCAGATTAT CTGATTTGAA AGATGCTTAT GAAATCTTAA AAGATAATGA AATTAATAAT 960 GATACTAGAT TACTAATTCT TCCAGCATCA GCTGAAATTT ACAAACAGGC AATCCATGAA 1020 GGATACATTG ATGCATTTAT AGATGCGGGA GCTATTATAT GCAATCCAGG ATGCGGACCA 1080 TGTCTTGGAG GACATATGGG AGTTTTATCT GAAGGTGAAA CCTGCCTATC CACAACAAAC 1140 AGGAATTTCA AAGGAAGAAT GGGGGATCCG AAATCCTCAG TATATCTGGC TAATTCAAAG 1200
GTTGTTGCTG CTTCAGCTAT TGAAGGAGTT ATTACAAATC CTAAAGATTT ATAG 1254 <210> 188 <211> 1254 <212> DNA <213> 萬氏甲烷球菌SB <220> <221> CDS <222> (1)..(1254) <400> 188 48 atg act ctg get gaa get ate ctg tee aag aaa ctg ggt aaa aac gtt Met Thr Leu Ala Glu Ala lie Leu Ser Lys Lys Leu Gly Lys Asn Val 1 5 10 15 96 tac get aaa gac tee gtt gaa ate gac gtt gat ctg gca atg act cac Tyr Ala Lys Asp Ser Val Glu lie Asp Val Asp Leu Ala Met Thr His 20 25 30 gac ggt act act ccg ctg acc gtt aaa geg ttc gaa gag ate tet gac 144
Asp Gly Thr Thr Pro Leu Thr Val Lys Ala Phe Glu Glu lie Ser Asp 35 40 45 ege gta ttc gac aac aag aag ate gtt ate gtt ttc gac cac aac ate 192 254 146966·序列表.doc 201037078
Arg Val Phe Asp Asn Lys Lys lie Val lie Val Phe Asp His Asn lie 50 55 60 ccg get aac acc tet aaa geg gca aac atg cag ate ate act ege gac 240
Pro Ala Asn Thr Ser Lys Ala Ala Asn Met Gin lie lie Thr Arg Asp 65 70 75 80 ttc ate aag aag cac gac ate aaa aac tac tac ctg gat ggt gaa ggt 288
Phe lie Lys Lys His Asp lie Lys Asn Tyr Tyr Leu Asp Gly Glu Gly 85 90 95 ate tgc cac cag att ctg ccg gaa aaa ggt cac gtt aag ccg aac atg 336 lie Cys His Gin lie Leu Pro Glu Lys Gly His Val Lys Pro Asn Met 100 105 110 gtt ate gtt ggt get gac tet cac act tgc act cac ggt geg ttc ggt 384
Val lie Val Gly Ala Asp Ser His Thr Cys Thr His Gly Ala Phe Gly 115 120 125 gca ttc gca act ggc ttc ggt get tet gac atg ggt tac gtt tac gca 432
Ala Phe Ala Thr Gly Phe Gly Ala Ser Asp Met Gly Tyr Val Tyr Ala 130 135 140 act ggt aaa acc tgg ttc ege gta ccg gaa acc ate ege gtt aac gta 480
Thr Gly Lys Thr Trp Phe Arg Val Pro Glu Thr lie Arg Val Asn Val 145 150 155 160 act ggt aaa aac gaa aac ate tee ggt aaa gat ate gtt ctg aaa act 528
Thr Gly Lys Asn Glu Asn lie Ser Gly Lys Asp lie Val Leu Lys Thr 165 170 175 tgc aaa gaa gtt ggt cgt tet ggt gca act tac atg geg ctg gaa tac 576
Cys Lys Glu Val Gly Arg Ser Gly Ala Thr Tyr Met Ala Leu Glu Tyr 180 185 190 ggt ggt tet get gtt aaa geg ctg aac atg gac gaa cgt atg gta ctg 624
Gly Gly Ser Ala Val Lys Ala Leu Asn Met Asp Glu Arg Met Val Leu 195 200 205 tgc aac atg get ate gaa atg ggt ggt aaa gtt ggt ctg att gaa get 672
Cys Asn Met Ala lie Glu Met Gly Gly Lys Val Gly Leu lie Glu Ala 210 215 220 gac cac acc act tac gac tac ctg aaa aac get ggc gta tet aac cag 720
Asp His Thr Thr Tyr Asp Tyr Leu Lys Asn Ala Gly Val Ser Asn Gin 225 230 235 240 gaa ate get gaa ctg cag cgt aac aag ate tee ate act gaa aac gaa 768
Glu lie Ala Glu Leu Gin Arg Asn Lys lie Ser lie Thr Glu Asn Glu -255- 146966-序列表.doc 201037078 245 250 255 gaa act tac ttc aaa acc gtt gag ttc gac ate act gac atg 816
Glu Thr Tyr Phe Lys Thr Val Glu Phe Asp lie Thr Asp Met 260 265 270 cag gtt get tgc ccg cac cac ccg gat aac gtt aaa ggt ate 864
Gin Val Ala Cys Pro His His Pro Asp Asn Val Lys Gly lie 275 280 285 gtt ctg ggt act ccg ate gac cag att ttc ate ggt tee tgc 912
Val Leu Gly Thr Pro lie Asp Gin lie Phe lie Gly Ser Cys 290 295 300 ggt cac ate ggc gat ctg cgt ate get geg aag att ctg aaa 960
Gly His lie Gly Asp Leu Arg lie Ala Ala Lys lie Leu Lys 305 310 315 320 tet ate aac aaa aac acc cgt ctg ate gtt ate ccg get tet 1008
Ser lie Asn Lys Asn Thr Arg Leu lie Val lie Pro Ala Ser 325 330 335 ate ctg aag cag geg ctg aac gaa ggt ctg ate gac ate ttc 1056 lie Leu Lys Gin Ala Leu Asn Glu Gly Leu lie Asp lie Phe 340 345 350 ttc ggt geg ctg ate tgc gca cca ggt tgc ggt ccg tgc ctg 1104
Phe Gly Ala Leu He Cys Ala Pro Gly Cys Gly Pro Cys Leu 355 360 365 cac gaa ggc gta ctg ggt gac ggt gaa gtt tgc ctg gca act 1152
His Glu Gly Val Leu Gly Asp Gly Glu Val Cys Leu Ala Thr 370 375 380 cgt aac ttc aaa ggt cgt atg ggt aac ate aac tet gaa gtt 1200
Arg Asn Phe Lys Gly Arg Met Gly Asn He Asn Ser Glu Val 385 390 395 400 tee tet ccg gca ate get get aaa tet get ate aaa ggt cac 1248
Ser Ser Pro Ala He Ala Ala Lys Ser Ala lie Lys Gly His 405 410 415 aac gag Asn Glu <210> 189 <211> 418
<212> PRT <213> 萬氏曱烷球菌SB 146966-序列表.doc •256· gaa gaa Glu Glu tet gaa Ser Glu acc aac Thr Asn ggt aag Gly Lys aaa tet Lys Ser gtt gac Val Asp ggt gca Gly Ala acc aac Thr Asn tac ctg Tyr Leu ate act lie Thr 1254 201037078 <400> 189
Met Thr Leu Ala Glu Ala lie Leu Ser Lys Lys Leu Gly Lys Asn Val 15 10 15
Tyr Ala Lys Asp Ser Val Glu lie Asp Val Asp Leu Ala Met Thr His 20 25 30
Asp Gly Thr Thr Pro Leu Thr Val Lys Ala Phe Glu Glu lie Ser Asp 35 40 45
Arg Val Phe Asp Asn Lys Lys lie Val lie Val Phe Asp His Asn lie 50 55 60
Pro Ala Asn Thr Ser Lys Ala Ala Asn Met Gin lie lie Thr Arg Asp 65 70 75 80
Phe lie Lys Lys His Asp lie Lys Asn Tyr Tyr Leu Asp Gly Glu Gly 85 90 95 lie Cys His Gin lie Leu Pro Glu Lys Gly His Val Lys Pro Asn Met 100 105 110
Val lie Val Gly Ala Asp Ser His Thr Cys Thr His Gly Ala Phe Gly 115 120 125
Ala Phe Ala Thr Gly Phe Gly Ala Ser Asp Met Gly Tyr Val Tyr Ala 130 135 140 ❹ 〇
Thr Gly Lys Thr Trp Phe Arg Val Pro Giu Thr lie Arg Val Asn Val 145 150 155 160
Thr Gly Lys Asn Glu Asn lie Ser Gly Lys Asp lie Val Leu Lys Thr 165 170 175
Cys Lys Glu Val Gly Arg Ser Gly Ala Thr Tyr Met Ala Leu Glu Tyr 180 185 190
Gly Gly Ser Ala Val Lys Ala Leu Asn Met Asp Glu Arg Met Val Leu 195 200 205
Cys Asn Met Ala lie Glu Met Gly Gly Lys Val Gly Leu lie Glu Ala 210 215 220
Asp His Thr Thr Tyr Asp Tyr Leu Lys Asn Ala Gly Val Ser Asn Gin 225 230 235 240
Glu lie Ala Glu Leu Gin Arg Asn Lys lie Ser lie Thr Glu Asn Glu 245 250 255
Glu Thr Tyr Phe Lys Thr Val Glu Phe Asp lie Thr Asp Met Glu Glu 260 265 270
Gin Val Ala Cys Pro His His Pro Asp Asn Val Lys Gly lie Ser Glu 275 280 285
Val Leu Gly Thr Pro lie Asp Gin lie Phe lie Gly Ser Cys Thr Asn 290 295 300
Gly His lie Gly Asp Leu Arg lie Ala Ala Lys lie Leu Lys Gly Lys 305 310 315 320
Ser lie Asn Lys Asn Thr Arg Leu lie Val lie Pro Ala Ser Lys Ser 325 330 335 lie Leu Lys Gin Ala Leu Asn Glu Gly Leu lie Asp lie Phe Val Asp 340 345 350
Phe Gly Ala Leu lie Cys Ala Pro Gly Cys Gly Pro Cys Leu Gly Ala 355 360 365
His Glu Gly Val Leu Gly Asp Gly Glu Val Cys Leu Ala Thr Thr Asn 370 375 380
Arg Asn Phe Lys Gly Arg Met Gly Asn lie Asn Ser Glu Val Tyr Leu 385 390 395 400
Ser Ser Pro Ala lie Ala Ala Lys Ser Ala lie Lys Gly His lie Thr 410 -257· 405
Asn Glu <210> 190 <211> 1257
<212> DNA
<213>萬氏甲烧球菌SB 146966-序列表.doc 415 201037078 <220> <223> 野生型 <400> 190
AAAATGTATA TGCTAAAGAT GAACTACGCC ATTGACTGTA AAAAAATAGT AATAGTTTTT TGCAGATTAT AACAAGAGAC GAGAAGGAAT ATGTCATCAA TCGTTGGTGC AGATAGCCAT TTGGTGCAAG CGACATGGGT AAACAATACG GGTGAATGTT TAAAAACCTG TAAAGAAGTT GAAGTGCTGT AAAAGCCCTA AAATGGGCGG AAAAGTTGGA ATGCAGGCGT TTCCAATCAA AAAATGAAGA AACATACTTT TTGCATGCCC ACATCATCCC TTGACCAAAT ATTTATCGGT CAAAAATTTT AAAAGGAAAA CTAAATCGAT ACTTAAACAG GTGCATTAAT CTGTGCTCCA GCGATGGTGA AGTATGTCTT TTAATTCAGA GGTATATTTA ATATTACAAA CGAATAA ATGACACTTG CTGAAGCAAT TCTTTCAAAA AAACTTGGAA 60 AGCGTTGAAA TAGACGTCGA CCTTGCAATG ACCCATGATG 120 AAAGCTTTCG AAGAAATTTC AGACAGGGTT TTTGATAATA 180 GACCATAATA TTCCAGCAAA TACGTCAAAA GCAGCAAATA 240 TTCATTAAAA AACACGACAT TAAAAACTAC TATCTTGATG 300 ATACTTCCTG AAAAAGGCCA TGTAAAGCCA AACATGGTAA 360 ACCTGTACCC ATGGGGCATT TGGGGCTTTT GCAACGGGTT 420 TACGTTTATG CTACTGGAAA AACATGGTTT AGAGTTCCTG 480 ACTGGTAAAA ACGAAAATAT TTCCGGAAAG GACATTGTTT 540 GGAAGAAGTG GTGCAACATA TATGGCATTG GAATACGGAG 600 AACATGGATG AAAGAATGGT TTTATGCAAT ATGGCAATTG 660
TTAATTGAAG CAGACCATAC AACTTACGAT TACCTTAAAA 720
GAAATAGCCG AATTACAAAG AAACAAAATT TCAATTACTG 780 AAAACTGTTG AATTTGATAT AACTGACATG GAAGAACAAG 840 GATAATGTAA AAGGTATTTC TGAGGTTTTA GGTACTCCTA 900 TCTTGTACAA ATGGGCATAT AGGCGACCTT AGAATTGCTG 960 TCCATAAATA AAAATACAAG GCTTATAGTA ATTCCTGCAT 1020 GCCCTAAATG AAGGATTAAT TGATATTTTT GTGGATTTTG 1080 GGATGCGGCC CTTGCCTTGG CGCACATGAA GGGGTACTTG 1140 GCAACTACAA ACCGCAACTT TAAAGGAAGA ATGGGAAACA 1200 TCTTCTCCAG CAATTGCTGC AAAAAGTGCA ATTAAAGGGC 1257 <210> 191 <211> 1263
<212> DNA <213> 雜色甲烧球菌Nankai-3 <220>
<221> CDS <222> (1)… (1263) <400> 191 atg act ctg get gaa gaa ate ctg tee aag aaa Met Thr Leu Ala Glu Glu lie Leu Ser Lys Lys 1 5 10 146966-序列表.doc •258- gtt ggt aag aaa gtt Val Gly Lys Lys Val 15 201037078 aaa gcg ggt gac gtt gtt gaa ate gat ate gac ctg geg atg act cac Lys Ala Gly Asp Val Val Glu lie Asp lie Asp Leu Ala Met Thr His 20 25 30 gac ggt act act ccg ctg tet gcg aaa gca ttc aag cag ate act gac 144
Asp Gly Thr Thr Pro Leu Ser Ala Lys Ala Phe Lys Gin lie Thr Asp 35 40 45 aaa gta tgg gat aac aag aaa ate gtt ate gtt ttc gac cac aac gtt 192
Lys Val Trp Asp Asn Lys Lys lie Val lie Val Phe Asp His Asn Val 50 55 60 ccg get aac acc ctg aaa get get aac atg cag aag ate act ege gaa 240
Pro Ala Asn Thr Leu Lys Ala Ala Asn Met Gin Lys lie Thr Arg Glu 65 70 75 80 ❹ ttc ate aaa gag cag aac ate ate aac cac tac ctg gac ggt gaa ggt 288
Phe lie Lys Glu Gin Asn lie lie Asn His Tyr Leu Asp Gly Glu Gly 85 90 95 gtt tgc cac cag gta ctg ccg gaa aac ggt cac att cag ccg aac atg 336
Val Cys His Gin Val Leu Pro Glu Asn Gly His lie Gin Pro Asn Met 100 105 110 gtt ate get ggc ggc gat tet cac acc tgt act tac ggc gca ttc ggt 384
Val lie Ala Gly Gly Asp Ser His Thr Cys Thr Tyr Gly Ala Phe Gly 115 120 125 gcg ttc get act ggc ttc ggt gca act gac atg ggt aac ate tac gca 432
Ala Phe Ala Thr Gly Phe Gly Ala Thr Asp Met Gly Asn lie Tyr Ala 130 135 140 act ggt aaa acc tgg ctg aaa gtt ccg aaa act att cgt ate aac gtt 480
Thr Gly Lys Thr Trp Leu Lys Val Pro Lys Thr lie Arg lie Asn Val 145 150 155 160 aac ggt gaa aac gac aag ate acc ggt aaa gac ate ate ctg aaa ate 528
Asn Gly Glu Asn Asp Lys lie Thr Gly Lys Asp lie lie Leu Lys lie 165 170 175 tgc aaa gaa gtt ggt cgt tet ggt gca act tac atg gcg ctg gaa tac 576
Cys Lys Glu Val Gly Arg Ser Gly Ala Thr Tyr Met Ala Leu Glu Tyr 180 185 190 ggt ggt gaa gca ate aag aaa ctg tet atg gac gaa cgt atg gtt ctg 624
Gly Gly Glu Ala lie Lys Lys Leu Ser Met Asp Glu Arg Met Val Leu 195 200 205 -259- 146966-序列表.doc 201037078 tct aac atg get ate gaa atg ggt ggt aaa gtt ggt ctg ate gaa get 672
Ser Asn Met Ala lie Glu Met Gly Gly Lys Val Gly Leu lie Glu Ala 210 215 220 gac gaa acc act tac aac tat ctg cgt aac gtt ggt att tct gaa gag 720
Asp Glu Thr Thr Tyr Asn Tyr Leu Arg Asn Val Gly lie Ser Glu Glu 225 230 235 240 aag ate ctg gaa ctg aag aaa aac cag ate act ate gac gaa aac aac 768
Lys lie Leu Glu Leu Lys Lys Asn Gin lie Thr lie Asp Glu Asn Asn 245 250 255 ate gac aac gac aac tac tac aaa ate ate aac ate gac ate act gac 816 lie Asp Asn Asp Asn Tyr Tyr Lys lie lie Asn lie Asp lie Thr Asp 260 265 270 atg gaa gaa cag gtt get tgc ccg cac cac ccg gat aac gtt aaa aac 864
Met Glu Glu Gin Val Ala Cys Pro His His Pro Asp Asn Val Lys Asn 275 280 285 ate tct gaa gtt aaa ggc gca cca ate aac cag gta ttc ate ggt tee 912 lie Ser Glu Val Lys Gly Ala Pro lie Asn Gin Val Phe lie Gly Ser 290 295 300 tgc acc aac ggt ege ctg aac gat ctg ege att get tct aaa tac ctg 960
Cys Thr Asn Gly Arg Leu Asn Asp Leu Arg lie Ala Ser Lys Tyr Leu 305 310 315 320 aaa ggt aag aaa gtt cac aac gac gta cgt ctg ate gtt ate ccg get 1008
Lys Gly Lys Lys Val His Asn Asp Val Arg Leu 工le Val lie Pro Ala 325 330 335 tee aag tct ate ttc aag cag geg ctg aaa gaa ggt ctg ate gac ate 1056
Ser Lys Ser lie Phe Lys Gin Ala Leu Lys Glu Gly Leu lie Asp lie 340 345 350 ttc gtt gac get ggc geg ctg ate tgc act ccg ggt tgc ggt ccg tgc 1104
Phe Val Asp Ala Gly Ala Leu lie Cys Thr Pro Gly Cys Gly Pro Cys 355 360 365 ctg ggt gca cac cag ggc gta ctg ggt gac ggt gaa gtt tgc ctg gca 1152
Leu Gly Ala His Gin Gly Val Leu Gly Asp Gly Glu Val Cys Leu Ala 370 375 380 act acc aac cgt aac ttc aaa ggt cgt atg ggt aac acc act get gaa 1200
Thr Thr Asn Arg Asn Phe Lys Gly Arg Met Gly Asn Thr Thr Ala Glu 385 390 395 400 •260- 146966-序列表.doc 201037078 ate tac ctg tee tet ccg gca ate get get aaa tet get ate aaa ggt 1248 lie Tyr Leu Ser Ser Pro Ala lie Ala Ala Lys Ser Ala lie Lys Gly 405 410 415 tac ate act aac gag 1263
Tyr lie Thr Asn Glu 420 <210> 192 <211> 421
<212> PRT <213>雜色甲烷球菌Nankai-3 <400> 192
Met Thr Leu Ala Glu Glu lie Leu Ser Lys Lys Val Gly Lys Lys Val 15 10 15
Lys Ala Gly Asp Val Val Glu lie Asp 工le Asp Leu Ala Met Thr His 20 25 30
Asp Gly Thr Thr Pro Leu Ser Ala Lys Ala Phe Lys Gin lie Thr Asp 35 40 45
Lys Val Trp Asp Asn Lys Lys lie Val lie Val Phe Asp His Asn Val 50 55 60
Pro Ala Asn Thr Leu Lys Ala Ala Asn Met Gin Lys lie Thr Arg Glu 65 70 75 80
Phe lie Lys Glu Gin Asn lie lie Asn His Tyr Leu Asp Gly Glu Gly 85 90 95
Val Cys His Gin Val Leu Pro Glu Asn Gly His 工le Gin Pro Asn Met 100 105 110
Val lie Ala Gly Gly Asp Ser His Thr Cys Thr Tyr Gly Ala Phe Gly 115 120 125
Ala Phe Ala Thr Gly Phe Gly Ala Thr Asp Met Gly Asn lie Tyr Ala 130 135 140
Thr Gly Lys Thr Trp Leu Lys Val Pro Lys Thr lie Arg lie Asn Val 145 150 155 160
Asn Gly Glu Asn Asp Lys lie Thr Gly Lys Asp lie lie Leu Lys lie 165 170 175
Cys Lys Glu Val Gly Arg Ser Gly Ala Thr Tyr Met Ala Leu Glu Tyr 180 185 190
Gly Gly Glu Ala lie Lys Lys Leu Ser Met Asp Glu Arg Met Val Leu 195 200 205
Ser Asn Met Ala lie Glu Met Gly Gly Lys Val Gly Leu lie Glu Ala 210 . 215 220
Asp Glu Thr Thr Tyr Asn Tyr Leu Arg Asn Val Gly lie Ser Glu Glu 225 230 235 240
Lys lie Leu Glu Leu Lys Lys Asn Gin lie Thr lie Asp Glu Asn Asn 245 250 255 lie Asp Asn Asp Asn Tyr Tyr Lys lie 工le Asn lie Asp lie Thr Asp 260 265 270
Met Glu Glu Gin Val Ala Cys Pro His His Pro Asp Asn Val Lys Asn 275 280 285 lie Ser Glu Val Lys Gly Ala Pro lie Asn Gin Val Phe lie Gly Ser 290 295 300
Cys Thr Asn Gly Arg Leu Asn Asp Leu Arg He Ala Ser Lys Tyr Leu 305 310 315 320
Lys Gly Lys Lys Val His Asn Asp Val Arg Leu lie Val lie Pro Ala 325 330 335
Ser Lys Ser lie Phe Lys Gin Ala Leu Lys Glu Gly Leu lie Asp lie 340 345 350
Phe Val Asp Ala Gly Ala Leu lie Cys Thr Pro Gly Cys Gly Pro Cys 355 360 365 •261 - 146966-序列表.doc 201037078
Leu Gly 370 Thr Thr 385 lie Tyr Tyr lie
Ala His Gin Gly Val 375 Asn Arg Asn Phe Lys 390 Leu Ser Ser Pro Ala 405 Thr Asn Glu 420
Leu Gly Asp Gly Glu Val Cys Leu Ala 380
Gly Arg Met Gly Asn Thr Thr Ala Glu 395 400 lie Ala Ala Lys Ser Ala lie Lys Gly 410 415 <210> 193 <211> 1266 <212> DNA <213> 雜色甲烷球菌Nankai-3 <220> <223> 野生型 <400> 193 ATGACATTGG CAGAGGAAAT ATTATCAAAA AAAGTAGGAA AAAAAGTAAA AGCAGGAGAT 60 GTTGTAGAAA TAGATATAGA TTTAGCAATG ACTCATGATG GAACAACACC ATTATCTGCA 120 AAGGCATTTA AACAGATAAC CGATAAGGTA TGGGACAATA AAAAAATAGT CATAGTATTT 180
TGCAAAAAAT TACACGAGAA GTGAAGGCGT ATGCCATCAA TAGCTGGTGG AGATAGTCAC TTGGGGCTAC CGACATGGGG GACCATAATG TCCCAGCAAA TACATTAAAA GCCGCAAATA 240
TTTATAAAAG AACAAAATAT AATAAATCAT TATTTGGATG 300
GTGCTACCTG AAAACGGACA TATACAACCA AACATGGTTA 360
ACATGCACCT ATGGGGCATT TGGGGCATTT GCAACAGGAT 420 AATATATATG 480 AATGGAGAAA 540 GGACGAAGTG 600 AGTATGGACG 660 CTTATCGAAG 720 AAAATATTAG 780
CAACAGGAAA ATGATAAAAT GAGCTACTTA AAAGAATGGT CCGATGAAAC AATTGAAAAA
AACTTGGTTA TACTGGAAAA CATGGCACTT TTTAAGCAAT CACATATAAT AAATCAAATA
AAAGTTCCAA GATATTATTT GAATACGGCG ATGGCTATCG TACCTTAGAA ACCATTGATG
AAACCATAAG TAAAAATTTG GGGAAGCAAT AAATGGGCGG ATGTAGGAAT AGAACAATAT
AATAAATGTT TAAAGAAGTT AAAAAAATTA AAAAGTTGGG TAGTGAAGAA AGATAATGAT AATTATTATA ΑΑΑΤΤΑΤΑΆΑ TATAGATATT ACAGACATGG AGGAGCAAGT GGCATGCCCT 840 CACCATCCCG ATAATGTTAA AAATATTTCG GAAGTAAAAG GAGCTCCCAT AAATCAGGTG 900 TTCATAGGTT CATGCACAAA TGGTAGGTTG AACGATTTAA GAATAGCCTC AAAATATTTA 960 AAAGGAAAAA AGGTTCATAA TGATGTTAGA TTAATAGTAA TACCTGCTTC AAAATCAATA 1020 TTTAAACAGG CATTAAAAGA AGGATTAATT GATATTTTTG TAGATGCTGG AGCTTTAATT 1080 TGCACCCCCG GATGCGGTCC TTGTTTGGGG GCCCACCAAG GAGTTTTAGG AGATGGTGAA 1140 GTTTGTTTAG CCACCACAAA TAGAAATTTC AAAGGAAGAA TGGGAAATAC GACAGCGGAA 1200 ATATATTTAT CCTCCCCCGC TATTGCCGCA AAAAGTGCAA TTAAAGGATA TATCACAAAT 1260 -262- 146966-序列表.doc 201037078 GAATAA 1266 <210> 194 <211> 1254 <212> DNA <213〉 海沼甲烷球菌S2 <220> <221> CDS <222> (1) .. (1254) <400> 194 atg act ctg get gag aag ate ate tee aaa aac gtt ggt aaa aac gtt
Met Thr Leu Ala Glu Lys lie lie Ser Lys Asn Val Gly Lys Asn Val 15 10 15 tac geg aaa gac tee gtt gaa ate tee gtt gac ate geg atg act cac
Tyr Ala Lys Asp Ser Val Glu lie Ser Val Asp lie Ala Met Thr His 20 25 30 gac ggt act act ccg ctg acc gtt aaa geg ttc gaa cag ate tet gac 144
Asp Gly Thr Thr Pro Leu Thr Val Lys Ala Phe Glu Gin lie Ser Asp 35 40 45 aaa gta tgg gat aac gag aag ate gtt ate ate ttc gac cac aac ate 192
Lys Val Trp Asp Asn Glu Lys He Val lie lie Phe Asp His Asn lie 50 55 60 ccg get aac acc tet aaa get get aac atg caa gtt ate act cgt gaa 240
Pro Ala Asn Thr Ser Lys Ala Ala Asn Met Gin Val lie Thr Arg Glu 65 70 75 80 ttc ate aag aag cag ggt ate aaa aac tac tac ctg gat ggt gaa ggt 288
Phe lie Lys Lys Gin Gly lie Lys Asn Tyr Tyr Leu Asp Gly Glu Gly 85 90 95 ate tgc cac cag gta ctg ccg gaa aaa ggt cac gtt aag ccg aac atg 336 lie Cys His Gin Val Leu Pro Glu Lys Gly His Val Lys Pro Asn Met 100 105 110 ate ate get ggc gca gac tet cac act tgc act cac ggt gca ttc ggt 384 lie lie Ala Gly Ala Asp Ser His Thr Cys Thr His Gly Ala Phe Gly 115 120 125 gca ttc get acc ggt ttc ggt gca act gac atg ggt tac gtt tac gca 432
Ala Phe Ala Thr Gly Phe Gly Ala Thr Asp Met Gly Tyr Val Tyr Ala 130 135 140 act ggt aaa acc tgg ctg ege gta ccg gaa acc att ege gtt aac gta 480
Thr Gly Lys Thr Trp Leu Arg Val Pro Glu Thr He Arg Val Asn Val 145 150 155 160 146966-序列表.doc 48 96 ❹ 263 - 201037078 act ggc gaa aac gaa aac ate tee ggt aaa gac ate ate ctg aaa act 528
Thr Gly Glu Asn Glu Asn lie Ser Gly Lys Asp lie lie Leu Lys Thr 165 170 175 tgc aaa gaa gtt ggt cgt ege ggt gca act tac atg tet ctg gaa tac 576
Cys Lys Glu Val Gly Arg Arg Gly Ala Thr Tyr Met Ser Leu Glu Tyr 180 185 190 ggt ggt aac get gtt cac aac ctg tet atg gac gaa cgt atg gtt ctg 624
Gly Gly Asn Ala Val His Asn Leu Ser Met Asp Glu Arg Met Val Leu 195 200 205 tet aac atg get ate gaa atg ggt ggt aaa get ggt ate ate gaa get 672
Ser Asn Met Ala lie Glu Met Gly Gly Lys Ala Gly lie lie Glu Ala 210 215 220 gac gac acc act tac ege tac ctg gaa aac get ggc gtt tee cgt gaa 720
Asp Asp Thr Thr Tyr Arg Tyr Leu Glu Asn Ala Gly Val Ser Arg Glu 225 230 235 240 gaa ate ctg gaa ctg aag aaa aac aag ate acc gtt gac gaa tet gaa 768
Glu lie Leu Glu Leu Lys Lys Asn Lys lie Thr Val Asp Glu Ser Glu 245 250 255 gaa gat tac tac aaa act att gaa ttc gac ate acc ggt atg gaa gaa 816
Glu Asp Tyr Tyr Lys Thr lie Glu Phe Asp lie Thr Gly Met Glu Glu 260 265 270 cag gtt get tgc cca cac cac ccg gac aac gtt aaa ggc gtt tet gaa 864
Gin Val Ala Cys Pro His His Pro Asp Asn Val Lys Gly Val Ser Glu 275 280 285 gtt gaa ggt act gaa ctg aac cag gta ttc ate ggt tee tgc acc aac 912
Val Glu Gly Thr Glu Leu Asn Gin Val Phe He Gly Ser Cys Thr Asn 290 295 300 ggt cgt ctg aac gat ctg cgt att get geg aaa tac ctg aaa ggt aag 960
Gly Arg Leu Asn Asp Leu Arg lie Ala Ala Lys Tyr Leu Lys Gly Lys 305 310 315 320 aaa gtt aac gaa aac acc cgt ctg ate gtt ate ccg gca tet aaa tet 1008
Lys Val Asn Glu Asn Thr Arg Leu lie Val lie Pro Ala Ser Lys Ser 325 330 335 ate ttc aaa gaa geg ctg aac gaa ggt ctg ate gac ate ttc gtt gac 1056 lie Phe Lys Glu Ala Leu Asn Glu Gly Leu lie Asp lie Phe Val Asp 340 345 350 -264 146966·序列表.doc 201037078 tcc 1104 ggt gca ctg ate tgc act ccg ggt tgc ggt ccg tgc ctg ggt gca Ser Gly Ala 355 Leu lie Cys Thr 360 Pro Gly Cys Gly Pro 365 Cys Leu Gly Ala cac 1152 cag ggc gtt ctg ggt gac ggt gaa gtt tgc ctg gca acc act aac His Gin 370 Gly Val Leu Gly Asp 375 Gly Glu Val Cys 380 Leu Ala Thr Thr Asn cgt 1200 aac ttc aaa ggt cgt atg ggt aac acc aac get cag gtt tac ctg Arg Asn 385 Phe Lys Gly Arg Met 390 Gly Asn Thr 395 Asn Ala Gin Val 400 Tyr Leu tcc 1248 tct ccg aag ate get geg aag tct geg gta aaa ggt tac ate act Ser Ser Pro Lys lie 405 Ala Ala Lys Ser 410 Ala Val Lys Gly Tyr 415 lie Thr aat Asn gag Glu 1254
<210> 195 <211> 418 <212> PRT <213> 海沼甲燒球菌S2 <400〉 195
Met Thr Leu Ala Glu Lys lie lie Ser Lys Asn Val Gly Lys Asn Val 15 10 15
Tyr Ala Lys Asp Ser Val Glu lie Ser Val Asp lie Ala Met Thr His 20 25 30
Asp Gly Thr Thr Pro Leu Thr Val Lys Ala Phe Glu Gin lie Ser Asp 35 40 45
Lys Val Trp Asp Asn Glu Lys lie Val lie lie Phe Asp His Asn lie 50 55 60
Pro Ala Asn Thr Ser Lys Ala Ala Asn Met Gin Val lie Thr Arg Glu 65 70 75 80
Phe lie Lys Lys Gin Gly lie Lys Asn Tyr Tyr Leu Asp Gly Glu Gly 85 90 95 lie Cys His Gin Val Leu Pro Glu Lys Gly His Val Lys Pro Asn Met 100 105 110 lie lie Ala Gly Ala Asp Ser His Thr Cys Thr His Gly Ala Phe Gly 115 120 125
Ala Phe Ala Thr Gly Phe Gly Ala Thr Asp Met Gly Tyr Val Tyr Ala 130 135 140
Thr Gly Lys Thr Trp Leu Arg Val Pro Glu Thr lie Arg Val Asn Val 145 150 155 160
Thr Gly Glu Asn Glu Asn lie Ser Gly Lys Asp lie lie Leu Lys Thr 165 170 175
Cys Lys Glu Val Gly Arg Arg Gly Ala Thr Tyr Met Ser Leu Glu Tyr 180 185 190
Gly Gly Asn Ala Val His Asn Leu Ser Met Asp Glu Arg Met Val Leu 195 200 205
Ser Asn Met Ala lie Glu Met Gly Gly Lys Ala Gly lie lie Glu Ala 210 215 220
Asp Asp Thr Thr Tyr Arg Tyr Leu Glu Asn Ala Gly Val Ser Arg Glu 225 230 235 240
Glu lie Leu Glu Leu Lys Lys Asn Lys lie Thr Val Asp Glu Ser Glu -265- 146966-序列表.doc 201037078 245 250 255
Glu Asp Tyr Tyr Lys Thr lie Glu Phe Asp lie Thr Gly Met Glu Glu 260 265 270
Gin Val Ala Cys Pro His His Pro Asp Asn Val Lys Gly Val Ser Glu 275 280 285
Val Glu Gly Thr Glu Leu Asn Gin Val Phe lie Gly Ser Cys Thr Asn 290 295 300
Gly Arg Leu Asn Asp Leu Arg lie Ala Ala Lys Tyr Leu Lys Gly Lys 305 310 315 320
Lys Val Asn Glu Asn Thr Arg Leu lie Val lie Pro Ala Ser Lys Ser 325 330 335 lie Phe Lys Glu Ala Leu Asn Glu Gly Leu lie Asp lie Phe Val Asp 340 345 350
Ser Gly Ala Leu lie Cys Thr Pro Gly Cys Gly Pro Cys Leu Gly Ala 355 360 365
His Gin Gly Val Leu Gly Asp Gly Glu Val Cys Leu Ala Thr Thr Asn 370 375 380
Arg Asn Phe Lys Gly Arg Met Gly Asn Thr Asn Ala Gin Val Tyr Leu 385 390 395 400
Ser Ser Pro Lys 工le Ala Ala Lys Ser Ala Val Lys Gly Tyr lie Thr 410 415 405
Asn Glu <210> 196 <211> 1257
<212> DNA <213> 海沼曱烷球菌S2 <220> <223> 野生型 <400> 196 ATGACACTTG CTGAGAAAAT CATTTCAAAA AATGTTGGAA AAAATGTTTA CGCCAAAGAC 60
AGCGTCGAAA TAAGCGTAGA TATTGCAATG ACACATGACG GGACCACCCC GCTTACGGTA 120 AAAGCCTTTG AGCAGATTTC AGATAAAGTA TGGGATAATG AAAAGATAGT CATTATTTTT 180 GACCACAATA TCCCTGCAAA CACGTCAAAA GCTGCAAATA TGCAGGTTAT AACGAGAGAA 240
TTCATAAAAA AGCAGGGGAT TAAAAATTAC TACCTCGATG GCGAAGGAAT ATGCCATCAA 300
GTTCTACCTG AAAAGGGCCA TGTAAAACCA AACATGATAA TTGCGGGAGC TGACAGCCAC 360 ACATGTACTC ATGGGGCCTT TGGAGCATTT GCTACAGGTT TTGGTGCTAC AGACATGGGT 420 TACGTCTATG CAACAGGAAA AACCTGGCTT AGAGTTCCTG AAACTATCCG CGTAAATGTA 480 ACTGGAGAAA ATGAAAATAT TTCTGGAAAA GACATTATTT TAAAAACTTG TAAGGAAGTT 540 GGAAGACGTG GGGCTACGTA CATGTCTTTA GAATACGGTG GAAATGCAGT CCACAATCTT 600
TCAATGGATG AAAGAATGGT TCTGTCAAAC ATGGCTATTG AAATGGGCGG AAAAGCAGGA 660
ATTATCGAAG CTGATGATAC TACATATAGA TATCTTGAAA ATGCAGGAGT TTCGCGCGAA 720 GAAATTCTTG AATTGAAAAA AAATAAAATA ACAGTTGATG AATCCGAAGA AGACTACTAC 780 AAAACAATTG AATTTGACAT AACCGGTATG GAAGAACAGG TTGCATGCCC TCACCACCCT 840 266· 146966-序列表.doc 201037078 GATAACGTAA AAGGAGTTTC AGAAGTTGAA GGAACAGAAT TAAACCAGGT ATTCATCGGT 900 TCATGCACAA ACGGAAGATT AAACGACTTA AGAATTGCTG CAAAATATTT GAAAGGAAAA 960 AAAGTTAATG AAAACACAAG ATTAATTGTA ATCCCTGCAT CAAAGTCAAT ATTTAAAGAA 1020 GCCCTAAATG AAGGATTAAT TGATATCTTT GTAGATTCCG GAGCATTAAT ATGTACCCCT 1080 GGATGCGGAC CATGTCTTGG AGCCCATCAG GGGGTTTTAG GTGATGGAGA AGTATGCCTT 1140 GCTACAACTA ACCGGAACTT TAAAGGAAGA ATGGGAAACA CGAACGCACA AGTTTACCTC 1200 TCTTCTCCAA AAATAGCTGC AAAATCTGCG GTTAAAGGCT ACATTACAAA TGAATAA 1257 <210> 197 <211> 483
<212> DNA <213〉 史氏曱烷短桿菌ATCC 35061 <220> <221> CDS <222> (1) . . (483) <400> 197 atg gac ate ate aaa ggt aaa acc tgg act ttc ggt gaa aac ate gac
Met Asp lie lie Lys Gly Lys Thr Trp Thr Phe Gly Glu Asn lie Asp 15 10 15 act gac gtt ate ate cca ggt cgt tac etc ege act ttc aac ccg cag
Thr Asp Val lie lie Pro Gly Arg Tyr Leu Arg Thr Phe Asn Pro Gin 20 25 30 gac ctg gca gac cac gta ctg gaa ggt gaa cgt ccg gac ttc acc aag 144
Asp Leu Ala Asp His Val Leu Glu Gly Glu Arg Pro Asp Phe Thr Lys 35 40 45 aac gtt aag aaa ggc gac ate ate gtt get gac gaa aac ttc ggt tgc 192
Asn Val Lys Lys Gly Asp lie lie Val Ala Asp Glu Asn Phe Gly Cys 50 55 60 ggt tet tet ege gaa cag gca ccg gtt get ate aaa act get ggc gtt 240
Gly Ser Ser Arg Glu Gin Ala Pro Val Ala lie Lys Thr Ala Gly Val 65 70 75 80 gat get ate gtt geg aag tet ttc gca cgt ate ttc tac cgt aac get 288
Asp Ala lie Val Ala Lys Ser Phe Ala Arg lie Phe Tyr Arg Asn Ala 85 90 95 ate aac ate ggt ctg ccg gtt ate gtt tgc gac att cag geg aaa gac 336 lie Asn lie Gly Leu Pro Val lie Val Cys Asp lie Gin Ala Lys Asp 100 105 110 ggt gac ate ate aac ate gac ctg tet aaa ggt att ctg act aac gaa 384 146966-序列表.doc 48 96
267- 201037078
Gly Asp lie lie Asn lie Asp Leu Ser Lys Gly lie Leu Thr Asn Glu 115 120 125 acc act ggc gaa tcc gta act ttc gaa ccg ttc aaa gag ttc atg ctg 432
Thr Thr Gly Glu Ser Val Thr Phe Glu Pro Phe Lys Glu Phe Met Leu 130 135 140 9 4 tops a 8 s 4 Aϊ—_ c t a ctg gaa gat aac ggt ctg gtt aac cac tac ctg aaa gaa aaa Leu Glu Asp Asn Gly Leu Val Asn His Tyr Leu Lys Glu Lys 150 155 160 cag 483
Gin <210> 198
<211> 161 <212> PRT <213> 史氏甲烷短桿菌ATCC 35061 <400> 198
Met Asp lie lie Lys Gly Lys Thr Trp Thr Phe Gly Glu Asn lie Asp 15 10 15
Thr Asp Val lie lie Pro Gly Arg Tyr Leu Arg Thr Phe Asn Pro Gin 20 25 30
Asp Leu Ala Asp His Val Leu Glu Gly Glu Arg Pro Asp Phe Thr Lys 35 40 45
Asn Val Lys Lys Gly Asp lie lie Val Ala Asp Glu Asn Phe Gly Cys 50 55 60
Gly Ser Ser Arg Glu Gin Ala Pro Val Ala 工le Lys Thr Ala Gly Val 65 70 75 80
Asp Ala lie Val Ala Lys Ser Phe Ala Arg lie Phe Tyr Arg Asn TVla 85 90 95 lie Asn lie Gly Leu Pro Val lie Val Cys Asp lie Gin Ala Lys Asp 100 105 110
Gly Asp lie lie Asn lie Asp Leu Ser Lys Gly lie Leu Thr Asn Glu 115 120 125
Thr Thr Gly Glu Ser Val Thr Phe Glu Pro Phe Lys Glu Phe Met Leu 130 135 140
Asp lie Leu Glu Asp Asn Gly Leu Val Asn His Tyr Leu Lys Glu Lys 145 150 155 160
Gin <210> 199 <211> 486
<212> DNA <213> 史氏甲烷短桿菌ATCC35061 <220> <223> 野生型 <400> 199 ATGGATATAA TTAAAGGAAA 7UVCATGGACT TTTGGTGAAA ATATTGATAC GGATGTTATT 60
ATTCCCGGAA GATATTTGAG AACCTTTAAT CCTCAAGATT TGGCTGATCA TGTACTTGAA 120 GGAGAACGTC CAGATTTTAC AAAGAATGTT AAAAAAGGAG ACATAATAGT AGCTGATGAA 180 -268- 146966-序列表.doc 201037078 AATTTTGGTT GCGGTTCATC AAGAGAGCAG GCTCCAGTAG CTATTAAAAC TGCAGGTGTA 240 GATGCTATTG TGGCTAAATC TTTCGCCAGA ATTTTTTACA GAAATGCAAT TAACATTGGA 300 TTGCCAGTAA TTGTTTGTGA TATTCAGGCA AAAGACGGAG ATATTATAAA CATTGATTTG 360 TCAAAAGGAA TATTAACTAA TGAAACTACT GGTGAATCAG TTACTTTTGA ACCTTTTAAA 420 GAGTTTATGT TAGATATTTT AGAAGATAAT GGTCTTGTTA ATCATTATCT AAAAGAAAAA 480 CAATAA 486
<210> 200 <211> 483 <212> DNA <213> 萬氏甲炫球菌SB o <220> <221> CDS <222> (1)..(483) <400> 200 atg aag ctg aaa ggt aaa gcg cac gta ttc tct gac gac gtt gat act Met Lys Leu Lys Gly Lys Ala His Val Phe Ser Asp Asp Val Asp Thr 195 200 205 48 gac gca ate att ccg ggt get tat ctg cgt act act gac gtt tac gaa Asp Ala lie lie Pro Gly Ala Tyr Leu Arg Thr Thr Asp Val Tyr Glu 210 215 220 96
ctg gca tct cac tgc atg get ggt ate gac gaa aac ttc ccg aag aaa 144 Leu Ala Ser His Cys Met Ala Gly lie Asp Glu Asn Phe Pro Lys Lys 225 230 235 240 gtt aac ctg ggt gac ttc ate gtt get ggc gaa aac ttc ggt tgc ggt 192 Val Asn Leu Gly Asp Phe lie Val Ala Gly Glu Asn Phe Gly Cys Gly 245 250 255 tct tcc cgt gaa cag gca ccg ate tcc ate aaa tac ctg ggt ate tee 240 Ser Ser Arg Glu Gin Ala Pro lie Ser lie Lys Tyr Leu Gly lie Ser 260 265 270 gca ate ate get gaa tcc ttc get ege att ttc tac cgt aac tcc ate 288 Ala lie lie Ala Glu Ser Phe Ala Arg lie Phe Tyr Arg Asn Ser lie 275 280 285 aac ctg ggt gtt ate ccg ate gaa tgc aaa aac ate tcc aag cac gtt 336 Asn Leu Gly Val lie Pro lie Glu Cys Lys Asn lie Ser Lys His Val 290 295 300 aaa act ggc gac ctg ate gaa ctg gat ctg gaa aac aag aaa ate ate 384 Lys Thr Gly Asp Leu lie Glu Leu Asp Leu Glu Asn Lys Lys lie lie 305 310 315 320 146966-序列表.doc -269· 201037078 ctg aaa gac ate gtt ctg gaa tgc acc gta ccg act ggt aaa geg aaa 432
Leu Lys Asp lie Val Leu Glu Cys Thr Val Pro Thr Gly Lys Ala Lys 325 330 335 gag ate ate gac ctg ggt ggt ctg ate aac tac get aaa geg cag atg 480
Glu lie lie Asp Leu Gly Gly Leu lie Asn Tyr Ala Lys Ala Gin Met 340 345 350 ggt 483
Gly
<210> 201 <211> 161 <212> PRT <213> 萬氏甲炫球菌SB <400> 201
Met Lys Leu Lys Gly Lys Ala His Val Phe Ser Asp Asp Val Asp Thr 195 200 205
Asp Ala lie lie Pro Gly Ala Tyr Leu Arg Thr Thr Asp Val Tyr Glu 210 215 220
Leu Ala Ser His Cys Met Ala Gly lie Asp Glu Asn Phe Pro Lys Lys 225 230 235 240
Val Asn Leu Gly Asp Phe lie Val Ala Gly Glu Asn Phe Gly Cys Gly 245 250 255
Ser Ser Arg Glu Gin Ala Pro lie Ser lie Lys Tyr Leu Gly lie Ser 260 265 270
Ala lie lie Ala Glu Ser Phe Ala Arg lie Phe Tyr Arg Asn Ser lie 275 280 285
Asn Leu Gly Val lie Pro lie Glu Cys Lys Asn lie Ser Lys His Val 290 295 300
Lys Thr Gly Asp Leu lie Glu Leu Asp Leu Glu Asn Lys Lys lie lie 305 310 315 320
Leu Lys Asp lie Val Leu Glu Cys Thr Val Pro Thr Gly Lys Ala Lys 325 330 335
Glu lie lie Asp Leu Gly Gly Leu lie Asn Tyr Ala Lys Ala Gin Met 340 345 350
Gly <210> 202 <211> 486
<212> DNA <213> 萬氏曱炫球菌SB <220> <223> 野生型 <400> 202 ATGAAATTAA AAGGCAAAGC ACACGTATTT TCAGATGATG TCGATACCGA TGCTATAATT 60 CCAGGAGCAT ATCTAAGAAC TACTGATGTT TATGAACTTG CATCGCACTG TATGGCGGGG 120 ATTGATGAGA ATTTTCCTAA AAAAGTAAAT TTGGGGGATT TTATTGTTGC AGGTGAAAAT 180 TTTGGCTGTG GAAGTTCAAG GGAACAGGCT CCAATTTCGA TAAAATATCT TGGAATAAGT 240 -270- 146966-序列表.doc
201037078 GCAATAATTG CGGAAAGTTT TGCAAGAATT TTTTATCGAA ATTCTATAAA CTTAGGTGTA 300 ATTCCAATTG AATGCAAAAA CATTTCAAAA CACGTTAAAA CAGGGGATTT AATAGAATTA 360 GACCTTGAAA ACAAAAAAAT CATTTTAAAA GACATTGTTT TAGAATGCAC GGTTCCAACT 420 GGAAAGGCAA AAGAAATAAT TGATTTAGGC GGGCTTATAA ATTATGCAAA AGCGCAAATG 480 GGCTAA 486 <210> 203
<211> 495 <212> DNA <213 > 雜色甲烷球菌Nankai-3 <220> <221> CDS <222> (1) .. (495) <400> 203 atg ate ate aaa ggt aac ate cac ctg ttc ggt gac gac ate gac act Met lie lie Lys Gly Asn lie His Leu Phe Gly Asp Asp lie Asp Thr 15 10 15 gac get ate ate cca ggt get tac ctg aaa acc act gac ccg aaa gag Asp Ala lie lie Pro Gly Ala Tyr Leu Lys Thr Thr Asp Pro Lys Glu 20 25 30 ctg gca tet cac tgc atg geg ggt ate gac gaa aaa ttc tet acc aaa 144
Leu Ala Ser His Cys Met Ala Gly lie Asp Glu Lys Phe Ser Thr Lys 35 40 45 48 96
gtt 192 Val tet 240 aaa gac ggt Lys Asp Gly 50 tee cgt gaa Ser Arg Glu gac ate ate Asp lie lie 55 cag gca ccg Gin Ala Pro 70 gtt get ggc Val Ala Gly 60 ate tee ate lie Ser lie 75 gaa aac ttc Glu Asn Phe aag cac acc Lys His Thr ggt tgc ggt Gly Cys Gly ggt ate aaa Gly lie Lys geg gtt gtt get gaa tee ttc get ege att ttc tac cgt aac tgc ate 288
Ala Val Val Ala Glu Ser Phe Ala Arg lie Phe Tyr Arg Asn Cys lie 85 90 95 aac ate ggt ctg ate ccg ate acc tgt gaa ggt ate aac gaa cag att 336
Asn lie Gly Leu lie Pro lie Thr Cys Glu Gly lie Asn Glu Gin lie 100 105 110 cag aac ctg aaa gac ggt gac acc ate gaa ate gat ctg cag aac gaa 384
Gin Asn Leu Lys Asp Gly Asp Thr lie Glu lie Asp Leu Gin Asn Glu 115 120 125 acc ate aag ate aac tee atg atg ctg aac tgc ggt gca ccg aaa ggt 432 271 · 146966-序列表.doc 201037078
Thr lie Lys lie Asn Ser Met Met Leu Asn Cys Gly Ala Pro Lys Gly 130 135 140 c o e 5 t 8 1 4 a 4 I 1 gaa aaa gaa ate ctg gat get ggc ggt ctg gta cag tac acc aag
Glu Lys Glu lie Leu Asp Ala Gly Gly Leu Val Gin Tyr Thr Lys 150 155 160 aac aag ctg aag aaa 495
Asn Lys Leu Lys Lys 165 <210> 204 <211> 165
<212> PRT <213> 雜色甲烧球菌Nankai-3 <400> 204
Met lie lie Lys Gly Asn lie His Leu Phe Gly Asp Asp lie Asp Thr 15 10 15
Asp Ala lie lie Pro Gly Ala Tyr Leu Lys Thr Thr Asp Pro Lys Glu 20 25 30
