CN109415417A - 通过含有编码糖磷酸转移酶***(pts)的基因的微生物发酵生产甲硫氨酸或其羟基类似物形式的方法 - Google Patents

通过含有编码糖磷酸转移酶***(pts)的基因的微生物发酵生产甲硫氨酸或其羟基类似物形式的方法 Download PDF

Info

Publication number
CN109415417A
CN109415417A CN201780040978.2A CN201780040978A CN109415417A CN 109415417 A CN109415417 A CN 109415417A CN 201780040978 A CN201780040978 A CN 201780040978A CN 109415417 A CN109415417 A CN 109415417A
Authority
CN
China
Prior art keywords
ala
leu
val
gly
glu
Prior art date
Legal status (The legal status 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 status listed.)
Pending
Application number
CN201780040978.2A
Other languages
English (en)
Inventor
G·贝斯泰尔-科尔
C·雷诺
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Evonik Operations GmbH
Metabolic Explorer SA
Original Assignee
Evonik Degussa GmbH
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 Evonik Degussa GmbH filed Critical Evonik Degussa GmbH
Publication of CN109415417A publication Critical patent/CN109415417A/zh
Pending legal-status Critical Current

Links

Classifications

    • 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.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • 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
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/24—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
    • C07K14/245—Escherichia (G)
    • 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
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • 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
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • 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
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • 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
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli
    • 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
    • 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.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1294—Phosphotransferases with paired acceptors (2.7.9)
    • 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
    • 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
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
    • C12P7/06—Ethanol, i.e. non-beverage
    • 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
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
    • C12P7/18—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic polyhydric
    • 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
    • C12P7/42—Hydroxy-carboxylic acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y207/00—Transferases transferring phosphorus-containing groups (2.7)
    • C12Y207/11—Protein-serine/threonine kinases (2.7.11)
    • C12Y207/11003—Dephospho-[reductase kinase] kinase (2.7.11.3)
    • 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
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1205—Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/185—Escherichia
    • C12R2001/19—Escherichia coli
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y207/00—Transferases transferring phosphorus-containing groups (2.7)
    • C12Y207/09—Phosphotransferases with paired acceptors (2.7.9)
    • C12Y207/09002—Pyruvate, water dikinase (2.7.9.2)
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00—Technologies for the production of fuel of non-fossil origin
    • Y02E50/10—Biofuels, e.g. bio-diesel

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Virology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Mycology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Botany (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

本发明涉及一种在发酵过程中通过转化碳源来生产甲硫氨酸或其羟基类似物形式的新方法，包括培养经遗传修饰的微生物以生产甲硫氨酸或其羟基类似物形式，其中所述微生物包括编码PTS碳水化合物利用***的功能基因，并且其中调节磷酸烯醇式丙酮酸合酶(PPS)表达的蛋白质的表达被下调。本发明还涉及用于本发明方法的经遗传修饰的微生物。