Leu Ala Ser His Cys Met Ala Gly lie Asp Glu Lys Phe Ser Thr Lys 35 40 45
Val Lys Asp Gly Asp lie lie Val Ala Gly Glu Asn Phe Gly Cys Gly 50 55 60
Ser Ser Arg Glu Gin Ala Pro lie Ser lie Lys His Thr Gly lie Lys 65 70 75 80
Ala Val Val Ala Glu Ser Phe Ala Arg lie Phe Tyr Arg Asn Cys lie 85 90 95
Asn lie Gly Leu lie Pro lie Thr Cys Glu Gly lie Asn Glu Gin lie 100 105 110
Gin Asn Leu Lys Asp Gly Asp Thr lie Glu lie Asp Leu Gin Asn Glu 115 120 125
Thr lie Lys lie Asn Ser Met Met Leu Asn Cys Gly Ala Pro Lys Gly 130 135 140 lie Glu Lys Glu lie Leu Asp Ala Gly Gly Leu Val Gin Tyr Thr Lys 145 150 155 160
Asn Lys Leu Lys Lys 165 <210> 205 <211> 498
<212> DNA <213> 雜色甲烧球菌Nankai-3 <220> <223> 野生型 <400> 205 ATGATAATAA AAGGAAATAT TCATTTATTT GGTGATGATA TTGATACCGA TGCCATAATT 60 CCCGGGGCCT ACCTTAAAAC AACAGACCCA AAGGAGTTGG CATCTCATTG CATGGCTGGA 120 ATTGATGAAA AATTTTCAAC AAAGGTAAAA GACGGCGATA TAATTGTTGC AGGTGAAAAT 180 TTTGGCTGTG GAAGTAGTAG GGAACAGGCA CCAATATCCA TAAAACACAC CGGAATAAAG 240 •272· 146966-序列表.doc 201037078 GCAGTAGTTG CTGAAAGTTT TGCACGGATA TTTTATAGAA ATTGTATTAA TATAGGATTA 300 ATACCTATAA CTTGCGAAGG AATAAATGAA CAAATCCAAA ACCTAAAAGA TGGCGACACA 360 ATAGAAATTG ATTTGCAAAA TGAAACAATA ΑΑΑΆΤΑΑΑΤΤ CTATGATGTT AAATTGTGGA 420 GCTCCCAAAG GGATAGAAAA AGAAATTTTA GATGCTGGTG GATTAGTACA ATATACAAAA 480 AATAAGTTAA AAAAATAA 498 <210> 206 <211> 483 <212> DNA <213> 海沼甲烷球菌S2 <220> <221> CDS <222> (1) .. (483)
<400> 206 atg aag ate acc ggt aaa gtt cac ctg ttc ggt gac gac ate gac act Met Lys lie Thr Gly Lys Val His Leu Phe Gly Asp Asp lie Asp Thr 15 10 15 48 gac get ate att ccg ggt get tac ctg aaa acc act gac gaa tac gaa Asp Ala lie lie Pro Gly Ala Tyr Leu Lys Thr Thr Asp Glu Tyr Glu 20 25 30 96 ctg get tet cac tgc atg geg ggt ate gac gaa aac ttc ccg gaa ege 144 Leu Ala Ser His Cys Met Ala Gly lie Asp Glu Asn Phe Pro Glu Arg 35 40 45 gtt gaa gat ggc gac ttc ctg gtt get ggc gaa aac ttc ggt tgc ggt 192 Val Glu Asp Gly Asp Phe Leu Val Ala Gly Glu Asn Phe Gly Cys Gly 50 55 60 tet tee cgt gaa cag gca ccg att get ate aaa tac tgc ggt ate aaa 240 Ser Ser Arg Glu Gin Ala Pro lie Ala lie Lys Tyr Cys Gly lie Lys 65 70 75 80 gca ate ate gtt gaa tee ttc gca cgt ate ttc tac cgt aac tgc ate 288 Ala lie lie Val Glu Ser Phe Ala Arg lie Phe Tyr Arg Asn Cys lie 85 90 95 aac ctg ggc gta ttc ccg ate gaa tgc aaa ggt ate tee aag cac gtt 336 Asn Leu Gly Val Phe Pro lie Glu Cys Lys Gly lie Ser Lys His Val 100 105 110 aaa gac ggt gac gtt ate gaa ctg gat ctg gaa gag aag aaa gtt ate 384 Lys Asp Gly Asp Val lie Glu Leu Asp Leu Glu Glu Lys Lys Val lie 115 120 125 ctg aaa gac acc gta ctg gac tgc aac etc ccg act ggt act geg aaa 432 146966-序列表.doe -273· 201037078
Leu Lys Asp Thr Val Leu Asp Cys Asn Leu Pro Thr Gly Thr Ala Lys 130 135 140 t o p 5 a 8 s 4 Q- 4 A 1 ate atg gac gaa ggt ggt ctg lie Met Asp Glu Gly Gly Leu 150 ate aac tac get aag aag cag aaa lie Asn Tyr Ala Lys Lys Gin Lys 155 160 aac 483
Asn <210> 207 <211> 161 <212> PRT <213> 海沼甲烷球菌S2 <400> 207
Met Lys 工le Thr Gly Lys Val His Leu Phe Gly Asp Asp lie Asp Thr 15 10 15
Asp Ala 工le lie Pro Gly Ala Tyr Leu Lys Thr Thr Asp Glu Tyr Glu 20 25 30
Leu Ala Ser His Cys Met Ala Gly lie Asp Glu Asn Phe Pro Glu Arg 35 40 45
Val Glu Asp Gly Asp Phe Leu Val Ala Gly Glu Asn Phe Gly Cys Gly 50 55 60
Ser Ser Arg Glu Gin Ala Pro lie Ala lie Lys Tyr Cys Gly lie Lys 65 70 75 80
Ala lie lie Val Glu Ser Phe Ala Arg lie Phe Tyr Arg Asn Cys lie 85 90 95
Asn Leu Gly Val Phe Pro lie Glu Cys Lys Gly lie Ser Lys His Val 100 105 110
Lys Asp Gly Asp Val lie Glu Leu Asp Leu Glu Glu Lys Lys Val lie 115 120 125
Leu Lys Asp Thr Val Leu Asp Cys Asn Leu Pro Thr Gly Thr Ala Lys 130 135 140
Asp lie Met Asp Glu Gly Gly Leu lie Asn Tyr Ala Lys Lys Gin Lys 145 150 155 160
Asn <210> 208 <211> 486 <212> DNA <213> 海沼甲烷球菌S2 <220> <223>野生型 <400> 208 ATGAAAATAA CTGGTAAGGT GCACTTATTT GGGGATGACA TCGATACTGA TGCGATAATT 60 CCCGGAGCTT ATTTAAAAAC GACTGATGAA TATGAGCTTG CATCGCACTG TATGGCAGGA 120 ATTGACGAAA ATTTTCCAGA AAGGGTCGAA GATGGTGACT TTTTAGTTGC AGGTGAAAAT 180 TTTGGATGCG GAAGTTCAAG GGAACAGGCC CCAATTGCCA TAAAATACTG CGGAATCAAG 240 GCAATAATTG TTGAGAGTTT TGCAAGGATA TTTTACAGAA ATTGCATAAA TTTAGGAGTA 300 274· 146966-序列表.doc 201037078 TTTCCAATTG 7\ATGCAAGGG AATATCAAAA CACGTCAAAG ATGGGGATGT AATAGAATTA 360 GATCTTGAAG AAAAAAAAGT TATCTTAAAA GACACGGTTC TTGACTGCAA TCTTCCGACA 420 GGGACTGCAA AAGATATAAT GGATGAAGGC GGGCTTATAA ATTACGCAAA GAAACAAAAA 480 AATTAA 486 <210> 209 <211> 999
<212> DNA <213> 史氏甲烷短桿菌ATCC35061 <220> <221> CDS <222> (1) . (999) <400> 209 48
atg tac cgc att get ate att ccg ggc gac ggt ate ggt aaa gaa gta Met Tyr Arg lie Ala lie lie Pro Gly Asp Gly lie Gly Lys Glu Val 15 10 15 atg gaa tee ggt gaa tat ctg ctg gat aaa ctg gat ctg aac ttc tet Met Glu Ser Gly Glu Tyr Leu Leu Asp Lys Leu Asp Leu Asn Phe Ser 96 20 25 30 ttc gaa tac ggt gaa gca ggc ttc gaa tgc tac aac aaa aac ggc gtt 144
Phe Glu Tyr Gly Glu Ala Gly Phe Glu Cys Tyr Asn Lys Asn Gly Val 35 40 45 act ctg ccg gaa gaa acc ate aaa att get aag aaa tet gac gca act 192
Thr Leu Pro Glu Glu Thr lie Lys lie Ala Lys Lys Ser Asp Ala Thr 50 55 60 ctg ttc ggt get tet act tet acc cca ggt cag ccg tcc ccg ate ate 240
Leu Phe Gly Ala Ser Thr Ser Thr Pro Gly Gin Pro Ser Pro lie lie 65 70 75 80 aac ctg aga aag gaa ctg gac gtt tac get aac etc cgt ccg ate aaa 288
Asn Leu Arg Lys Glu Leu Asp Val Tyr Ala Asn Leu Arg Pro lie Lys 85 90 95 tet tac cgt ggt gtt cgt tet ate tet gac aac ate gac ttc ctg ate 336
Ser Tyr Arg Gly Val Arg Ser lie Ser Asp Asn lie Asp Phe Leu lie 100 105 110 gtt cgt gaa aac act gaa ggt ctg tac tet cag ate gaa tac gag cag 384
Val Arg Glu Asn Thr Glu Gly Leu Tyr Ser Gin lie Glu Tyr Glu Gin 115 120 125 gac aac aaa gtt ate get cag cgc gtt ate act cgt cgt get tet gag 432
Asp Asn Lys Val lie Ala Gin Arg Val lie Thr Arg Arg Ala Ser Glu 130 135 140 146966·序列表.doc •275- 201037078 aag ate get aaa gtt gca ttc gaa cag tgc ate geg aag cag aag cag 480
Lys lie Ala Lys Val Ala Phe Glu Gin Cys lie Ala Lys Gin Lys Gin 145 150 155 160 aaa gtt act tgc gta cac aaa tet aac gtt ctg aag aaa act gac ggc 528
Lys Val Thr Cys Val His Lys Ser Asn Val Leu Lys Lys Thr Asp Gly 165 170 175 gta ttc aaa gag tet ttc tac aaa ate get gaa aac tac ccg aac ate 576
Val Phe Lys Glu Ser Phe Tyr Lys lie Ala Glu Asn Tyr Pro Asn lie 180 185 190 gaa tee aac gac ttc tac gtt gac gca act geg atg tac ctg ate act 624
Glu Ser Asn Asp Phe Tyr Val Asp Ala Thr Ala Met Tyr Leu lie Thr 195 200 205 cag ccg cag aac ttc gac gtt ate gtt act tet aac ctg ttc ggt gac 672
Gin Pro Gin Asn Phe Asp Val lie Val Thr Ser Asn Leu Phe Gly Asp 210 215 220 ate ctg tet gac gaa ggt geg ggt ctg gtt ggt ggt ctg ggt ctg gca 720 lie Leu Ser Asp Glu Gly Ala Gly Leu Val Gly Gly Leu Gly Leu Ala 225 230 235 240 ccg tet ggt aac ate ggt gac gac cac ggt ctg ttc gaa ccg gtt cac 768
Pro Ser Gly Asn lie Gly Asp Asp His Gly Leu Phe Glu Pro Val His 245 250 255 ggt tet gca ccg gat ate get ggt aaa ggt ate get aac ccg tgc tee 816
Gly Ser Ala Pro Asp lie Ala Gly Lys Gly lie Ala Asn Pro Cys Ser 260 265 270 atg ate ctg act ate geg atg atg ctg gac tac ctg aaa gaa tac gaa 864
Met lie Leu Thr lie Ala Met Met Leu Asp Tyr Leu Lys Glu Tyr Glu 275 280 285 ate tee aac aag ate aac aaa geg gta gaa aac gtt gtt tet get ggt 912 lie Ser Asn Lys lie Asn Lys Ala Val Glu Asn Val Val Ser Ala Gly 290 295 300 aaa act ctg act ccg gac ctg ggt ggt aac tee acc act tet gaa ctg 960
Lys Thr Leu Thr Pro Asp Leu Gly Gly Asn Ser Thr Thr Ser Glu Leu 305 310 315 320 acc aag tet ate ate gac gaa ate ctg gaa ggc gat tac Thr Lys Ser lie lie Asp Glu lie Leu Glu Gly Asp Tyr 325 330 -276· 146966-序列表.doc 999 201037078 <210> 210 <211> 333
<212> PRT <213> 史氏甲烷短桿菌ATCC 35061 <400> 210
Met Tyr Arg lie Ala lie lie Pro Gly Asp Gly lie Gly Lys Glu Val 15 10 15
Met Glu Ser Gly Glu Tyr Leu Leu Asp Lys Leu Asp Leu Asn Phe Ser 20 25 30
Phe Glu Tyr Gly Glu Ala Gly Phe Glu Cys Tyr Asn Lys Asn Gly Val 35 40 45
Thr Leu Pro Glu Glu Thr lie Lys lie Ala Lys Lys Ser Asp Ala Thr 50 55 60
Leu Phe Gly Ala Ser Thr Ser Thr Pro Gly Gin Pro Ser Pro lie lie 65 70 75 80
Asn Leu Arg Lys Glu Leu Asp Val Tyr Ala Asn Leu Arg Pro lie Lys 85 90 95
Ser Tyr Arg Gly Val Arg Ser lie Ser Asp Asn lie Asp Phe Leu lie 100 105 110
Val Arg Glu Asn Thr Glu Gly Leu Tyr Ser Gin lie Glu Tyr Glu Gin 115 120 125
Asp Asn Lys Val lie Ala Gin Arg Val lie Thr Arg Arg Ala Ser Glu 130 135 140
Lys lie Ala Lys Val Ala Phe Glu Gin Cys lie Ala Lys Gin Lys Gin 145 150 155 160
Lys Val Thr Cys Val His Lys Ser Asn Val Leu Lys Lys Thr Asp Gly 165 170 175
Val Phe Lys Glu Ser Phe Tyr Lys lie Ala Glu Asn Tyr Pro Asn lie 180 185 190
Glu Ser Asn Asp Phe Tyr Val Asp Ala Thr Ala Met Tyr Leu lie Thr 195 200 205
Gin Pro Gin Asn Phe Asp Val lie Val Thr Ser Asn Leu Phe Gly Asp 210 215 220 lie Leu Ser Asp Glu Gly Ala Gly Leu Val Gly Gly Leu Gly Leu Ala 225 230 235 240
Pro Ser Gly Asn lie Gly Asp Asp His Gly Leu Phe Glu Pro Val His 245 250 255
Gly Ser Ala Pro Asp lie Ala Gly Lys Gly lie Ala Asn Pro Cys Ser
260 Met lie Leu Thr 275 lie Ser Asn Lys 290 Lys Thr Leu Thr 305 265 270 lie Ala Met Met Leu Asp Tyr Leu Lys Glu Tyr Glu 280 285 工le Asn Lys Ala Val Glu Asn Val Val Ser Ala Gly 295 300 Pro Asp Leu Gly Gly Asn Ser Thr Thr Ser Glu Leu 310 315 320
Thr Lys Ser lie lie Asp Glu lie Leu Glu Gly Asp Tyr 325 330
<210> 211 <211> 1002 <212> DNA <213> 史氏曱烷短桿菌ATCC 35061 <220> <223> 野生型 <400> 211 ATGTACAGAA TTGCAATAAT ACCCGGAGAT GGAATTGGAA AGGAAGTAAT GGAATCTGGC 60 -277- 146966-序列表.doc 201037078 GAATATTTAC TGGACAAACT TGATTTGAAT TTCAGTTTTG AATATGGGGA AGCAGGTTTT 120 GAATGTTATA ACAAAAATGG AGTTACATTA CCTGAAGAAA CCATTAAAAT AGCTAAAAAA 180 TCAGATGCAA CATTATTTGG AGCCAGTACA AGTACCCCCG GCCAGCCAAG TCCAATTATA 240 AATCTTAGAA AAGAACTTGA TGTTTATGCA AATTTAAGGC CGATAAAATC CTATAGAGGC 300 GTCAGATCAA TAAGTGATAA TATTGACTTT TTAATAGTTC GTGAAAATAC TGAGGGACTT 360 TACAGCCAGA TTGAATATGA ACAGGACAAT AAAGTAATTG CCCAAAGAGT CATTACAAGA 420 AGAGCCAGTG AAAAAATAGC CAAAGTTGCT TTTGAACAAT GTATCGCCAA ACAAAAACAA 480 AAGGTAACCT GTGTACATAA AAGCAATGTC CTAAAAAAAA CCGACGGAGT CTTTAAAGAA 540 AGTTTTTACA AAATAGCTGA AAATTACCCC AATATTGAAA GCAATGATTT TTATGTTGAT 600 GCAACAGC7\A TGTATCTTAT TACACAACCG CAAAACTTTG ATGTAATTGT AACCAGCAAC 660 CTGTTTGGAG ATATACTGTC TGATGAAGGT GCCGGTCTTG TAGGGGGACT TGGACTTGCC 720 CCTTCAGGAA ATATTGGAGA TGATCATGGA TTATTTGAAC CTGTTCACGG ATCTGCACCA 780 GATATTGCAG GTAAAGGTAT AGCTAACCCA TGCTCCATGA TTTTAACAAT AGCTATGATG 840
CTGGATTACT TAAAAGAATA TGAAATAAGT AATAAAATAA ATAAAGCTGT TGAAAATGTT 900
GTAAGTGCAG GCAAAACACT GACTCCTGAT TTAGGAGGAA ACAGCACTAC ATCAGAACTT 960 ACCAAATCTA TCATAGATGA AATACTTGAA GGTGACTACT AA 1002 o gt 4 t ev a Μ 1 > o c r c e t s
a T tct aac aag aaa gac aac aaa tac cag ate get gtt att Ser Asn Lys Lys Asp Asn Lys Tyr Gin lie Ala Val lie 5 10 15 48 <210> 212 <211> 1008 <212> DNA <213> 史氏甲烷短桿菌ATCC 35061 <220> <221> CDS <222> (1)..(1008) 96 ccg ggt gac ggt ate ggt aaa gaa gta atg gaa geg act ate tee gta
Pro Gly Asp Gly lie Gly Lys Glu Val Met Glu Ala Thr lie Ser Val 20 25 30 ctg gat gaa ctg gac gtt gac ttc gac tac ate tac ggt ate get ggc 144
Leu Asp Glu Leu Asp Val Asp Phe Asp Tyr lie Tyr Gly lie Ala Gly 35 40 45 gac gag tgc aac gaa gag cac ggt act ccg ctg ccg cag gaa acc ate 192
Asp Glu Cys Asn Glu Glu His Gly Thr Pro Leu Pro Gin Glu Thr lie 50 55 60 -278 146966-序列表.doc 201037078 gac ate gtt ege gac tet gac get tgc ctg ttc ggt gca get ggc gaa 240
Asp lie Val Arg Asp Ser Asp Ala Cys Leu Phe Gly Ala Ala Gly Glu 65 70 75 80 act gca get gac gtt ate gtt aag ate cgt cag gaa atg aag atg ttc 288
Thr Ala Ala Asp Val lie Val Lys lie Arg Gin Glu Met I»ys Met Phe 85 90 95 get aat ctg cgt ccg gta aaa tet tac ccg aac acc aag tet ctg ttc 336
Ala Asn Leu Arg Pro Val Lys Ser Tyr Pro Asn Thr Lys Ser Leu Phe 100 105 110 gaa aac gtt gac ttc atg ate gtt cgt gaa aac act gaa ggt ctg tac 384
Glu Asn Val Asp Phe Met lie Val Arg Glu Asn Thr Glu Gly Leu Tyr 115 120 125
ate get gac cag gaa gaa gaa act gaa gat ggc gca ate geg aag ege 432 lie Ala Asp Gin Glu Glu Glu Thr Glu Asp Gly Ala lie Ala Lys Arg 130 135 140 gtt ate act cgt gaa gca gaa gag cgt ate ate gac tac gca ttc cag 480
Val lie Thr Arg Glu Ala Glu Glu Arg lie lie Asp Tyr Ala Phe Gin 145 150 155 160 tac get aaa gac aac aac ege act aaa gtt act gca gta cac aaa get 528
Tyr Ala Lys Asp Asn Asn Arg Thr Lys Val Thr Ala Val His Lys Ala 165 170 175 aac gtt ctg aag aaa act gac ggt ctg ttc aag aaa ate ttc tac gaa 576
Asn Val Leu Lys Lys Thr Asp Gly Leu Phe Lys Lys lie Phe Tyr Glu 180 185 190 gtt ggt gaa aaa tac ccg gat ate gac act gaa gat ttc tac gtt gat 624
Val Gly Glu Lys Tyr Pro Asp lie Asp Thr Glu Asp Phe Tyr Val Asp 195 200 205 gca act geg atg tac ctg gta act cag ccg cag gaa ttc cag gtt gtt 672
Ala Thr Ala Met Tyr Leu Val Thr Gin Pro Gin Glu Phe Gin Val Val 210 215 220 gtt acc act aac ctg ttc ggt gac ate ctg tet gac gaa ggt get ggt 720
Val Thr Thr Asn Leu Phe Gly Asp lie Leu Ser Asp Glu Gly Ala Gly 225 230 235 240 ctg gtt ggt ggt ctg ggt ctg att ccg tet get aac ate ggt get gac 768
Leu Val Gly Gly Leu Gly Leu lie Pro Ser Ala Asn lie Gly Ala Asp 245 250 255 -279- 146966-序列表.doc 201037078 ggt geg ctg ttc gaa ccg gtt cac ggt tct gca ccg gat ate get ggt 816
Gly Ala Leu Phe Glu Pro Val His Gly Ser Ala Pro Asp lie Ala Gly 260 265 270 cag cag aaa geg aac cca ate geg atg atg ctg tct gca ate atg atg 864
Gin Gin Lys Ala Asn Pro lie Ala Met Met Leu Ser Ala lie Met Met 275 280 285 ctg cgt tac ctg ggt gaa aac gac gca get gac aaa ttc gac get get 912
Leu Arg Tyr Leu Gly Glu Asn Asp Ala Ala Asp Lys Phe Asp Ala Ala 290 295 300 c o e 5 t 6 1 o a 9 I 3 ctg aaa gtt ctg tct gaa ggt aaa act ctg act ggc gac ctg ggt
Leu Lys Val Leu Ser Glu Gly Lys Thr Leu Thr Gly Asp Leu Gly 310 315 320 ggt tct gca act acc atg gaa gtt get cag geg gtt aaa aac geg tta 1008
Gly Ser Ala Thr Thr Met Glu Val Ala Gin Ala Val Lys Asn Ala Leu 325 330 335 <210> 213 <211> 336
<212> PRT <213〉史氏甲烷短桿菌ATCC 35061 <400> 213
Met Ser Thr Ser Asn Lys Lys Asp Asn Lys Tyr Gin lie Ala Val lie 15 10 15
Pro Gly Asp Gly lie Gly Lys Glu Val Met Glu Ala Thr lie Ser Val 20 25 30
Leu Asp Glu Leu Asp Val Asp Phe Asp Tyr lie Tyr Gly lie Ala Gly 35 40 45
Asp Glu Cys Asn Glu Glu His Gly Thr Pro Leu Pro Gin Glu Thr lie 50 55 60
Asp lie Val Arg Asp Ser Asp Ala Cys Leu Phe Gly Ala Ala Gly Glu 65 70 75 80
Thr Ala Ala Asp Val lie Val Lys lie Arg Gin Glu Met Lys Met Phe 85 90 95
Ala Asn Leu Arg Pro Val Lys Ser Tyr Pro Asn Thr Lys Ser Leu Phe 100 105 110
Glu Asn Val Asp Phe Met lie Val Arg Glu Asn Thr Glu Gly Leu Tyr 115 120 125 lie Ala Asp Gin Glu Glu Glu Thr Glu Asp Gly Ala lie Ala Lys Arg 130 135 140
Val lie Thr Arg Glu Ala Glu Glu Arg lie lie Asp Tyr Ala Phe Gin 145 150 155 160
Tyr Ala Lys Asp Asn Asn Arg Thr Lys Val Thr Ala Val His Lys Ala 165 170 175
Asn Val Leu Lys Lys Thr Asp Gly Leu Phe Lys Lys lie Phe Tyr Glu 180 185 190
Val Gly Glu Lys Tyr Pro Asp lie Asp Thr Glu Asp Phe Tyr Val Asp 195 200 205
Ala Thr Ala Met Tyr Leu Val Thr Gin Pro Gin Glu Phe Gin Val Val 210 215 220
Val Thr Thr Asn Leu Phe Gly Asp lie Leu Ser Asp Glu Gly Ala Gly •280- 146966-序列表.doc 201037078 240 Ala Asn lie Gly Ala Asp 255 Ala Pro Asp lie Ala Gly 270 Leu Ser Ala lie Met Met 285 225 230 235
Leu Val Gly Gly Leu Gly Leu lie Pro Ser 245 250
Gly Ala Leu Phe Glu Pro Val His Gly Ser 260 265
Gin Gin Lys Ala Asn Pro lie Ala Met Met 275 280
Leu Arg Tyr 290 lie Leu Lys 305 Gly Ser Ala
Leu Gly Glu Asn Asp Ala Ala Asp Lys Phe Asp Ala Ala 295 300
Val Leu Ser Glu Gly Lys Thr Leu Thr Gly Asp Leu Gly 310 315 320
Thr Thr Met Glu Val Ala Gin Ala Val Lys Asn Ala Leu 325 330 335 <210> 214 <211> 1011 <212> DNA <213> 史氏甲烷短桿菌ATCC 35061 <220> <223> 野生型 <400> 214
G TTGAGTACCT CAAACAAAAA AGACAATAAA TATCAGATAG CTGTTATTCC AGGTGATGGA 60 ATAGGTAAAG AAGTTATGGA AGCAACAATC TCTGTTTTAG ACGAATTAGA TGTTGATTTT 120 GATTATATTT ATGGTATAGC AGGTGACGAA TGCAATGAAG AACATGGAAC ACCTCTTCCT 180 CAGGAAACTA TCGATATTGT AAGGGATTCT GATGCATGTT TATTTGGAGC TGCCGGAGAA 240 ACTGCAGCAG ATGTTATTGT AAAAATCCGT CAGGAAATGA AAATGTTTGC TAATTTAAGA 300 CCTGTCAAAT CTTATCCTAA CACCAAATCA TTATTTGAAA ATGTTGACTT TATGATTGTA 360 AGGGAAAATA CAGAAGGATT ATACATTGCA GACCAGGAAG AAGAAACCGA AGATGGTGCA 420 ATAGCTAAAC GTGTTATTAC AAGAGAAGCT GAAGAACGTA TTATTGATTA TGCTTTTCAA 480 TATGCAAAGG ACAACAACAG AACTAAAGTT ACTGCAGTTC ACAAAGCTAA TGTTCTTAAA 540 AAAACCGACG GATTATTTAA AAAAATATTC TACGAAGTCG GCGAAAAATA TCCGGATATT 600 GATACTGAAG ACTTTTACGT TGATGCAACA GCAATGTATC TTGTAACACA GCCTCAGGAA 660 TTCCAGGTTG TTGTAACTAC CAATTTATTT GGAGATATTT TATCTGATGA AGGTGCAGGA 720 CTTGTTGGCG GACTTGGTTT AATTCCATCA GCAAATATTG GAGCAGATGG TGCTTTATTT 780 GAACCTGTTC ATGGTTCAGC ACCGGATATT GCAGGTCAGC AAAAAGCAAA TCCAATAGCT 840
ATGATGCTCT CAGCAATTAT GATGCTTAGA TACCTTGGTG AAAATGATGC TGCTGACAAA 900
TTTGATGCTG CTATTTTAAA AGTATTAAGT GAAGGTAAAA CCTTAACTGG TGATTTAGGC 960
GGCAGTGCAA CTACCATGGA AGTAGCACAA GCTGTTAAAA ATGCATTATA A 1011 <210> 215 281 - 146966-序列表.doc 48 201037078 <211> 1011
<212> DNA
<213>萬氏曱烷球菌SB <220>
<221> CDS <222> (1)..(1011) <400> 215 atg ggt tac atg ccg aaa ate tgc gtt ate act ggc gac ggt ate ggt
Met Gly Tyr Met Pro Lys lie Cys Val lie Thr Gly Asp Gly lie Gly 15 10 15 aaa gaa gtt gtt ccg gaa act ctg ege gta ctg aac gaa gtt cac gac
Lys Glu Val Val Pro Glu Thr Leu Arg Val Leu Asn Glu Val His Asp 20 25 30 ttc gaa tac ate gaa gca cac geg ggt tac gag tgc ttc aag ege tgc 144
Phe Glu Tyr lie Glu Ala His Ala Gly Tyr Glu Cys Phe Lys Arg Cys 35 40 45 ggt gaa tee ate ccg gaa tee act att cag act geg aaa aac tet gac 192
Gly Glu Ser lie Pro Glu Ser Thr lie Gin Thr Ala Lys Asn Ser Asp 50 55 60 tee ate ctg ttc ggt tet gtt acc act ccg aaa cca act gaa ctg aaa 240
Ser lie Leu Phe Gly Ser Val Thr Thr Pro Lys Pro Thr Glu Leu Lys 65 70 75 80 aac aag ccg tac ege tet ccg att ctg act ctg cgt cag gaa ctg gat 288
Asn Lys Pro Tyr Arg Ser Pro lie Leu Thr Leu Arg Gin Glu Leu Asp 85 90 95 ctg tac get aac ate cgt ccg act tac aac ttc aaa gac ctg gac ttc 336
Leu Tyr Ala Asn lie Arg Pro Thr Tyr Asn Phe Lys Asp Leu Asp Phe 100 105 110 gtt ate ate cgt gaa aac act gaa tgc ctg tac gtt aag cgt gaa tac 384
Val lie lie Arg Glu Asn Thr Glu Cys Leu Tyr Val Lys Arg Glu Tyr 115 120 125 tac gac gaa ate aac gaa gtt get ate get gaa cgt ate ate tee aag 432
Tyr Asp Glu lie Asn Glu Val Ala lie Ala Glu Arg lie lie Ser Lys 130 135 140 aaa ggt tet gaa cgt ate ate aaa ttc get ttc gaa tac gca cgt ctg 480
Lys Gly Ser Glu Arg lie lie Lys Phe Ala Phe Glu Tyr Ala Arg Leu 145 150 155 160 aac aac cgt aag aaa gtt tee tgc ate cac aaa get aac gta ctg ege 528
Asn Asn Arg Lys Lys Val Ser Cys lie His Lys Ala Asn Val Leu Arg 165 170 175 •282 - 96 146966-序列表.doc 201037078 gta act gac ggt ctg ttc ctg gaa ate ttc gag aag ate geg aaa ctg 576
Val Thr Asp Gly Leu Phe Leu Glu lie Phe Glu Lys lie Ala Lys Leu 180 185 190 tac gaa aac ttc ggt ate tet tet aac gac tac ctg ate gac gca act 624
Tyr Glu Asn Phe Gly lie Ser Ser Asn Asp Tyr Leu lie Asp Ala Thr 195 200 205 gca atg tac ctg ate aaa aac ccg tac atg ttc gac gta atg gtt acc 672
Ala Met Tyr Leu 工le Lys Asn Pro Tyr Met Phe Asp Val Met Val Thr 210 215 220 act aac ctg ttc ggt gac ate ctg tet gac gaa get get ggt ctg ate 720
Thr Asn Leu Phe Gly Asp lie Leu Ser Asp Glu Ala Ala Gly Leu lie 225 230 235 240
ggt ggt ctg ggt atg tet ccg tet get aac ate ggt gac aac ctg ggt 768
Gly Gly Leu Gly Met Ser Pro Ser Ala Asn lie Gly Asp Asn Leu Gly 245 250 255 ctg ttc gaa ccg gtt cac ggt tet gca ccg gat ate get ggt aaa ggt 816
Leu Phe Glu Pro Val His Gly Ser Ala Pro Asp lie Ala Gly Lys Gly 260 265 270 ate tee aac ccg ate geg act ate ctg tet geg tet atg atg ctg gat 864 lie Ser Asn Pro 工le Ala Thr lie Leu Ser Ala Ser Met Met Leu Asp 275 280 285 cac ctg aaa atg aac aag aaa gca gaa ate ate cgt aac get gtt aag 912
His Leu Lys Met Asn Lys Lys Ala Glu lie lie Arg Asn Ala Val Lys 290 295 300 aaa act ate aac aac ggt tac ctg act ccg gac ctg ggt ggt tet ctg 960
Lys Thr lie Asn Asn Gly Tyr Leu Thr Pro Asp Leu Gly Gly Ser Leu 305 310 315 320 aaa act tet gaa gtt gtt aac aaa gtt ate gaa ttc ate ege gac gag 1008
Lys Thr Ser Glu Val Val Asn Lys Val lie Glu Phe lie Arg Asp Glu 325 330 335 att 1011 lie <210> 216 <211> 337
<212> PRT <213> 萬氏甲烧球菌SB <400> 216 -283- 146966-序列表.doc 201037078
Met Gly Tyr Met Pro Lys lie Cys Val lie Thr Gly Asp Gly lie Gly 15 10 15
Lys Glu Val Val Pro Glu Thr Leu Arg Val Leu Asn Glu Val His Asp 20 25 30
Phe Glu Tyr lie Glu Ala His Ala Gly Tyr Glu Cys Phe Lys Arg Cys 35 40 45
Gly Glu Ser lie Pro Glu Ser Thr lie Gin Thr Ala Lys Asn Ser Asp 50 55 60
Ser lie Leu Phe Gly Ser Val Thr Thr Pro Lys Pro Thr Glu Leu Lys 65 70 75 80
Asn Lys Pro Tyr Arg Ser Pro lie Leu Thr Leu Arg Gin Glu Leu Asp 85 90 95
Leu Tyr Ala Asn lie Arg Pro Thr Tyr Asn Phe Lys Asp Leu Asp Phe 100 105 110
Val lie lie Arg Glu Asn Thr Glu Cys Leu Tyr Val Lys Arg Glu Tyr 115 120 125
Tyr Asp Glu lie Asn Glu Val Ala lie Ala Glu Arg lie lie Ser Lys 130 135 140
Lys Gly Ser Glu Arg lie lie Lys Phe Ala Phe Glu Tyr Ala Arg Leu 145 150 155 160
Asn Asn Arg Lys Lys Val Ser Cys lie His Lys Ala Asn Val Leu Arg 165 170 175
Val Thr Asp Gly Leu Phe Leu Glu lie Phe Glu Lys lie Ala Lys Leu 180 185 190
Tyr Glu Asn Phe Gly lie Ser Ser Asn Asp Tyr Leu lie Asp Ala Thr 195 200 205
Ala Met Tyr Leu lie Lys Asn Pro Tyr Met Phe Asp Val Met Val Thr 210 215 220
Thr Asn Leu Phe Gly Asp lie Leu Ser Asp Glu Ala Ala Gly Leu lie 225 230 235 240
Gly Gly Leu Gly Met Ser Pro Ser Ala Asn lie Gly Asp Asn Leu Gly 245 250 255
Leu Phe Glu Pro Val His Gly Ser Ala Pro Asp lie Ala Gly Lys Gly 260 265 270 lie Ser Asn Pro 工le Ala Thr lie Leu Ser Ala Ser Met Met Leu Asp 275 280 285
His Leu Lys Met Asn Lys Lys Ala Glu lie lie Arg Asn Ala Val Lys 290 295 300
Lys Thr lie Asn Asn Gly Tyr Leu Thr Pro Asp Leu Gly Gly Ser Leu 305 310 315 320
Lys Thr Ser Glu Val Val Asn Lys Val lie Glu Phe lie Arg Asp Glu 325 330 335 lie <210> 217 <211> 1014
<212> DNA
<213> 萬氏甲烷球菌SB <220> <223> 野生型 <400> 217 ATGGGCTATA TGCCAAAAAT CTGTGTCATA ACTGGTGATG GAATTGGAAA AGAAGTCGTG 60 CCTGAAACAT TAAGAGTTTT AAACGAAGTT CACGACTTTG AATATATTGA AGCCCATGCT 120 GGGTACGAGT GTTTTAAGAG ATGCGGAGAA TCAATACCTG AAAGCACGAT TCAAACAGCT 180 -284- 146966-序列表.doc Ο
201037078 AAAAATTCTG ATTCAATTCT TTTTGGTTCA GTAACTACTC CAAAACCAAC TGAATTAAAA 240 AATAAGCCAT ATCGTAGTCC AATACTTACG TTAAGGCAGG AATTAGACCT TTATGCAAAC 300 ATACGGCCCA CTTATAATTT TAAAGACTTG GATTTTGTCA TAATACGGGA AAATACTGAG 360 TGTCTTTACG TAAAAAGGGA GTATTATGAC GAAATAAACG AAGTAGCAAT TGCCGAAAGA 420 ATAATTTCAA AAAAGGGAAG CGAAAGAATA ATAAAATTTG CATTTGAATA TGCAAGGTTA 480 AATAATCGAA AAAAAGTGTC TTGCATACAC AAAGCGAATG TATTAAGAGT AACTGACGGA 540 TTATTTTTGG AAATTTTCGA AAAAATAGCT AAACTTTATG AAAACTTTGG TATATCGAGC 600 AATGATTACT TAATAGATGC AACAGCAATG TACCTTATTA AAAATCCATA TATGTTTGAT 660 GTAATGGTTA CAACAAACCT TTTTGGAGAT ATTTTATCTG ATGAGGCCGC AGGACTTATT 720 GGGGGTCTTG GAATGTCGCC TTCTGCAAAT ATTGGGGATA ATTTAGGATT ATTTGAGCCT 780 GTTCATGGTT CAGCCCCAGA TATTGCTGGA AAAGGAATAT CTAATCCGAT TGCGACAATT 840 TTAAGTGCTT CAATGATGCT TGACCATTTA AAAATGAATA AAAAGGCGGA AATTATAAGA 900 AATGCAGTTA AAAAAACGAT AAATAATGGT TATTTGACAC CCGATCTTGG TGGAAGCCTG 960 AAAACTTCCG AAGTTGTAAA TAAAGTTATA GAATTTATTC GGGATGAAAT CTAA 1014 <210> 218 <211> 1029
<212> DNA <213> 雜色曱烷球菌Nankai-3 <220> <221> CDS <222> (1)..(1029) <400> 218 atg aag ate ccg aaa ate tgc gtt ate gaa ggt gac ggt ate ggt aaa Met Lys lie Pro Lys lie Cys Val lie Glu Gly Asp Gly lie Gly Lys 15 10 15 48 gaa gtt ate cca Glu Val lie Pro 20 gaa ttc ate tac 144 Glu Phe lie Tyr 35 gac get ate ccg 192 Asp Ala lie Pro 50 gaa acc gtt ege Glu Thr Val Arg 25 gaa cac get ggt Glu His Ala Gly 40 gag aaa act ctg Glu Lys Thr Leu 55 att ctg aaa gaa lie Leu Lys Glu 30 tac gaa tgc ttc Tyr Glu Cys Phe 45 aaa act geg aaa Lys Thr Ala Lys60 ate ggt gac ttc lie Gly Asp Phe aag ege tgc ggt Lys Arg Cys Gly gag tgc gac get Glu Cys Asp Ala 96 ate ctg ttc ggt geg gta tet act ccg aaa ctg gac gaa act gaa cgt 240 lie Leu Phe Gly Ala Val Ser Thr Pro Lys Leu Asp Glu Thr Glu Arg 285- 146966-序列表.doc 201037078 65 70 75 80 aag ccg tac aaa tct ccg att ctg act ctg cgt aaa gaa ctg gat ctg 288
Lys Pro Tyr Lys Ser Pro lie Leu Thr Leu Arg Lys Glu Leu Asp Leu 85 90 95 tac get aac gtt cgt ccg ate cac aaa ctg gat aac tct gac tcc tcc 336
Tyr Ala Asn Val Arg Pro lie His Lys Leu Asp Asn Ser Asp Ser Ser 100 105 110 aac aac ate gac ttc ate ate ate cgt gaa aac act gaa ggt ctg tac 384
Asa Asn lie Asp Phe lie lie lie Arg Glu Asn Thr Glu Gly Leu Tyr 115 120 125 tcc ggt gtt gaa tac tac gac gaa gaa aaa gaa ctg gca ate tct gaa 432
Ser Gly Val Glu Tyr Tyr Asp Glu Glu Lys Glu Leu Ala lie Ser Glu 130 135 140 cgt cac ate tcc aag aaa ggt tcc aag ege ate ate aaa ttc gca ttc 480
Arg His lie Ser Lys Lys Gly Ser Lys Arg lie lie Lys Phe Ala Phe 145 150 155 160 gaa tac get gtt aag cac cac cgt aag aaa gtt tcc tgc ate cac aag 528
Glu Tyr Ala Val Lys His His Arg Lys Lys Val Ser Cys lie His Lys 165 170 175 tct aac ate ctg cgt ate act gac ggt ctg ttc ctg aac ate ttc aac 576
Ser Asn lie Leu Arg lie Thr Asp Gly Leu Phe Leu Asn lie Phe Asn 180 185 190 gaa ttc aaa gaa aaa tac aaa aac gaa tac aac ate gaa ggt aac gac 624
Glu Phe Lys Glu Lys Tyr Lys Asn Glu Tyr Asn lie Glu Gly Asn Asp 195 200 205 tac ctg gtt gac gca act geg atg tac ate ctg aaa tct ccg cag atg 672
Tyr Leu Val Asp Ala Thr Ala Met Tyr lie Leu Lys Ser Pro Gin Met 210 215 220 ttc gac gtt ate gtt act acc aac ctg ttc ggt gac att ctg tct gac 720
Phe Asp Val lie Val Thr Thr Asn Leu Phe Gly Asp lie Leu Ser Asp 225 230 235 240 gaa geg tct ggt ctg ctg ggt ggt ctg ggt ctg geg ccg tct get aac 768
Glu Ala Ser Gly Leu Leu Gly Gly Leu Gly Leu Ala Pro Ser Ala Asn 245 250 255 ate ggt gac aac tac ggt ctg ttc gaa ccg gtt cac ggt tct gca ccg 816 lie Gly Asp Asn Tyr Gly Leu Phe Glu Pro Val His Gly Ser Ala Pro 260 265 270 •286· 146966-序列表.doc 201037078 gat ate get ggt aaa ggc gtt get aac ccg ate get gca gta ctg tet 864
Asp lie Ala Gly Lys Gly Val Ala Asn Pro lie Ala Ala Val Leu Ser 275 280 285 get tet atg atg ctg tac tac ctg gat atg aaa gag aag tet ege ctg 912
Ala Ser Met Met Leu Tyr Tyr Leu Asp Met Lys Glu Lys Ser Arg Leu 290 295 300 ctg aaa gac get gtt aaa cag gta ctg gca cac aaa gac ate act ccg 960
Leu Lys Asp Ala Val Lys Gin Val Leu Ala His Lys Asp lie Thr Pro 305 310 315 320 gac ctg ggt ggt aac ctg aaa acc aaa gaa gtt tet gac aag ate ate 1008
Asp Leu Gly Gly Asn Leu Lys Thr Lys Glu Val Ser Asp Lys lie lie 325 330 335 1029
❹ gaa gaa ctg cgt aag ate teg Glu Glu Leu Arg Lys lie Ser 340 <210> 219 <211> 343
<212> PRT <213>雜色甲烷球菌Nankai-3 <400> 219
Met Lys lie Pro Lys He Cys Val lie Glu Gly Asp Gly lie Gly Lys 15 10 15
Glu Val lie Pro Glu Thr Val Arg lie Leu Lys Glu lie Gly Asp Phe 20 25 30
Glu Phe lie Tyr Glu His Ala Gly Tyr Glu Cys Phe Lys Arg Cys Gly 35 40 45
Asp Ala lie Pro Glu Lys Thr Leu Lys Thr Ala Lys Glu Cys Asp Ala 50 55 60 lie Leu Phe Gly Ala Val Ser Thr Pro Lys Leu Asp Glu Thr Glu Arg 65 70 75 80
Lys Pro Tyr Lys Ser Pro lie Leu Thr Leu Arg Lys Glu Leu Asp Leu 85 90 95
Tyr Ala Asn Val Arg Pro lie His Lys Leu Asp Asn Ser Asp Ser Ser 100 105 110
Asn Asn lie Asp Phe lie lie 工le Arg Glu Asn Thr Glu Gly Leu Tyr 115 120 125
Ser Gly Val Glu Tyr Tyr Asp Glu Glu Lys Glu Leu Ala lie Ser Glu 130 135 140
Arg His lie Ser Lys Lys Gly Ser Lys Arg lie lie Lys Phe Ala Phe 145 150 155 160
Glu Tyr Ala Val Lys His His Arg Lys Lys Val Ser Cys lie His Lys 165 170 175
Ser Asn lie Leu Arg lie Thr Asp Gly Leu Phe Leu Asn lie Phe Asn 180 185 190
Glu Phe Lys Glu Lys Tyr Lys Asn Glu Tyr Asn lie Glu Gly Asn Asp 195 200 205
Tyr Leu Val Asp Ala Thr Ala Met Tyr lie Leu Lys Ser Pro Gin Met 210 215 220
Phe Asp Val lie Val Thr Thr Asn Leu Phe Gly Asp lie Leu Ser Asp -287- 146966-序列表.doc 201037078 225 230 235 240
Glu Ala Ser Gly Leu Leu Gly Gly Leu Gly Leu Ala Pro Ser Ala Asn 245 250 255 lie Gly Asp Asn Tyr Gly Leu Phe Glu Pro Val His Gly Ser Ala Pro 260 265 270
Asp lie Ala Gly Lys Gly Val Ala Asn Pro lie Ala Ala Val Leu Ser 275 280 285
Ala Ser Met Met Leu Tyr Tyr Leu Asp Met Lys Glu Lys Ser Arg Leu 290 295 300
Leu Lys Asp Ala Val Lys Gin Val Leu Ala His Lys Asp lie Thr Pro 305 310 315 320
Asp Leu Gly Gly Asn Leu Lys Thr Lys Glu Val Ser Asp Lys lie lie 325 330 335
Glu Glu Leu Arg Lys lie Ser 340 <210> 220 <211> 1032 <212> DNA <213> 雜色曱烷球菌Nankai-3 <220> <223> 野生型 <400> 220 TTGAAAATAC CTAAAATATG TGTGATAGAA GGGGACGGCA TAGGAAAAGA AGTAATACCT 60 GAAACAGTCC GCATATTAAA AGAT^ATAGGG GACTTTGAAT TCATATATGA ACATGCGGGA 120 TATGAATGTT TTAAACGGTG CGGTGATGCA ATACCCGAAA AAACATTAAA AACTGCAAAA 180 GAATGCGATG CCATATTATT TGGTGCAGTT AGCACCCCTA AATTAGATGA AACGGAGCGA 240 AAACCATATA AAAGCCCCAT ATTAACACTT AGAAAAGAAT TAGACCTATA TGCAAATGTA 300 AGACCAATAC ATAAATTAGA TAATTCCGAT AGTTCAAATA ATATAGATTT CATAATAATT 360 CGAGAAAACA CAGAAGGACT ATATTCTGGT GTGGAATATT ATGACGAAGA AAAAGAGTTG 420 GCAATATCGG AAAGGCATAT ATCTAAAAAA GGAAGCAAAA GAATTATTAA ATTTGCATTT 480 GAATATGCTG TGAAACATCA CAGAAAAAAA GTATCCTGTA TCCATAAATC AAATATTTTA 540 AGAATTACGG ACGGTCTTTT TTTAAACATA TTCAATGAAT TTAAAGAAAA ATATAAAAAT 600 GAATACAATA TAGAAGGAAA CGATTATTTA GTAGATGCTA CGGCAATGTA TATTTTA7\AA 660
AGCCCACAAA TGTTCGATGT AATTGTAACT ACAAATTTAT TTGGTGACAT ATTATCCGAT 720 GAAGCATCGG GATTATTGGG AGGTTTAGGC TTGGCTCCTT CAGCAAATAT TGGGGATAAT 780 TATGGATTAT TTGAGCCTGT GCATGGTTCC GCCCCAGACA TAGCTGGAAA AGGTGTTGCA 840 AATCCGATAG CTGCGGTTCT TAGTGCTTCG ATGATGCTAT ATTACCTAGA TATGAAGGAG 900 AAAAGTAGGC TATTAAAAGA TGCCGTAAAA CAGGTGTTGG CCCATAAAGA TATAACTCCT 960 GATTTAGGGG GCAATTTAAA AACAAAGGAA GTAAGCGATA AAATTATTGA AGAATTACGG 1020 AAAATATCCT AA 1032 288- 146966-序列表.doc 48 201037078 <210> 221 <211> 1017
<212> DNA <213> 海沼甲烷球菌S2 <220> <221> CDS <222> (1).·(1017) <400> 221 atg cgt aac act ccg aaa ate tgc gtt ate aac ggt gac ggt ate ggt
Met Arg Asn Thr Pro Lys lie Cys Val lie Asn Gly Asp Gly lie Gly 15 10 15 96 aac gaa gtt gtt ccg gaa acc gtt ege gta ctg aac gaa ctg ggt gac
Asn Glu Val Val Pro Glu Thr Val Arg Val Leu Asn Glu Leu Gly Asp 20 25 30 ttc gaa ttc ate cac geg cac get ggt tac gaa tgc ttc aag ege tgc 144
Phe Glu Phe lie His Ala His Ala Gly Tyr Glu Cys Phe Lys Arg Cys 35 40 45 ggt gac get ate ccg gaa aac acc ate gaa ate get aaa gag tet gac 192
Gly Asp Ala lie Pro Glu Asn Thr lie Glu lie Ala Lys Glu Ser Asp 50 55 60 tgc ate ctg ttc ggt tet gta act act ccg aaa cca act gaa ctg aaa 240
Cys 工le Leu Phe Gly Ser Val Thr Thr Pro Lys Pro Thr Glu Leu Lys 65 70 75 80 aac aag tet tac ege tet ccg att ctg act ctg cgt aaa gag ctg gat 288
Asn Lys Ser Tyr Arg Ser Pro lie Leu Thr Leu Arg Lys Glu Leu Asp 85 90 95 ctg tac get aac ate cgt ccg act tac aac ttc gac aac ctg gat ttc 336
Leu Tyr Ala Asn lie Arg Pro Thr Tyr Asn Phe Asp Asn Leu Asp Phe 100 105 110 gtt ate ate cgt gaa aac act gaa ggt ctg tac gtt aag aaa gaa tac 384
Val lie lie Arg Glu Asn Thr Glu Gly Leu Tyr Val Lys Lys Glu Tyr 115 120 125 tac gac gag aaa aac gaa gtt get ate get gaa cgt ate ate tee aag 432 *
Tyr Asp Glu Lys Asn Glu Val Ala lie Ala Glu Arg lie lie Ser Lys 130 135 140 ttc ggt tet tet ege ate gtt aaa ttc gca ttc gac tac gca gta cag 480
Phe Gly Ser Ser Arg lie Val Lys Phe Ala Phe Asp Tyr Ala Val Gin 145 150 155 160 -289· 146966-序列表.doc 201037078 aac aac cgt aag aaa gtt tcc tgc ate cac aaa geg aac gtt ctg ege 528
Asn Asn Arg Lys Lys Val Ser Cys lie His Lys Ala Asn Val Leu Arg 165 170 175 gta act gac ggt ctg ttc ctg gaa gtt ttc gaa gaa atg tec aag cac 576
Val Thr Asp Gly Leu Phe Leu Glu Val Phe Glu Glu Met Ser Lys His 180 185 190 tac gaa aaa ctg ggt ate aaa tet gac gac tac ctg ate gac gca act 624
Tyr Glu Lys Leu Gly lie Lys Ser Asp Asp Tyr Leu lie Asp Ala Thr 195 200 205 geg atg tac ctg ate cgt aac ccg cag atg ttc gac gtt ctg gtt act 672