Description

通过含有编码糖磷酸转移酶***(PTS)的基因的微生物发酵 生产甲硫氨酸或其羟基类似物形式的方法
技术领域
本发明涉及在发酵过程中通过碳源的转化来生产甲硫氨酸或其羟基类似物形式的新方法，包含培养经遗传修饰的微生物以产生所述感兴趣的分子，其中所述微生物包含编码PTS碳水化合物利用***的功能基因，并且其中调节磷酸烯醇式丙酮酸合酶(PPS)表达的蛋白质的表达被下调。本发明还涉及用于本发明方法的遗传修饰的微生物。
背景技术
在细菌中，外源碳水化合物(糖)被转运到细胞中并被磷酸烯醇式丙酮酸：糖磷酸转移酶***(PTS)磷酸化。磷酸烯醇式丙酮酸(PEP)是中心代谢的关键分子。在许多微生物中，支持生长的碳水化合物被摄取，并且与此同时通过PTS消耗一分子PEP/分子碳水化合物被磷酸化(Postma&Roseman 1976)。PTS由两种细胞质蛋白质酶I(EI)和HPr以及可变数量的特异于待被摄取的碳水化合物的膜蛋白复合体(酶II，EII)组成。总之，这些EI、HPr和EII蛋白充当PEP和碳水化合物之间的磷酰基转移链，所述碳水化合物在它穿过细胞膜时被磷酸化：
除了作为PTS的磷酸供体的作用外，PEP还参与通过丙酮酸激酶生成丙酮酸的糖酵解的最后一步(Kornberg&Malcovati 1973)：
此外，PEP通过生成草酰乙酸的添补反应连接糖酵解和柠檬酸循环，并由PEP羧化酶催化(Canovas&Kornberg 1965)：
PEP也是通过分支酸途径的芳香族氨基酸、醌类和C1代谢物的前体(Pittard&Wallace 1966)：
一些研究组已经研发出了提高PEP可用性的策略，以增强所需产物的产生和产率：失活PTS和/或丙酮酸激酶(Gosset等，1996，Meza等，2012)，失活整体的调节子CsrA(Tatarko&Romeo 2001)，过表达糖异生酶PEP羧激酶(Kim等2004)或PEP合酶(Patnaik等1992)。
PEP合酶(PPS，EC 2.7.9.2)催化将丙酮酸磷酸化为PEP并伴随ATP水解为AMP(Cooper&Kornberg，1965)：
在许多微生物中，PPS受到属于DUF299家族的PPS调节蛋白(PRPP)介导的磷酸化/去磷酸化机制的调节(Burnell，2010)。
Burnell研究的目的是表征蛋白质DUF299和编码所述蛋白质的基因的结构和功能。然而，该文章没有提出调节该蛋白质的表达以获得特定效果例如增加甲硫氨酸或其羟基类似物形式的产生的可能性。
发明内容
申请人惊奇地发现，调节PPS表达的蛋白质的表达的失活允许产生甲硫氨酸或其羟基类似物形式，其通常是通过待增加的微生物中的发酵过程产生的。
本发明人的发现是有利的，因为它克服了已知用于增加代谢产物产生的其他现有技术方法(例如专利申请WO2004033471中提出的)的许多缺点。
实际上，为了增加甲硫氨酸或其羟基类似物形式的产生，通常需要通过进行几种遗传修饰来改善生产者微生物中的碳源摄取。然而，参与碳源摄取的基因，更具体而言是参与碳水化合物输入的基因，参与复杂的调节***(Gabor等，2011；Kotrba等，2001)。因此，这种遗传修饰导致不可预测的后果，并且获得的菌株可能是不稳定的。此外，这些方法具有高成本。
因此，需要提供新的方法使得使用稳定的微生物菌株以低成本生产感兴趣的分子。
根据本发明，可以通过仅使PPS调节蛋白(PRPP)失活来增加所需产物的产生。
关于第一方面，本发明因此涉及通过在发酵过程中转化碳源来生产甲硫氨酸或其羟基类似物形式的方法，包含以下步骤：
-在包含以碳水化合物作为碳源的适当培养基中培养经遗传修饰的微生物以生产甲硫氨酸或其羟基类似物形式；以及
-从培养基中回收甲硫氨酸或其羟基类似物形式，其中所述经遗传修饰的微生物包含编码PTS碳水化合物利用***的功能基因，并且其中在所述经遗传修饰的微生物中，调节磷酸烯醇式丙酮酸合酶(PPS)表达的双功能ADP依赖性激酶-Pi-依赖性焦磷酸化酶蛋白的表达降低。
用于本发明方法的微生物具有特定的特征，例如具有编码PTS碳水化合物利用***的功能基因和降低的调节PPS表达的双功能ADP依赖性激酶-Pi-依赖性焦磷酸化酶蛋白表达。该微生物可被认为是特异的和令人惊讶的，因为获得经遗传修饰的微生物并不是明显的，其中磷酸烯醇式丙酮酸合酶(PPS)的表达受到影响而不影响整个碳水化合物摄取级联的功能性。
关于第二方面，本发明因此涉及经遗传修饰的微生物，其用于增强从以碳水化合物作为碳源的甲硫氨酸或其羟基类似物形式的生产，其中所述经遗传修饰的微生物包含编码PTS碳水化合物利用***的功能基因和降低的调节PPS表达的双功能ADP依赖性激酶-Pi-依赖性焦磷酸化酶蛋白表达。
具体实施方式