Ala Met Tyr Leu lie Arg Asn Pro Gin Met Phe Asp Val Leu Val Thr 210 215 220 acc aac ctg ttc ggt gac ate ctg tet gac gaa gca get ggt ctg att 720
Thr Asn Leu Phe Gly Asp lie Leu Ser Asp Glu Ala Ala Gly Leu lie 225 230 235 240 ggt ggt ctg ggt atg tet ccg tet get aac ate ggt gac aaa aac ggt 768
Gly Gly Leu Gly Met Ser Pro Ser Ala Asn lie Gly Asp Lys Asn Gly 245 250 255 ctg ttc gaa ccg gtt cac ggt tet gca ccg gat ate get ggt aaa ggt 816
Leu Phe Glu Pro Val His Gly Ser Ala Pro Asp lie Ala Gly Lys Gly 260 265 270 ate tec aac ccg ate geg act ate ctg tet get gca atg atg ctg gat 864
He Ser Asn Pro 工le Ala Thr lie Leu Ser Ala Ala Met Met Leu Asp 275 280 285 cac ctg aaa atg aac aaa gaa get gaa tac ate cgt aaa geg gtt aag 912
His Leu Lys Met Asn Lys Glu Ala Glu Tyr lie Arg Lys Ala Val Lys 290 295 300 aaa acc gtt gaa tgc aaa tac ctg act ccg gac ctg ggt ggt aac ctg 960
Lys Thr Val Glu Cys Lys Tyr Leu Thr Pro Asp Leu Gly Gly Asn Leu 305 310 315 320 aaa act ttc gaa gtt act gaa aag ate ate gaa tec ate cgt tet cag 1008
Lys Thr Phe Glu Val Thr Glu Lys lie lie Glu Ser lie Arg Ser Gin 325 330 335 atg att caa 1017
Met He Gin <210> 222 <211> 339 •290· 146966-序列表.doe 201037078
<212> PRT <213>海沼甲烷球菌S2 <400> 222
Met Arg Asn Thr Pro Lys lie Cys Val lie Asn Gly Asp Gly lie Gly 15 10 15
Asn Glu Val Val Pro Glu Thr Val Arg Val Leu Asn Glu Leu Gly Asp 20 25 30
Phe Glu Phe lie His Ala His Ala Gly Tyr Glu Cys Phe Lys Arg Cys 35 40 45
Gly Asp Ala lie Pro Glu Asn Thr lie Glu lie Ala Lys Glu Ser Asp 50 55 60
Cys lie Leu Phe Gly Ser Val Thr Thr Pro Lys Pro Thr Glu Leu Lys 65 70 75 80
Asn Lys Ser Tyr Arg Ser Pro lie Leu Thr Leu Arg Lys Glu Leu Asp 85 90 95
Leu Tyr Ala Asn 工le Arg Pro Thr Tyr Asn Phe Asp Asn Leu Asp Phe 100 105 110
Val lie 工le Arg Glu Asn Thr Glu Gly Leu Tyr Val Lys Lys Glu Tyr 115 120 125
Tyr Asp Glu Lys Asn Glu Val Ala lie Ala Glu Arg lie lie Ser Lys 130 135 140
Phe Gly Ser Ser Arg. lie Val Lys Phe Ala Phe Asp Tyr Ala Val Gin 145 150 155 160
Asn Asn Arg Lys Lys Val Ser Cys lie His Lys Ala Asn Val Leu Arg 165 170 175
Val Thr Asp Gly Leu Phe Leu Glu Val Phe Glu Glu Met Ser Lys His 180 185 190
Tyr Glu Lys Leu Gly lie Lys Ser Asp Asp Tyr Leu lie Asp Ala Thr 195 200 205
Ala Met Tyr Leu lie Arg Asn Pro Gin Met Phe Asp Val Leu Val Thr 210 215 220
Thr Asn Leu Phe Gly Asp lie Leu Ser Asp Glu Ala Ala Gly Leu lie 225 230 235 240
Gly Gly Leu Gly Met Ser Pro Ser Ala Asn lie Gly Asp Lys Asn Gly 245 250 255
Leu Phe Glu Pro Val His Gly Ser Ala Pro Asp lie Ala Gly Lys Gly 260 265 270 lie Ser Asn Pro lie Ala Thr lie Leu Ser Ala Ala Met Met Leu Asp 275 280 285
His Leu Lys Met Asn Lys Glu Ala Glu Tyr lie Arg Lys Ala Val Lys 290 295 300
Lys Thr Val Glu Cys Lys Tyr Leu Thr Pro Asp Leu Gly Gly Asn Leu 305 310 315 320
Lys Thr Phe Glu Val Thr Glu Lys lie lie Glu Ser lie Arg Ser Gin 325 330 335
Met He Gin <210> 223 <211〉 1020 <212> DNA <213> 海沼甲烷球菌S2 <220> <223> 野生型 <400> 223 ATGAGAAACA CTCCCAAAAT TTGTGTTATC AATGGTGACG GTATTGGAAA CGAAGTAGTT 60 291 - 146966·序列表.doc 201037078
CCTGAAACGG TGCGAGTTTT AAATGAACTT GGTGACTTCG AATTCATTCA TGCCCATGCA120 GGTTACGAAT GTTTTAAAAG ATGTGGCGAT GCGATACCAG AAAACACAAT TGAAATTGCA 180 AAAGAATCTG ATTGTATTTT ATTTGGATCA GTTACCACTC CAAAACCGAC TGAATTAAAA 240 AATAAATCAT ATAGAAGTCC AATATTAACT TTAAGAAAAG AACTTGACCT TTATGCAAAT 300 ATTAGGCCAA CTTATAACTT TGATAATCTT GATTTTGTTA TAATTCGAGA AAATACTGAA 360 GGACTCTATG ΤΑΆΑΑΑΑΛΘΑ ATATTACGAC GAAAAAAACG AAGTTGCAAT TGCTGAGCGA 420 ATAATTTCAA AATTTGGAAG TTCTAGAATT GTAAAATTTG CTTTTGATTA TGCGGTTCAA 480 AATAACAGAA AAAAAGTATC CTGCATACAT AAAGCAAACG TATTACGGGT TACTGACGGA 540 TTATTTTTAG AAGTTTTCGA AGAAATGTCT AAACATTACG AAAAATTAGG AATAAAGTCT 600 GATGACTACC TAATTGACGC GACAGCAATG TATTTGATTA GAAACCCGCA AATGTTTGAT 660 GTATTGGTTA CT^VCAAATCT TTTTGGAGAT ATTTTATCTG ATGAAGCTGC AGGACTTATT 720 GGCGGACTTG GAATGTCTCC TTCAGCAAAC ATTGGTGATA AAAACGGATT ATTTGAGCCA 780 GTTCATGGAT CTGCACCAGA CATTGCTGGA AAAGGAATTT CAAACCCGAT TGCAACAATA 840 TTGAGTGCTG CAATGATGCT TGACCATTTA AAAATGAATA AAGAAGCCGA ATACATTAGA 900 AAAGCGGTTA AAAAAACGGT TGAATGTAAA TATTTAACTC CTGATCTTGG GGGAAACTTA 960 AAAACTTTTG AAGTTACGGA AAAAATCATT G7VATCCATAA GGTCTCAGAT GATTCAGTGA 1020 <210> 224 <211> 498 <212> DNA <213> 醋酸甲烷八聯球菌C2A <220> <221> CDS <222> (1) - . (498) <400> 224 atg atg aaa aac ccg ate ate ggt ege gta tgg aaa ttc ggt gac gac Met Met Lys Asn Pro lie lie Gly Arg Val Trp Lys Phe Gly Asp Asp 1 5 10 15 48 ate gac act gac gtt ate ate cca ggt aaa tat ctg cgt act aaa gac lie Asp Thr Asp Val lie lie Pro Gly Lys Tyr Leu Arg Thr Lys Asp 20 25 30 96 atg cag ate ttc get gca cac geg atg gaa ggt att get ccg gaa ttc 144 Met Gin lie Phe Ala Ala His Ala Met Glu Gly lie Ala Pro Glu Phe 35 40 45 acc aag aaa geg aag 192 Thr Lys Lys Ala Lys 50 ccg ggc gac ate ate gtt get ggc gaa aac ttc Pro Gly Asp lie lie Val Ala Gly Glu Asn Phe 55 60 146966·序列表.doc -292 201037078 ggt tgc ggt tct tcc cgt gag cag gca ccg ctg gca ate aag cac get 240
Gly Cys Gly Ser Ser Arg Glu Gin Ala Pro Leu Ala lie Lys His Ala 65 70 75 80 ggt ate get tgc gtt gtt get aaa tct ttc gca cgt ate ttc ttc cgt 288
Gly lie Ala Cys Val Val Ala Lys Ser Phe Ala Arg lie Phe Phe Arg 85 90 95 aac gca ate aac gtt ggt ctg ccg ctg atg gaa get gac ate gaa tgc 336
Asn Ala lie Asn Val Gly Leu Pro Leu Met Glu Ala Asp lie Glu Cys 100 105 110 cag gaa ggt gac gaa ate gaa gtt gat ctg ctg aaa ggt gaa gtt ege 384
Gin Glu Gly Asp Glu lie Glu Val Asp Leu Leu Lys Gly Glu Val Arg 115 120 125
o gtt ccg ggt aaa ggc gta ttc gtt ggt aac aag ctg ccg gac ttc ctg 432
Val Pro Gly Lys Gly Val Phe Val Gly Asn Lys Leu Pro Asp Phe Leu 130 135 140 ctg gat ate ctg act gac ggt ggt ctg gtt tct cac cgt aag aaa geg 480
Leu Asp lie Leu Thr Asp Gly Gly Leu Val Ser His Arg Lys Lys Ala 145 150 155 160 cag aac gaa cag aaa gaa 498
Gin Asn Glu Gin Lys Glu 165 <210> 225
<211> 166 <212> PRT <213> 醋酸甲烷八聯球菌C2A <400> 225
Met Met Lys Asn Pro lie lie Gly Arg Val Trp Lys Phe Gly Asp Asp 15 10 15 lie Asp Thr Asp Val lie lie Pro Gly Lys Tyr Leu Arg Thr Lys Asp 20 25 30
Met Gin lie Phe Ala Ala His Ala Met Glu Gly lie Ala Pro Glu Phe 35 40 45
Thr Lys Lys Ala Lys Pro Gly Asp lie lie Val Ala Gly Glu Asn Phe 50 55 60
Gly Cys Gly Ser Ser Arg Glu Gin Ala Pro Leu Ala lie Lys His Ala 65 70 75 80
Gly lie Ala Cys Val Val Ala Lys Ser Phe Ala Arg lie Phe Phe Arg 85 90 95
Asn Ala lie Asn Val Gly Leu Pro Leu Met Glu Ala Asp lie Glu Cys 100 105 110
Gin Glu Gly Asp Glu lie Glu Val Asp Leu Leu Lys Gly Glu Val Arg 115 120 125
Val Pro Gly Lys Gly Val Phe Val Gly Asn Lys Leu Pro Asp Phe Leu 130 135 140
Leu Asp lie Leu Thr Asp Gly Gly Leu Val Ser His Arg Lys Lys Ala •293 - 146966·序列表.doc 201037078 145 150 155 160
Gin Asn Glu Gin Lys Glu 165
<210> 226 <211> 501 <212> DNA <213>醋酸甲烷八聯球菌C2A <220> <223>野生型 <400> 226 GTGATGAAAA ACCCCATCAT AGGCCGAGTC TGGAAATTCG GAGACGACAT AGACACTGAT 60 GTGATTATCC CGGGAAAATA CCTGCGGACA AAGGATATGC AGATTTTTGC AGCTCATGCG 120 ATGGAAGGTA TTGCTCCTGA GTTTACGAAA AAGGCAAAGC CCGGAGATAT TATTGTTGCA 180 GGAGAGAATT TCGGGTGCGG TTCCTCAAGG GAACAGGCTC CTCTTGCTAT AAAACATGCG 240 GGAATAGCCT GCGTTGTGGC AAAGTCCTTT GCAAGAATCT TTTTCAGAAA TGCGATTAAC 300 GTGGGGCTGC CCCTGATGGA GGCTGATATT GAGTGTCAGG AGGGGGACGA GATAGAAGTC 360 GACCTGCTCA AAGGGGAGGT CAGGGTTCCC GGAAAAGGGG TGTTTGTTGG AAACAAACTG 420 CCTGACTTCC TGCTCGATAT ACTCACAGAC GGCGGACTTG TTTCCCATAG AAAAAAAGCG 480 CAAAACGAGC AGAAAGAATA A 501 <210> 227 <211> 486
<212> DNA <213> 亨氏甲烷螺菌JF-1 <220> <221> CDS <222> (1) .. (486) <400> 227 atg act geg ctg aaa ggt tet ggt ccg get gtt tgc ate ggt gaa gat 48 Met Thr Ala Leu Lys Gly Ser Gly Pro Ala Val Cys lie Gly Glu Asp 195 200 205 ate gac act gac ctg gtt ate get ggt cgt tat ctg cgt act aaa gac 96 lie Asp Thr Asp Leu Val lie Ala Gly Arg Tyr Leu Arg Thr Lys Asp 210 215 220 tgg tet ttc tgg gca cag cac gta ttc gaa gat ctg gac ccg tee ctg 144 Trp Ser Phe Trp Ala Gin His Val Phe Glu Asp Leu Asp Pro Ser Leu 225 230 235 240 gca gaa cgt ctg aaa ggc gca gta ctg gtt get ggt aag aac atg ggc 192
Ala Glu Arg Leu Lys Gly Ala Val Leu Val Ala Gly Lys Asn Met Gly 245 250 255 -294 146966-序列表.doc 201037078 tgc ggt tct tct cgc gaa cag get gca cgt geg ctg cac gaa get ggc 240
Cys Gly Ser Ser Arg Glu Gin Ala Ala Arg Ala Leu His Glu Ala Gly 260 265 270 gtt ctg geg gtt ate get ccg tct ttc get cgt ate ttc ttc cgt aac 288
Val Leu Ala Val lie Ala Pro Ser Phe Ala Arg lie Phe Phe Arg Asn 275 280 285 tgc ate aac gtt ggt ctg ccg ctg ctg gaa tgc gac ctg act ggc tgc 336
Cys lie Asn Val Gly Leu Pro Leu Leu Glu Cys Asp Leu Thr Gly Cys 290 295 300 act gac ggt atg ate ate act ttc gac tgc act gaa ggc tgg gtt gaa 384
Thr Asp Gly Met lie lie Thr Phe Asp Cys Thr Glu Gly Trp Val Glu 305 310 315 320
gtt gac ggt act cgc tac ctg ttc cgt ccg ctg tct ccg cgt atg cag 432
Val Asp Gly Thr Arg Tyr Leu Phe Arg Pro Leu Ser Pro Arg Met Gin 325 330 335 gaa ate ctg tct acc ggt ggt ctg att gaa tac tgg aag cgc cgt aaa 480
Glu lie Leu Ser Thr Gly Gly Leu lie Glu Tyr Trp Lys Arg Arg Lys 340 345 350 gag cgc 486
Glu Arg
<210> 228 <211> 162 <212> PRT <213> 亨氏甲烷螺菌JF-1 <400> 228
Met Thr Ala Leu Lys Gly Ser Gly Pro Ala Val Cys lie Gly Glu Asp 195 200 205 lie Asp Thr Asp Leu Val lie Ala Gly Arg Tyr Leu Arg Thr Lys Asp 210 215 220
Trp Ser Phe Trp Ala Gin His Val Phe Glu Asp Leu Asp Pro Ser Leu 225 230 235 240
Ala Glu Arg Leu Lys Gly Ala Val Leu Val Ala Gly Lys Asn Met Gly 245 250 255
Cys Gly Ser Ser Arg Glu Gin Ala Ala Arg Ala Leu His Glu Ala Gly 260 265 270
Val Leu Ala Val lie Ala Pro Ser Phe Ala Arg lie Phe Phe Arg Asn 275 280 285
Cys lie Asn Val Gly Leu Pro Leu Leu Glu Cys Asp Leu Thr Gly Cys 290 295 300
Thr Asp Gly Met lie lie Thr Phe Asp Cys Thr Glu Gly Trp Val Glu 305 310 315 320
Val Asp Gly Thr Arg Tyr Leu Phe Arg Pro Leu Ser Pro Arg Met Gin 325 330 335
Glu lie Leu Ser Thr Gly Gly Leu lie Glu Tyr Trp Lys Arg Arg Lys 340 345 350
Glu Arg -295 - 146966-序列表.doc 201037078 <210> 229
<211> 489 <212> DNA <213>亨氏曱烷螺菌JF-1 <220> <223>野生型 <400> 229 ATGACTGCTC TCAAAGGCTC CGGCCCTGCA GTCTGTATCG GAGAGGATAT CGATACGGAT 60 CTGGTGATTG CCGGACGATA CCTGAGAACA AAAGACTGGA GTTTTTGGGC ACAGCATGTG 120 TTTGAGGATC TTGATCCTTC TCTTGCCGAA AGACTGAAAG GAGCAGTGCT TGTCGCCGGA 180 AAGAACATGG GTTGCGGGTC TTCACGGGAA CAGGCGGCCC GGGCTCTGCA TGAGGCAGGA 240 GTTCTTGCTG TCATCGCCCC TTCGTTTGCC AGAATTTTTT TCCGGAACTG CATCAATGTC 300 GGGCTTCCGT TACTGGAATG TGATCTGACC GGATGCACCG ATGGCATGAT CATCACTTTT 360 GATTGCACTG AAGGGTGGGT GGAGGTGGAT GGGACGCGGT ATCTCTTCCG GCCCCTCTCT 420 CCACGGATGC AGGAGATTCT CAGCACCGGC GGTCTGATTG AGTACTGGAA ACGGAGAAAA 480 GAGCGATGA 489 <210> 230 <211> 489
<212> DNA <213> 思氏曱烷球形菌DSM 3091 <220> <221> CDS <222> (1)..(489) <400> 230 atg gac tcc atg aaa ggt aaa gta tgg act ttc cgc gac tgc ate gac
Met Asp Ser Met Lys Gly Lys Val Trp Thr Phe Arg Asp Cys lie Asp 15 10 15 act gac gtt ate att get ggt cgt tat ctg cgt act ttc aac ccg gaa
Thr Asp Val lie lie Ala Gly Arg Tyr Leu Arg Thr Phe Asn Pro Glu 20 25 30 gat ctg geg gca cac gta atg gaa gca gaa gat ccg gaa ttc tet tec 144
Asp Leu Ala Ala His Val Met Glu Ala Glu Asp Pro Glu Phe Ser Ser 35 40 45 aaa gtt ggt aaa ggc gac ate ate gtt ggc ggc tgg aac ttc ggt tgc 192
Lys Val Gly Lys Gly Asp lie lie Val Gly Gly Trp Asn Phe Gly Cys 50 55 60 ggt tet tec cgt gag cag gca ccg gtt get ate aaa act get ggc gta 240
Gly Ser Ser Arg Glu Gin Ala Pro Val Ala lie Lys Thr Ala Gly Val 146966·序列表.doc 48 96 296- 201037078 65 70 75 80 tct gcg gtt ate geg aag tet ttc get ege att ttc tac cgt aac get 288
Ser Ala Val lie Ala Lys Ser Phe Ala Arg lie Phe Tyr Arg Asn Ala 85 90 95 ate aac ate ggt ctg ccg gtt ate act gca gac ate gaa gtt gac gaa 336 lie Asn lie Gly Leu Pro Val lie Thr Ala Asp lie Glu Val Asp Glu 100 105 110 ggt gac ate ctg gaa gtt aac ate gaa gat ggt ate ate ate aac gaa 384
Gly Asp lie Leu Glu Val Asn lie Glu Asp Gly lie lie lie Asn Glu 115 120 125 act acc aag aaa act ttc aaa ate aag ccg ttc gac get gaa atg ctg 432
Thr Thr Lys Lys Thr Phe Lys lie Lys Pro Phe Asp Ala Glu Met Leu 130 135 140 gac ate ctg gaa aac ggt ggt ctg gtt aac cag tac ctg aaa aac aag 480
Asp lie Leu Glu Asn Gly Gly Leu Val Asn Gin Tyr Leu Lys Asn Lys 145 150 155 160 aaa gaa gtt 489
Lys Glu Val <210> 231 <211> 163
<212> PRT <213> 思氏曱烷球形菌DSM3091 <400> 231
Met Asp Ser Met Lys Gly Lys Val Trp Thr Phe Arg Asp Cys lie Asp 1 5 10 15
Thr Asp Val lie lie Ala Gly Arg Tyr Leu Arg Thr Phe Asn Pro Glu 20 25 30
Asp Leu Ala Ala His Val Met Glu Ala Glu Asp Pro Glu Phe Ser Ser 35 40 45
Lys Val Gly Lys Gly Asp lie lie Val Gly Gly Trp Asn Phe Gly Cys 50 55 60
Gly Ser Ser Arg Glu Gin Ala Pro Val Ala lie Lys Thr Ala Gly Val 65 70 75 80
Ser Ala Val lie Ala Lys Ser Phe Ala Arg lie Phe Tyr Arg Asn Ala 85 90 95 lie Asn lie Gly Leu Pro Val lie Thr Ala Asp lie Glu Val Asp Glu 100 105 110
Gly Asp lie Leu Glu Val Asn lie Glu Asp Gly lie lie lie Asn Glu 115 120 125
Thr Thr Lys Lys Thr Phe Lys lie Lys Pro Phe Asp Ala Glu Met Leu 130 135 140
Asp lie Leu Glu Asn Gly Gly Leu Val Asn Gin Tyr Leu Lys Asn Lys 145 150 155 160
Lys Glu Val <210> 232 •297· 146966-序列表.doc 201037078 <211> 492
<212> DNA <213> 思氏曱烷球形菌DSM 3091 <220> <223> 野生型 <400> 232 ATGGATAGTA TGAAAGGAAA AGTATGGACA TTTAGAGATT GTATAGATAC TGATGTAATT 60 ATTGCTGGAA GATATTTAAG AACATTTAAT CCAGAAGATT TAGCAGCACA TGTAATGGAA 120 GCTGAAGATC CAGAATTTTC AAGTAAAGTT GGAAAGGGAG ATATAATTGT TGGTGGATGG 180 AATTTTGGTT GTGGATCTTC AAGGGAACAG GCACCAGTTG CAATAAAGAC AGCAGGAGTA 240 TCAGCAGTAA TAGCTAAATC ATTTGCCAGA ATTTTTTATA GAAATGCTAT TAATATTGGC 300 TTACCTGTTA TAACAGCAGA TATTGAAGTG GATGAAGGGG ATATTTTAGA AGTAAATATT 360 GAAGATGGAA TTATTATAAA TGAAACAACT AAAAAAACAT TTAAAATTAA GCCATTTGAT 420 GCTGAAATGT TGGATATTTT AGAAAATGGT GGTCTTGTTA ATCAATATTT AAAAAATAAG 480 AAGGAGGTAT AG 492 <210> 233 <211> 483
<212> DNA <213>嗜熱曱烷鬃菌PT <220> <221> CDS <222> (1)·.(483) <400> 233 atg ate ege ggt cgt gca tgg gta ttc ggt gac gac gtt gac act gac
Met lie Arg Gly Arg Ala Trp Val Phe Gly Asp Asp Val Asp Thr Asp 195 200 205 gtt ate ate cca ggt aaa tat ctg cgt act aaa gat ctg cgt gaa ctg
Val lie lie Pro Gly Lys Tyr Leu Arg Thr Lys Asp Leu Arg Glu Leu 210 215 220 gca gaa cac gta atg gaa ggt ctg gat ccg tet ttc tet tee aaa gtt 144
Ala Glu His Val Met Glu Gly Leu Asp Pro Ser Phe Ser Ser Lys Val 225 230 235 240 cag ccg ggc gac gtt ate gtt get ggt aaa aac ttc ggt tgc ggt tet 192
Gin Pro Gly Asp Val lie Val Ala Gly Lys Asn Phe Gly Cys Gly Ser 245 250 255 tee cgt gag cag gca ccg ctg geg ctg aaa ctg geg ggt ate tee tgc 240
Ser Arg Glu Gin Ala Pro Leu Ala Leu Lys Leu Ala Gly lie Ser Cys 260 265 270 146966-序列表.doc 48 96 •298 201037078 ate gtt geg cgt tet ttc get ege att ttc tac cgt aac get ate aac 288 工le Val Ala Arg Ser Phe Ala Arg lie Phe Tyr Arg Asn Ala lie Asn 275 280 285 ate ggt ctg ccg ctg ate gaa get gac gta atg tgc gaa ccg ggt gaa 336 lie Gly Leu Pro Leu lie Glu Ala Asp Val Met Cys Glu Pro Gly Glu 290 295 300 gtt gtt gaa gtt gac ctg tet ege ggt ege gtt ate get ggc ggt cgt 384
Val Val Glu Val Asp Leu Ser Arg Gly Arg Val lie Ala Gly Gly Arg 305 310 315 320 gaa tac acc ggt cgt aag ctg ccg gac ttc ctg atg gaa ate ctg aac 432
Glu Tyr Thr Gly Arg Lys Leu Pro Asp Phe Leu Met Glu lie Leu Asn 325 330 335
gac ggt ggt ctg gtt get cac ege cgt aag cag cgt gaa cgt ggt ctg 480
Asp Gly Gly Leu Val Ala His Arg Arg Lys Gin Arg Glu Arg Gly Leu 340 345 350 ate 483 lie <210> 234
<211> 161 <212> PRT <213>嗜熱甲烷鬃菌PT <400> 234
Met lie Arg Gly Arg Ala Trp Val Phe Gly Asp Asp Val Asp Thr Asp 195 200 205
Val lie lie Pro Gly Lys Tyr Leu Arg Thr Lys Asp Leu Arg Glu Leu 210 215 220
Ala Glu His Val Met Glu Gly Leu Asp Pro Ser Phe Ser Ser Lys Val 225 230 235 240
Gin Pro Gly Asp Val lie Val Ala Gly Lys Asn Phe Gly Cys Gly Ser 245 250 255
Ser Arg Glu Gin Ala Pro Leu Ala Leu Lys Leu Ala Gly lie Ser Cys 260 265 270 lie Val Ala Arg Ser Phe Ala Arg lie Phe Tyr Arg Asn Ala lie Asn 275 280 285 lie Gly Leu Pro Leu lie Glu Ala Asp Val Met Cys Glu Pro Gly Glu 290 295 300
Val Val Glu Val Asp Leu Ser Arg Gly Arg Val lie Ala Gly Gly Arg 305 310 315 320
Glu Tyr Thr Gly Arg Lys Leu Pro Asp Phe Leu Met Glu lie Leu Asn 325 330 335
Asp Gly Gly Leu Val Ala His Arg Arg Lys Gin Arg Glu Arg Gly Leu 340 345 350 lie <210> 235 <211> 486
<212> DNA -299- 146966-序列表.doc 201037078 <213>嗜熱甲烷鬃菌PT <220> <223> 野生型 <400> 235 ATGATCAGGG GCAGGGCATG GGTCTTTGGG GATGATGTTG ATACAGATGT AATAATTCCA 60 GGGAAATACC TCAGGACAAA GGATCTGCGC GAGCTGGCGG AGCACGTCAT GGAGGGTCTG 120 GATCCGTCGT TCTCATCAAA GGTCCAGCCA GGGGATGTTA TAGTAGCAGG CAAGAACTTC 180 GGGTGCGGCT CTTCCAGGGA GCAGGCCCCG CTTGCCCTGA AGCTCGCAGG AATATCATGC 240 ATCGTCGCCC GATCCTTCGC CAGGATATTT TACAGGAACG CCATAAACAT CGGTCTCCCT 300 CTCATTGAGG CAGATGTCAT GTGCGAGCCG GGGGAGGTCG TGGAGGTAGA TCTCTCCCGG 360 GGGAGGGTCA TCGCAGGCGG AAGGGAATAT ACCGGCAGAA AACTTCCGGA CTTCCTGATG 420 GAGATCCTGA ACGATGGAGG GCTCGTGGCC CACAGAAGAA AGCAAAGGGA GCGCGGTCTG 480 ATTTGA 486 <210> 236 <211> 1272
<212> DNA
<213> 醋酸曱烷八聯球菌C2A <220> <221> CDS <222> (1).·(1272) <400> 236 atg ctg ttc gac tac cgt aaa ggt ggt ctg ttc ctg ggt act ate tet
Met Leu Phe Asp Tyr Arg Lys Gly Gly Leu Phe Leu Gly Thr lie Ser 15 10 15 gag aag ate ttc tet ege get get ggt act gaa geg aaa get aac gac
Glu Lys lie Phe Ser Arg Ala Ala Gly Thr Glu Ala Lys Ala Asn Asp 20 25 30 ttc gtt ctg get gac gtt gac tac gca atg geg cac gac ggt act tet 144
Phe Val Leu Ala Asp Val Asp Tyr Ala Met Ala His Asp Gly Thr Ser 35 40 45 gta ctg geg gtt aac gca ttc aaa gaa atg gaa atg gaa aaa gta tgg 192
Val Leu Ala Val Asn Ala Phe Lys Glu Met Glu Met Glu Lys Val Trp 50 55 60 gac ccg tee ege ate gtt gtt cog ttc gac cac ate geg ccg gca aac 240
Asp Pro Ser Arg lie Val Val Pro Phe Asp His lie Ala Pro Ala Asn 65 70 75 80 aac gaa act tet gca act ctg cag cgt gaa ate cgt gaa tgg gtt aaa 288
Asn Glu Thr Ser Ala Thr Leu Gin Arg Glu lie Arg Glu Trp Val Lys 146966·序列表.doe -300- 48 96 201037078 85 90 95 gag cag ggt ate ccg aac ttc tac gaa gtt ggt gaa ggt ate tgc cac 336
Glu Gin Gly lie Pro Asn Phe Tyr Glu Val Gly Glu Gly lie Cys His 100 105 110 cag gtt ctg ccg gaa aac ggc ttc geg ctg cca ggt aag ctg gtt gtt 384
Gin Val Leu Pro Glu Asn Gly Phe Ala Leu Pro Gly Lys Leu Val Val 115 120 125 ggt get gac tet cac tee tgc acc tac ggt geg ttc ggt geg ttc gca 432
Gly Ala Asp Ser His Ser Cys Thr Tyr Gly Ala Phe Gly Ala Phe Ala 130 135 140 act ggc gtt ggt gca act gac atg get gaa ate ttc gca act ggt aag 480
Thr Gly Val Gly Ala Thr Asp Met Ala Glu lie Phe Ala Thr Gly Lys 145 150 155 160 ctg tgg ttc aaa gtt ccg gaa tee ttc cgt atg acc gtt gaa ggt tet 528
Leu Trp Phe Lys Val Pro Glu Ser Phe Arg Met Thr Val Glu Gly Ser 165 170 175 ctg cgt aaa ggc gtt tac get aaa gac ctg act ctg tac ctg att ggt 576
Leu Arg Lys Gly Val Tyr Ala Lys Asp Leu Thr Leu Tyr Leu lie Gly 180 185 190 aag act ggt att get ggc gca act tac aaa geg gtt gaa ttc tac ggt 624
Lys Thr Gly lie Ala Gly Ala Thr Tyr Lys Ala Val Glu Phe Tyr Gly 195 200 205 cag get ate cgt gaa ctg acc gtt get ggt cgt atg act ctg tgc aac 672
Gin Ala lie Arg Glu Leu Thr Val Ala Gly Arg Met Thr Leu Cys Asn 210 215 220 atg get ate gaa atg ggt geg aaa acc ggt ate gtt ccg ccg gac gaa 720
Met Ala lie Glu Met Gly Ala Lys Thr Gly lie Val Pro Pro Asp Glu 225 230 235 240 aag act ttc gaa ttc ctg aaa aac ege get get gca act tac gaa ccg 768
Lys Thr Phe Glu Phe Leu Lys Asn Arg Ala Ala Ala Thr Tyr Glu Pro 245 250 255 gtt tac get gac ccg gat get gtt tac ctg gaa gag ttc act tac gac 816
Val Tyr Ala Asp Pro Asp Ala Val Tyr Leu Glu Glu Phe Thr Tyr Asp 260 265 270 get gat gac ate gaa cca cag gtt get tgc ccg cac cag gtt gac aac 864
Ala Asp Asp lie Glu Pro Gin Val Ala Cys Pro His Gin Val Asp Asn 275 280 285 -301 - 146966-序列表.doc 201037078 gtt aag ccg gtt ggt gaa gtt gaa ggt act cac gtt gac cag gta ttc 912
Val Lys Pro Val Gly Glu Val Glu Gly Thr His Val Asp Gin Val Phe 290 295 300 ate ggt act tgc acc aac ggt cgt ctg gaa gat ctg gaa gtt get get 960 lie Gly Thr Cys Thr Asn Gly Arg Leu Glu Asp Leu Glu Val Ala Ala 305 310 315 320 geg gta ctg aaa ggt aaa cag gtt get gtt cgt act ate gtt ate ccg 1008
Ala Val Leu Lys Gly Lys Gin Val Ala Val Arg Thr lie Val lie Pro 325 330 335 gca tet ege act act ctg ctg gca get ate gaa aac ggc acc atg gaa 1056
Ala Ser Arg Thr Thr Leu Leu Ala Ala lie Glu Asn Gly Thr Met Glu 340 345 350 act ctg ctg aaa get ggc gta act ctg gca act ccg ggt tgc ggt ccg 1104
Thr Leu Leu Lys Ala Gly Val Thr Leu Ala Thr Pro Gly Cys Gly Pro 355 360 365 tgc ctg ggt gca cac cag ggc gta ctg ggt gaa ggt gaa gtt tgc gta 1152
Cys Leu Gly Ala His Gin Gly Val Leu Gly Glu Gly Glu Val Cys Val 370 375 380 tet act get aac cgt aac ttc aaa ggt cgt atg ggt aaa ggc ggc ttc 1200
Ser Thr Ala Asn Arg Asn Phe Lys Gly Arg Met Gly Lys Gly Gly Phe 385 390 395 400 ate tac ctg get tet ccg gca act geg gca get tet geg ctg act ggc 1248
He Tyr Leu Ala Ser Pro Ala Thr Ala Ala Ala Ser Ala Leu Thr Gly 405 410 415 gaa ate act gat ccg ege act gtc 1212
Glu lie Thr Asp Pro Arg Thr Val 420 <210> 237 <211> 424
<212> PRT <213> 醋酸甲烷八聯球g C2A <400> 237
Met Leu Phe Asp Tyr Arg Lys Gly Gly Leu Phe Leu Gly Thr lie Ser 15 10 15
Glu Lys lie Phe Ser Arg Ala Ala Gly Thr Glu Ala Lys Ala Asn Asp 20 25 30
Phe Val Leu Ala Asp Val Asp Tyr Ala Met Ala His Asp Gly Thr Ser 35 40 45
Val Leu Ala Val Asn Ala Phe Lys Glu Met Glu Met Glu Lys Val Trp 50 55 60
Asp Pro Ser Arg He Val Val Pro Phe Asp His He Ala Pro Ala Asn 302- 146966·序列表.doc 201037078 65 70 75 80
Asn Glu Thr Ser Ala Thr Leu Gin Arg Glu lie Arg Glu Trp Val Lys 85 90 95
Glu Gin Gly lie Pro Asn Phe Tyr Glu Val Gly Glu Gly lie Cys His 100 105 110
Gin Val Leu Pro Glu Asn Gly Phe Ala Leu Pro Gly Lys Leu Val Val 115 120 125
Gly Ala Asp Ser His Ser Cys Thr Tyr Gly Ala Phe Gly Ala Phe Ala 130 135 140
Thr Gly Val Gly Ala Thr Asp Met Ala Glu lie Phe Ala Thr Gly Lys 145 150 155 160
Leu Trp Phe Lys Val Pro Glu Ser Phe Arg Met Thr Val Glu Gly Ser 165 170 175
Leu Arg Lys Gly Val Tyr Ala Lys Asp Leu Thr Leu Tyr Leu lie Gly 180 185 190
Lys Thr Gly lie Ala Gly Ala Thr Tyr Lys Ala Val Glu Phe Tyr Gly 195 200 205
Gin Ala 工le Arg Glu Leu Thr Val Ala Gly Arg Met Thr Leu Cys Asn 210 215 220
Met Ala lie Glu Met Gly Ala Lys Thr Gly lie Val Pro Pro Asp Glu 225 230 235 240
Lys Thr Phe Glu Phe Leu Lys Asn Arg Ala Ala Ala Thr Tyr Glu Pro 245 250 255
Val Tyr Ala Asp Pro Asp Ala Val Tyr Leu Glu Glu Phe Thr Tyr Asp 260 265 270
Ala Asp Asp lie Glu Pro Gin Val Ala Cys Pro His Gin Val Asp Asn 275 280 285
Val Lys Pro Val Gly Glu Val Glu Gly Thr His Val Asp Gin Val Phe 290 295 300
He Gly Thr Cys Thr Asn Gly Arg Leu Glu Asp Leu Glu Val Ala Ala 305 310 315 320
Ala Val Leu Lys Gly Lys Gin Val Ala Val Arg Thr He Val lie Pro 325 330 335
Ala Ser Arg Thr Thr Leu Leu Ala Ala lie Glu Asn Gly Thr Met Glu 340 345 350
Thr Leu Leu Lys Ala Gly Val Thr Leu Ala Thr Pro Gly Cys Gly Pro 355 360 365
Cys Leu Gly Ala His Gin Gly Val Leu Gly Glu Gly Glu Val Cys Val 370 375 380
Ser Thr Ala Asn Arg Asn Phe Lys Gly Arg Met Gly Lys Gly Gly Phe 385 390 395 400 lie Tyr Leu Ala Ser Pro Ala Thr Ala Ala Ala Ser Ala Leu Thr Gly 405 410 415
Glu lie Thr Asp Pro Arg Thr Val 420 <210> 238 <211> 1275
<212> DNA <213> 醋酸曱烷八聯球菌C2A <220> <223> 野生型 <400> 238 TTGTTATTTG ATTATCGGAA AGGAGGCTTA TTTCTGGGAA CAATCAGCGA GAAAATCTTT 60 TCCCGGGCAG CGGGAACAGA GGCAAAAGCT AACGATTTTG TGCTGGCAGA TGTGGACTAT 120 303- 146966·序列表.doc 201037078 GCAATGGCGC ATGACGGCAC ATCGGTACTT GCCGTGAATG CTTTTAAGGA AATGGAGATG 180 GAAAAGGTCT GGGACCCTTC AAGGATCGTA GTCCCCTTTG ACCACATCGC GCCTGCGAAT 240 AATGAGACCT CAGCCACCCT GCAGAGAGAG ATCAGGGAAT GGGTAAAGGA GCAGGGGATC 300 CCTAACTTCT ATGAGGTCGG GGAAGGGATC TGTCACCAGG TCCTTCCGGA AAACGGTTTT 360 GCATTGCCCG GAAAGCTGGT TGTTGGGGCT GATTCGCATT CCTGCACGTA CGGGGCTTTT 420 GGAGCTTTTG CT^ACAGGGGT AGGGGCTACC GATATGGCTG AAATCTTTGC TACGGGAAAA 480 CTCTGGTTTA AGGTTCCGGA AAGCTTCAGG ATGACGGTTG AAGGGAGCCT TCGGAAAGGG 540 GTCTATGCAA AGGATCTGAC TCTTTACCTG ATCGGAAAGA CCGGAATTGC CGGGGCGACC 600 TATAAAGCAG TTGAGTTTTA CGGGCAGGCA ATCCGCGAAC TTACGGTTGC TGGCAGGATG 660
ACGCTCTGCA ATATGGCAAT TGAGATGGGC GCAAAGACCG GGATCGTCCC TCCGGACGAA 720
AAGACCTTCG AGTTCCTGAA AAACAGGGCA GCAGCTACTT ATGAACCTGT CTATGCCGAC 780 CCGGATGCTG TTTATCTGGA AGAGTTCACT TATGATGCCG ACGATATCGA ACCCCAGGTT 840
GCCTGCCCGC ATCAGGTGGA TAATGTAAAG CCCGTAGGAG AGGTTGAAGG CACTCATGTA 900
GACCAGGTCT TTATCGGGAC GTGCACGAAC GGCAGGCTTG AAGACCTCGA GGTCGCAGCA 960 GCAGTCCTGA AAGGAAAACA GGTTGCAGTC AGGACAATTG TGATCCCTGC CTCTCGCACC 1020 ACCCTCCTTG CAGCAATCGA AAACGGGACA ATGGAAACTC TGCTGAAAGC CGGGGTAACC 1080 CTTGCAACCC CGGGCTGCGG ACCCTGCCTC GGTGCCCATC AGGGGGTGCT CGGAGAAGGC 1140 GAGGTCTGCG TTTCAACCGC AAACAGGAAC TTCAAAGGCA GGATGGGAAA AGGCGGTTTT 1200 ATTTATCTTG CATCTCCAGC AACTGCAGCA GCCTCGGCCC TGACAGGAGA AATCACCGAT 1260 CCGAGGACAG TTTGA 1275 <210> 239
<211> 1212 <212> DNA <213> 亨氏曱烷螺菌JF-1 <220> <221> CDS <222> (1)..(1212) <400> 239 atg gtt act Met Val Thr 1 ctg Leu 5 tct gag aag Ser Glu Lys ate ctg ggt gca ccg gca ggc act tac lie Leu Gly Ala Pro Ala Gly Thr Tyr 10 15 48 ate gac cgt cac lie Asp Arg His 20 ate gac ege gca lie Asp Arg Ala 25 ttc tgc cac gac Phe Cys His Asp 30 ggt act ggt att Gly Thr Gly He cag geg aag ate ate tac gac geg atg ggc gca ccg ggt ate get aac 144 •304 146966-序列表.doc 96 201037078
Gin Ala Lys lie lie Tyr Asp Ala Met Gly Ala Pro Gly lie Ala Asn 35 40 45 ccg gac tcc gtt tac ate ate tac gac cac att get ccg get aac aac ' 192
Pro Asp Ser Val Tyr lie lie Tyr Asp His lie Ala Pro Ala Asn Asn 50 55 60 tet cag act gca gaa ctg cag get gaa ctg cgt act ctg geg cgt gaa 240
Ser Gin Thr Ala Glu Leu Gin Ala Glu Leu Arg Thr Leu Ala Arg Glu 65 70 75 80 tgc ggt gtt cac ttc tgg gat ate ggt tet ggt ate tgc cac cag gta 288
Cys Gly Val His Phe Trp Asp lie Gly Ser Gly lie Cys His Gin Val 85 90 95 atg get gaa ggt cag gtt get ccg ggt gaa gtt gtt ate ggt get gac 336
Ο
Met Ala Glu Gly Gin Val Ala Pro Gly Glu Val Val lie Gly Ala Asp 100 105 110 tet cac tec tgc act ctg ggt geg ctg ggt gca ttc get acc ggt gtt 384
Ser His Ser Cys Thr Leu Gly Ala Leu Gly Ala Phe Ala Thr Gly Val 115 120 125 ggt gca tet gac atg get ggt ate tgg gtt tet ggt gaa acc tgg ctg 432
Gly Ala Ser Asp Met Ala Gly lie Trp Val Ser Gly Glu Thr Trp Leu 130 135 140 ege gtt ccg gac tec ate ggt ate cac ctg tet ggt tet ctg aag cag 480
Arg Val Pro Asp Ser lie Gly lie His Leu Ser Gly Ser Leu Lys Gin 145 150 155 160 ggc gtt gag tgg aaa gac gtt geg ctg act tac gtt get ege ctg ggt 528
Gly Val Glu Trp Lys Asp Val Ala Leu Thr Tyr Val Ala Arg Leu Gly 165 170 175 atg gac ggt gca act tac get geg ctg gaa ttc ate ggt gaa tec act 576
Met Asp Gly Ala Thr Tyr Ala Ala Leu Glu Phe lie Gly Glu Ser Thr 180 185 190 ccg tet gta ccg atg gaa ggt cgt ctg act ctg tgc aac atg get gtt 624
Pro Ser Val Pro Met Glu Gly Arg Leu Thr Leu Cys Asn Met Ala Val 195 200 205 gaa gca ggc geg aaa acc ggt ctg ttc tac get gac aaa gaa act gaa 672
Glu Ala Gly Ala Lys Thr Gly Leu Phe Tyr Ala Asp Lys Glu Thr Glu 210 215 220 cgt tac ctg get gaa tac tec gta cca tgc ccg atg caa gtt ctg gaa 720
Arg Tyr Leu Ala Glu Tyr Ser Val Pro Cys Pro Met Gin Val Leu Glu -305 - 146966-序列表.doc 201037078 225 230 235 240 aac cca gac tac gtt cag gac tgc tac ctg gat ctg get gac ate 768
Asn Pro Asp Tyr Val Gin Asp Cys Tyr Leu Asp Leu Ala Asp lie 245 250 255 ccg gtt tgc gca gta ccg cac ege gtt gac acc gta cag ccg gtt 816
Pro Val Cys Ala Val Pro His Arg Val Asp Thr Val Gin Pro Val 260 265 270 gca ctg geg ggt act cac ctg gat cag gtt ttc ate ggt act tgc 864
Ala Leu Ala Gly Thr His Leu Asp Gin Val Phe lie Gly Thr Cys 275 280 285 aac ggt cgt ttc gaa gat ctg get ege get geg cgt ate ctg aaa 912
Asn Gly Arg Phe Glu Asp Leu Ala Arg Ala Ala Arg lie Leu Lys 290 295 300 cgt ege gtt aaa gtt cgt act ate gtt gtt ccg gca tet gaa cgt 960
Arg Arg Val Lys Val Arg Thr lie Val Val Pro Ala Ser Glu Arg 305 310 315 320 ttc ctg aaa gca ate ctg tet ggt gtt get get gac ctg gta cag 1008
Phe Leu Lys Ala lie Leu Ser Gly Val Ala Ala Asp Leu Val Gin 325 330 335 ggc tgc acc ate ggt ccg ccg ggt tgc ggt ccg tgc ctg ggt geg 1056
Gly Cys Thr lie Gly Pro Pro Gly Cys Gly Pro Cys Leu Gly Ala 340 345 350 atg ggc gtt ctg ggt gaa ggt gaa gtt geg ctg tet act gca aac 1104
Met Gly Val Leu Gly Glu Gly Glu Val Ala Leu Ser Thr Ala Asn 355 360 365 aac ttc aaa aac cgt atg ggc gta ggt get tet tac tac ctg tgc 1152
Asn Phe Lys Asn Arg Met Gly Val Gly Ala Ser Tyr Tyr Leu Cys 370 375 380 ccg tet act get get gca tet get ate tac ggt gaa ate act gac 1200
Pro Ser Thr Ala Ala Ala Ser Ala lie Tyr Gly Glu lie Thr Asp 385 390 395 400 cgt gaa gtc gtt Arg Glu Val Val <210> 240 <211> 404
<212> PRT <213> 亨氏曱烷螺菌JF-1 146966-序列表.doc 306- gaa Glu ccg Pro acc Thr ggt Gly gac Asp get Ala cac His cgt Arg tet Ser ccg Pro 1212 201037078 <400> 240
Met Val Thr Leu Ser Glu Lys lie Leu Gly Ala Pro Ala Gly Thr Tyr 15 10 15 lie Asp Arg His lie Asp Arg Ala Phe Cys His Asp Gly Thr Gly lie 20 25 30
Gin Ala Lys lie lie Tyr Asp Ala Met Gly Ala Pro Gly lie Ala Asn 35 40 45
Pro Asp Ser Val Tyr lie lie Tyr Asp His lie Ala Pro Ala Asn Asn 50 55 60
Ser Gin Thr Ala Glu Leu Gin Ala Glu Leu Arg Thr Leu Ala Arg Glu 65 70 75 80
Cys Gly Val His Phe Trp Asp lie Gly Ser Gly lie Cys His Gin Val 85 90 95
Met Ala Glu Gly Gin Val Ala Pro Gly Glu Val Val lie Gly Ala Asp 100 105 110
Ser His Ser Cys Thr Leu Gly Ala Leu Gly Ala Phe Ala Thr Gly Val 115 120 125
Gly Ala Ser Asp Met Ala Gly lie Trp Val Ser Gly Glu Thr Trp Leu 130 135 140
Arg Val Pro Asp Ser lie Gly lie His Leu Ser Gly Ser Leu Lys Gin 145 150 155 160 Ο
Gly Val Glu Trp Lys Asp Val Ala Leu Thr Tyr Val Ala Arg Leu Gly 165 170 175
Met Asp Gly Ala Thr Tyr Ala Ala Leu Glu Phe lie Gly Glu Ser Thr 180 185 190
Pro Ser Val Pro Met Glu Gly Arg Leu Thr Leu Cys Asn Met Ala Val 195 200 205
Glu Ala Gly Ala Lys Thr Gly Leu Phe Tyr Ala Asp Lys Glu Thr Glu 210 215 220
Arg Tyr Leu Ala Glu Tyr Ser Val Pro Cys Pro Met Gin Val Leu Glu 225 230 235 240
Asn Pro Asp Tyr Val Gin Asp Cys Tyr Leu Asp Leu Ala Asp lie Glu 245 250 255
Pro Val Cys Ala Val Pro His Arg Val Asp Thr Val Gin Pro Val Pro 260 265 270
Ala Leu Ala Gly Thr His Leu Asp Gin Val Phe lie Gly Thr Cys Thr 275 280 285
Asn Gly Arg Phe Glu Asp Leu Ala Arg Ala Ala Arg lie Leu Lys Gly 290 295 300
Arg Arg Val Lys Val Arg Thr lie Val Val Pro Ala Ser Glu Arg Asp 305 310 315 320
Phe Leu Lys Ala lie Leu Ser Gly Val Ala Ala Asp Leu Val Gin Ala 325 330 335
Gly Cys Thr lie Gly Pro Pro Gly Cys Gly Pro Cys Leu Gly Ala His 340 345 350
Met Gly Val Leu Gly Glu Gly Glu Val Ala Leu Ser Thr Ala Asn Arg 355 360 365
Asn Phe Lys Asn Arg Met Gly Val Gly Ala Ser Tyr Tyr Leu Cys Ser 370 375 380
Pro Ser Thr Ala Ala Ala Ser Ala lie Tyr Gly Glu lie Thr Asp Pro 385 390 395 400
Arg Glu Val Val <210> 241 <211> 1215
<212> DNA <213>亨氏甲烷螺菌JF-l <220> <223> 野生型 -307- 146966-序列表.doc 201037078 <400> 241 ATGGTGACAT TATCGGAGAA GATTCTAGGT GCCCCGGCCG GCACGTATAT TGATCGACAT 60 ATTGACCGGG CATTCTGCCA TGACGGGACC GGGATTCAGG CAAAGATAAT CTATGATGCA 120 ATGGGGGCTC CGGGTATTGC AAATCCGGAT TCTGTCTACA TTATTTATGA TCATATTGCT 180 CCGGCGAACA ACTCCCAGAC TGCAGAACTC CAGGCAGAGC TCCGGACCCT TGCCAGAGAG 240 TGCGGGGTGC ATTTCTGGGA TATCGGCTCT GGTATATGTC ATCAGGTGAT GGCAGAAGGG 300 CAGGTGGCAC CAGGGGAGGT TGTCATCGGA GCTGATTCTC ATTCATGCAC ACTCGGTGCT 360 CTCGGGGCGT TTGCAACCGG CGTCGGGGCA AGTGACATGG CAGGTATCTG GGTGTCTGGT 420 GAGACCTGGC TGAGGGTTCC TGATTCAATC GGCATTCATC TCTCCGGATC CTTAAAGCAA 480 GGCGTTGAGT GGAAGGATGT TGCGCTGACC TATGTGGCAC GGCTCGGGAT GGACGGAGCA 540 ACCTATGCAG CCCTGGAGTT CATTGGTGAA TCCACTCCGT CGGTTCCCAT GGAAGGACGA 600 CTGACATTAT GCAATATGGC GGTTGAGGCC GGGGCAAAGA CCGGGCTCTT TTATGCAGAC 660 AAGGAGACAG AGCGATATCT CGCAGAGTAT TCTGTTCCCT GTCCGATGCA GGTTTTGGAA 720 AACCCGGACT ATGTGCAGGA CTGCTATCTT GATCTCGCAG ATATTGAGCC GGTATGTGCG 780 GTTCCTCACC GGGTGGATAC TGTCCAACCG GTTCCGGCCC TTGCAGGTAC CCATCTTGAT 840 CAGGTCTTTA TCGGGACCTG TACGAACGGC CGGTTTGAAG ATCTCGCACG TGCTGCCAGG 900 ATTCTGAAAG GACGCCGGGT AAAAGTCAGA ACCATTGTTG TTCCGGCATC AGAGCGTGAT 960 TTCCTCAAGG CTATTTTGTC AGGGGTTGCT GCTGACCTCG TGCAGGCAGG ATGTACCATC 1020 GGACCGCCCG GCTGTGGTCC CTGCCTTGGT GCCCATATGG GTGTTCTGGG TGAGGGTGAA 1080 GTAGCCCTTT CAACCGCAAA CCGGAACTTT AAGAACCGGA TGGGTGTGGG GGCTTCGTAT 1140 TATCTCTGTT CACCCTCCAC TGCTGCAGCA AGTGCCATAT ATGGTGAGAT TACTGATCCA 1200 AGGGAGGTTG TATGA 1215 <210> 242 <211> 1245
<212> DNA <213> 思氏甲烷球形菌DSM 3091 <220> <221> CDS <222> (1)..(1245) <400> 242 gaa Glu 48 atg aac ate tet gaa aag att Met Asn lie Ser Glu Lys lie 1 5 ctg geg aaa geg tet aac aaa gaa Leu Ala Lys Ala Ser Asn Lys Glu 10 15 gtt tet ccg ggc gac acc ate Val Ser Pro Gly Asp Thr lie act get aac ate gac gtt gca atg tet Thr Ala Asn He Asp Val Ala Met Ser -308· 146966-序列表.doc 96 201037078 20 25 30 cac gac ggt act tct ccg cca acc ate aaa gta ttc gag aag ate get 144
His Asp Gly Thr Ser Pro Pro Thr lie Lys Val Phe Glu Lys lie Ala 35 40 45 gac aaa gta tgg gat ccg gaa aag ate gtt ctg gta ttc gac cac gtt 192
Asp Lys Val Trp Asp Pro Glu Lys lie Val Leu Val Phe Asp His Val 50 55 60 ate ccg get aac acc ate ggt tct get gaa ttc caa cag gtt gtt cgt 240 lie Pro Ala Asn Thr lie Gly Ser Ala Glu Phe Gin Gin Val Val Arg 65 70 75 80 gaa ttc ggt aag aaa cag aaa ate ccg aac atg tac att cag ggc gaa 288
Glu Phe Gly Lys Lys Gin Lys lie Pro Asn Met Tyr lie Gin Gly Glu 85 90 95
ggt gtt tgc cac gaa gtt ctg ccg gac tac ggt cac gtt aag ccg tct 336
Gly Val Cys His Glu Val Leu Pro Asp Tyr Gly His Val Lys Pro Ser 100 105 110 acc gtt ate gtt ggt get gac tct cac acc tgt act tac ggc gca ttc 384
Thr Val lie Val Gly Ala Asp Ser His Thr Cys Thr Tyr Gly Ala Phe 115 120 125 ggt geg ttc tct acc ggt ctg ggt gca act gac ctg gca atg gtt tac 432
Gly Ala Phe Ser Thr Gly Leu Gly Ala Thr Asp Leu Ala Met Val Tyr 130 135 140 gca act ggt cag acc tgg ttc aac gta ccg gaa tct ctg aaa ate aac 480
Ala Thr Gly Gin Thr Trp Phe Asn Val Pro Glu Ser Leu Lys 工le Asn 145 150 155 160 gtt aac ggt act ctg aac gaa aac gtt tac tcc aaa gac gtt ate ctg 528
Val Asn Gly Thr Leu Asn Glu Asn Val Tyr Ser Lys Asp Val lie Leu 165 170 175 aaa ate ate aaa gaa ctg ggt gca tac ggt gca act tac aaa tct ctg 576
Lys 工le lie Lys Glu Leu Gly Ala Tyr Gly Ala Thr Tyr Lys Ser Leu 180 185 190 gaa ttc cac ggt gac acc ate gac aac atg tcc gtt get tct ege ctg 624
Glu Phe His Gly Asp Thr lie Asp Asn Met Ser Val Ala Ser Arg Leu 195 200 205 acc atg act aac atg get ate gaa tgc ggt geg aaa aac ggt ate atg 672
Thr Met Thr Asn Met Ala lie Glu Cys Gly Ala Lys Asn Gly lie Met 210 215 220 -309- 146966-序列表.doc 201037078 gtt ccg aac aag cag act aaa gaa tac ctg tct cag cgt ggt ate act 720
Val Pro Asn Lys Gin Thr Lys Glu Tyr Leu Ser Gin Arg Gly lie Thr 225 230 235 240 gac tac acc ate acc act geg tct aaa gac get gaa tac gag aag ate 768
Asp Tyr Thr lie Thr Thr Ala Ser Lys Asp Ala Glu Tyr Glu Lys lie 245 250 255 tac gac ttc gac gtt gac gat ctg cag ccg cag ate get tgc ccg cac 816
Tyr Asp Phe Asp Val Asp Asp Leu Gin Pro Gin lie Ala Cys Pro His 260 265 270 aac gtt gac aac gtt gaa gat ate gac aaa gtt get ggt act cac ate 864
Asn Val Asp Asn Val Glu Asp lie Asp Lys Val Ala Gly Thr His lie 275 280 285 gac cag geg gta ctg ggt tee tgc acc aac ggt cgt tac gaa gat ctg 912
Asp Gin Ala Val Leu Gly Ser Cys Thr Asn Gly Arg Tyr Glu Asp Leu 290 295 300 ctg cag get gca gaa gtt ate gaa ggt cac aaa ate cac gaa gat gtt 960
Leu Gin Ala Ala Glu Val lie Glu Gly His Lys lie His Glu Asp Val 305 310 315 320 gaa ctg ctg gta ttc ccg gca tct ege cac gtt tac gaa aaa get ate 1008
Glu Leu Leu Val Phe Pro Ala Ser Arg His Val Tyr Glu Lys Ala lie 325 330 335 gaa acc ggt gtt ate cag act ctg ctg aaa tct aac geg att ate tgc 1056
Glu Thr Gly Val lie Gin Thr Leu Leu Lys Ser Asn Ala lie lie Cys 340 345 350 aac cca ggt tgc ggt ccg tgc ctg ggt gca cac atg ggc gta atg act 1104
Asn Pro Gly Cys Gly Pro Cys Leu Gly Ala His Met Gly Val Met Thr 355 360 365 gac gac atg act tgc ate tee acc act aac cgt aac ttc ctg ggt cgt 1152
Asp Asp Met Thr Cys lie Ser Thr Thr Asn Arg Asn Phe Leu Gly Arg 370 375 380 atg ggt tct geg aag tct tac gtt tac ctg tct aac ccg get gtt gtt 1200
Met Gly Ser Ala Lys Ser Tyr Val Tyr Leu Ser Asn Pro Ala Val Val 385 390 395 400 get get tct get ate aaa ggt gaa ate act aac ccg tct gag ate Ala Ala Ser Ala lie Lys Gly Glu lie Thr Asn Pro Ser Glu lie 405 410 415 -310- 146966-序列表.doc 1245 201037078 <210> 243 <211> 415
<212> PRT <213> 思氏甲烷球形菌DSM3091 <400> 243
Met Asn lie Ser Glu Lys lie Leu Ala Lys Ala Ser Asn Lys Glu Glu 15 10 15
Val Ser Pro Gly Asp Thr lie Thr Ala Asn lie Asp Val Ala Met Ser 20 25 30
His Asp Gly Thr Ser Pro Pro Thr lie Lys Val Phe Glu Lys lie Ala 35 40 45
Asp Lys Val Trp Asp Pro Glu Lys lie Val Leu Val Phe Asp His Val 50 55 60 lie Pro Ala Asn Thr lie Gly Ser Ala Glu Phe Gin Gin Val Val Arg 65 70 75 80
Glu Phe Gly Lys Lys Gin Lys lie Pro Asn Met Tyr lie Gin Gly Glu 85 90 95
Gly Val Cys His Glu Val Leu Pro Asp Tyr Gly His Val Lys Pro Ser 100 105 110
G
Thr Val lie Val Gly Ala Asp Ser His Thr Cys Thr Tyr Gly Ala Phe 115 120 125
Gly Ala Phe Ser Thr Gly Leu Gly Ala Thr Asp Leu Ala Met Val Tyr 130 135 140
Ala Thr Gly Gin Thr Trp Phe Asn Val Pro Glu Ser Leu Lys lie Asn 145 150 155 160
Val Asn Gly Thr Leu Asn Glu Asn Val Tyr Ser Lys Asp Val lie Leu 165 170 175
Lys lie 工le Lys Glu Leu Gly Ala Tyr Gly Ala Thr Tyr Lys Ser Leu 180 185 190
Glu Phe His Gly Asp Thr lie Asp Asn Met Ser Val Ala Ser Arg Leu 195 200 205
Thr Met Thr Asn Met Ala lie Glu Cys Gly Ala Lys Asn Gly lie Met 210 215 220
Val Pro Asn Lys Gin Thr Lys Glu Tyr Leu Ser Gin Arg Gly lie Thr 225 230 235 240
Asp Tyr Thr lie Thr Thr Ala Ser Lys Asp Ala Glu Tyr Glu Lys lie 245 250 255
Tyr Asp Phe Asp Val Asp Asp Leu Gin Pro Gin lie Ala Cys Pro His 260 265 270
Asn Val Asp Asn Val Glu Asp lie Asp Lys Val Ala Gly Thr His lie 275 280 285
Asp Gin Ala Val Leu Gly Ser Cys Thr Asn Gly Arg Tyr Glu Asp Leu 290 295 300
Leu Gin Ala Ala Glu Val lie Glu Gly His Lys lie His Glu Asp Val 305 310 315 320
Glu Leu Leu Val Phe Pro Ala Ser Arg His Val Tyr Glu Lys Ala lie 325 330 335
Glu Thr Gly Val lie Gin Thr Leu Leu Lys Ser Asn Ala lie lie Cys 340 345 350
Asn Pro Gly Cys Gly Pro Cys Leu Gly Ala His Met Gly Val Met Thr 355 360 365
Asp Asp Met Thr Cys lie Ser Thr Thr Asn Arg Asn Phe Leu Gly Arg 370 375 380
Met Gly Ser Ala Lys Ser Tyr Val Tyr Leu Ser Asn Pro Ala Val Val 385 390 395 400
Ala Ala Ser Ala lie Lys Gly Glu lie Thr Asn Pro Ser Glu lie 405 410 415 <210> 244 -311- 146966-序列表.doc 201037078 <211> 1248 <212> DNA <213> 思氏曱烷球形菌DSM 3091 <220> <223> 野生型 <400> 244
ATGAATATAA GTGAAAAAAT ACTTGCAAAA GCATCAAATA AGGAAGAAGT AAGTCCTGGA 60 GATACT^VTTA CTGCAAATAT AGATGTTGCA ATGAGTCATG ATGGAACAAG TCCTCCTACA 120 ATAAAAGTAT TTGAAAAAAT AGCTGATAAA GTATGGGATC CAGAAAAAAT AGTTCTTGTA 180 TTCGATCATG TGATTCCTGC AAACACAATC GGATCTGCTG AATTTCAACA GGTAGTAAGA 240 GAATTTGGTA AAAAACAAAA GATTCCAAAC ATGTATATTC AAGGAGA7VGG AGTTTGTCAT 300 GAAGTACTTC CAGACTATGG TCATGTT^lAAA CCTTCAACAG TAATAGTTGG AGCAGATTCA 360
CATACATGTA CATATGGTGC TTTTGGTGCA TTCTCAACAG GACTTGGAGC TACAGATCTT 420
GCAATGGTAT ATGCAACTGG ACAAACATGG TTTAATGTTC CAGAATCCTT AAAAATAAAT 480 GTTAATGGTA CATTAAATGA AAATGTATAC TCTAAAGATG TAATTCTAAA AATAATTAAG 540 GAATTAGGTG CCTATGGTGC TACTTATAAA AGTCTTGAAT TCCATGGTGA TACAATTGAT 600 AATATGTCTG TAGCTAGTAG ATTAACAATG ACAAATATGG CAATTGAATG TGGAGCTAAA 660 AATGGTATAA TGGTACCTAA TAAACAGACA AAGGAATATC TCTCACAAAG AGGAATAACT 720 GATTATACTA TTACTACTGC TAGTAAAGAT GCAGAATATG AGAAAATTTA TGACTTTGAT 780 GTTGATGATC TTCAACCACA AATTGCATGT CCACATAATG TAGATAATGT TGAAGATATA 840 GATAAAGTAG CAGGTACTCA TATAGATCAA GCAGTACTTG GTTCATGTAC CAATGGACGT 900 TATGAAGATT TACTTCAAGC AGCTGAAGTT ATTGAAGGAC ATAAAATACA TGAAGATGTT 960 GAATTACTAG TATTCCCAGC ATCAAGACAT GTGTATGAAA AAGCAATAGA AACAGGTGTT 1020 ATTCAAACAT TACTTAAATC AAATGCAATT ATATGTAATC CTGGATGTGG ACCATGTCTT 1080 GGAGCACATA TGGGTGTAAT GACAGACGAT ATGACTTGTA TTTCCACTAC TAATAGAAAT 1140 TTCCTTGGTA GAATGGGTAG TGCTAAGTCA TATGTATATC TATCAAATCC AGCAGTTGTT 1200
GCAGCTTCAG CAATAAAAGG TGAAATTACA AATCCAAGTG AGATATAA 1248 <210> 245 <211> 1140 <212> DNA <213> 嗜熱甲烷鬃菌PT <220> <221> CDS <222> (1)-.(1140) 312- 146966·序列表.doc 48 201037078 <400> 245 atg tct cac gac ggt act tct gta ctg gca ate aaa geg ttc cgt gaa
Met Ser His Asp Gly Thr Ser Val Leu Ala lie Lys Ala Phe Arg Glu 15 10 15 96 atg ggt tct gaa aaa gta tgg gac aag tct ege ate gtt ate ccg ttc
Met Gly Ser Glu Lys Val Trp Asp Lys Ser Arg lie Val lie Pro Phe 20 25 30 gac cac ate gtt ccg get aac aac gaa act get gca act ctg cag get 144
Asp His lie Val Pro Ala Asn Asn Glu Thr Ala Ala Thr Leu Gin Ala 35 40 45 gaa gtt cgt ege tgg geg cgt get cag ggt ate gaa aac ttc tac gac 192
Glu Val Arg Arg Trp Ala Arg Ala Gin Gly lie Glu Asn Phe Tyr Asp 50 55 60
Ο tgc ggt cac ggt ate tgc cac cag gta ttc tgc gaa atg ggc ttc gca 240
Cys Gly His Gly lie Cys His Gin Val Phe Cys Glu Met Gly Phe Ala 65 70 75 80 ctg ccg ggt geg ctg gtt gtt ggt get gat tct cac tee tgc acc tac 288
Leu Pro Gly Ala Leu Val Val Gly Ala Asp Ser His Ser Cys Thr Tyr 85 90 95 ggt geg ctg ggt gca ttc ggt act ggc gtt ggt gca act gac atg get 336
Gly Ala Leu Gly Ala Phe Gly Thr Gly Val Gly Ala Thr Asp Met Ala 100 105 110 gaa ate tac tct ege ggt cgt ctg tgg ttc ege gta ccg gaa acc ate 384
Glu lie Tyr Ser Arg Gly Arg Leu Trp Phe Arg Val Pro Glu Thr lie 115 120 125 tgc atg cgt ctg gaa ggt act ctg ggt gac atg gtt tct geg aaa gac 432
Cys Met Arg Leu Glu Gly Thr Leu Gly Asp Met Val Ser Ala Lys Asp 130 135 140 ctg geg ctg ttc gtt gtt aaa gag atg ggt get gac ggt get aac tac 480
Leu Ala Leu Phe Val Val Lys Glu Met Gly Ala Asp Gly Ala Asn Tyr 145 150 155 160 atg tee gtt gag ttc gtt ggt ggt get gtt gaa cgt ctg tct ate tee 528
Met Ser Val Glu Phe Val Gly Gly Ala Val Glu Arg Leu Ser lie Ser 165 170 175 ggt cgt atg act ctg tgc aac atg ggc gtt gaa atg ggt geg aaa get 576
Gly Arg Met Thr Leu Cys Asn Met Gly Val Glu Met Gly Ala Lys Ala 180 185 190 get ate gtt ccg ccg gac gaa tee gtt gac get tac ctg gca cgt cgt 624 •313· 146966-序列表.doc 201037078
Ala lie Val Pro Pro Asp Glu Ser Val Asp Ala Tyr Leu Ala Arg Arg 195 200 205 get ege cgt cog tac act cac att cac tet gat ccg ggt tet tac tac 672
Ala Arg Arg Pro Tyr Thr His lie His Ser Asp Pro Gly Ser Tyr Tyr 210 215 220 cgt gaa ate gaa tac gac gtt tet gac att ccg ccg atg ate get gca 720
Arg Glu lie Glu Tyr Asp Val Ser Asp lie Pro Pro Met 工le Ala Ala 225 230 235 240 ccg tac ege gtt gac aac gtt cac ccg gta cgt gac ctg get ggt ate 768
Pro Tyr Arg Val Asp Asn Val His Pro Val Arg Asp Leu Ala Gly lie 245 250 255 gaa gtt gat cag gta ttc ate ggt act tgc acc aac ggt cgt ctg gaa 816
Glu Val Asp Gin Val Phe lie Gly Thr Cys Thr Asn Gly Arg Leu Glu 260 265 270 gat ctg gaa atg get get ege ate gtt aaa ggt aag ege gtt aag ate 864
Asp Leu Glu Met Ala Ala Arg lie Val Lys Gly Lys Arg Val Lys lie 275 280 285 cgt act ctg gtt ate ccg gca tet ege gaa ate tac ctg ggt geg ctg 912
Arg Thr Leu Val lie Pro Ala Ser Arg Glu lie Tyr Leu Gly Ala Leu 290 295 300 cgt tet ggt gtt ate gaa act ctg gtt gaa geg ggt geg atg ate ggt 960
Arg Ser Gly Val lie Glu Thr Leu Val Glu Ala Gly Ala Met lie Gly 305 310 315 320 ccg ccg ggt tgc ggt ccg tgc ctg ggt gca cac atg ggc gta ctg ggt 1008
Pro Pro Gly Cys Gly Pro Cys Leu Gly Ala His Met Gly Val Leu Gly 325 330 335 gac ggt gaa gtt tgc ctg tet act tet aac cgt aac ttc cca ggt cgt 1056
Asp Gly Glu Val Cys Leu Ser Thr Ser Asn Arg Asn Phe Pro Gly Arg 340 345 350 atg ggt cgt aac ggt aaa gtt tac ctg get tet ccg gca act get gca 1104
Met Gly Arg Asn Gly Lys Val Tyr Leu Ala Ser Pro Ala Thr Ala Ala 355 360 365 gca act get ate acc ggt aaa ate act gac cct egg Ala Thr Ala lie Thr Gly Lys lie Thr Asp Pro Arg 370 375 380 <210> 246 <211> 380
<212> PRT -314- 146966-序列表.doc 1140 201037078 <213>嗜熱甲烷鬃菌FT <400> 246
Met Ser His Asp Gly Thr Ser Val Leu Ala lie Lys Ala Phe Arg Glu 15 10 15
Met Gly Ser Glu Lys Val Trp Asp Lys Ser Arg lie Val lie Pro Phe 20 25 30
Asp His lie Val Pro Ala Asn Asn Glu Thr Ala Ala Thr Leu Gin Ala 35 40 45
Glu Val Arg Arg Trp Ala Arg Ala Gin Gly lie Glu Asn Phe Tyr Asp 50 55 60
Cys Gly His Gly lie Cys His Gin Val Phe Cys Glu Met Gly Phe Ala 65 70 75 80
Leu Pro Gly Ala Leu Val Val Gly Ala Asp Ser His Ser Cys Thr Tyr 85 90 95
Gly Ala Leu Gly Ala Phe Gly Thr Gly Val Gly Ala Thr Asp Met Ala 100 105 110
Glu lie Tyr Ser Arg Gly Arg Leu Trp Phe Arg Val Pro Glu Thr lie 115 120 125
G
Cys Met Arg Leu Glu Gly Thr Leu Gly Asp Met Val Ser Ala Lys Asp 130 135 140
Leu Ala Leu Phe Val Val Lys Glu Met Gly Ala Asp Gly Ala Asn Tyr 145 150 155 160
Met Ser Val Glu Phe Val Gly Gly Ala Val Glu Arg Leu Ser lie Ser 165 170 175
Gly Arg Met Thr Leu Cys Asn Met Gly Val Glu Met Gly Ala Lys Ala 180 185 190
Ala lie Val Pro Pro Asp Glu Ser Val Asp Ala Tyr Leu Ala Arg Arg 195 200 205
Ala Arg Arg Pro Tyr Thr His lie His Ser Asp Pro Gly Ser Tyr Tyr 210 215 220
Arg Glu lie Glu Tyr Asp Val Ser Asp lie Pro Pro Met lie Ala Ala 225 230 235 240
Pro Tyr Arg Val Asp Asn Val His Pro Val Arg Asp Leu Ala Gly lie 245 250 255
Glu Val Asp Gin Val Phe lie Gly Thr Cys Thr Asn Gly Arg Leu Glu 260 265 270
Asp Leu Glu Met Ala Ala Arg lie Val Lys Gly Lys Arg Val Lys lie 275 280 285
Arg Thr Leu Val lie Pro Ala Ser Arg Glu lie Tyr Leu Gly Ala Leu 290 295 300
Arg Ser Gly Val lie Glu Thr Leu Val Glu Ala Gly Ala Met lie Gly 305 310 315 320
Pro Pro Gly Cys Gly Pro Cys Leu Gly Ala His Met Gly Val Leu Gly 325 330 335
Asp Gly Glu Val Cys Leu Ser Thr Ser Asn Arg Asn Phe Pro Gly Arg 340 345 350
Met Gly Arg Asn Gly Lys Val Tyr Leu Ala Ser Pro Ala Thr Ala Ala 355 360 365
Ala Thr Ala lie Thr Gly Lys lie Thr Asp Pro Arg 370 375 380 <210> 247 <211> 1149
<212> DNA
<213>嗜熱曱烷鬃菌PT <220> <223>野生型 315- 146966-序列表.doc 201037078 <400> 247 ATGTCGCACG ACGGCACAAG CGTCCTTGCC ATAAAGGCAT TCAGGGAGAT GGGGTCAGAG 60 AAGGTCTGGG AT7^AAAGCAG GATAGTAATA CCGTTCGATC ACATCGTGCC CGCAAACAAT 120 GAGACCGCTG CGACGCTTCA GGCGGAGGTG AGAAGATGGG CGAGGGCTCA GGGGATTGAG 180 AACTTCTACG ACTGCGGTCA TGGCATATGC CACCAGGTCT TCTGCGAGAT GGGTTTCGCT 240 CTTCCTGGGG CGCTTGTCGT GGGCGCCGAC TCTCATTCCT GTACTTATGG TGCACTCGGC 300 GCATTCGGAA CAGGTGTGGG CGCCACGGAC ATGGCTGAGA TCTATTCCCG CGGGAGGCTA 360 TGGTTCAGAG TGCCGGAGAC GATATGCATG CGCCTTGAGG GCACTCTGGG TGATATGGTA 420 TCAGCAAAGG ATCTCGCCCT CTTCGTGGTG AAGGAGATGG GCGCGGATGG CGCCAACTAC 480 ATGTCCGTGG AGTTCGTCGG CGGGGCTGTG GAGAGGCTGA GCATATCAGG CAGGATGACT 540 CTGTGCAACA TGGGTGTTGA GATGGGAGCA AAGGCTGCGA TCGTCCCGCC GGATGAGAGC 600 GTCGACGCAT ACCTCGCTAG AAGAGCCAGA CGTCCATACA CGCACATCCA CTCAGACCCG 660 GGATCATACT ACAGAGAGAT CGAGTACGAT GTGAGCGATA TTCCTCCAAT GATTGCGGCT 720 CCATACCGCG TTGACAATGT TCATCCAGTC AGGGATCTGG CAGGCATCGA GGTGGACCAG 780 GTATTCATCG GCACATGTAC CAACGGAAGG CTGGAGGATC TGGAGATGGC AGCCCGGATC 840 GTGAAGGGCA AAAGGGTTAA GATCAGAACG CTTGTGATCC CCGCCTCCAG AGAGATATAT 900 CTTGGTGCTC TGAGATCTGG GGTAATTGAG ACCCTTGTCG AGGCCGGCGC GATGATCGGC 960 CCGCCGGGAT GCGGTCCATG CCTTGGCGCA CACATGGGAG TTCTGGGCGA CGGAGAGGTC 1020 TGTTTGTCCA CATCAAACAG AAACTTCCCG GGAAGGATGG GCAGA7VACGG AAAGGTCTAC 1080 CTGGCATCGC CTGCAACTGC CGCAGCCACG GCGATCACAG GAAAGATCAC AGATCCAAGG 1140 GACGTATGA 1149 <210> 248 <211> 993
<212> DNA
<213> 醋酸曱烷八聯球菌C2A <220> <221〉 (993) <222> <400> 248 atg aag ctg geg gtt ate gaa ggt gac ggt ate ggt cgt gaa gtt ate 48
Met Lys Leu Ala Val lie Glu Gly Asp Gly lie Gly Arg Glu Val lie 15 10 15 ccg get get gtt gaa gtt ctg gac gca ttc ggt ctg gaa Pro Ala Ala Val Glu Val Leu Asp Ala Phe Gly Leu Glu
a u 3 Tx g G
as a y a L 6 9 20 25 30 •316. 146966-序列表.doc 201037078 gtt tct ctg gaa ctg ggt tac get ege tgg gaa cgt act ggt tet geg 144
Val Ser Leu Glu Leu Gly Tyr Ala Arg Trp Glu Arg Thr Gly Ser Ala 35 40 45 att tct gaa gaa gat ate gca act ate aaa ggc tgc gac gca gta ctg 192 lie Ser Glu Glu Asp lie Ala Thr lie Lys Gly Cys Asp Ala Val Leu 50 55 60 ttc ggt geg gta act acc gtt ccg gat ccg tct tac aaa tct gta ctg 240
Phe Gly Ala Val Thr Thr Val Pro Asp Pro Ser Tyr Lys Ser Val Leu 65 70 75 80 ctg act att cgt aaa gag ctg gac ctg tac get aac gta cgt ccg gtt 288
Leu Thr lie Arg Lys Glu Leu Asp Leu Tyr Ala Asn Val Arg Pro Val 85 90 95
aag ccg ctg cca ggt ate acc ggt gtt acc ggt cgt aac gac ttc gac 336
Lys Pro Leu Pro Gly lie Thr Gly Val Thr Gly Arg Asn Asp Phe Asp 100 105 110 ttc ate ate gtt cgt gaa aac act gaa ggt ctg tac tee ggt ate gaa 384
Phe lie lie Val Arg Glu Asn Thr Glu Gly Leu Tyr Ser Gly lie Glu 115 120 125 gaa ate ggt ccg gac ctg tee tgg acc aag ege gtt gtt act cgt aaa 432
Glu lie Gly Pro Asp Leu Ser Trp Thr Lys Arg Val Val Thr Arg Lys 130 135 140 ggt tct gaa ege gtt get gaa tac get tgc aaa ctg geg aag cag cgt 480
Gly Ser Glu Arg Val Ala Glu Tyr Ala Cys Lys Leu Ala Lys Gin Arg 145 150 155 160 aaa aac aaa ctg act ate gtt cac aag tct aac gta ctg aaa tct gac 528
Lys Asn Lys Leu Thr lie Val His Lys Ser Asn Val Leu Lys Ser Asp 165 170 175 aag ctg ttc ctg gac gtt tgc cgt cag gtt get tct tct cac ggt gtt 576
Lys Leu Phe Leu Asp Val Cys Arg Gin Val Ala Ser Ser His Gly Val 180 185 190 gaa tac ggt gac atg ctg gtt gat tee atg get tac tct ctg atg atg 624
Glu Tyr Gly Asp Met Leu Val Asp Ser Met Ala Tyr Ser Leu Met Met 195 200 205 cgt ccg gac aaa tac gac gtt gtt gtt acc act aac ate ttc ggt gac 672
Arg Pro Asp Lys Tyr Asp Val Val Val Thr Thr Asn lie Phe Gly Asp 210 215 220 -317- 146966-序列表.doc 201037078 ate ctg tet gat atg tgc gca gca ctg gtt ggt tet ctg ggt ctg gtt 720 lie Leu Ser Asp Met Cys Ala Ala Leu Val Gly Ser Leu Gly Leu Val 225 230 235 240 ccg tet get aac ate ggt gaa aaa tac gca ttc ttc gaa ccg gtt cac 768
Pro Ser Ala Asn lie Gly Glu Lys Tyr Ala Phe Phe Glu Pro Val His 245 250 255 ggt tet gca ccg gac ate get ggt aaa ggt ate get aac ccg ctg get 816
Gly Ser Ala Pro Asp lie Ala Gly Lys Gly lie Ala Asn Pro Leu Ala 260 265 270 get ate ctg tgc gta aaa atg ctg ctg gaa tgg tet ggt ege ggt aaa 864
Ala lie Leu Cys Val Lys Met Leu Leu Glu Trp Ser Gly Arg Gly Lys 275 280 285 gaa geg gca ctg ate gac ege get ate tet tac ate ctg gag aag aaa 912
Glu Ala Ala Leu lie Asp Arg Ala lie Ser Tyr lie Leu Glu Lys Lys 290 295 300 ctg ate act ccg gac ctg ggt ggt gtt tet acc act gca gaa gtt ggt 960
Leu lie Thr Pro Asp Leu Gly Gly Val Ser Thr Thr Ala Glu Val Gly 305 310 315 320 cag act gtt geg aaa tac gta atg cag aac ctg Gin Thr Val Ala Lys Tyr Val Met Gin Asn Leu 325 330 <210> 249 <211> 331
<212> PRT <213> 醋酸甲烷八聯球菌C2A <400> 249
Met Lys Leu Ala Val lie Glu Gly Asp Gly lie Gly Arg Glu Val lie 15 10 15
Pro Ala Ala Val Glu Val Leu Asp Ala Phe Gly Leu Glu Leu Glu Lys 20 25 30
Val Ser Leu Glu Leu Gly Tyr Ala Arg Trp Glu Arg Thr Gly Ser Ala 35 40 45 lie Ser Glu Glu Asp lie Ala Thr lie Lys Gly Cys Asp Ala Val Leu 50 55 60
Phe Gly Ala Val Thr Thr Val Pro Asp Pro Ser Tyr Lys Ser Val Leu 65 70 75 80
Leu Thr lie Arg Lys Glu Leu Asp Leu Tyr Ala Asn Val Arg Pro Val 85 90 95
Lys Pro Leu Pro Gly lie Thr Gly Val Thr Gly Arg Asn Asp Phe Asp 100 105 110
Phe lie lie Val Arg Glu Asn Thr Glu Gly Leu Tyr Ser Gly lie Glu 115 120 125
Glu lie Gly Pro Asp Leu Ser Trp Thr Lys Arg Val Val Thr Arg Lys 130 135 140
Gly Ser Glu Arg Val Ala Glu Tyr Ala Cys Lys Leu Ala Lys Gin Arg 145 150 155 160 318- 993 146966-序列表.doc 201037078
Lys Asn Lys Leu Thr lie Val His Lys Ser Asn Val Leu Lys Ser Asp 165 170 175
Lys Leu Phe Leu Asp Val Cys Arg Gin Val Ala Ser Ser His Gly Val 180 185 190
Glu Tyr Gly Asp Met Leu Val Asp Ser Met Ala Tyr Ser Leu Met Met 195 200 205
Arg Pro Asp Lys Tyr Asp Val Val Val Thr Thr Asn lie Phe Gly Asp 210 215 220 lie Leu Ser Asp Met Cys Ala Ala Leu Val Gly Ser Leu Gly Leu Val 225 230 235 240
Pro Ser Ala Asn lie Gly Glu Lys Tyr Ala Phe Phe Glu Pro Val His 245 250 255
Gly Ser Ala Pro Asp lie Ala Gly Lys Gly lie Ala Asn Pro Leu Ala 260 265 270
Ala lie Leu Cys Val Lys Met Leu Leu Glu Trp Ser Gly Arg Gly Lys 275 280 285
Glu Ala Ala Leu lie Asp Arg Ala lie Ser Tyr lie Leu Glu Lys Lys 290 295 300
Leu lie Thr Pro Asp Leu Gly Gly Val Ser Thr Thr Ala Glu Val Gly 305 310 315 320
Gin Thr Val Ala Lys Tyr Val Met Gin Asn Leu 325 330 <210> 250 <211> 996
<212> DNA
<213>醋酸曱烷八聯球菌C2A <220> <223> 野生型 <400> 250 ATGAAACTCG CGGTAATCGA AGGTGATGGG ATCGGCAGGG AAGTAATTCC CGCAGCTGTT 60 GAAGTCCTGG ACGCGTTCGG ACTTGAGCTT GAGAAAGTGT CTCTGGAACT GGGCTATGCA 120 AGATGGGAAC GGACAGGGAG TGCGATCTCG GAAGAAGATA TAGCCACAAT AAAAGGGTGC 180 GACGCTGTTC TTTTCGGGGC GGTTACCACA GTACCTGACC CGAGTTACAA AAGTGTGCTT 240 CTGACAATCC GAAAGGAACT TGACCTCTAT GCCAATGTGA GGCCCGTAAA ACCTCTCCCC 300 GGCATA^CGG GCGTTACCGG GAGAAATGAT TTCGATTTTA TTATTGTCAG GGAAAATACG 360 GAAGGACTCT ACTCCGGGAT TGAGGAAATA GGCCCTGACC TTTCCTGGAC AAAAAGAGTA 420 GTTACCCGAA AAGGCTCTGA GAGGGTTGCA GAATACGCCT GCAAGCTTGC AAAACAGAGA 480 AAGAACAAAC TTACCATTGT CCACAAATCC AACGTCCTGA AGTCCGACAA ACTCTTCCTT 540 GATGTCTGCC GGCAGGTTGC CAGCTCCCAT GGGGTGGAAT ATGGCGACAT GCTGGTCGAT 600 TCTATGGCTT ATAGCCTTAT GATGCGCCCC GATAAGTACG ATGTTGTGGT TACTACAAAC 660 ATTTTTGGAG ATATCCTGAG CGACATGTGC GCAGCCCTTG TGGGAAGCCT GGGGCTTGTC 720 CCGAGCGCCA ATATAGGGGA GAAATACGCC TTTTTCGAGC CCGTACACGG GAGCGCTCCG 780 GACATCGCAG GAAAAGGTAT TGCAAACCCT CTTGCGGCAA TCCTCTGCGT GAAGATGCTG 840 319- 146966·序列表.doe 201037078 CTTGAATGGA GTGGCAGGGG GAAAGAAGCT GCTCTTATTG ACAGAGCTAT CAGTTACATA 900 CTGGAAAAGA AGCTCATCAC TCCTGACCTC GGGGGCGTCA GCACCACAGC AGAAGTCGGG 960 CAGACGGTAG CGAAGTATGT AATGCAGAAT CTTTAA 996 <210> 251 <211〉 987
<212> DNA <213>亨氏甲烷螺菌JF_1 <220> <221> CDS <222> (1)..(987) <400> 251 atg aag cgc gtt gtt gtt get ccg ggc gac ggt ate ggt ccg gaa gtt
Met Lys Arg Val Val Val Ala Pro Gly Asp Gly lie Gly Pro Glu Val 15 10 15 ate ccg tet gca ctg gaa gtt ctg cgc ttc ttc cac ccg gaa tgg gaa lie Pro Ser Ala Leu Glu Val Leu Arg Phe Phe His Pro Glu Trp Glu 20 25 30 tac ate ccg gtt tac ctg ggt tac gaa tgc tgg aaa cgt act ggc gac 144
Tyr lie Pro Val Tyr Leu Gly Tyr Glu Cys Trp Lys Arg Thr Gly Asp 35 40 45 gca ctg tet cag cgt act ctg gaa act ctg aag aaa get gac ctg ate 192
Ala Leu Ser Gin Arg Thr Leu Glu Thr Leu Lys Lys Ala Asp Leu lie 50 55 60 ctg ttc ggt get ate acc act ccg cca gac ccg aaa tac cac tet gtt 240
Leu Phe Gly Ala lie Thr Thr Pro Pro Asp Pro Lys Tyr His Ser Val 65 70 75 80 gtt ctg cgt ate cgt aaa gag ctg gat ctg tac get aac etc cgt ccg 288
Val Leu Arg lie Arg Lys Glu Leu Asp Leu Tyr Ala Asn Leu Arg Pro 85 90 95 gta ttc ggt gaa ggc ttc gac ate ctg ate gtt cgt gaa aac act gaa 336
Val Phe Gly Glu Gly Phe Asp lie Leu 工le Val Arg Glu Asn Thr Glu 100 105 110 ggt ctg tac tee ggt ate gag tgg cag gaa aaa gac cgc get tgc act 384
Gly Leu Tyr Ser Gly lie Glu Trp Gin Glu Lys Asp Arg Ala Cys Thr 115 120 125 ctg cgc gtt gtt tet gaa geg ggt tet cgc cgt ate get cgc ttc get 432
Leu Arg Val Val Ser Glu Ala Gly Ser Arg Arg lie Ala Arg Phe Ala 130 135 140 146966-序列表.doc -320- 48 96 201037078 tgc ggt tgc gcg aag cgt cgt cgt cgt cac ctg act ate ggt aac aaa 480
Cys Gly Cys Ala Lys Arg Arg Arg Arg His Leu Thr lie Gly Asn Lys 145 150 155 160 get aac gtt ctg aaa tet gac tee tac ttc ctg gat ate tgc atg gaa 528
Ala Asn Val Leu Lys Ser Asp Ser Tyr Phe Leu Asp lie Cys Met Glu 165 170 175 gaa gca gaa aaa get ggt ate tee ate gac aag aaa tac ate gac tet 576
Glu Ala Glu Lys Ala Gly lie Ser lie Asp Lys Lys Tyr lie Asp Ser 180 185 190 ctg gta ctg gac gta ctg cag cac cca ggt cgt tac gac gtt ate gtt 624
Leu Val Leu Asp Val Leu Gin His Pro Gly Arg Tyr Asp Val lie Val 195 200 205
act acc aac ate ttc ggt gac ate ctg tet gac gca gca gca ttc ctg 672
Thr Thr Asn lie Phe Gly Asp lie Leu Ser Asp Ala Ala Ala Phe Leu 210 215 220 gaa ggt ggt ctg ggt atg ctg ccg tet get aac ate ggt cgt cac cag 720
Glu Gly Gly Leu Gly Met Leu Pro Ser Ala Asn lie Gly Arg His Gin 225 230 235 240 gca ctg ttc gaa ccg gtt cac ggt tet gcg ccg gat ate get ggt aaa 768
Ala Leu Phe Glu Pro Val His Gly Ser Ala Pro Asp lie Ala Gly Lys 245 250 255 ggt ctg get aac ccg ate get gca ate ege tgc gta tet ctg ctg ctg 816
Gly Leu Ala Asn Pro lie Ala Ala lie Arg Cys Val Ser Leu Leu Leu 260 265 270 aaa tac gtt ggc gag aag ccg tet gcg att gaa gtt gaa aaa gcg att 864
Lys Tyr Val Gly Glu Lys Pro Ser Ala lie Glu Val Glu Lys Ala lie 275 280 285 cag aaa act ate get gac ggt ate aaa act ccg gat ctg ggt ggt act 912
Gin Lys Thr He Ala Asp Gly He Lys Thr Pro Asp Leu Gly Gly Thr 290 295 300 gcg acc act gaa gaa gtt ggt cgt gcg gta ctg tet cac ctg get tet 960
Ala Thr Thr Glu Glu Val Gly Arg Ala Val Leu Ser His Leu Ala Ser 305 310 315 320 tee aag aaa gca gaa ggt ccg gac etc Ser Lys Lys Ala Glu Gly Pro Asp Leu 325 <210〉 252 -321 - 146966-序列表.doe 987 201037078 <211> 329
<212> PRT <213> 亨氏甲烷螺菌JF-1 <400> 252
Met Lys Arg Val Val Val Ala Pro Gly Asp Gly lie Gly Pro Glu Val 1 5 10 15 lie Pro Ser Ala Leu Glu Val Leu Arg Phe Phe His Pro Glu Trp Glu 20 25 30
Tyr lie Pro Val Tyr Leu Gly Tyr Glu Cys Trp Lys Arg Thr Gly Asp 35 40 45
Ala Leu Ser Gin Arg Thr Leu Glu Thr Leu Lys Lys Ala Asp Leu lie 50 55 60
Leu Phe Gly Ala lie Thr Thr Pro Pro Asp Pro Lys Tyr His Ser Val 65 70 75 80
Val Leu Arg lie Arg Lys Glu Leu Asp Leu Tyr Ala Asn Leu Arg Pro 85 90 95
Val Phe Gly Glu Gly Phe Asp lie Leu lie Val Arg Glu Asn Thr Glu 100 105 110
Gly Leu Tyr Ser Gly lie Glu Trp Gin Glu Lys Asp Arg Ala Cys Thr 115 120 125
Leu Arg Val Val Ser Glu Ala Gly Ser Arg Arg lie Ala Arg Phe Ala 130 135 140
Cys Gly Cys Ala Lys Arg Arg Arg Arg His Leu Thr lie Gly Asn Lys 145 150 155 160
Ala Asn Val Leu Lys Ser Asp Ser Tyr Phe Leu Asp lie Cys Met Glu 165 170 175
Glu Ala Glu Lys Ala Gly lie Ser lie Asp Lys Lys Tyr lie Asp Ser 180 185 190
Leu Val Leu Asp Val Leu Gin His Pro Gly Arg Tyr Asp Val lie Val 195 200 205
Thr Thr Asn lie Phe Gly Asp lie Leu Ser Asp Ala Ala Ala Phe Leu 210 215 220
Glu Gly Gly Leu Gly Met Leu Pro Ser Ala Asn lie Gly Arg His Gin 225 230 235 240
Ala Leu Phe Glu Pro Val His Gly Ser Ala Pro Asp lie Ala Gly Lys 245 250 255
Gly Leu Ala Asn Pro lie Ala Ala lie Arg Cys Val Ser Leu Leu Leu 260 265 270
Lys Tyr Val Gly Glu Lys Pro Ser Ala lie Glu Val Glu Lys Ala lie 275 280 285
Gin Lys Thr lie Ala Asp Gly lie Lys Thr Pro Asp Leu Gly Gly Thr 290 295 300
Ala Thr Thr Glu Glu Val Gly Arg Ala Val Leu Ser His Leu Ala Ser 305 310 315 320
Ser Lys Lys Ala Glu Gly Pro Asp Leu 325 <210> 253 <211> 990
<212> DNA <213>亨氏f烷螺菌JF-1 <220> <223> 野生型 <400> 253
ATGAAGCGGG TTGTTGTGGC TCCCGGTGAT GGTATCGGAC CTGAAGTAAT ACCTTCTGCA 60 •322- 146966-序列表.doc
201037078 CTTGAGGTGC TTCGGTTCTT TCATCCTGAG TGGGAGTATA TTCCGGTATA TTTGGGGTAT 120 GAGTGCTGGA AGAGAACCGG AGATGCATTG TCACAAAGGA CACTTGAAAC GCTCAAAAAG 180 GCGGATCTAA TCCTGTTTGG TGCGATTACA ACACCTCCTG ATCCGAAGTA TCACAGTGTG 240 GTGCTACGGA TTCGAAAGGA ACTGGATCTC TATGCAAATC TCCGCCCGGT ATTCGGAGAA 300 GGGTTTGATA TTTTGATCGT GCGGGAGAAT ACCGAGGGGC TCTATTCAGG TATCGAATGG 360 CAGGAGAAGG ATCGTGCCTG TACCCTCCGT GTTGTCAGTG AAGCAGGATC GCGTCGGATT 420 GCCAGATTTG CTTGTGGCTG TGCCAAGCGG AGACGAAGAC ATCTGACAAT CGGCAATAAG 480 GCAAATGTCC TGAAGTCTGA CTCTTATTTC CTTGATATCT GCATGGAGGA GGCTGAAAAG 540 GCAGGAATAT CCATAGATAA AAAGTATATC GACTCACTGG TTCTTGACGT CCTGCAGCAC 600 CCAGGCCGGT ATGATGTCAT TGTGACGACG AACATCTTTG GGGACATCCT TTCAGATGCG 660 GCTGCATTTC TGGAAGGGGG CCTTGGGATG CTTCCCAGCG CAAATATTGG ACGGCATCAG 720 GCACTCTTCG AACCAGTTCA TGGCAGTGCC CCTGATATTG CCGGGAAGGG TCTTGCAAAT 780 CCCATCGCAG CCATCAGATG TGTATCCCTG CTTCTTAAGT ATGTCGGAGA GAAGCCCAGT 840
GCTATTGAAG TGGAAAAGGC GATTCAAAAG ACCATCGCCG ACGGGATAAA GACTCCTGAT 900 CTCGGAGGTA CTGCAACAAC TGAGGAAGTG GGACGGGCAG TCCTATCACA TCTTGCATCA 960 TCAAAAAAGG CAGAGGGTCC GGACCTCTGA 990 <210> 254 <211> 969
<212> DNA <213> 思氏甲烷球形菌DSM 3091 <220> <221> CDS <222> (1) .. (969) <400> 254 atg tac aaa ate acc gtt ate cca ggt gac ggt ate ggt cag gaa gta Met Tyr Lys lie Thr Val lie Pro Gly Asp Gly lie Gly Gin Glu Val 15 10 15 atg cag ccg act ate gac ate ctg gaa act ctg aac tee aag ttc gaa Met Gin Pro Thr lie Asp lie Leu Glu Thr Leu Asn Ser Lys Phe Glu 20 25 30 ttc ate ccg aaa gaa get ggt aaa gag tgc tac cag aaa tac gat acc 144
Phe lie Pro Lys Glu Ala Gly Lys Glu Cys Tyr Gin Lys Tyr Asp Thr 35 40 45 aat ctg ccg gaa gaa acc ate gtt cag tgc ege gaa tet gac tee act 192
Asn Leu Pro Glu Glu Thr lie Val Gin Cys Arg Glu Ser Asp Ser Thr 50 55 60 146966-序列表.doc 48 96 323 - 201037078 ctg ttc ggt gcg gta act tcc ate ccg cag cag aag tet gca ate gtt 240
Leu Phe Gly Ala Val Thr Ser lie Pro Gin Gin Lys Ser Ala lie Val 65 70 75 80 act ctg cgt aaa gag ctg gat ctg tac gtt aac cag cgt ccg ate cac 288
Thr Leu Arg Lys Glu Leu Asp Leu Tyr Val Asn Gin Arg Pro lie His 85 90 95 tet tac acc aac ccg gat ate gac ttc act ate ate cgt gaa aac tet 336
Ser Tyr Thr Asn Pro Asp lie Asp Phe Thr lie lie Arg Glu Asn Ser 100 105 110 gaa ggt ctg tac tet cac ate gaa gaa tec act ggc gac gaa get ate 384
Glu Gly Leu Tyr Ser His lie Glu Glu Ser Thr Gly Asp Glu Ala lie 115 120 125 get ate cgt aag ate act tac aaa gcg tet gaa cgt ate ate aac tac 432
Ala lie Arg Lys lie Thr Tyr Lys Ala Ser Glu Arg lie lie Asn Tyr 130 135 140 gca ttc aac tac gca ctg aaa act gaa aaa tet aaa gtt act get tet 480
Ala Phe Asn Tyr Ala Leu Lys Thr Glu Lys Ser Lys Val Thr Ala Ser 145 150 155 160 cac aaa get aac gtt ctg ccg gta act gac ggt ate ttc aag aac acc 528
His Lys Ala Asn Val Leu Pro Val Thr Asp Gly lie Phe Lys Asn Thr 165 170 175 ttc tac aaa gtt get tet aac tac ccg act ate aag tet aac gac tac 576
Phe Tyr Lys Val Ala Ser Asn Tyr Pro Thr lie Lys Ser Asn Asp Tyr 180 185 190 tac ate gac gca atg gcg atg tac ctg ate act aac ccg gca cag ttc 624
Tyr lie Asp Ala Met Ala Met Tyr Leu lie Thr Asn Pro Ala Gin Phe 195 200 205 gac ate ate gtt act acc aac ctg ttc ggt gac ate ctg tet gac gaa 672
Asp lie lie Val Thr Thr Asn Leu Phe Gly Asp lie Leu Ser Asp Glu 210 215 220 ggt ggt ggt ctg gtt ggt act ctg ggt ctg att ccg tet get aac ate 720
Gly Gly Gly Leu Val Gly Thr Leu Gly Leu lie Pro Ser Ala Asn lie 225 230 235 240 ggt gac aaa acc ggt ctg ttc gaa ccg gtt cac ggt tet gca ccg gac 768
Gly Asp Lys Thr Gly Leu Phe Glu Pro Val His Gly Ser Ala Pro Asp 245 250 255 •324· 146966-序列表.doc 201037078 ate get ggt ctg aac aaa get aac ccg ate geg atg ate etc tet tee 816 lie Ala Gly Leu Asn Lys Ala Asn Pro lie Ala Met lie Leu Ser Ser 260 265 270 tgc ctg atg ctg gaa tac ctg ggt ctg tac gac gac geg aag ege att 864
Cys Leu Met Leu Glu Tyr Leu Gly Leu Tyr Asp Asp Ala Lys Arg lie 275 280 285 cag aac get gtt gaa gaa acc att tet gaa tee aaa gtt aaa act ccg 912
Gin Asn Ala Val Glu Glu Thr lie Ser Glu Ser Lys Val Lys Thr Pro 290 295 300 gat atg ggt ggt cac aac aac act cag gac gtt get aac aac ate ctg 960
Asp Met Gly Gly His Asn Asn Thr Gin Asp Val Ala Asn Asn lie Leu 305 310 315 320 Ο
cac aga eta 969
His Arg Leu <210> 255 <211> 323
<212> PRT <213> 思氏曱烷球形菌DSM 3091 <400> 255
Met Tyr Lys lie Thr Val lie Pro Gly Asp Gly lie Gly Gin Glu Val 15 10 15
Met Gin Pro Thr lie Asp lie Leu Glu Thr Leu Asn Ser Lys Phe Glu 20 25 .30
Phe lie Pro Lys Glu Ala Gly Lys Glu Cys Tyr Gin Lys Tyr Asp Thr 35 40 45
Asn Leu Pro Glu Glu Thr lie Val Gin Cys Arg Glu Ser Asp Ser Thr 50 55 60
Leu Phe Gly Ala Val Thr Ser lie Pro Gin Gin Lys Ser Ala lie Val 65 70 75 80
Thr Leu Arg Lys Glu Leu Asp Leu Tyr Val Asn Gin Arg Pro lie His 85 90 95
Ser Tyr Thr Asn Pro Asp 工le Asp Phe Thr lie lie Arg Glu Asn Ser 100 105 110
Glu Gly Leu Tyr Ser His lie Glu Glu Ser Thr Gly Asp Glu Ala lie 115 120 125
Ala 工le Arg Lys lie Thr Tyr Lys Ala Ser Glu Arg lie lie Asn Tyr 130 135 140
Ala Phe Asn Tyr Ala Leu Lys Thr Glu Lys Ser Lys Val Thr Ala Ser 145 150 155 160
His Lys Ala Asn Val Leu Pro Val Thr Asp Gly lie Phe Lys Asn Thr 165 170 175
Phe Tyr Lys Val Ala Ser Asn Tyr Pro Thr lie Lys Ser Asn Asp Tyr 180 185 190
Tyr lie Asp Ala Met Ala Met Tyr Leu lie Thr Asn Pro Ala Gin Phe 195 200 205
Asp lie lie Val Thr Thr Asn Leu Phe Gly Asp lie Leu Ser Asp Glu 210 215 220
Gly Gly Gly Leu Val Gly Thr Leu Gly Leu lie Pro Ser Ala Asn lie 225 230 235 240 -325- 146966-序列表.doc 201037078
Gly Asp Lys Thr Gly 245 lie Ala Gly Leu Asn 260
Leu Phe Glu Pro 250 Lys Ala Asn Pro 265
Val His Gly Ser Ala Pro Asp 255 lie Ala Met lie Leu Ser Ser 270
Cys Leu Met Leu Glu 275 Gin Asn Ala Val Glu 290
Tyr Leu Gly Leu 280 Glu Thr lie Ser 295
Tyr Asp Asp Ala Lys Arg 285 Glu Ser Lys Val Lys Thr 300 lie Pro
Asp Met Gly Gly His Asn Asn Thr Gin Asp Val Ala Asn Asn lie Leu 305 310 315 320
His Arg Leu <210> 256 <211> 972 <212> DNA <213> 思氏甲烷球形菌DSM 3091 <220> <223> 野生型 <400> 256 TTGTATAAAA TAACAGTAAT ACCTGGTGAT GGTATAGGAC AAGAAGTTAT GCAACCAACA 60 ATAGATATTC TTGAAACATT AAATTCTAAA TTTGAATTTA TACCAAAAGA AGCAGGAAAA 120 GAATGTTATC AAAAATATGA TACAAATCTA CCTGAAGAGA CTATTGTTCA ATGTAGAGAA 180 AGTGATAGTA CACTCTTTGG AGCAGTAACA TCAATTCCAC AACAAAAAAG TGCAATAGTG 240 ACATTACGTA AAGAACTTGA TTTATATGTA AATCAAAGAC CAATACATTC ATATACAAAT 300 CCAGATATAG ATTTTACAAT AATACGTGAA AATTCTGAAG GACTATATTC ACATATTGAA 360 GAAAGTACTG GTGATGAAGC AATAGCCATT AGAAAAATAA CATACAAAGC TTCAGAAAGA 420 ATAATAAATT ATGCTTTTAA CTATGCACTT AAAACAGAAA AAAGTAAAGT AACAGCATCA 480 CATAAAGCAA ATGTACTACC AGTAACTGAT GGAATATTTA AAAATACATT CTATAAAGTA 540 GCAAGTAATT ATCCAACAAT CAAATCCAAT GATTATTATA TAGATGCTAT GGCAATGTAT 600 CTTATAACAA ATCCTGCTCA ATTTGATATT ATAGTAACAA CAAATCTCTT TGGAGACATA 660 TTATCTGATG AAGGTGGAGG ATTAGTTGGA ACTCTTGGAC TTATACCTTC AGCAAATATT 720 GGGGATAAAA CTGGATTATT TGAACCAGTA CATGGATCAG CACCAGATAT AGCAGGACTT 780 AATAAGGCAA ATCCAATAGC AATGATCTTA TCAAGTTGTT TAATGCTTGA ATATTTAGGA 840 TTATATGATG ATGCAAAAAG AATTCAAAAT GCAGTAGAAG AAACAATTTC TGAAAGTAAA 900
GTTAAAACAC CAGATATGGG TGGACATAAC AATACACAAG ATGTTGCT^AA TAATATACTC 960 CATAGATTAT AA 972 <210> <211> <212> <213> 257 969
DNA
嗜熱甲烷鬃菌PT -326· 146966-序列表.doc 48 201037078 <220> <221> CDS <222> ⑴..（969) <400> 257 atg aag aag ate get ctg gtt ccg ggc gac ggt ate ggt ccg gaa gtt
Met Lys Lys lie Ala Leu Val Pro Gly Asp Gly lie Gly Pro Glu Val 15 10 15 96 ate tet tee gca ctg ege gta ctg aac gca gca ggc ttc gac ggt gaa lie Ser Ser Ala Leu Arg Val Leu Asn Ala Ala Gly Phe Asp Gly Glu 20 25 30 ctg gtt gaa ttc gac ate ggt tac ggt ege tgg cgt ege gac ggt aaa 144
Leu Val Glu Phe Asp lie Gly Tyr Gly Arg Trp Arg Arg Asp Gly Lys 35 40 45 geg ate act gac gac gat ate gaa cgt atg aaa gac tgc gac tgc ate 192
Ala lie Thr Asp Asp Asp lie Glu Arg Met Lys Asp Cys Asp Cys lie 50 55 60 ctg ttc ggt gca ate acc act ccg ccg gat ccg ggt tac cgt tet gta 240
Leu Phe Gly Ala lie Thr Thr Pro Pro Asp Pro Gly Tyr Arg Ser Val 65 70 75 80 ctg ate ege att cgt aaa gag ctg gat ctg tac get aac ate cgt ccg 288
Leu lie Arg lie Arg Lys Glu Leu Asp Leu Tyr Ala Asn lie Arg Pro 85 90 95 ctg cgt tet tet ege ate gac gtt ate ate gtt cgt gaa aac act gaa 336
Leu Arg Ser Ser Arg lie Asp Val lie lie Val Arg Glu Asn Thr Glu 100 105 110 ggt ctg tac tee ggt ctg gaa atg ctg ggt gac gaa gaa geg cgt acc 384
Gly Leu Tyr Ser Gly Leu Glu Met Leu Gly Asp Glu Glu Ala Arg Thr 115 120 125 gtt ege gtt ate act cgt aaa ggt tet cag cgt att get gaa gtt get 432
Val Arg Val lie Thr Arg Lys Gly Ser Gin Arg lie Ala Glu Val Ala 130 135 140 tgc cgt ate get tet gaa cgt aag cac ctg act ate ate cac aaa get 480
Cys Arg lie Ala Ser Glu Arg Lys His Leu Thr lie lie His Lys Ala 145 150 155 160 aac gta ctg aaa tet gac gtt ctg ttc ctg aaa act tgc cgt gaa gtt 528
Asn Val Leu Lys Ser Asp Val Leu Phe Leu Lys Thr Cys Arg Glu Val 165 170 175 get gaa cgt tac ggt ate ege tac gac gac atg ctg gtt gac get get 576 -327- 146966-序列表.doe 201037078
Ala Glu Arg Tyr Gly lie Arg Tyr Asp Asp Met Leu Val Asp Ala Ala 180 185 190 get tac aac atg gtt ate cgt ccg gaa atg ttc gac gta atg gtt act 624
Ala Tyr Asn Met Val lie Arg Pro Glu Met Phe Asp Val Met Val Thr 195 200 205 acc aac ctg ttc ggt gac ate ctg tet gac gaa ggt geg ggt ate gtt 672
Thr Asn Leu Phe Gly Asp lie Leu Ser Asp Glu Gly Ala Gly lie Val 210 215 220 ggt tet ctg ggt ctg tgc gca tet get aac ctg ggt gac ege tgg geg 720
Gly Ser Leu Gly Leu Cys Ala Ser Ala Asn Leu Gly Asp Arg Trp Ala 225 230 235 240 ctg ttc gaa cca ate cac ggt tet gca ccg gat ate get ggt aac ggt 768
Leu Phe Glu Pro lie His Gly Ser Ala Pro Asp lie Ala Gly Asn Gly 245 250 255 att get aac ccg gtt ggt gca ate cgt tet gca gca atg atg ctg gag 816 lie Ala Asn Pro Val Gly Ala lie Arg Ser Ala Ala Met Met Leu Glu 260 265 270 tgg ttc ggt gaa atg gaa cgt get cag tet ate cac cag geg gtt gac 864
Trp Phe Gly Glu Met Glu Arg Ala Gin Ser lie His Gin Ala Val Asp 275 280 285 cgt act ctg tet aaa ggc gtt aaa act ccg gat ctg ggt ggt ctg tgc 912
Arg Thr Leu Ser Lys Gly Val Lys Thr Pro Asp Leu Gly Gly Leu Cys 290 295 300 acc act tet gaa ttc act gac geg gtt ate gac gaa atg cgt ege get 960
Thr Thr Ser Glu Phe Thr Asp Ala Val lie Asp Glu Met Arg Arg Ala 305 310 315 320 ggc geg tgt 969
Gly Ala Cys <210> 258 <211> 323
<212> PRT <213>嗜熱甲烷鬃菌PT <400> 258
Met Lys Lys lie Ala Leu Val Pro Gly Asp Gly lie Gly Pro Glu Val 15 10 15 lie Ser Ser Ala Leu Arg Val Leu Asn Ala Ala Gly Phe Asp Gly Glu 20 25 30
Leu Val Glu Phe Asp lie Gly Tyr Gly Arg Trp Arg Arg Asp Gly Lys 35 40 45
Ala lie Thr Asp Asp Asp lie Glu Arg Met Lys Asp Cys Asp Cys lie -328· 146966-序列表.doc 201037078 50 55 60
Leu Phe Gly Ala 工le Thr Thr Pro Pro Asp Pro Gly Tyr Arg Ser Val 65 70 75 80
Leu lie Arg lie Arg Lys Glu Leu Asp Leu Tyr Ala Asn lie Arg Pro 85 90 95
Leu Arg Ser Ser Arg lie Asp Val lie lie Val Arg Glu Asn Thr Glu 100 105 110
Gly Leu Tyr Ser Gly Leu Glu Met Leu Gly Asp Glu Glu Ala Arg Thr 115 120 125
Val Arg Val lie Thr Arg Lys Gly Ser Gin Arg lie Ala Glu Val Ala 130 135 140
Cys Arg 工le Ala Ser Glu Arg Lys His Leu Thr lie lie His Lys Ala 145 150 155 160
Asn Val Leu Lys Ser Asp Val Leu Phe Leu Lys Thr Cys Arg Glu Val 165 170 175
Ala Glu Arg Tyr Gly lie Arg Tyr Asp Asp Met Leu Val Asp Ala Ala 180 185 190
Ala Tyr Asn Met Val lie Arg Pro Glu Met Phe Asp Val Met Val Thr 195 200 205 〇
Thr Asn Leu Phe Gly Asp lie Leu Ser Asp Glu Gly Ala Gly lie Val 210 215 220
Gly Ser Leu Gly Leu Cys Ala Ser Ala Asn Leu Gly Asp Arg Trp Ala 225 230 235 240
Leu Phe Glu Pro lie His Gly Ser Ala Pro Asp 工le Ala Gly Asn Gly 245 250 255 lie Ala Asn Pro Val Gly Ala lie Arg Ser Ala Ala Met Met Leu Glu 260 265 270
Trp Phe Gly Glu Met Glu Arg Ala Gin Ser lie His Gin Ala Val Asp 275 280 285
Arg Thr Leu Ser Lys Gly Val Lys Thr Pro Asp Leu Gly Gly Leu Cys 290 295 300
Thr Thr Ser Glu Phe Thr Asp Ala Val lie Asp Glu Met Arg Arg Ala 305 310 315 320
Gly Ala Cys > > > > 0 12 3 -—-1 -—I ϊ—I 2 2 2 2 < < < < PT 菌 鬃 烷 甲 9 2认熱2597§嗜 <220> <223> 野生型 <400> 259 ATGAAGAAAA TCGCTCTTGT GCCGGGCGAT GGCATCGGGC CTGAGGTCAT ATCGAGCGCT 60 CTTCGTGTCT TGAATGCAGC AGGATTCGAT GGGGAGCTCG TAGAGTTCGA TATAGGCTAT 120 GGCAGATGGA GGCGTGATGG AAAGGCCATA ACAGATGATG ATATAGAAAG GATGAAGGAT 180 TGCGACTGCA TACTCTTCGG GGCGATCACG ACTCCTCCTG ATCCTGGTTA TAGGAGTGTC 240 CTGATAAGGA TAAGAAAGGA GCTGGACCTG TATGCGAACA TACGCCCCCT GCGCTCGAGC 300 AGGATCGATG TGATCATTGT GAGGGAGAAC ACCGAGGGGC TGTATTCAGG TCTGGAGATG 360 CTCGGCGATG AAGAGGCCAG GACCGTGCGT GTCATAACGA GAAAGGGGAG CCAGAGGATA 420 GCCGAGGTGG CGTGCAGGAT CGCATCGGAA CGCAAACATC TCACAATAAT ACACAAGGCG 480 329 - 146966-序列表.doc 201037078 AACGTCCTGA AGAGCGACGT GCTGTTTTTA AAAACATGCA GGGAGGTCGC TGAAAGGTAT 540 GGAATCAGGT AT GAT GAT AT GCTGGTTGAT GCCGCGGCAT ACAACATGGT GATAAGGCCG 600 GAGATGTTTG ATGTGATGGT GACGACAAAT CTCTTCGGGG ACATTCTCAG CGATGAGGGC 660 GCCGGCATTG TCGGCAGCCT GGGGCTCTGC GCCAGCGCGA ATCTGGGCGA TCGCTGGGCG 720 CTCTTCGAGC CGATACACGG CAGCGCGCCC GACATTGCAG GCAATGGCAT AGCAAACCCG 780 GTGGGAGCCA TACGGAGCGC TGCGATGATG CTCGAGTGGT TCGGTGAGAT GGAGAGGGCG 840 CAGTCGATCC ATCAGGCGGT GGACCGCACG CTCTCGAAAG GGGTGAAAAC GCCGGATCTG 900 GGCGGCTTGT GCACGACATC TGAGTTCACA GATGCGGTCA TCGATGAGAT GAGGCGCGCC 960 GGAGCATGTT GA 972 <210> 260 <211> 1215
<212> DNA
<213> 醋酸甲烷八聯球菌C2A <220> <221> CDS <222> (1)..(1215) <400> 260 atg tct gaa tct gaa cag tac tcc cgt aac acc ctg atg gac ttc ate
Met Ser Glu Ser Glu Gin Tyr Ser Arg Asn Thr Leu Met Asp Phe lie 15 10 15 gaa tac cgt ccg ctg gat ate gaa ate tgc gac gta act ctg cgt gac
Glu Tyr Arg Pro Leu Asp lie Glu lie Cys Asp Val Thr Leu Arg Asp 20 25 30 ggt gaa cag act ccg ggc gta gta ttc acc aaa gag cag aaa ctg geg 144
Gly Glu Gin Thr Pro Gly Val Val Phe Thr Lys Glu Gin Lys Leu Ala 35 40 45 gtt get tct gaa ctg gac tcc atg ggt ate gaa gtt ate gaa gca ggc 192
Val Ala Ser Glu Leu Asp Ser Met Gly lie Glu Val lie Glu Ala Gly 50 55 60 ttc ccg gtt gtt tct get tac gaa aaa gaa ate gtt aaa gaa ate get 240
Phe Pro Val Val Ser Ala Tyr Glu Lys Glu lie Val Lys Glu lie Ala 65 70 75 80 aac cag ggt tac gac tcc cgt ate tgc tgc ctg tct ege get gtt aaa 288
Asn Gin Gly Tyr Asp Ser Arg lie Cys Cys Leu Ser Arg Ala Val Lys 85 90 95 ggc gac gtt gac gca gca ctg gac tgc gac gtt gat ate gtt tcc ate 336
Gly Asp Val Asp Ala Ala Leu Asp Cys Asp Val Asp lie Val Ser lie 100 105 110 146966-序列表.doc -330 48 96 201037078 ttc ate geg atg tet gac atg cac ctg aaa tac aaa tac cac cgt act 384
Phe lie Ala Met Ser Asp Met His Leu Lys Tyr Lys Tyr His Arg Thr 115 120 125 ctg gaa gat atg ctg ggt tgc gca aaa gaa get ate gaa tac gca act 432
Leu Glu Asp Met Leu Gly Cys Ala Lys Glu Ala lie Glu Tyr Ala Thr 130 135 140 gac cac ggt ctg aac gtt ege ttc get gca gaa gat get tee cgt act 480
Asp His Gly Leu Asn Val Arg Phe Ala Ala Glu Asp Ala Ser Arg Thr 145 150 155 160 ccg ate gac ege ctg aag cag gca ttc aaa gaa gtt gaa aac gaa tac 528
Pro 工le Asp Arg Leu Lys Gin Ala Phe Lys Glu Val Glu Asn Glu Tyr 165 170 175
Ο aaa gtt cag tac gtt tet ctg get gac acc ate ggt att ctg aac ccg 576
Lys Val Gin Tyr Val Ser Leu Ala Asp Thr lie Gly lie Leu Asn Pro 180 185 190 act acc act cac tac ctg gtt tet gaa ate ttc aaa tgc gta aac acc 624
Thr Thr Thr His Tyr Leu Val Ser Glu lie Phe Lys Cys Val Asn Thr 195 200 205 tet ate tgc ate cac tgc cac gac gac ctg ggt atg gca act geg aac 672
Ser lie Cys 工le His Cys His Asp Asp Leu Gly Met Ala Thr Ala Asn 210 215 220 acc ctg gca get get gaa get ggc geg aag cag ctg cac acc acc gtt 720
Thr Leu Ala Ala Ala Glu Ala Gly Ala Lys Gin Leu His Thr Thr Val 225 230 235 240 aac ggt ate ggt gaa ege get ggt aac get tet ctg gaa gaa atg ctg 768
Asn Gly lie Gly Glu Arg Ala Gly Asn Ala Ser Leu Glu Glu Met Leu 245 250 255 gtt get ctg ege gta cag tac ggt ate gaa cgt tac gac act acc aag 816
Val Ala Leu Arg Val Gin Tyr Gly lie Glu Arg Tyr Asp Thr Thr Lys 260 265 270 ctg act geg ctg tet cgt atg ate tet gaa tac tee aac ate act ccg 864
Leu Thr Ala Leu Ser Arg Met lie Ser Glu Tyr Ser Asn lie Thr Pro 275 280 285 tet gtt aac aaa geg gtt gtt ggt cag aac gca ttc act cac gaa tee 912
Ser Val Asn Lys Ala Val Val Gly Gin Asn Ala Phe Thr His Glu Ser 290 295 300 -331 146966·序列表.doc 201037078 ggt ate cac gtt get geg att ctg gaa gaa ccg cgt act tac gaa ctg 960
Gly lie His Val Ala Ala lie Leu Glu Glu Pro Arg Thr Tyr Glu Leu 305 310 315 320 ttc ctg cca gaa atg gtt ggt ggt aag cgt aac ctg gtt gtt ggt aag 1008
Phe Leu Pro Glu Met Val Gly Gly Lys Arg Asn Leu Val Val Gly Lys 325 330 335 cac act ggc acc aaa geg ctg aaa ggt ate ate aac tee ate ggc ttc 1056
His Thr Gly Thr Lys Ala Leu Lys Gly lie lie Asn Ser lie Gly Phe 340 345 350 tgc ctg gaa cgt gaa gaa ctg tgc gca ctg ate gaa aaa gtt aaa gtt 1104
Cys Leu Glu Arg Glu Glu Leu Cys Ala Leu lie Glu Lys Val Lys Val 355 360 365 tgc act gac gag aag ege cgt tet ate tet ege gag cag ctg gaa aaa 1152
Cys Thr Asp Glu Lys Arg Arg Ser lie Ser Arg Glu Gin Leu Glu Lys 370 375 380 ctg ate get cag gta cgt cag gaa cag aag ccg tet get tet gaa aaa 1200
Leu lie Ala Gin Val Arg Gin Glu Gin Lys Pro Ser Ala Ser Glu Lys 385 390 395 400 gag aag ttc tee att 1215
Glu Lys Phe Ser lie 405 <210> 261 <211> 405
<212> PRT <213〉醋酸曱烷八聯球菌C2A <400> 261
Met Ser Glu Ser Glu Gin Tyr Ser Arg Asn Thr Leu Met Asp Phe lie 15 10 15
Glu Tyr Arg Pro Leu Asp lie Glu lie Cys Asp Val Thr Leu Arg Asp 20 25 30
Gly Glu Gin Thr Pro Gly Val Val Phe Thr Lys Glu Gin Lys Leu Ala 35 40 45
Val Ala Ser Glu Leu Asp Ser Met Gly lie Glu Val 工le Glu Ala Gly 50 55 60
Phe Pro Val Val Ser Ala Tyr Glu Lys Glu lie Val Lys Glu 工le Ala 65 70 75 80
Asn Gin Gly Tyr Asp Ser Arg lie Cys Cys Leu Ser Arg Ala Val Lys 85 90 95
Gly Asp Val Asp Ala Ala Leu Asp Cys Asp Val Asp lie Val Ser lie 100 105 110
Phe lie Ala Met Ser Asp Met His Leu Lys Tyr Lys Tyr His Arg Thr 115 120 125
Leu Glu Asp Met Leu Gly Cys Ala Lys Glu Ala lie Glu Tyr Ala Thr 130 135 140
Asp His Gly Leu Asn Val Arg Phe Ala Ala Glu Asp Ala Ser Arg Thr 145 150 155 160 -332- 146966·序列表.doc 201037078
Pro lie Asp Arg Leu Lys Gin Ala Phe Lys Glu Val Glu Asn Glu Tyr 165 170 175
Lys Val Gin Tyr Val Ser Leu Ala Asp Thr lie Gly lie Leu Asn Pro 180 185 190
Thr Thr Thr His Tyr Leu Val Ser Glu lie Phe Lys Cys Val Asn Thr 195 200 205
Ser lie Cys lie His Cys His Asp Asp Leu Gly Met Ala Thr Ala Asn 210 215 220
Thr Leu Ala Ala Ala Glu Ala Gly Ala Lys Gin Leu His Thr Thr Val 225 230 235 240
Asn Gly 工le Gly Glu Arg Ala Gly Asn Ala Ser Leu Glu Glu Met Leu 245 250 255
Val Ala Leu Arg Val Gin Tyr Gly lie Glu Arg Tyr Asp Thr Thr Lys 260 265 270
Leu Thr Ala Leu Ser Arg Met lie Ser Glu Tyr Ser Asn lie Thr Pro 275 280 285
Ser Val Asn Lys Ala Val Val Gly Gin Asn Ala Phe Thr His Glu Ser 290 295 300
Gly lie His Val Ala Ala lie Leu Glu Glu Pro Arg Thr Tyr Glu Leu 305 310 315 320
Phe Leu Pro Glu Met Val Gly Gly Lys Arg Asn Leu Val Val Gly Lys 325 330 335
His Thr Gly Thr Lys Ala Leu Lys Gly lie lie Asn Ser lie Gly Phe 340 345 350
Cys Leu Glu Arg Glu Glu Leu Cys Ala Leu lie Glu Lys Val Lys Val 355 360 365
Cys Thr Asp Glu Lys Arg Arg Ser lie Ser Arg Glu Gin Leu Glu Lys 370 375 380
Leu lie Ala Gin Val Arg Gin Glu Gin Lys Pro Ser Ala Ser Glu Lys 385 390 395 400
Glu Lys Phe Ser lie 405
<210> 262 <211> 1218 <212> DNA
<213> 醋酸甲烷八聯球菌C2A <220> <223> 野生型 <400> 262 ATGTCAGAGA GCGAGCAGTA CTCCAGAAAT ACACTTATGG ACTTCATCGA ATATCGCCCC 60
CTCGATATCG AAATCTGTGA CGTGACCCTG CGCGATGGGG AACAGACCCC TGGTGTTGTG 120 TTCACGAAAG AACAGAAGCT GGCAGTAGCC AGCGAGCTTG ATTCCATGGG TATTGAGGTT 180 ATAGAAGCCG GCTTTCCGGT AGTTTCCGCA TACGAAAAAG AAATCGTAAA GGAAATCGCA 240 AACCAGGGTT ATGATTCCAG GATCTGCTGC CTCTCAAGAG CAGTAAAGGG AGATGTTGAT 300 GCTGCCCTTG ATTGTGACGT CGATATCGTC AGCATTTTCA TTGCAATGTC CGACATGCAC 360 CTCAAATACA AGTACCACAG GACTCTTGAG GATATGCTGG GCTGCGCTAA GGAAGCTATT 420 GAATATGCAA CTGACCACGG TTTAAACGTA CGTTTTGCAG CCGAGGATGC AAGCCGCACT 480 CCGATTGACC GCCTCAAGCA GGCTTTCAAG GAAGTTGAGA ACGAGTATAA AGTGCAGTAC 540 -333 - 146966-序列表.doc 201037078 GTAAGCCTGG CAGATACAAT CGGTATCCTG AACCCGACCA CCACCCATTA CCTTGTAAGT 600 GAAATCTTCA AGTGCGTAAA CACTTCAATC TGCATCCACT GCCACGATGA CCTCGGGATG 660 GCTACGGCTA ACACCCTTGC AGCCGCCGAA GCCGGAGCAA AGCAGCTCCA TACGACAGTC 720 AACGGGATTG GAGAAAGGGC CGGAAACGCT TCTCTTGAAG AAATGCTGGT TGCCCTTAGG 780 GTACAGTACG GAATTGAACG TTATGATACA ACAAAGCTGA CCGCGCTCTC CAGAATGATT 840 TCGGAATACT CAAACATCAC CCCCTCAGTA AACAAAGCCG TTGTAGGCCA GAACGCCTTT 900 ACCCATGAGT CCGGAATTCA CGTCGCTGCA ATCCTGGAAG AGCCGCGCAC TTACGAACTC 960 TTCCTTCCCG AGATGGTAGG TGGAAAGCGC AACCTTGTTG TCGGGAAGCA TACCGGGACA 1020 AAAGCCCTGA AAGGGATCAT CAACAGTATC GGTTTCTGCC TTGAACGGGA AGAACTCTGC 1080 GCCCTGATCG AAAAAGTCAA GGTCTGTACC GACGAAAAGC GAAGAAGCAT TTCAAGGGAG 1140 CAGCTTGAAA AACTGATAGC TCAGGTCAGG CAGGAGCAGA AACCTTCCGC CAGCGAAAAA 1200 GAAAAGTTTT CTATCTGA 1218 <210> 263 <211> 1179
<212> DNA <213> 亨氏甲烷螺菌JF-1 <220> <221> CDS <222> (1) . . (1179) <400> 263 atg aag ccg tgg cac ate gaa ate tgc gac gta act ctg cgt gac ggt
Met Lys Pro Trp His lie Glu lie Cys Asp Val Thr Leu Arg Asp Gly 15 10 15 gaa cag act ccg ggc gtt tet ttc act get gac gag aag aaa gac ate
Glu Gin Thr Pro Gly Val Ser Phe Thr Ala Asp Glu Lys Lys Asp lie 20 25 30 get tet ege ctg gat get ate ggt ate gaa gtt ate gaa geg ggt ttc 144
Ala Ser Arg Leu Asp Ala lie Gly lie Glu Val lie Glu Ala Gly Phe 35 40 45 ccg ate gtt tet gca cac gaa aaa gaa atg gtt cgt aac ate tee cgt 192
Pro lie Val Ser Ala His Glu Lys Glu Met Val Arg Asn lie Ser Arg 50 55 60 ctg ggt ctg tet geg aaa ate tgc ggt ctg tet ege get tgc ege gag 240
Leu Gly Leu Ser Ala Lys lie Cys Gly Leu Ser Arg Ala Cys Arg Glu 65 70 75 80 gac gtt gac get gca ctg gat get gaa gtt gac atg ate ggt ctg ttc 288
Asp Val Asp Ala Ala Leu Asp Ala Glu Val Asp Met lie Gly Leu Phe 146966-序列表.doc -334· 48 96 201037078 85 90 95 ate get ccg tet gat ctg cac ctg aaa tac aaa cac aag aag ccg cgt 336 工le Ala Pro Ser Asp Leu His Leu Lys Tyr Lys His Lys Lys Pro Arg 100 105 110 gac gtt gtt gtt get aac geg ctg gaa cag ctg gac tac get gtt gac 384
Asp Val Val Val Ala Asn Ala Leu Glu Gin Leu Asp Tyr Ala Val Asp 115 120 125 cac ggt ctg ate gtt cgt ttc ggt gca gaa gat get tee cgt act gac 432
His Gly Leu lie Val Arg Phe Gly Ala Glu Asp Ala Ser Arg Thr Asp 130 135 140 ccg gac ate ctg gtt gac ate tac cgt cag gca get gac cac aaa geg 480 Ο
Pro Asp lie Leu Val Asp lie Tyr Arg Gin Ala Ala Asp His Lys Ala 145 150 155 160 act tac gtt act tac get gac acc act ggc tgc ctg act ccg ctg gaa 528
Thr Tyr Val Thr Tyr Ala Asp Thr Thr Gly Cys Leu Thr Pro Leu Glu 165 170 175 gtt gca acc gta atg aaa gac ctg gtt ccg aaa act ccg att ccg ate 576
Val Ala Thr Val Met Lys Asp Leu Val Pro Lys Thr Pro lie Pro lie 180 185 190 geg atg cac geg cac aac gac ctg ggt tgc gca act get aac acc ctg 624
Ala Met His Ala His Asn Asp Leu Gly Cys Ala Thr Ala Asn Thr Leu 195 200 205 ate get gca gaa ctg ggt get tac cag ctg cac acc acc gtt aac ggt 672
He Ala Ala Glu Leu Gly Ala Tyr Gin Leu His Thr Thr Val Asn Gly 210 215 220 ctg ggt gaa cgt geg ggt aac geg cgt ctg gaa gaa gtt ctg gtt tet 720
Leu Gly Glu Arg Ala Gly Asn Ala Arg Leu Glu Glu Val Leu Val Ser 225 230 235 240 ctg gta ctg aaa ggt ggt ate act ege tac gac ctg act gaa ate ccg 768
Leu Val Leu Lys Gly Gly He Thr Arg Tyr Asp Leu Thr Glu He Pro 245 250 255 gca ctg tet gaa ege gta cag cag tac acc ggt ate ttc atg ccg gca 816
Ala Leu Ser Glu Arg Val Gin Gin Tyr Thr Gly He Phe Met Pro Ala 260 265 270 acc aag ccg gtt gtt ggt get aac gca ttc get cac gaa tee ggt ate 864
Thr Lys Pro Val Val Gly Ala Asn Ala Phe Ala His Glu Ser Gly He 275 280 285 •335- 146966-序列表.doc 201037078 cac att get gca ate ctg gaa aac ccg gaa act tac gaa ttc ate ccg 912
His lie Ala Ala 工le Leu Glu Asn Pro Glu Thr Tyr Glu Phe lie Pro 290 295 300 cca gaa ctg ctg ggt ctg gaa cgt ege ttc ate ctg ggt aag cac acc 960
Pro Glu Leu Leu Gly Leu Glu Arg Arg Phe lie Leu Gly Lys His Thr 305 310 315 320 ggt aaa cgt geg ctg gtt cac att ctg act tet ttc ggt tac cac ate 1008
Gly Lys Arg Ala Leu Val His lie Leu Thr Ser Phe Gly Tyr His lie 325 330 335 act gac gaa cag gta atg caa gtt ctg gaa ctg ate aaa gac aaa tet 1056
Thr Asp Glu Gin Val Met Gin Val Leu Glu Leu 工le Lys Asp Lys Ser 340 345 350 gaa gat aaa tgc tcc ate aac cag cgt gta ctg gca gaa ate ate tet 1104
Glu Asp Lys Cys Ser lie Asn Gin Arg Val Leu Ala Glu lie lie Ser 355 360 365 cag gtt acc ggt gaa gat aaa ate aac ggt gac ate ate ggc gag gac 1152
Gin Val Thr Gly Glu Asp Lys lie Asn Gly Asp 工le lie Gly Glu Asp 370 375 380 tec ege tgc cca ggt cgt cac gtt tac 1179
Ser Arg Cys Pro Gly Arg His Val Tyr 385 390 <210> 264 <211> 393
<212> PRT <213> 亨氏甲烷螺菌JF-1 <400> 264
Met Lys Pro Trp His lie Glu lie Cys Asp Val Thr Leu Arg Asp Gly 15 10 15
Glu Gin Thr Pro Gly Val Ser Phe Thr Ala Asp Glu Lys Lys Asp lie 20 25 30
Ala Ser Arg Leu Asp Ala lie Gly lie Glu Val lie Glu Ala Gly Phe 35 40 45
Pro lie Val Ser Ala His Glu Lys Glu Met Val Arg Asn lie Ser Arg 50 55 60
Leu Gly Leu Ser Ala Lys lie Cys Gly Leu Ser Arg Ala Cys Arg Glu 65 70 75 80
Asp Val Asp Ala Ala Leu Asp Ala Glu Val Asp Met lie Gly Leu Phe 85 90 95 lie Ala Pro Ser Asp Leu His Leu Lys Tyr Lys His Lys Lys Pro Arg 100 105 110
Asp Val Val Val Ala Asn Ala Leu Glu Gin Leu Asp Tyr Ala Val Asp 115 120 125
His Gly Leu lie Val Arg Phe Gly Ala Glu Asp Ala Ser Arg Thr Asp 130 135 140
Pro Asp lie Leu Val Asp lie Tyr Arg Gin Ala Ala Asp His Lys Ala - 336- 146966-序列表.doc 201037078 145 150 155 160
Thr Tyr Val Thr Tyr Ala Asp Thr Thr Gly Cys Leu Thr Pro Leu Glu 165 170 175
Val Ala Thr Val Met Lys Asp Leu Val Pro Lys Thr Pro lie Pro lie 180 185 190
Ala Met His Ala His Asn Asp Leu Gly Cys Ala Thr Ala Asn Thr Leu 195 200 205 lie Ala Ala Glu Leu Gly Ala Tyr Gin Leu His Thr Thr Val Asn Gly 210 215 220
Leu Gly Glu Arg Ala Gly Asn Ala Arg Leu Glu Glu Val Leu Val Ser 225 230 235 240
Leu Val Leu Lys Gly Gly lie Thr Arg Tyr Asp Leu Thr Glu lie Pro 245 250 255
Ala Leu Ser Glu Arg Val Gin Gin Tyr Thr Gly lie Phe Met Pro Ala 260 265 270
Thr Lys Pro Val Val Gly Ala Asn Ala Phe Ala His Glu Ser Gly lie 275 280 285
His lie Ala Ala lie Leu Glu Asn Pro Glu Thr Tyr Glu Phe lie Pro 290 295 300
Pro Glu Leu Leu Gly Leu Glu Arg Arg Phe 工le Leu Gly Lys His Thr 305 310 315 320
Gly Lys Arg Ala Leu Val His lie Leu Thr Ser Phe Gly Tyr His lie 325 330 335
Thr Asp Glu Gin Val Met Gin Val Leu Glu Leu lie Lys Asp Lys Ser 340 345 350
Glu Asp Lys Cys Ser lie Asn Gin Arg Val Leu Ala Glu lie lie Ser 355 360 365
Gin Val Thr Gly Glu Asp Lys lie Asn Gly Asp lie lie Gly Glu Asp 370 375 380
Ser Arg Cys Pro Gly Arg His Val Tyr 385 390
<210> 265 <211> 1182 <212> DNA <213> 亨氏甲烷螺菌JF-1 <220> <223> 野生型 <400> 265 ATGAAACCAT GGCACATTGA AATTTGTGAT GTCACTCTTC GGGACGGTGA ACAGACCCCT 60 GGAGTATCAT TCACTGCTGA TGAGAAGAAG GATATCGCAT CCCGGCTTGA TGCAATCGGA 120 ATTGAGGTGA TTGAAGCCGG ATTTCCTATT GTATCCGCCC ATGAAAAGGA GATGGTCCGG 180 AATATCTCCC GGCTGGGTCT CTCGGCAAAG ATATGTGGTC TGTCACGGGC CTGCAGGGAG 240 GATGTGGATG CAGCGCTTGA TGCTGAAGTT GACATGATCG GGCTTTTTAT TGCCCCTTCT 300 GATCTGCATC TGAAATACAA GCATAAAAAG CCCCGGGATG TTGTTGTTGC AAATGCTCTT 360 GAACAGCTTG ATTATGCGGT GGATCATGGT CTTATCGTCC GGTTTGGTGC TGAAGATGCA 420 TCCCGTACCG ACCCTGATAT CCTCGTTGAC ATCTACCGGC AGGCAGCAGA TCACAAGGCG 480 ACCTATGTCA CATATGCAGA TACGACCGGG TGTCTGACCC CTCTTGAGGT CGCTACGGTC 540 337- 146966·序列表.doc 201037078 ATGAAGGATC TTGTCCCCAA AACGCCCATC CCGATTGCCA TGCATGCTCA TAATGATCTC 600 GGGTGTGCGA CGGCAAATAC ACTCATTGCC GCAGAACTGG GAGCGTATCA GTTGCATACC 660 ACGGTGAACG GCCTTGGTGA ACGGGCAGGG AATGCACGGC TAGAGGAGGT TTTGGTATCA 720 CTCGTGCTCA AAGGCGGGAT TACCCGGTAT GATCTCACAG AGATTCCGGC TCTTTCAGAA 780 CGGGTTCAAC AGTATACCGG GATTTTTATG CCTGCAACGA AACCGGTGGT CGGTGCGAAT 840 GCGTTTGCCC ATGAGAGTGG GATTCATATT GCAGCTATTC TGGAAAATCC GGAGACCTAC 900 GAGTTCATAC CCCCAGAGCT CCTGGGACTG GAACGGCGGT TTATCCTGGG GAAGCATACC 960 GGAAAGCGTG CTCTGGTGCA TATTCTGACT TCTTTTGGCT ATCACATCAC CGATGAGCAG 1020 GTTATGCAGG TTCTTGAATT GATAAAGGAT AAAAGTGAGG ATAAATGCAG TATCAACCAG 1080 CGGGTGCTTG CGGAGATCAT CTCCCAGGTA ACCGGGGAGG ATAAGATCAA TGGTGACATT 1140
ATCGGAGAAG ATTCTAGGTG CCCCGGCCGG CACGTATATT GA <210> 266 <211> 837
<212> DNA <213> 思氏曱烷球形菌DSM3091 <220> <221> CDS <222> (1)..(837) <400> 266 atg ggt ctg tet gat ctg cac ctg gaa gtt aaa ate aac aag ccg ege
Met Gly Leu Ser Asp Leu His Leu Glu Val Lys lie Asn Lys Pro Arg 15 10 15 gac gtt gtt aac cag ate tgc atg gac get ate gac tac ggt aaa gac
Asp Val Val Asn Gin lie Cys Met Asp Ala lie Asp Tyr Gly Lys Asp 20 25 30 cac ggt ctg ttc gta gca ttc tet gca gaa gat gca act cgt act gaa 144
His Gly Leu Phe Val Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Glu 35 40 45
g 2 u t 9 e c 1 L 9 c c o r p aaa ctg ctg gac gtt tac aag cag gcg cag gac cac ggt get Lys Leu Leu Asp Val Tyr Lys Gin Ala Gin Asp His Gly Ala 55 60 gac ege att cac att get gac acc act ggt tet ate aac ccg tac gca 240
Asp Arg lie His lie Ala Asp Thr Thr Gly Ser lie Asn Pro Tyr Ala 65 70 75 80 act cag tac ctg gtt aaa aac ate aag aaa gag ate gac act gaa ate 288
Thr Gin Tyr Leu Val Lys Asn lie Lys Lys Glu lie Asp Thr Glu lie 85 90 95 -338 146966-序列表.doc 201037078 gcg ctg cac tgc cac aac gac ttc ggt ttc get gtt get aac tee ate 336
Ala Leu His Cys His Asn Asp Phe Gly Phe Ala Val Ala Asn Ser lie 100 105 110 get ggt ctg ttc gaa ggt gca act get ate tee act acc gtt aac ggt 384
Ala Gly Leu Phe Glu Gly Ala Thr Ala lie Ser Thr Thr Val Asn Gly 115 120 125 ate ggt gaa cgt get ggt aac gca tet ctg gaa gag ctg ate atg tet 432 lie Gly Glu Arg Ala Gly Asn Ala Ser Leu Glu Glu Leu lie Met Ser 130 135 140 ctg aaa ctg ctg tac aac aaa gac ctg ggc ttc aaa act gaa gtt ate 480
Leu Lys Leu Leu Tyr Asn Lys Asp Leu Gly Phe Lys Thr Glu Val lie 145 150 155 160 tac gaa ctg tet cag ctg gta tet aaa tac tee aag ate cca ate tet
528
Tyr Glu Leu Ser Gin Leu Val Ser Lys Tyr Ser Lys lie Pro lie Ser 165 170 175 gac tee aaa gcg att gtt ggt aac aac gta ttc cgt cac gaa tee ggt 576
Asp Ser Lys Ala lie Val Gly Asn Asn Val Phe Arg His Glu Ser Gly 180 185 190 ate cac gta gac get ate gtt aaa aac ccg ctg get tac gaa ccg ttc 624 lie His Val Asp Ala lie Val Lys Asn Pro Leu Ala Tyr Glu Pro Phe 195 200 205 ate ccg gaa atg ate ggc acc aag cgt cag ate gtt ctg ggt aaa cac 672 lie Pro Glu Met lie Gly Thr Lys Arg Gin lie Val Leu Gly Lys His 210 215 220 tet ggt aaa tet gcg gtt ate gaa aaa ctg gat act ctg aac ate aaa 0 720
Ser Gly Lys Ser Ala Val lie Glu Lys Leu Asp Thr Leu Asn lie Lys 225 230 235 240 gtt gat gac act cag ctg tet cag ate gtt tet ctg gtt aag cag gaa 768
Val Asp Asp Thr Gin Leu Ser Gin lie Val Ser Leu Val Lys Gin Glu 245 250 255 cgt gaa cgt ggt gaa gaa ate act aac aac aag ttc gac gaa ate ctg 816
Arg Glu Arg Gly Glu Glu lie Thr Asn Asn Lys Phe Asp Glu lie Leu 260 265 270 gaa aaa gtt aac ate aaa agg i
Glu Lys Val Asn lie Lys Arg 275 <210> 267 339- 146966·序列表.doc 201037078 <211> 279
<212> PRT <213>思氏甲烷球形菌DSM 3091 <400> 267
Met Gly Leu Ser Asp Leu His Leu Glu Val Lys lie Asn Lys Pro Arg 15 10 15
Asp Val Val Asn Gin lie Cys Met Asp Ala lie Asp Tyr Gly Lys Asp 20 25 30
His Gly Leu Phe Val Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Glu 35 40 45
Leu Pro Lys Leu Leu Asp Val Tyr Lys Gin Ala Gin Asp His Gly Ala 50 55 60
Asp Arg lie His lie Ala Asp Thr Thr Gly Ser lie Asn Pro Tyr Ala 65 70 75 80
Thr Gin Tyr Leu Val Lys Asn lie Lys Lys Glu lie Asp Thr Glu lie 85 90 95
Ala Leu His Cys His Asn Asp Phe Gly Phe Ala Val Ala Asn Ser lie 100 105 110
Ala Gly Leu Phe Glu Gly Ala Thr Ala lie Ser Thr Thr Val Asn Gly 115 120 125 lie Gly Glu Arg Ala Gly Asn Ala Ser Leu Glu Glu Leu lie Met Ser 130 135 140
Leu Lys Leu Leu Tyr Asn Lys Asp Leu Gly Phe Lys Thr Glu Val lie 145 150 155 160
Tyr Glu Leu Ser Gin Leu Val Ser Lys Tyr Ser Lys lie Pro lie Ser 165 170 175
Asp Ser Lys Ala lie Val Gly Asn Asn Val Phe Arg His Glu Ser Gly 180 185 190 lie His Val Asp Ala lie Val Lys Asn Pro Leu Ala Tyr Glu Pro Phe 195 200 205 lie Pro Glu Met lie Gly Thr Lys Arg Gin lie Val Leu Gly Lys His 210 215 220
Ser Gly Lys Ser Ala Val lie Glu Lys Leu Asp Thr Leu Asn lie Lys 225 230 235 240
Val Asp Asp Thr Gin Leu Ser Gin lie Val Ser Leu Val Lys Gin Glu 245 250 255
Arg Glu Arg Gly Glu Glu lie Thr Asn Asn Lys Phe Asp Glu lie Leu 260 265 270
Glu Lys Val Asn lie Lys Arg 275 <210> 268 <211> 840
<212> DNA <213> 思氏甲烷球形菌DSM 3091 <220> <223> 野生型 <400> 268
ATGGGATTAT CAGACCTACA TCTAGAAGTT AAAAT7VAACA AACCACGAGA TGTAGTAAAT 60 CAAATATGTA TGGATGCAAT AGATTATGGT AAGGATCATG GACTATTTGT AGCATTCTCT 120 GCTGAAGATG CAACAAGAAC AGAACTTCCA AAACTTCTTG ATGTATATAA ACAAGCACAA 180 GATCATGGAG CAGACAGAAT TCATATAGCA GATACAACAG GTTCAATAAA TCCATATGCA 240 •340· 146966-序列表.doc 201037078 ACACAGTACT TAGTAAAAAA TATTAAAAAA GAAATAGATA CAGAAATAGC ACTACACTGT 300 CATAATGACT TTGGATTTGC AGTTGCAAAT TCCATTGCAG GATTATTTGA AGGCGCAACT 360 GCAATATCTA CAACAGTAAA TGGTATAGGT GAAAGAGCAG GAAATGCCTC TCTAGAAGAA 420 TTAATAATGT CATTAAAACT ATTATATAAT AAGGATTTAG GATTTAAAAC AGAAGTAATC 480 TATGAACTAT CACAACTTGT ATCAAAATAT AGTAAAATTC CAATATCAGA TAGTAAAGCT 540 ATAGTTGGAA ACAATGTATT TAGACATGAA TCAGGAATAC ATGTAGATGC AATAGTAAAA 600 AATCCATTAG CATATGAACC ATTCATTCCA GAAATGATTG GAACAAAAAG ACAAATAGTT 660 CTAGGAAAAC ATTCAGGTAA ATCAGCAGTA ATAGAAAAAT TAGACACATT AAATATAAAA 720 GTAGATGACA CACAATTATC ACAAATAGTT TCCCTAGTAA AACAAGAACG TGAAAGAGGA 780
GAAGAAATAA CAAATAATAA ATTCGATGAA ATACTAGAAA AAGTTAATAT TAAAAGATGA 840 <210> 269 <211> 1143
<212> DNA <213>肺炎克雷伯氏桿菌 <220> <221> CDS <222> (1)..(1143) <400> 269 atg gag cgc gtt ctg ate aac gac acc act ctg ege gac ggc gag cag
Met Glu Arg Val Leu lie Asn Asp Thr Thr Leu Arg Asp Gly Glu Gin 15 10 15 tet ccg ggc gta geg ttc cgt act tet gaa aaa gtt get ate get gaa
Ser Pro Gly Val Ala Phe Arg Thr Ser Glu Lys Val Ala lie Ala Glu 20 25 30 geg ctg tac get get ggt ate act gca atg gaa gtt ggt act ccg gca 144
Ala Leu Tyr Ala Ala Gly lie Thr Ala Met Glu Val Gly Thr Pro Ala 35 40 45 atg ggc gac gaa gag ate get cgc att cag ctg gtt cgt cgt cag ctg 192
Met Gly Asp Glu Glu lie Ala Arg lie Gin Leu Val Arg Arg Gin Leu 50 55 60 ccg gac gca act ctg atg acc tgg tgc cgt atg aac geg ctg gaa ate 240
Pro Asp Ala Thr Leu Met Thr Trp Cys Arg Met Asn Ala Leu Glu lie 65 70 75 80 cgt cag tet gca gac ctg ggt ate gac tgg gtt gat ate tee att ccg 288
Arg Gin Ser Ala Asp Leu Gly lie Asp Trp Val Asp lie Ser lie Pro 85 90 95 146966·序列表 _doc 48 96
-341 - 201037078 gca tct gac aag ctg cgt cag tac aaa ctg cgt gaa ccg ctg gcg gta 336
Ala Ser Asp Lys Leu Arg Gin Tyr Lys Leu Arg Glu Pro Leu Ala Val 100 105 110 ctg ctg gaa cgt ctg gcg atg ttc ate cac ctg gcg cac act ctg ggt 384
Leu Leu Glu Arg Leu Ala Met Phe lie His Leu Ala His Thr Leu Gly 115 120 125 ctg aaa gtt tgc ate ggt tgc gaa gat get tct ege get tct ggt cag 432
Leu Lys Val Cys lie Gly Cys Glu Asp Ala Ser Arg Ala Ser Gly Gin 130 135 140 act ctg ege get ate get gaa gtt get cag aac gcg ccg get get ege 480
Thr Leu Arg Ala lie Ala Glu Val Ala Gin Asn Ala Pro Ala Ala Arg 145 150 155 160 ctg cgt tac get gac acc gtt ggt ctg ctg gat ccg ttc acc act gca 528
Leu Arg Tyr Ala Asp Thr Val Gly Leu Leu Asp Pro Phe Thr Thr Ala 165 170 175 get cag ate tee gca ctg ege gac gta tgg tct ggt gaa ate gaa atg 576
Ala Gin lie Ser Ala Leu Arg Asp Val Trp Ser Gly Glu lie Glu Met 180 185 190 cac gcg cac aac gac ctg ggt atg gcg act get aac acc ctg gca get 624
His Ala His Asn Asp Leu Gly Met Ala Thr Ala Asn Thr Leu Ala Ala 195 200 205 gtt tct get ggc gca act tct gtt aac acc acc gtt ctg ggt ctg ggt 672
Val Ser Ala Gly Ala Thr Ser Val Asn Thr Thr Val Leu Gly Leu Gly 210 215 220 gaa cgt get ggt aac gca get gca tgg aaa ccg tct gca ctg ggt ctg 720
Glu Arg Ala Gly Asn Ala Ala Ala Trp Lys Pro Ser Ala Leu Gly Leu 225 230 235 240 gaa cgt tgc ctg ggt gtt gaa act ggc gtt cac ttc tct gca ctg ccg 768
Glu Arg Cys Leu Gly Val Glu Thr Gly Val His Phe Ser Ala Leu Pro 245 250 255 gca ctg tgc cag ege gtt get gaa gca get cag cgt get ate gac ccg 816
Ala Leu Cys Gin Arg Val Ala Glu Ala Ala Gin Arg Ala lie Asp Pro 260 265 270 cag cag ccg ctg gtt ggt gaa ctg gta ttc act cac gaa tee ggt gtt 864
Gin Gin Pro Leu Val Gly Glu Leu Val Phe Thr His Glu Ser Gly Val 275 280 285 -342· 146966-序列表.doc 201037078 cac gtt get geg ctg ctg ege gac tet gaa tee tac cag tet ate geg 912
His Val Ala Ala Leu Leu Arg Asp Ser Glu Ser Tyr Gin Ser lie Ala 290 295 300 cca age ctg atg ggt cgt tet tac cgt ctg gtt ctg ggt aag cac tet 960
Pro Ser Leu Met Gly Arg Ser Tyr Arg Leu Val Leu Gly Lys His Ser 305 310 315 320 ggt cgt cag get gtt aac ggc gta ttc gac cag atg ggt tac cac ctg 1008
Gly Arg Gin Ala Val Asn Gly Val Phe Asp Gin Met Gly Tyr His Leu 325 330 335 aac get geg cag ate aac cag ctg ctg ccg gca ate cgt ege ttc get 1056
Asn Ala Ala Gin lie Asn Gin Leu Leu Pro Ala lie Arg Arg Phe Ala 340 345 350 Ο
gaa aac tgg aaa cgt tet ccg aaa gac tac gaa ctg gtt get ate tac 1104
Glu Asn Trp Lys Arg Ser Pro Lys Asp Tyr Glu Leu Val Ala lie Tyr 355 360 365 1143 gac gaa ctg tgc ggt gaa tee gca ctg cgt geg cgt gga Asp Glu Leu Cys Gly Glu Ser Ala Leu Arg Ala Arg Gly 370 375 380 <210> 270
<211> 381 <212> PRT <213>肺炎克雷伯氏桿菌 <400> 270
Met Glu Arg Val Leu lie Asn Asp Thr Thr Leu Arg Asp Gly Glu Gin 1 5 10 15
Ser Pro Gly Val Ala Phe Arg Thr Ser Glu Lys Val Ala He Ala Glu 20 25 30 TVla Leu Tyr Ala Ala Gly He Thr Ala Met Glu Val Gly Thr Pro Ala 35 40 45
Met Gly Asp Glu Glu He Ala Arg He Gin Leu Val Arg Arg Gin Leu 50 55 60
Pro Asp Ala Thr Leu Met Thr Trp Cys Arg Met Asn Ala Leu Glu He 65 70 75 80
Arg Gin Ser Ala Asp Leu Gly lie Asp Trp Val Asp lie Ser lie Pro 85 90 95
Ala Ser Asp Lys Leu Arg Gin Tyr Lys Leu Arg Glu Pro Leu Ala Val 100 105 no
Leu Leu Glu Arg Leu Ala Met Phe He His Leu Ala His Thr Leu Gly 115 120 125
Leu Lys Val Cys 工le Gly Cys Glu Asp Ala Ser Arg Ala Ser Gly Gin 130 135 140
Thr Leu Arg Ala 工le Ala Glu Val Ala Gin Asn Ala Pro Ala Ala Arg 145 150 155 160
Leu Arg Tyr Ala Asp Thr Val Gly Leu Leu Asp Pro Phe Thr Thr Ala 165 170 175
Ala Gin lie Ser Ala Leu Arg Asp Val Trp Ser Gly Glu lie Glu Met 180 185 190
His Ala His Asn Asp Leu Gly Met Ala Thr Ala Asn Thr Leu Ala Ala - 343 - 146966-序列表.doc 201037078 195 200 205
Val Ser Ala Gly Ala Thr Ser Val Asn Thr Thr Val Leu Gly Leu Gly 210 215 220
Glu Arg Ala Gly Asn Ala Ala Ala Trp Lys Pro Ser Ala Leu Gly Leu 225 230 235 240
Glu Arg Cys Leu Gly Val Glu Thr Gly Val His Phe Ser Ala Leu Pro 245 250 255
Ala Leu Cys Gin Arg Val Ala Glu Ala Ala Gin Arg Ala lie Asp Pro 260 265 270
Gin Gin Pro Leu Val Gly Glu Leu Val Phe Thr His Glu Ser Gly Val 275 280 285
His Val Ala Ala Leu Leu Arg Asp Ser Glu Ser Tyr Gin Ser lie Ala 290 295 300
Pro Ser Leu Met Gly Arg Ser Tyr Arg Leu Val Leu Gly Lys His Ser 305 310 315 320
Gly Arg Gin Ala Val Asn Gly Val Phe Asp Gin Met Gly Tyr His Leu 325 330 335
Asn Ala Ala Gin lie Asn Gin Leu Leu Pro Ala lie Arg Arg Phe Ala 340 345 350
Glu Asn Trp Lys Arg Ser Pro Lys Asp Tyr Glu Leu Val Ala lie Tyr 355 360 365
Asp Glu Leu Cys Gly Glu Ser Ala Leu Arg Ala Arg Gly 370 375 380 <210> 271 <211> 1146
<212> DNA <213>肺炎克雷伯氏桿菌 <220> <223> 野生型 <400> 271 ATGGAACGCG TGCTGATTAA CGATACCACC CTGCGCGACG GCGAGCAGAG CCCCGGCGTC 60 GCCTTTCGCA CCAGCGAAAA GGTCGCCATT GCCGAGGCGC TTTACGCCGC AGGAATAACG 120 GCGATGGAGG TCGGCACCCC GGCGATGGGC GACGAGGAGA TCGCGCGGAT CCAGCTGGTG 180 CGTCGCCAGC TGCCCGACGC GACCCTGATG ACCTGGTGTC GGATGAACGC GCTGGAGATC 240 CGCCAGAGCG CCGATCTGGG CATCGACTGG GTGGATATCT CGATTCCGGC TTCGGATAAG 300 CTGCGGCAGT ACAAACTGCG CGAGCCGCTG GCGGTGCTGC TGGAGCGGCT GGCGATGTTT 360 ATCCATCTTG CGCATACCCT CGGCCTGAAG GTATGCATCG GCTGCGAGGA CGCCTCGCGG 420 GCCAGCGGCC AGACCCTGCG CGCTATCGCC GAGGTCGCGC AGAATGCGCC GGCCGCCCGC 480 CTGCGCTATG CCGATACGGT CGGCCTGCTC GACCCTTTTA CCACCGCGGC GCAAATCTCG 540 GCCCTGCGCG ACGTCTGGTC CGGCGAAATC GAAATGCATG CCCATAACGA TCTGGGTATG 600 GCGACCGCCA ATACGCTGGC GGCGGTAAGC GCCGGGGCCA CCAGCGTGAA TACGACGGTC 660 CTCGGTCTCG GCGAGCGGGC GGGCAACGCG GCCGCATGGA AACCGTCCGC GCTGGGCCTT 720 GAACGCTGCC TGGGCGTGGA GACCGGCGTG CATTTTTCGG CGCTGCCCGC GCTCTGTCAG 780 344- 146966-序列表.doc 201037078 AGGGTCGCGG AAGCCGCGCA GCGCGCCATC GACCCGCAGC AGCCGCTGGT CGGCGAGCTG 840 GTGTTTACCC ATGAGTCAGG TGTCCACGTG GCGGCGCTGC TGCGCGACAG CGAGAGCTAC 900 CAGTCCATCG CCCCTTCCCT GATGGGCCGC AGCTACCGGC TGGTGCTGGG CAAACACTCC 960 GGGCGTCAGG CGGTCAACGG CGTTTTTGAC CAGATGGGCT ATCACCTCAA CGCCGCGCAG 1020 ATTAACCAGC TGCTGCCCGC CATCCGCCGC TTCGCCGAGA ACTGGAAGCG CAGCCCGAAA 1080 GATTACGAGC TGGTGGCTAT CTACGACGAG CTGTGCGGTG AATCCGCTCT GCGGGCGAGG 1140 GGGTAA 1146 <210> 272 <211> 1158
<212> DNA
<213> 萬氏曱烷球菌SB O<220> <221> CDS <222> (1)..(1158) <400> 272 atg gac tgg aaa gaa gtt tct cag tac aac ccg aaa ctg gac ctg aaa
Met Asp Trp Lys Glu Val Ser Gin Tyr Asn Pro Lys Leu Asp Leu Lys 15 10 15 gag tgc tac gtt tac gac acc act ctg cgt gac ggt gaa cag act ccg
Glu Cys Tyr Val Tyr Asp Thr Thr Leu Arg Asp Gly Glu Gin Thr Pro 20 25 30 ggc gtt tgc ttc acc ggt aac cag aaa ctg gaa ate geg aag aaa ctg 144
Gly Val Cys Phe Thr Gly Asn Gin Lys Leu Glu lie Ala Lys Lys Leu 35 40 45 gat gac ctg ggt ate aag cag att gaa get ggc ttc ccg acc gtt tct 192
Asp Asp Leu Gly lie Lys Gin lie Glu Ala Gly Phe Pro Thr Val Ser 50 55 60 gaa aac gaa cgt aag tgc ate aaa tct ate tct tct gaa ggt ctg aac 240
Glu Asn Glu Arg Lys Cys lie Lys Ser lie Ser Ser Glu Gly Leu Asn 65 70 75 80 get gac ate ctg geg ctg tct ege gta ctg aaa gaa gat ate gac ege 288
Ala Asp lie Leu Ala Leu Ser Arg Val Leu Lys Glu Asp lie Asp Arg 85 90 95 get ate gag tgc gac gtt gac ggt ate ate act ttc gtt gca act tct 336
Ala lie Glu Cys Asp Val Asp Gly lie He Thr Phe Val Ala Thr Ser 100 105 110 ccg atg cac ctg aaa tac aaa ctg cac aaa tct ttc gaa gaa gtt gaa 384
Pro Met His Leu Lys Tyr Lys Leu His Lys Ser Phe Glu Glu Val Glu 146966·序列表.doc 48 96 o -345 · 201037078 115 120 125 gag atg ggt atg aaa gca ate gaa tac get aaa gac cac ggt ctg ttc 432
Glu Met Gly Met Lys Ala lie Glu Tyr Ala Lys Asp His Gly Leu Phe 130 135 140 gtt gca ttc tet get gaa gat gca act cgt act tee ate gaa aac ate 480
Val Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Ser lie Glu Asn lie 145 150 155 160 ate aaa ate cac aaa aac get gaa gat tac ggt get gac ege gtt cac 528 lie Lys lie His Lys Asn Ala Glu Asp Tyr Gly Ala Asp Arg Val His 165 170 175 ate get gac act ctg ggt tgc gca act ccg cag tet atg tac cag ate 576 lie Ala Asp Thr Leu Gly Cys Ala Thr Pro Gin Ser Met Tyr Gin lie 180 185 190 tgc tet gaa ctg aac aag tet ctg aag aaa geg cac ate ggt gtt cac 624
Cys Ser Glu Leu Asn Lys Ser Leu Lys Lys Ala His lie Gly Val His 195 200 205 tgc cac aac gac ttc ggt ttc get get ate aac tee ate tac ggt ctg 672
Cys His Asn Asp Phe Gly Phe Ala Ala lie Asn Ser lie Tyr Gly Leu 210 215 220 atg ggt ggt geg aaa geg gtt tee act acc gtt aac ggt ate ggt gaa 720
Met Gly Gly Ala Lys Ala Val Ser Thr Thr Val Asn Gly lie Gly Glu 225 230 235 240 cgt get ggt aac get gca ctg gaa gaa gtt gta atg geg ctg aaa gtt 768
Arg Ala Gly Asn Ala Ala Leu Glu Glu Val Val Met Ala Leu Lys Val 245 250 255 ctg tac aac tac gac atg ggt ctg aac act gaa ctg ate atg gaa act 816
Leu Tyr Asn Tyr Asp Met Gly Leu Asn Thr Glu Leu lie Met Glu Thr 260 265 270 tet aaa ctg gtt gaa act tac tee aaa ate aaa gtt ccg gaa aac aag 864
Ser Lys Leu Val Glu Thr Tyr Ser Lys lie Lys Val Pro Glu Asn Lys 275 280 285 ccg ctg gtt ggt gaa atg gta ttc tac cac gaa tee ggt att cac gtt 912
Pro Leu Val Gly Glu Met Val Phe Tyr His Glu Ser Gly lie His Val 290 295 300 gac gca gta ctg gaa aac ccg ctg act tac gaa ccg ttc ctg cca gag 960
Asp Ala Val Leu Glu Asn Pro Leu Thr Tyr Glu Pro Phe Leu Pro Glu 305 310 315 320 • 346- 146966-序列表.doc 201037078 aag ate ggt cag aag cgt aag ate gtt ctg ggt aaa cac tet ggt tgc 1008
Lys lie Gly Gin Lys Arg Lys He Val Leu Gly Lys His Ser Gly Cys 325 330 335 cgt get gtt get tac cgt ctg aac gaa ctg ggc ttc gaa gca act ege 1056
Arg Ala Val Ala Tyr Arg Leu Asn Glu Leu Gly Phe Glu Ala Thr Arg 340 345 350 gac gaa ctg tgg gaa ate gtt aag aaa acc aaa gaa act cgt gaa cag 1104
Asp Glu Leu Trp Glu lie Val Lys Lys Thr Lys Glu Thr Arg Glu Gin 355 360 365 ggt act gaa ate tet gac gaa gta ttc aaa aac ate gtt act cac att 1152
Gly Thr Glu lie Ser Asp Glu Val Phe Lys Asn lie Val Thr His lie 370 375 380 ett aac 1158 Ο
Leu Asn 385 <210> 273 <211> 386
<212> PRT <213〉萬氏甲炫球菌SB <400> 273
Met Asp Trp Lys Glu Val Ser Gin Tyr Asn Pro Lys Leu Asp Leu Lys 15 10 15
Glu Cys Tyr Val Tyr Asp Thr Thr Leu Arg Asp Gly Glu Gin Thr Pro 20 25 30
Gly Val Cys Phe Thr Gly Asn Gin Lys Leu Glu lie Ala Lys Lys Leu 35 40 45
Asp Asp Leu Gly lie Lys Gin lie Glu Ala Gly Phe Pro Thr Val Ser 50 55 60
Glu Asn Glu Arg Lys Cys lie Lys Ser lie Ser Ser Glu Gly Leu Asn 65 70 * 75 80
Ala Asp lie Leu Ala Leu Ser Arg Val Leu Lys Glu Asp lie Asp Arg 85 90 95
Ala lie Glu Cys Asp Val Asp Gly lie lie Thr Phe Val Ala Thr Ser 100 105 110
Pro Met His Leu Lys Tyr Lys Leu His Lys Ser Phe Glu Glu Val Glu 115 120 125
Glu Met Gly Met Lys Ala lie Glu Tyr Ala Lys Asp His Gly Leu Phe 130 135 140
Val Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Ser lie Glu Asn lie 145 150 155 160 lie Lys lie His Lys Asn Ala Glu Asp Tyr Gly Ala Asp Arg Val His 165 170 175 lie Ala Asp Thr Leu Gly Cys Ala Thr Pro Gin Ser Met Tyr Gin lie 180 185 190
Cys Ser Glu Leu Asn Lys Ser Leu Lys Lys Ala His lie Gly Val His 195 200 205
Cys His Asn Asp Phe Gly Phe Ala Ala lie Asn Ser lie Tyr Gly Leu 210 215 220
Met Gly Gly Ala Lys Ala Val Ser Thr Thr Val Asn Gly lie Gly Glu -347- 146966-序列表.doc 201037078 225 230 235 240
Arg Ala Gly Asn Ala Ala Leu Glu Glu Val Val Met Ala Leu Lys Val 245 250 255
Leu Tyr Asn Tyr Asp Met Gly Leu Asn Thr Glu Leu lie Met Glu Thr 260 265 270
Ser Lys Leu Val Glu Thr Tyr Ser Lys lie Lys Val Pro Glu Asn Lys 275 280 285
Pro Leu Val Gly Glu Met Val Phe Tyr His Glu Ser Gly lie His Val 290 295 300
Asp Ala Val Leu Glu Asn Pro Leu Thr Tyr Glu Pro Phe Leu Pro Glu 305 310 315 320
Lys lie Gly Gin Lys Arg Lys lie Val Leu Gly Lys His Ser Gly Cys 325 330 335
Arg Ala Val Ala Tyr Arg Leu Asn Glu Leu Gly Phe Glu Ala Thr Arg 340 345 350
Asp Glu Leu Trp Glu lie Val Lys Lys Thr Lys Glu Thr Arg Glu Gin 355 360 365
Gly Thr Glu lie Ser Asp Glu Val Phe Lys Asn lie Val Thr His lie 370 375 380
Leu Asn 385
<210> 274 <211> 1161 <212> DNA <213> 萬氏甲烷球菌SB <220> <223> 野生型 <400> 274 ATGGATTGGA AAGAAGTTTC TCAATATAAT CCTAAATTAG ACTTAAAAGA ATGTTATGTA 60 TATGATACTA CATTAAGGGA TGGTGAACAG ACTCCTGGAG TTTGTTTTAC GGGTAATCAA 120 AAGCTTGAAA TTGCTAAAAA ATTAGATGAC CTTGGAATAA AACAAATTGA AGCCGGTTTT 180 CCAACGGTTT CTGAAAATGA AAGAAAATGT ATTAAATCAA TTTCCTCTGA AGGACTTAAT 240 GCGGATATTT TGGCGCTATC AAGAGTTTTA AAGGAAGATA TCGATAGGGC GATTGAATGC 300 GATGTTGATG GAATAATCAC TTTTGTTGCA ACTTCGCCTA TGCACCTAAA ATATAAGCTA 360 CATAAGTCTT TTGAAGAAGT TGAAGAAATG GGAATGAAAG CTATCGAGTA TGCAAAAGAT 420 CATGGCCTTT TTGTTGCGTT TTCTGCAGAA GATGCAACAA GAACCTCGAT TGAAAATATC 480 ATAAAAATTC ATAAAAATGC AGAAGATTAC GGTGCAGATA GGGTCCATAT TGCAGATACA 540 CTTGGATGTG CAACACCTCA ATCAATGTAC CAAATATGTT CTGAACTGAA TAAAAGCCTG 600 AAAAAAGCAC ATATTGGAGT TCATTGCCAT AATGATTTTG GATTTGCAGC CATAAATTCA 660 ATATACGGAC TTATGGGTGG TGCAAAGGCA GTTTCAACTA CTGTTAATGG CATTGGAGAA 720 AGGGCCGGAA ATGCAGCACT AGAAGAGGTA GTAATGGCTT TAAAAGTACT TTATAATTAC 780 GATATGGGGC TAAATACTGA ACTTATAATG GAAACTTCAA AGTTAGTTGA AACATATTCA 840 -348- 146966-序列表.doc
Ο 201037078 AAAATCAAGG TTCCAGAAAA TAAGCCTCTT GTTGGCGAAA TGGTATTTTA TCATGAAAGC 900 GGAATACACG TTGATGCAGT TTTAGAAAAT CCTTTAACCT ATGAGCCGTT TTTACCTGAA 960 AAAATCGGTC 7VAAAACGAAA AATTGTGCTT GGAAAACATT CTGGATGCAG AGCCGTTGCA 1020 TATAGATTAA ATGAACTTGG ATTTGAAGCG ACTCGTGATG AACTCTGGGA AATTGTTAAA 1080 AAAACTAAGG AAACGAGAGA ACAAGGAACC GAAATAAGTG ACGAAGTATT TAAAAACATT 1140 GTAACACATA TTTTAAATTA A 1161 <210> 275 <211> 1170
<212> DNA <213> 史氏曱烷短桿菌ATCC 35061 <220> <221> CDS <222> (1)..(1170) <400> 275 atg cag tac tac ate tet cac tac aac aaa gag ccg gaa ctg aac ttc
Met Gin Tyr Tyr lie Ser His Tyr Asn Lys Glu Pro Glu Leu Asn Phe 15 10 15 ccg gat gaa ate acc gtt tac gac acc act ctg cgt gac ggt gaa cag
Pro Asp Glu lie Thr Val Tyr Asp Thr Thr Leu Arg Asp Gly Glu Gin 20 25 30 act ccg ggc gtt tgc ttc tet ccg gaa gag aag ctg gaa ate geg aag 144
Thr Pro Gly Val Cys Phe Ser Pro Glu Glu Lys Leu Glu lie Ala Lys 35 40 45 aaa ctg gat gaa gtt aaa ate aag cag ate gaa gca ggc ttc ccg ate 192
Lys Leu Asp Glu Val Lys lie Lys Gin lie Glu Ala Gly Phe Pro lie 50 55 60 gtt tee aag aaa gag cag gaa tee gtt aaa gca ate act tet gaa ggt 240
Val Ser Lys Lys Glu Gin Glu Ser Val Lys Ala lie Thr Ser Glu Gly 65 70 75 80 ctg aac gca cag ate ate tet ctg tet cgt acc aag aaa gaa gat ate 288
Leu Asn Ala Gin lie lie Ser Leu Ser Arg Thr Lys Lys Glu Asp lie 85 90 95 gac get gca ctg gac tgc gac gtt gac ggc gtt ate act ttc atg ggt 336
Asp Ala Ala Leu Asp Cys Asp Val Asp Gly Val lie Thr Phe Met Gly 100 105 110 act tet gac ate cac ctg gaa cac aaa atg cac att ggt cgt cag gaa 384
Thr Ser Asp lie His Leu Glu His Lys Met His lie Gly Arg Gin Glu 115 120 125 146966-序列表.doe 48 96 -349- 201037078 gcg ctg aac act tgc atg aac get ate gaa tac get aaa gac cac ggt 432
Ala Leu Asn Thr Cys Met Asn Ala lie Glu Tyr Ala Lys Asp His Gly 130 135 140 ctg ttc gtt gca ttc tet gca gaa gat gca act cgt act gac ctg gac 480
Leu Phe Val Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Asp Leu Asp 145 150 155 160 ttc ctg aaa cgt ate tac aac aaa get gaa tee tac ggt get gac ege 528
Phe Leu Lys Arg lie Tyr Asn Lys Ala Glu Ser Tyr Gly Ala Asp Arg 165 170 175 gtt cac att get gac ace act ggc gca ate act ccg cag ggt ate act 576
Val His lie Ala Asp Thr Thr Gly Ala lie Thr Pro Gin Gly lie Thr 180 185 190 tac ctg gtt aaa gaa ctg aag aaa gac gta aac ate gac ate gcg ctg 624
Tyr Leu Val Lys Glu Leu Lys Lys Asp Val Asn lie Asp lie Ala Leu 195 200 205 cac tgc cac aac gac ttc ggt ctg gcg gtt ate aac tee ate tee ggt 672
His Cys His Asn Asp Phe Gly Leu Ala Val lie Asn Ser lie Ser Gly 210 215 220 gtt ctg gcg ggt get aac ggt ate tee act acc gta aac ggt ate ggt 720
Val Leu Ala Gly Ala Asn Gly lie Ser Thr Thr Val Asn Gly lie Gly 225 230 235 240 gaa ege get ggt aac get tet ctg gaa gaa gtt ate atg tet ctg aaa 768
Glu Arg Ala Gly Asn Ala Ser Leu Glu Glu Val lie Met Ser Leu Lys 245 250 255 ctg ctg tac ggt aaa gac ctg ggc ttc aaa acc aag cac ate aaa gaa 816
Leu Leu Tyr Gly Lys Asp Leu Gly Phe Lys Thr Lys His lie Lys Glu 260 265 270 ctg tet gaa ctg gtt tet aaa gcg tet ggt ctg ccg gtt ccg tac aac 864
Leu Ser Glu Leu Val Ser Lys Ala Ser Gly Leu Pro Val Pro Tyr Asn 275 280 285 aag ccg gtt gtt ggt aac aac gta ttc cgt cac gaa tee ggt att cac 912
Lys Pro Val Val Gly Asn Asn Val Phe Arg His Glu Ser Gly lie His 290 295 300 gtt gat get gtt ate gaa gag ccg ctg tgc tac gaa ccg tac ate cca 960
Val Asp Ala Val lie Glu Glu Pro Leu Cys Tyr Glu Pro Tyr lie Pro 305 310 315 320 -350- 146966·序列表.doc 201037078 gaa ctg gtt ggt cag aaa cgt cag ctg gta ctg ggt aaa cac tct ggt 1008
Glu Leu Val Gly Gin Lys Arg Gin Leu Val Leu Gly Lys His Ser Gly 325 330 335 tgc cgc get gta ege get aaa ctg aac gaa tgc gac ctg gac gtt tct 1056
Cys Arg Ala Val Arg Ala Lys Leu Asn Glu Cys Asp Leu Asp Val Ser 340 345 350 gac gac act ctg ate gaa ate gtt aag aaa gtt aag aaa tct cgt gaa 1104
Asp Asp Thr Leu lie Glu lie Val Lys Lys Val Lys Lys Ser Arg Glu 355 360 365 gaa ggt act tac ate aac gac gac gta ttc aaa gaa ate gtt aaa tee 1152
Glu Gly Thr Tyr lie Asn Asp Asp Val Phe Lys Glu lie Val Lys Ser 370 375 380 tgc aac tac aag aaa gaa 1170
Cys Asn Tyr Lys Lys Glu 385 390 <210> 276
<211> 390 <212> PRT <213> 史氏甲烷短桿菌ATCC 35061 <400> 276
Met Gin Tyr Tyr lie Ser His Tyr Asn Lys Glu Pro Glu Leu Asn Phe 1 5 10 15
Pro Asp Glu lie Thr Val Tyr Asp Thr Thr Leu Arg Asp Gly Glu Gin 20 25 30
Thr Pro Gly Val Cys Phe Ser Pro Glu Glu Lys Leu Glu lie Ala Lys 35 40 45
Lys Leu Asp Glu Val Lys lie Lys Gin lie Glu Ala Gly Phe Pro lie 50 55 60
Val Ser Lys Lys Glu Gin Glu Ser Val Lys Ala lie Thr Ser Glu Gly 65 70 75 80
Leu Asn Ala Gin lie lie Ser Leu Ser Arg Thr Lys Lys Glu Asp lie 85 90 95
Asp Ala Ala Leu Asp Cys Asp Val Asp Gly Val lie Thr Phe Met Gly 100 105 110
Thr Ser Asp lie His Leu Glu His Lys Met His lie Gly Arg Gin Glu 115 120 125
Ala Leu Asn Thr Cys Met Asn Ala lie Glu Tyr Ala Lys Asp His Gly 130 135 140
Leu Phe Val Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Asp Leu Asp 145 150 155 160
Phe Leu Lys Arg lie Tyr Asn Lys Ala Glu Ser Tyr Gly Ala Asp Arg 165 170 175
Val His lie Ala Asp Thr Thr Gly Ala lie Thr Pro Gin Gly lie Thr 180 185 190
Tyr Leu Val Lys Glu Leu Lys Lys Asp Val Asn lie Asp lie Ala Leu 195 200 205
His Cys His Asn Asp Phe Gly Leu Ala Val lie Asn Ser lie Ser Gly 210 215 220
Val Leu Ala Gly Ala Asn Gly lie Ser Thr Thr Val Asn Gly lie Gly 225 230 235 240 -351 - 146966-序列表.doc 201037078
Glu Arg Ala Gly Asn Ala Ser Leu Glu Glu Val lie Met Ser Leu Lys 245 250 255
Leu Leu Tyr Gly Lys Asp Leu Gly Phe Lys Thr Lys His lie Lys Glu 260 265 270
Leu Ser Glu Leu Val Ser Lys Ala Ser Gly Leu Pro Val Pro Tyr Asn 275 280 285
Lys Pro Val Val Gly Asn Asn Val Phe Arg His Glu Ser Gly lie His 290 295 300
Val Asp Ala Val 工le Glu Glu Pro Leu Cys Tyr Glu Pro Tyr lie Pro 305 310 315 320
Glu Leu Val Gly Gin Lys Arg Gin Leu Val Leu Gly Lys His Ser Gly 325 330 335
Cys Arg Ala Val Arg Ala Lys Leu Asn Glu Cys Asp Leu Asp Val Ser 340 345 350
Asp Asp Thr Leu lie Glu lie Val Lys Lys Val Lys Lys Ser Arg Glu 355 360 365
Glu Gly Thr Tyr lie Asn Asp Asp Val Phe Lys Glu lie Val Lys Ser 370 375 380
Cys Asn Tyr Lys Lys Glu 385 390 <210> 277 <211> 1173
<212> DNA <213> 史氏甲烷短桿菌ATCC 35061 <220> <223> 野生型 <400> 277
TTGCAATATT ATATCAGTCA TTATAATAAA GAACCGGAA.T TAAATTTTCC TGATGAAATT 60 ACAGTTTATG ATACAACTTT AAGAGACGGT GAACAAACCC CAGGAGTTTG TTTTAGTCCT 120 GAAGAAAAGT TAGAAATTGC TAAAAAATTA GATGAAGTTA AAATTAAACA AATTGAAGCA 180 GGTTTTCCTA TAGTTTCCAA AAAAGAACAG GAATCAGTGA AAGCCATAAC TTCAGAAGGC 240 CTTAATGCTC AAATCATTTC TTTATCTAGA ACAAAAAAAG AAGACATTGA TGCTGCTCTT 300 GATTGTGATG TTGACGGTGT TATTACATTT ATGGGAACTT CAGACATCCA TTTAGAACAC 360 AAAATGCACA TCGGACGTCA GGAAGCACTG AATACATGTA TGAACGCTAT TGAATATGCT 420 AAAGATCACG GATTATTTGT TGCATTTTCT GCAGAAGACG CAACCAGAAC TGATTTGGAT 480 TTCTTAAAAA GAATATACAA CAAGGCTGAA AGCTATGGAG CAGACAGAGT ACACATAGCA 540 GACACTACCG GTGCTATAAC TCCGCAGGGA ATTACTTACC TGGTAAAAGA ACTTAAAAAA 600 GATGTAAATA TAGATATTGC TCTACATTGT CACAATGATT TCGGTCTGGC AGTTATAAAT 660 TCCATATCCG GAGTACTGGC AGGAGCTAAT GGAATTTCCA CAACAGTAAA CGGTATCGGA 720 GAACGTGCAG GAAACGCATC TCTTGAAGAA GTAATTATGT CTCTTAAATT ATTATACGGT 780 AAAGATTTAG GATTTAAAAC CAAACATATT AAAGAACTTT CCGAACTTGT ATCAAAAGCT 840 AGCGGATTGC CTGTTCCCTA TAACAAACCT GTAGTAGGAA ACAACGTATT CAGACACGAA 900 352 - 146966-序列表.doc 201037078 TCCGGAATTC ATGTTGATGC AGTTATTGAA GAGCCGTTAT GTTATGAACC ATATATTCCA 960 GAACTGGTAG GACAAAAAAG ACAGCTTGTA TTAGGTAAAC ATTCAGGCTG CAGAGCAGTC 1020 AGAGCCAAAT TAAATGAATG TGACCTTGAT GTAAGCGACG ATACCCTTAT TGAAATTGTT 1080 AAAAAAGTTA AAAAAAGCAG AGAAGAAGGA ACATACATTA ACGATGATGT ATTCAAAGAA 1140 ATTGTAAAAA GTTGTAACTA TAAAAAAGAA TAG 1173
<210> 278 <211> 1161 <212> DNA <213 > 雜色甲烷球菌Nankai-3 <220> <221> CDS <222> (1)..(1161)
<400> 278 atg aac tgg aaa gaa gtt tgc cag tac aac ccg aaa ctg aac ctg gaa Met Asn Trp Lys Glu Val Cys Gin Tyr Asn Pro Lys Leu Asn Leu Glu 48 1 5 10 15 96 gat tgc tac ate tac gat acc act ctg ege gac ggt gaa cag act ccg Asp Cys Tyr lie Tyr Asp Thr Thr Leu Arg Asp Gly Glu Gin Thr Pro 20 25 30 ggc gtt tgc ttc tee atg gag cag aaa ctg gac ate get aag aaa ctg 144
Gly Val Cys Phe Ser Met Glu Gin Lys Leu Asp lie Ala Lys Lys Leu 35 40 45 gac gaa ctg ggt gtt aag cag ate gaa geg ggt ttc ccg get gtt tet 192
Asp Glu Leu Gly Val Lys Gin lie Glu Ala Gly Phe Pro Ala Val Ser 50 55 60 aaa tet gaa ate gaa aac gtt aag aaa ate get aac gaa ggt ctg aac 240
Lys Ser Glu lie Glu Asn Val Lys Lys lie Ala Asn Glu Gly Leu Asn 65 70 75 80 get gaa ate ctg gca ctg tet ege gca ctg cag ggc gac ate gac aaa 288
Ala Glu lie Leu Ala Leu Ser Arg Ala Leu Gin Gly Asp lie Asp Lys 85 90 95 geg ctg tee tgc gac gtt gac ggt ate ate act ttc ate geg gca tet 336
Ala Leu Ser Cys Asp Val Asp Gly lie lie Thr Phe lie Ala Ala Ser 100 105 110 ccg ctg cac ctg aaa tac aaa ctg cac aaa tet ate gaa gaa gtt gaa 384
Pro Leu His Leu Lys Tyr Lys Leu His Lys Ser lie Glu Glu Val Glu 115 120 125 gag atg ggt atg aaa geg gta gaa tac get aaa gac cac ggt ctg ttc 432 -353- 146966-序列表.doc 201037078
Glu Met Gly Met Lys Ala Val Glu Tyr Ala Lys Asp His Gly Leu Phe 130 135 140 gtt gca ttc tct gca gaa gat gca act cgt act ccg ate gaa gat ctg 480
Val Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Pro lie Glu Asp Leu 145 150 155 160 gta cgt att cac aaa aac get gaa gag cac ggt get gac ege gtt cac 528
Val Arg lie His Lys Asn Ala Glu Glu His Gly Ala Asp Arg Val His 165 170 175 att get gac acc act ggc tgc ggt act ccg cag tee att cag tac ate 576 lie Ala Asp Thr Thr Gly Cys Gly Thr Pro Gin Ser lie Gin Tyr lie 180 185 190 tgc tct gaa ctg tct aac aac ctg aag aaa get cac ate ggc gta cac 624
Cys Ser Glu Leu Ser Asn Asn Leu Lys Lys Ala His lie Gly Val His 195 200 205 tgc cac aac gac ttc ggt ctg geg gtt ate aac tee ate tac ggt ctg 672
Cys His Asn Asp Phe Gly Leu Ala Val lie Asn Ser lie Tyr Gly Leu 210 215 220 ctg ggt ggt geg aaa get get tct act acc gtt aac ggt ate ggt gaa 720
Leu Gly Gly Ala Lys Ala Ala Ser Thr Thr Val Asn Gly lie Gly Glu 225 230 235 240 cgt get ggt aac gca ccg ctg gaa gaa ctg ctg ctg acc atg aac gta 768
Arg Ala Gly Asn Ala Pro Leu Glu Glu Leu Leu Leu Thr Met Asn Val 245 250 255 ctg tac gac gtt aaa act gac ctg aac ate tee ate ate aaa gaa ctg 816
Leu Tyr Asp Val Lys Thr Asp Leu Asn lie Ser lie lie Lys Glu Leu 260 265 270 tct acc atg gtt gaa aac tac tee ggt ate aaa ate ccg gtt aac aag 864
Ser Thr Met Val Glu Asn Tyr Ser Gly lie Lys lie Pro Val Asn Lys 275 280 285 cca ate gtt ggt gac aaa gta ttc tac cac gaa tee ggt ate cac gtt 912
Pro lie Val Gly Asp Lys Val Phe Tyr His Glu Ser Gly lie His Val 290 295 300 gac get gtt ate gaa aac ccg ctg act tac gaa ccg ttc ctg ccg gaa 960
Asp Ala Val lie Glu Asn Pro Leu Thr Tyr Glu Pro Phe Leu Pro Glu 305 310 315 320 cgt ate ggt cag aag cgt gaa ate gtt ctg ggt aaa cac tct ggt tgc 1008
Arg lie Gly Gin Lys Arg Glu lie Val Leu Gly Lys His Ser Gly Cys -354· 146966·序列表.doc 201037078 325 330 335 tcc gca gta gaa tcc aag ctg aaa gag ctg ggt ctg gaa gtt ccg aaa 1056
Ser Ala Val Glu Ser Lys Leu Lys Glu Leu Gly Leu Glu Val Pro Lys 340 345 350 gac cgt ate tgg gac ctg gtt aag aaa gtt aaa acc act cgt gaa ggt 1104
Asp Arg lie Trp Asp Leu Val Lys Lys Val Lys Thr Thr Arg Glu Gly 355 360 365 ggt gaa gat ate gac gac gaa atg ttc ate aag ate gtt gac ate ate 1152
Gly Glu Asp lie Asp Asp Glu Met Phe lie Lys lie Val Asp lie lie 370 375 380 aac aag cag 1161
Asn Lys Gin 385
<210> 279 <211> 387
<212> PRT
<213 >雜色曱燒球菌NankaW <400> 279
Met Asn Trp Lys Glu Val Cys Gin Tyr Asn Pro Lys Leu Asn Leu Glu 15 10 15
Asp Cys Tyr lie Tyr Asp Thr Thr Leu Arg Asp Gly Glu Gin Thr Pro 20 25 30
Gly Val Cys Phe Ser Met Glu Gin Lys Leu Asp lie Ala Lys Lys Leu 35 40 45
Asp Glu Leu Gly Val Lys Gin lie Glu Ala Gly Phe Pro Ala Val Ser 50 55 60
Lys Ser Glu lie Glu Asn Val Lys Lys lie Ala Asn Glu Gly Leu Asn 65 70 75 80
Ala Glu lie Leu Ala Leu Ser Arg Ala Leu Gin Gly Asp lie Asp Lys 85 90 95
Ala Leu Ser Cys Asp Val Asp Gly lie 工le Thr Phe lie Ala Ala Ser 100 105 110
Pro Leu His Leu Lys Tyr Lys Leu His Lys Ser lie Glu Glu Val Glu 115 120 125
Glu Met Gly Met Lys Ala Val Glu Tyr Ala Lys Asp His Gly Leu Phe 130 135 140
Val Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Pro lie Glu Asp Leu 145 150 155 160
Val Arg lie His Lys Asn Ala Glu Glu His Gly Ala Asp Arg Val His 165 170 175 lie Ala Asp Thr Thr Gly Cys Gly Thr Pro Gin Ser lie Gin Tyr lie 180 185 190
Cys Ser Glu Leu Ser Asn Asn Leu Lys Lys Ala His lie Gly Val His 195 200 205
Cys His Asn Asp Phe Gly Leu Ala Val lie Asn Ser lie Tyr Gly Leu 210 215 220
Leu Gly Gly Ala Lys Ala Ala Ser Thr Thr Val Asn Gly lie Gly Glu 225 230 235 240
Arg Ala Gly Asn Ala Pro Leu Glu Glu Leu Leu Leu Thr Met Asn Val 245 250 255
Leu Tyr Asp Val Lys Thr Asp Leu Asn lie Ser lie lie Lys Glu Leu •355 - 146966-序列表.doc 201037078 260 265 270
Ser Thr Met Val Glu Asn Tyr Ser Gly lie Lys lie Pro Val Asn Lys 275 280 285
Pro lie Val Gly Asp Lys Val Phe Tyr His Glu Ser Gly lie His Val 290 295 300
Asp Ala Val lie Glu Asn Pro Leu Thr Tyr Glu Pro Phe Leu Pro Glu 305 310 315 320
Arg lie Gly Gin Lys Arg Glu lie Val Leu Gly Lys His Ser Gly Cys 325 330 335
Ser Ala Val Glu Ser Lys Leu Lys 340 345
Asp Arg lie Trp Asp Leu Val Lys 355 360
Gly Glu Asp lie Asp Asp Glu Met 370 375
Asn Lys Gin 385
Glu Leu Gly Leu Glu Val Pro Lys 350
Lys Val Lys Thr Thr Arg Glu Gly 365
Phe lie Lys lie Val Asp lie lie 380 <210> 280 <211> 1164
<212> DNA <213> 雜色曱烧球菌Nankai-3 <220> <223> 野生型 <400> 280
ATGAATTGGA AAGAAGTTTG TCAATATAAT CCAAAGTTGA ATTTGGAGGA TTGCTATATT 60 TATGATACTA CACTTAGAGA TGGGGAGCAA ACTCCGGGAG TTTGTTTTTC AATGGAACAA 120
AAATTGGATA TTGCTAAAAA ATTAGATGAA TTGGGGGTAA AGCAGATTGA AGCAGGTTTT 180
CCAGCCGTAT CTAAATCAGA AATAGAAAAT GTAAAAAAAA TAGCAAACGA AGGACTAAAC 240 GCTGAAATAT TGGCGTTATC TCGGGCACTA CAAGGAGATA TTGATAAGGC ATTATCCTGT 300 GATGTAGATG GGATTATTAC ATTTATAGCA GCTTCTCCTC TTCATTTAAA ATATAAACTT 360 CATAAAAGCA TCGAGGAAGT AGAAGAGATG GGAATGAAGG CAGTGGAATA TGCAAAAGAC 420 CACGGGCTAT TTGTAGCTTT TTCGGCTGAA GATGCTACAA GGACACCAAT TGAGGACCTT 480 GTAAGAATTC ATAAAAATGC AGAGGAACAC GGAGCAGATA GGGTTCATAT AGCAGATACT 540 ACGGGCTGTG GAACACCACA ATCCATACAA TATATATGTT CAGAACTTAG CAATAATTTA 600 AAAAAGGCAC ATATTGGAGT TCATTGCCAT AATGATTTTG GACTTGCCGT AATAAATTCT 660 ATTTATGGAC TATTGGGTGG TGCAAAAGCA GCATCTACCA CAGTAAATGG AATTGGGGAA 720 AGAGCTGGAA ATGCACCTTT GGAGGAGCTC CTATTAACTA TGAATGTATT ATATGATGTA 780 AAAACTGATT TAAATATCTC AATAATAAAG GAATTATCGA CAATGGTTGA AAACTATTCG 840 GGGATAAAAA TACCAGTAAA TAAGCCGATA GTAGGGGATA AAGTATTTTA TCATGAAAGC 900 GGAATTCATG TTGATGCAGT AATTGAAAAT CCACTAACTT ATGAACCATT TTTGCCGGAA 960 •356- 146966-序列表.doc 201037078 AGAATCGGGC AAAAACGAGA AATTGTATTG GGAAAACATT CTGGTTGTAG TGCAGTCGAA 1020 TCAAAATTAA AAGAGCTTGG CTTAGAAGTT CCAAAAGATA GAATATGGGA TTTAGTTAAA 1080 AAAGTAAAAA CCACCAGAGA AGGCGGGGAA GATATAGACG ATGAGATGTT TATAAAAATA 1140 GTAGATATAA TAAATAAACA ATAA 1164 <210> 281 <211> 1158
<212> DNA <213>海沼甲燒球菌S2 <220> <221> CDS <222> (1) . . (1158) <400> 281 atg gac tgg aaa gcg gtt tct ccg tac aac ccg aaa ctg gat ctg aaa
Met Asp Trp Lys Ala Val Ser Pro Tyr Asn Pro Lys Leu Asp Leu Lys O 1 5 10 15 gac tgc tac ctg tac gac acc act ctg cgt gac ggt gaa cag act ccg
Asp Cys Tyr Leu Tyr Asp Thr Thr Leu Arg Asp Gly Glu Gin Thr Pro 20 25 30 ggc gtt tgc ttc get ggc gac cag aaa ctg gaa ate get aag aaa ctg 144
Gly Val Cys Phe Ala Gly Asp Gin Lys Leu Glu lie Ala Lys Lys Leu 35 40 45 gac gaa ctg aaa ate aag cag ate gaa gca ggc ttc ccg ate gtt tct 192
Asp Glu Leu Lys lie Lys Gin lie Glu Ala Gly Phe Pro lie Val Ser 50 55 60 gaa aac gaa cgt aaa gca ate aaa tct ate act ggc gaa ggt ctg aac 240
Glu Asn Glu Arg Lys Ala lie Lys Ser lie Thr Gly Glu Gly Leu Asn 65 70 75 80 〇 get cag att ctg gca ctg tct ege gta ctg aaa gaa gat ate gac aaa 288
Ala Gin lie Leu Ala Leu Ser Arg Val Leu Lys Glu Asp lie Asp Lys 85 90 95 gcg att gag tgc gac gtt gac ggt ate ate act ttc ate get act tct 336
Ala lie Glu Cys Asp Val Asp Gly lie 工le Thr Phe lie Ala Thr Ser 100 105 110 ccg atg cac ctg aaa tac aaa ctg cac aaa aac ctg gat gaa gtt gaa 384
Pro Met His Leu Lys Tyr Lys Leu His Lys Asn Leu Asp Glu Val Glu 115 120 125 gag atg ggt atg aaa gcg gtt gaa tac get aaa gac cac ggt ctg ttc 432
Glu Met Gly Met Lys Ala Val Glu Tyr Ala Lys Asp His Gly Leu Phe 130 135 140 146966-序列表.doc -357- 48 96 201037078 gtt gca ttc tct gca gaa gat gca act cgt act ccg ctg gaa gat ate 480
Val Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Pro Leu Glu Asp He 145 150 155 160 ate cgt ate cac aaa aac get gaa gaa cac ggt get gac ege gtt cac 528
He Arg He His Lys Asn Ala Glu Glu His Gly Ala Asp Arg Val His 165 170 175 att get gac act ctg ggt tgc gca act ccg cag gca atg tac cac ate 576 lie Ala Asp Thr Leu Gly Cys Ala Thr Pro Gin Ala Met Tyr His He 180 185 190 tgc tct gaa ctg tee aag cac ctg aag aaa geg cac ate ggt gtt cac 624
Cys Ser Glu Leu Ser Lys His Leu Lys Lys Ala His He Gly Val His 195 200 205 tgc cac aac gac ttc ggt ttc get gtt ate aac tee ate tac ggt ctg 672
Cys His Asn Asp Phe Gly Phe Ala Val lie Asn Ser lie Tyr Gly Leu 210 215 220 ate ggt ggt geg aaa get gtt tee act acc gtt aac ggt ate ggt gaa 720 lie Gly Gly Ala Lys Ala Val Ser Thr Thr Val Asn Gly lie Gly Glu 225 230 235 240 cgt get ggt aac get get ate gaa gaa ate geg atg geg ctg aaa gtt 768
Arg Ala Gly Asn Ala Ala lie Glu Glu lie Ala Met Ala Leu Lys Val 245 250 255 ctg tac gac cac gac atg ggt ctg aac act gaa ate ctg act gaa ate 816
Leu Tyr Asp His Asp Met Gly Leu Asn Thr Glu lie Leu Thr Glu lie 260 265 270 tee aaa ctg gtt gaa aac tac tee aag ate aag ate cca gaa aac aag 864
Ser Lys Leu Val Glu Asn Tyr Ser Lys lie Lys lie Pro Glu Asn Lys 275 280 285 ccg ctg gtt ggt gaa atg gta ttc tac cac gaa tee ggt ate cac gtt 912
Pro Leu Val Gly Glu Met Val Phe Tyr His Glu Ser Gly lie His Val 290 295 300 gac gca gta ctg gaa aac ccg ctg act tac gaa ccg ttc ctg ccg gaa 960
Asp Ala Val Leu Glu Asn Pro Leu Thr Tyr Glu Pro Phe Leu Pro Glu 305 310 315 320 aag ate ggt cag aaa cgt aaa ate ate ctg ggt aaa cac tct ggt tgc 1008
Lys lie Gly Gin Lys Arg Lys lie lie Leu Gly Lys His Ser Gly Cys 325 330 335 •358-
146966-序列表.doc 201037078 cgc get gtt get cac cgt ctg cag gaa ctg ggt ctg gaa gca tet ege 1056
Arg Ala Val Ala His Arg Leu Gin Glu Leu Gly Leu Glu Ala Ser Arg 340 345 350 gac gaa ctg tgg gaa ate gtt aag aaa acc aaa gaa act cgt gag gac 1104
Asp Glu Leu Trp Glu lie Val Lys Lys Thr Lys Glu Thr Arg Glu Asp 355 360 365 ggt act gaa ate tet gac gaa gta ttc aaa aac ate get gag aag ate 1152
Gly Thr Glu lie Ser Asp Glu Val Phe Lys Asn lie Ala Glu Lys lie 370 375 380 ate aag 1158 lie Lys 385 b
<210> 282 <211> 386 <212> PRT <213〉海沼甲烷球菌S2 <400> 282
Met Asp Trp Lys Ala Val Ser Pro Tyr Asn Pro Lys Leu Asp Leu Lys 15 10 15
Asp Cys Tyr Leu Tyr Asp Thr Thr Leu Arg Asp Gly Glu Gin Thr Pro 20 25 30
Gly Val Cys Phe Ala Gly Asp Gin Lys Leu Glu lie Ala Lys Lys Leu 35 40 45
Asp Glu Leu Lys lie Lys Gin lie Glu Ala Gly Phe Pro lie Val Ser 50 55 60
Glu Asn Glu Arg Lys Ala lie Lys Ser lie Thr Gly Glu Gly Leu Asn 65 70 75 80
Ala Gin lie Leu Ala Leu Ser Arg Val Leu Lys Glu Asp lie Asp Lys 85 90 95
Ala lie Glu Cys Asp Val Asp Gly lie lie Thr Phe lie Ala Thr Ser 100 105 110
Pro Met His Leu Lys Tyr Lys Leu His Lys Asn Leu Asp Glu Val Glu 115 120 125
Glu Met Gly Met Lys Ala Val Glu Tyr Ala Lys Asp His Gly Leu Phe 130 135 140
Val Ala Phe Ser Ala Glu Asp Ala Thr Arg Thr Pro Leu Glu Asp lie 145 150 155 160 lie Arg lie His Lys Asn Ala Glu Glu His Gly Ala Asp Arg Val His 165 170 175 lie Ala Asp Thr Leu Gly Cys Ala Thr Pro Gin Ala Met Tyr His lie 180 185 190
Cys Ser Glu Leu Ser Lys His Leu Lys Lys Ala His lie Gly Val His 195 200 205
Cys His Asn Asp Phe Gly Phe Ala Val lie Asn Ser lie Tyr Gly Leu 210 215 220 lie Gly Gly Ala Lys Ala Val Ser Thr Thr Val Asn Gly lie Gly Glu 225 230 235 240
Arg Ala Gly Asn Ala Ala lie Glu Glu lie Ala Met Ala Leu Lys Val 245 250 255
Leu Tyr Asp His Asp Met Gly Leu Asn Thr Glu lie Leu Thr Glu lie -359- 146966-序列表.doc 201037078 260 265 270
Ser Lys Leu Val Glu Asn Tyr Ser Lys lie Lys lie Pro Glu Asn Lys 275 280 285
Pro Leu Val Gly Glu Met Val Phe Tyr His Glu Ser Gly lie His Val 290 295 300
Asp Ala Val Leu Glu Asn Pro Leu Thr Tyr Glu Pro Phe Leu Pro Glu 305 310 315 320
Lys lie Gly Gin Lys Arg Lys lie lie Leu Gly Lys His Ser Gly Cys 325 330 335
Arg Ala Val TVla His Arg Leu Gin Glu Leu Gly Leu Glu Ala Ser Arg 340 345 350
Asp Glu Leu Trp Glu lie Val Lys Lys Thr Lys Glu Thr Arg Glu Asp 355 360 365
Gly Thr Glu lie Ser Asp Glu Val Phe Lys Asn lie Ala Glu Lys lie 370 375 380 lie Lys 385 c <210> 283 <211〉 1161 <212> DNA <213> 海沼曱烷球菌S2 <220> <223> 野生型 <400> 283 ATGGACTGGA AAGCTGTATC TCCGTACAAC CCAAAATTAG ATTTAAAAGA CTGTTATTTG 60 TATGATACAA CATTGAGAGA TGGTGAACAG ACCCCTGGAG TTTGTTTTGC AGGAGACCAA 120 AAACTTGAAA TTGCTAAGAA ACTGGATGAA CTTAAAATTA AACAGATTGA AGCAGGTTTT 180 CCAATTGTTT CTGAAAATGA AAGAAAAGCC ATCAAATCAA TTACTGGAGA AGGCTTAAAT 240 GCGCAGATTT TAGCATTATC AAGAGTTTTA AAAGAAGATA TTGATAAAGC CATCGAATGT 300 GACGTTGACG GGATAATTAC ATTTATTGCA ACTTCTCCAA TGCATTTGAA ATATAAATTA 360 CATAAGAATC TTGATGAAGT AGAAGAAATG GGTATGAAAG CCGTAGAATA TGCAAAAGAT 420 CACGGCCTTT TTGTAGCATT CTCCGCAGAA GATGCAACGA GAACCCCTCT TGAAGACATA 480 ATTAGAATTC ATAAAAATGC AGT^GAACAC GGCGCTGATA GGGTCCATAT TGCAGATACT 540 CTTGGATGTG CAACACCACA GGCAATGTAT CATATATGTT CTGAACTAAG CAAACACCTG 600 AAAAAAGCAC ATATTGGAGT ACACTGCCAC AATGACTTTG GATTTGCAGT TATAAACTCA 660 ATATACGGTT TAATCGGTGG TGCAAAAGCT GTATCTACCA CAGTTAATGG TATAGGTGAA 720 AGAGCAGGAA ATGCTGCAAT TGAAGAAATT GCAATGGCCT TAAAAGTACT TTACGACCAC 780 GATATGGGAT TAAATACTGA AATACTAACT GAAATATCTA AACTCGTTGA AAACTATTCA 840 AAAATCAAAA TTCCTGA7WV TAAACCGCTT GTTGGAGAAA TGGTATTCTA CCATGAAAGC 900 GGAATACACG TCGATGCCGT TTTAGAAAAC CCATTAACAT ATGAACCATT TTTGCCCGAA 960 360- 146966-序列表.doc 201037078 AAAATCGGTC AGAAAAGAAA AATTATCCTT GGAAAACATT CCGGTTGTAG GGCTGTCGCA 1020 CACAGACTAC AGGAACTCGG ACTTGAAGCT TCAAGAGACG AATTATGGGA AATTGTAAAG 1080 7\AAACTAAAG AAACTAGAGA AGACGGTACT GA7\ATAAGCG ACGAAGTGTT TAAAAACATT 1140 GCAGAAAAAA TCATAAAATA A 1161 〇 361- 146966-序列表.d〇c

Claims (1)

  1. 201037078 七、申請專利範圍： 1. -種製備α-酮庚二酸之方法，其包含將α·酮戊二酸轉化 為&剩己二酸及將α-酮己二酸轉化為庚二酸，其中 使用異源生物催化劑進行此等轉化之至少—者。 2. 士明求項1之方法，其中α-酮戊二酸係以生物催化方式自 碳源’尤其自碳水化合物製備。 〇 如叫求項1或2之方法，其中該異源生物催化劑包含催化 α_酮戊二酸之Cl_伸長為α_酮己二酸及/或α-酮己二酸之 Ci-伸長為α_酮庚二酸之異源生物催化劑。 4. 如請求項3之方法，其中該異源生物催化劑包含 a. 具有高⑷擰檬酸活性之AksA酶或其同系物； b. 至少一種選自以下之群的酶：具有高^烏頭酸酶活 性之AksD酶、具有高n-烏頭酸酶活性之AkSE酶、該等 AksD酶之同系物及該等AksE酶之同系物；及 c•具有高n-異擰檬酸脫氫酶之AksF酶或其同系物。 5. 如請求項3或4之方法，其中該異源酶系統來源於選自產 曱烧古生菌（archae)之群’較佳選自以下之群的有機體： 曱院球菌屬（Mei/z⑽oeocews)、甲烧暖球菌屬 (Mei/z⑽oca/i/ococcw)、曱烷八聯球菌屬（Μβί/ϊα„〇_ •sarcha)、甲烧熱桿菌屬、曱烧球 形菌屬（Mei/zawow/zaera)、甲烷火菌屬（Mei/mwopjrMs)及 甲烧短桿菌屬。 6. 如前述請求項十任一項之方法，其中該異源生物催化劑 包含催化α-酮戊二酸轉化為α-酮己二酸之酶系統，其中 146966 201037078 該酶系統形成離胺酸生物合成之胺基己二酸途徑之一部 分。 7. 如咕求項6之方法，其中該酶系統來自於選自酵母、真 菌、古生菌及細菌之群，尤其選自以下之群的有機體： 青徽菌屬、頭孢黴屬（c咖―—"·_)、擬 青黴菌 M (Paelicomyces)、髮癬蛰屬（Trich〇phytum)、魏 菌屬（Aperg///㈣、平革菌屬（户心似購心价）、裸胞殼屬 CEwerzce//<3)、黑穗病菌屬、裂殖酵母菌屬 、酵母菌屬（心cc；mr〇w;；ceiS)、假《） 、、-糸酵母函屬（Cawi/ic/of)、耶氏酵母菌屬、畢赤 酵母函屬（_P/cA/a)、克魯維酵母菌屬（_^wj；er〇w^ce>sl)、樓 熱包屬（Thermus)、奇球菌屬（Deinococcus)、火球菌屬 (P_yrococcw)、硫化葉菌屬（心//o/o6w)、熱球菌屬 (TT^rmococcw)、甲烷球菌屬、甲烷暖球菌屬、曱烷球 形菌屬、甲烷火菌屬、曱烷短桿菌屬、甲烷八聯球菌屬 及曱烷熱桿菌屬。 8. 如請求項1-6中任一項之方法，其中該異源生物催化劑包 ◎ 含催化α-酮戊二酸轉化為α-酮己二酸之酶系統，其中該 S#系統之至少一種6#來源於選自藍細菌（cyanobacteria) 、 根瘤菌 （rhizobiales) 、 γ-變形菌 （γ-proteobacteria) 及放線菌（actinobacteria)之群，尤其選自念珠藻屬 、微囊藻屬（Microti…）、集胞藻屬（办 /zoc;^沿）、根瘤菌屬（Λ/^zohwm)、慢生根瘤菌屬 (5ra办r/nzo6z’Mm)、假單胞菌屬（Psewi/omowas)、固氮菌 146966 201037078 屬㈣、克雷伯氏桿_屬叫及弗蘭克 氏菌屬（Frcmh’a)之群的固氮細菌。 9. 10. 11.
    -種製備5-甲醯基戊酸之方法’其包含以生物催化方式 使如前述請求項中任—項之方法中所製備之㈣庚二酸 脫羧基，藉此形成5_甲醯基戊酸。 -種製備6-胺基己酸之方法，其包含將如請求項9之方法 中所製備之5-甲醯基戊酸轉化為6_胺基己酸。 如請求項ίο之方法’其中該5_甲醯基戊酸轉化為6胺基 己酸包含轉胺或還原胺化。 12. 13. Ο 一種製備6-胺基己酸之方法’其包含將如請求項卜8中任 項之方法中所製備之α_酮庚二酸轉化為心胺基庚二 酸’及將《•絲庚二酸轉化為6胺基己酸，該等轉化較 佳以生物催化方式進行。 一種自如請求们_12中任—項之方法_所製備之^同庚 二酸製備辛二酸之方法，其包含使用對伸長具有 催化活性之生物催化劑，尤其包含以下之生物催化劑， 使該α_酮庚二酸經Ci_伸長： d·具有高⑷檸檬酸活性之AksA_或其同系物； e. 至少-種選自以下之群的酶：具有高n_烏頭酸酶活 性之AksD酶具有鬲n-烏頭酸酶活性之AksE酶、該等 AksD酶之同系物及該等AksE酶之同系物；及 f. 具有南異檸檬酸脫氫酶之AksF_或其同系物。 14.如凊求項13之方法’其中該等酶各獨立地來源於選自產 甲炫古生菌之群’較佳選自以下之群的有機體：甲院球 146966 201037078 菌屬、曱烷暖球菌屬、甲烷八聯球菌屬、曱烷熱桿菌 屬、甲烧球形菌屬、曱烷火菌屬及甲烷短桿菌屬。 15· 一種製備7_胺基庚酸之方法，其包含使如請求項13或14 之方法中所製備之α-酮辛二酸轉化。 16. 如前述請求項中任一項之方法，其中該方法在醱酵條件 下進行。 17. —種異源細胞’其包含一或多個異源核酸序列，編碼一 或多種在由α-酮戊二酸製備α酮庚二酸的至少一個反應步 驟中具有催化活性之異源酶。 18 ·如請求項17之異源細胞，其中該細胞不含能夠催化α_酮 己二酸轉化為α-胺基己二酸之胺基轉移酶。 19 ·如請求項17或18之異源細胞，其包含至少一個編碼以下 序列ID之任一者表示之酶或其同系物的核酸序列：4_ 77 261 、 264 、 267 、 270 、 273 、 276 、 279 ' 282 、 186 、 189、192、195、225、228、231、234 ' 198、201、 204、207、237、240、243、246、210、213、216、 219 、 222 、 249 、 252 、 255 、 258 。 20_如請求項17-19中任一項之異源細胞，其包含編碼對心酮 庚二酸脫缓形成5 -甲醯基戊酸具有催化活性之酶的核酸 序列，此類酶尤其選自脫羧酶（E.C. 4.M)之群，更尤其 選自以下之群：麩胺酸脫羧酶（EC 4.1.1.15)、二胺基庚 二酸脫羧酶（EC 4.1.1.20)、天冬胺酸1·脫竣酶（EC 4.1.1.11)、分支鍵α·酮酸脫缓酶、α-酮異戊酸脫叛酶、α_ 酮戊二酸脫綾酶、丙酮酸脫羧酶（EC 4.1· 1.1)及草醯乙酸 146966 201037078 脫羧酶（E.C.4.1.1.3)。 21 ·如請求項17-20中任一項之異源細胞，其中該細胞來自於 選自以下之群的有機體：產黃青徽 50#««/«)、黑麴菌Wggr)、玉米黑穗病菌 (Ustilago 、釀酒酵母cereW-
    «siae)、乳酸克魯維酵母/acib)、甲醇酵 母（Pichia pastoris) ' 多形漢森酵母（/iansenw/a polymorha)、大腸桿菌（五5c；2erz_c/n-a co/z·)、棕色固氮菌 (Azotobacter vinelandii)、施氏假單胞菌（pseudomonas siwizerz·/)、肺炎克雷伯氏桿菌（尺、 而t幸昌射奇球菌（De/wococcw*? raii/oMraws)、嗜熱奇球菌 (DeMococcwi 、嗜熱棲熱菌（772erWM5 thermophilus)、海、，召甲烧球菌（Mei/ianococcMS warz>a/w£^)、醋酸曱烷八聯球菌（似 acetivorans) f 氏 Ψ、底螺·壤{Methanospirillum hunga- iez·)、嗜熱甲烧鬚函（Λ/"βί/；α«(?ίββί<3 i/zerwop/n7<3)、史氏甲 烧短桿菌办reW^cier «smii/n·/)、萬氏甲烧球菌 (Mei/z㈣ococcws va”/^em)、雜色甲烷球菌⑽0_ coccw·? ⑽）及詹氏曱烷暖球菌紿训oca/办c〇ccw5 jannashii) ° 22·如請求項17-21中任一項之異源細胞，其包含至少一個選 自以下之群之任一序列表示的核酸序列：SEQ ID NO 149 ; SEQ ID NO 145、146、147、148 ; SEQ ID NO 167 、 168 、 169 、 170 、 171 、 172 、 173 、 174 ; SEQ ID 146966 ¥ 201037078 NO 177 、 178 、 179 、 180 、 181 、 182 、 183 、 184 ID NO 224 、 226 、 236 、 238 、 248 、 250 、 260 、 SEQ ID NO 227、229、239、241、251、253、 265 ； SEQ ID NO 194、196、206、208、221、 281 ' 283 ； SEQ ID NO 188、190、200、202、 217、272、274及其功能類似物。 23· —種如請求項1 7-22中任一項之異源細胞的用途， 於製備己内醯胺、6-胺基己酸或二胺基己烷。 6- ；SEQ 262 ； 263 ' 223 ' 215、 其係用 146966 201037078 四、指定代表圖： (一) 本案指定代表圖為：（無） (二) 本代表圖之元件符號簡單說明： 五、本案若有化學式時，請揭示最能顯示發明特徵的化學式： (無） 146966
TW099107114A 2009-03-11 2010-03-11 Preparation of alpha-ketopimelic acid TW201037078A (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
EP09154840 2009-03-11
EP09170092 2009-09-11
EP09180441 2009-12-22

Publications (1)

Publication Number Publication Date
TW201037078A true TW201037078A (en) 2010-10-16

Family

ID=42728978

Family Applications (1)

Application Number Title Priority Date Filing Date
TW099107114A TW201037078A (en) 2009-03-11 2010-03-11 Preparation of alpha-ketopimelic acid

Country Status (9)

Country Link
US (2) US20120028320A1 (zh)
EP (2) EP2406385A2 (zh)
JP (2) JP2012520069A (zh)
CN (2) CN102348805A (zh)
AU (2) AU2010221863A1 (zh)
BR (2) BRPI1009192A2 (zh)
EA (2) EA201101311A1 (zh)
TW (1) TW201037078A (zh)
WO (2) WO2010104391A2 (zh)

Families Citing this family (34)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
HUE035611T2 (en) * 2008-03-27 2018-05-28 Genomatica Inc Microorganisms suitable for the production of adipic acid and other compounds
WO2011031147A1 (en) * 2009-09-11 2011-03-17 Dsm Ip Assets B.V. Preparation of a compound comprising an amine group from an alpha-keto acid
WO2012031910A2 (en) * 2010-09-10 2012-03-15 Dsm Ip Assets B.V. Method for preparing alpha-ketopimelic acid by c1-elongation
US20130237698A1 (en) * 2010-09-10 2013-09-12 Dsm Ip Assets B.V. Preparation of 6-aminocaproic acid from alpha-ketopimelic acid
WO2012131059A1 (en) 2011-04-01 2012-10-04 Dsm Ip Assets B.V. Preparation of 5-formyl valeric acid from alpha-ketopimelic acid
MY166083A (en) 2011-06-17 2018-05-24 Invista Tech Sarl Use of hydrolases to increase monomer content in waste stream
EP2721161A2 (en) * 2011-06-17 2014-04-23 Invista Technologies S.à.r.l. Methods of making nylon intermediates from glycerol
JP2014525741A (ja) 2011-06-30 2014-10-02 インビスタ テクノロジーズ エス．アー．エール．エル． ナイロン−７、ナイロン−７，７、およびポリエステルを生産するための生物変換方法
US9102958B2 (en) 2011-12-16 2015-08-11 Invista North America S.á.r.l. Methods of producing 6-carbon chemicals via CoA-dependent carbon chain elongation associated with carbon storage
US9102960B2 (en) 2011-12-16 2015-08-11 Invista North America S.á.r.l. Methods of producing 6-carbon chemicals via CoA-dependent carbon chain elongation associated with carbon storage
JP2015500663A (ja) * 2011-12-21 2015-01-08 インビスタ テクノロジーズ エス．アー．エール．エル． ナイロン−７、ナイロン−７，７、およびポリエステルを生産するための生物変換方法
US8691960B2 (en) * 2012-02-29 2014-04-08 Duke University Oxidoreductases for enantioselective reactions
US9790525B2 (en) 2012-12-14 2017-10-17 Invista North America S.A.R.L. Methods of producing 7-carbon chemicals via CoA-dependent carbon chain elongation associated with carbon storage
EP2938732A1 (en) 2012-12-31 2015-11-04 Invista Technologies S.à.r.l. Methods of producing 7-carbon chemicals from long chain fatty acids via oxidative cleavage
WO2014105797A2 (en) 2012-12-31 2014-07-03 Invista North America S.A.R.L. Methods of producing 7-carbon chemicals via pyruvate and succinate semialdehyde aldol condensation
EP2938731A2 (en) 2012-12-31 2015-11-04 Invista Technologies S.A R.L. Methods of producing 7-carbon chemicals via carbon chain elongation associated with cyclohexane carboxylate synthesis
EP2938736A2 (en) 2012-12-31 2015-11-04 Invista Technologies S.A R.L. Methods of producing 7-carbon chemicals via c1 carbon chain elongation associated with coenzyme b synthesis
WO2014105805A2 (en) 2012-12-31 2014-07-03 Invista North America S.A.R.L. Methods of producing 6-carbon chemicals via methyl-ester shielded carbon chain elongation
WO2014105796A2 (en) 2012-12-31 2014-07-03 Invista North America S.A.R.L. Methods of producing 7-carbon chemicals via aromatic compounds
WO2014105794A2 (en) 2012-12-31 2014-07-03 Invista North America S.A.R.L. Methods of producing 7-carbon chemicals via methyl-ester shielded carbon chain elongation
WO2015176026A1 (en) 2014-05-15 2015-11-19 Invista North America S.á.r.l. Methods of producing 6-carbon chemicals using 2,6-diaminopimelate as precursor to 2-aminopimelate
BR112016029388A2 (pt) 2014-06-16 2017-10-17 Invista Tech Sarl métodos, reagentes e células para biossintetizar compostos
EP3155108A1 (en) 2014-06-16 2017-04-19 Invista Technologies S.à.r.l. Methods, reagents and cells for biosynthesizing compounds
US9938543B2 (en) 2014-06-16 2018-04-10 Invista North America S.A.R.L. Methods, reagents and cells for biosynthesizing glutarate methyl ester
EP3155107A1 (en) 2014-06-16 2017-04-19 Invista Technologies S.à.r.l. Methods, reagents and cells for biosynthesizing compounds
CN104195125B (zh) * 2014-07-01 2017-01-11 中国科学院广州能源研究所 一种α-酮酸脱羧酶KIVD-LL及其编码基因和应用
KR101839595B1 (ko) 2015-04-13 2018-04-26 한국과학기술원 락탐의 제조방법
CN109355220A (zh) * 2018-11-02 2019-02-19 三峡大学 一种微生物混合菌群及其应用
CA3121136A1 (en) * 2018-11-29 2020-08-27 Zymergen Inc. Engineered biosynthetic pathways for production of 2-oxoadipate by fermentation
EP3766982A1 (en) 2019-07-18 2021-01-20 Delft Advanced Biofuels B.V. Integrated system for biocatalytically producing and recovering an organic substance
CN110724680A (zh) * 2019-10-29 2020-01-24 天津大学 一种3-苯基马来酸异构酶及其应用
CN111662892B (zh) * 2020-07-29 2022-07-01 上海市农业科学院 β-酮己二酸代谢相关三个基因的结构优化与应用
WO2022210708A1 (ja) 2021-03-30 2022-10-06 旭化成株式会社 組換え微生物及びｃ６化合物の製造方法
CN113185589B (zh) * 2021-04-23 2023-05-30 东北农业大学 一种抗草甘膦相关蛋白及其编码基因与应用

Family Cites Families (26)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS6058068A (ja) * 1983-09-08 1985-04-04 Kao Corp 新規なアミンデヒドロゲナーゼｍ
DK122686D0 (da) 1986-03-17 1986-03-17 Novo Industri As Fremstilling af proteiner
AT399149B (de) 1993-06-07 1995-03-27 Chemie Linz Gmbh Verfahren zur herstellung primärer amine aus aldehyden
KR100516986B1 (ko) 1997-02-19 2005-09-26 코닌클리즈케 디에스엠 엔.브이. 6-아미노카프론산 유도체를 과열 수증기와 접촉시킴으로써 촉매 없이 카프로락탐을 제조하는 방법
EP1452553A1 (en) * 1999-05-20 2004-09-01 Daicel Chemical Industries, Ltd. UV absorbing polyurethane and spandex fibre derived therefrom
RU2201454C2 (ru) * 1999-07-09 2003-03-27 Закрытое акционерное общество "Научно-исследовательский институт Аджиномото-Генетика" Мутантная альфа-изопропилмалат синтаза (ipms), днк, кодирующая мутантную ipms, способ получения штамма escherichia coli, способ получения l-лейцина
DE60202478T3 (de) 2001-07-23 2010-01-07 Dsm Ip Assets B.V. Verfahren zur herstellung von polynukleotidvarianten
US20030167513A1 (en) * 2001-11-30 2003-09-04 Mourad George S. Selection and use of isopropylmalate synthase (IPMS) mutants desensitized in L-leucine negative feedback control
US7314974B2 (en) * 2002-02-21 2008-01-01 Monsanto Technology, Llc Expression of microbial proteins in plants for production of plants with improved properties
US7491520B2 (en) 2004-01-19 2009-02-17 Dsm Ip Assets B.V. Biochemical synthesis of 6-amino caproic acid
JP2005296010A (ja) * 2004-03-19 2005-10-27 Osaka Univ ２−イソプロピルリンゴ酸シンターゼ活性を有する新規耐熱性タンパク質
US7223873B2 (en) 2004-03-30 2007-05-29 Daisco Co., Ltd Process for preparing amines
JP2007530065A (ja) 2004-04-02 2007-11-01 ディーエスエム アイピー アセッツ ビー．ブイ． 改善された相同的組換え効率を有する糸状菌変異体
WO2006076423A2 (en) * 2005-01-12 2006-07-20 Monsanto Technology, Llc Genes and uses for plant improvement
GB2433260A (en) * 2005-12-16 2007-06-20 Mologic Ltd A selectable decarboxylase marker
WO2008034648A1 (en) * 2006-04-05 2008-03-27 Metanomics Gmbh Process for the production of a fine chemical
BRPI0713795B1 (pt) 2006-06-29 2018-03-20 Dsm Ip Assets B.V. Método de otimização de uma sequência de nucleotídeos codificadora que codifica uma sequência de aminoácidos predeterminada
CN100537511C (zh) * 2006-08-30 2009-09-09 中国石油天然气股份有限公司 连续酯化生产己二酸二甲酯的方法及设备
US20080295196A1 (en) * 2006-12-06 2008-11-27 Abad Mark S Genes and uses for plant improvement
BRPI0807235A2 (pt) * 2007-02-09 2015-05-26 Univ California Microorganismo recombinante, e, métodos para produzir um microorganismo recombinante, e para produzir um álcool
US9695426B2 (en) * 2007-02-09 2017-07-04 The Regents Of The University Of California Biofuel production by recombinant microorganisms
CA2698125A1 (en) * 2007-08-30 2009-03-12 Plant Sensory System, Llc. Alternative methods for the biosynthesis of gaba
HUE037878T2 (hu) 2008-03-11 2018-09-28 Genomatica Inc Adipátészter vagy tioészter szintézis
TW200951103A (en) * 2008-03-11 2009-12-16 Dsm Ip Assets Bv Preparation of 6-aminocaproic acid from α-ketopimelic acid
EP2346991B1 (en) * 2008-10-18 2016-12-28 The Regents of The University of California Production of c5-c8 alcohols using evolved enzymes and metabolically engineered microorganisms
RU2496880C2 (ru) * 2008-12-12 2013-10-27 СЕЛЕКСИОН, ЭлЭлСи Биологический синтез дифункциональных алканов из альфа-кетокислот

Also Published As

Publication number Publication date
WO2010104390A3 (en) 2011-04-21
EA201101312A1 (ru) 2012-03-30
AU2010221863A1 (en) 2011-10-06
EP2406384A2 (en) 2012-01-18
US20120156737A1 (en) 2012-06-21
EP2406385A2 (en) 2012-01-18
JP2012520070A (ja) 2012-09-06
BRPI1009192A2 (pt) 2015-09-22
JP2012520069A (ja) 2012-09-06
WO2010104391A3 (en) 2011-06-30
WO2010104391A2 (en) 2010-09-16
EA201101311A1 (ru) 2012-04-30
CN102892893A (zh) 2013-01-23
AU2010221862A1 (en) 2011-10-06
CN102348805A (zh) 2012-02-08
BRPI1009197A2 (pt) 2015-09-15
US20120028320A1 (en) 2012-02-02
WO2010104390A2 (en) 2010-09-16

Similar Documents

Publication Publication Date Title
TW201037078A (en) Preparation of alpha-ketopimelic acid
AU2009224089B2 (en) Preparation of 6-aminocaproic acid from 5 -formyl valeri C acid
US20120231512A1 (en) Preparation of alpha-ketopimelic acid
JP7127987B2 (ja) ジアミンの製造および処理方法
DK2252698T3 (en) ADIPATESTER- OR -THIOESTER-SYNTHESIS
WO2011031147A1 (en) Preparation of a compound comprising an amine group from an alpha-keto acid
US20120264179A1 (en) Microorganisms for the production of adipic acid and other compounds
TW201217536A (en) Method for preparing alpha-ketopimelic acid by C1-elongation
US8728771B2 (en) L-succinylaminoacylase and process for producing L-amino acid using it
TW201033369A (en) Preparation of 6-aminocaproic acid from α-ketopimelic acid
CN116783281A (zh) 用于制备酰胺化合物的方法和组合物
TW201033370A (en) Adipate (ester or thioester) synthesis