在详细描述本发明之前，应理解本发明不限于具体示例的方法，并且可或当然可变化。还应理解，本文使用的术语仅用于描述本发明的具体实施方案的目的，而不旨在限制，本发明仅受所附权利要求限制。
本文引用的所有出版物、专利和专利申请，无论上文或下文，均通过引用整体并入本文。然而，引用本文提及的出版物是为了描述和公开在出版物中报道的并且可以与本发明结合使用的方案、试剂和载体的目的。
此外，除非另有说明，本发明的实践采用本领域技术范围内的常规微生物和分子生物学技术。这些技术对于技术人员来说是熟知的，并且在文献中有充分的解释。
必须注意，如本文和所附权利要求中所使用的，单数形式“一种”、“一个”和“该”包括复数指代，除非上下文另有明确说明。因此，例如，提及“一种微生物”包括多种这样的微生物，提及“一种酶”是指一种或多种酶，等等。除非另外定义，否则本文使用的所有技术和科学术语具有与本发明所属领域的普通技术人员通常理解的含义相同的含义。尽管与本文描述的那些材料和方法类似或等价的任何材料和方法可用于实践或测试本发明，但现在描述优选的材料和方法。
如本文所用，以下术语可用于解释权利要求和说明书。
在随后的权利要求中和在本发明的前述描述中，除非上下文由于表述语言或必要的含义而另外要求，否则在包含意义上使用词语“包含”或变体例如“包括”或“含有”的，即指定所述特征的存在但不排除在本发明的各种实施方案中存在或添加其他特征。
在本发明的描述中，使用大肠杆菌(E.Coli)中相应基因的名称鉴定基因和蛋白质。然而，除非另有说明，否则这些名称的使用根据本发明具有更一般的含义并涵盖其他生物(尤其是微生物)中的所有相应基因和蛋白质。
PFAM(比对和隐马尔科夫模型的蛋白质家族数据库)展示了大量的蛋白质序列比对。每个PFAM都可以可视化多个比对、查看蛋白质结构域、评估生物体之间的分布、获得对其他数据库的访问、以及可视化已知的蛋白质结构。
通过比较来自代表38种主要***发生系的66个完全测序的基因组的蛋白质序列，获得COG(直系同源蛋白质组的簇)。每个COG由至少三个系定义，这允许鉴定以前的保守结构域。
鉴定同源序列及其同源性百分比的方法是本领域技术人员熟知的，特别是BLAST程序(Altschul等，1990)。然后可以使用例如程序CLUSTALW或MULTALIN来利用(例如，比对)所获得的序列。
使用GenBank上给出的已知基因的参考，本领域技术人员能够确定其他生物、细菌菌株、酵母、真菌、哺乳动物、植物等的等价基因。有利地，这种常规工作可以使用共有序列来完成，所述共有序列通过与衍生自其他微生物的基因进行序列比对来确定，并设计简并探针以克隆另一生物中的相应基因。这些分子生物学的常规方法是本领域技术人员熟知的，并且例如在Sambrook等(2001)中要求保护。
如上所述，本发明的方法允许通过在发酵过程中转化碳源来生产甲硫氨酸或其羟基类似物形式，包括以下步骤：
-在包含以碳水化合物作为碳源的适当培养基中培养经遗传修饰的微生物以生产甲硫氨酸或其羟基类似物形式，以及
-从培养基中回收甲硫氨酸或其羟基类似物形式，其中所述经遗传修饰的微生物包含编码PTS碳水化合物利用***的功能基因，并且其中在所述经遗传修饰的微生物中，调节磷酸烯醇式丙酮酸合酶(PPS)表达的双功能ADP依赖性激酶-Pi-依赖性焦磷酸化酶蛋白的表达降低。
术语“发酵过程”、“发酵”或“培养”在本文中可互换使用以表示微生物的生长。发酵通常在发酵罐中进行，所述发酵罐具有适合于所用微生物的已知的确定组成的无机培养基，其含有至少一种简单碳源，并且如果需要，还包含产生代谢物所必需的共底物。特别地，用于大肠杆菌的无机培养基可以与M9培养基(Anderson，1946)、M63培养基(Miller，1992)或Schaefer等(1999)定义的培养基具有相同或相似的组成。
在本发明的上下文中，“发酵转化”是指当在合适的发酵条件下培养微生物时，碳源转化为甲硫氨酸或其羟基类似物形式。
“培养基”在本文中是指包含对微生物的维持和/或生长必需或有益的营养物的培养基(例如，无菌液体培养基)，例如碳源或碳底物；氮源，例如蛋白胨、酵母提取物、肉提取物、麦芽提取物、尿素、硫酸铵、氯化铵、硝酸铵和磷酸铵；磷源，例如磷酸二氢钾或磷酸氢二钾；微量元素(例如金属盐)，例如镁盐、钴盐和/或锰盐；以及生长因子如氨基酸和维生素。
根据本发明的术语“碳的来源”、“碳源”或“碳底物”是指微生物能够代谢并且含有至少一个碳原子的任何分子。根据本发明的优选碳源的实例包括但不限于碳水化合物。
在本发明的一个优选实施方案中，碳源衍生自可再生原料。可再生原料被定义为某些工业过程所需的原料，其可以在短暂的延迟内再生，并且其量充足，以使其转化为所需的产物。预处理或未预处理的植物生物质是特别优选的可再生碳源。
术语“碳水化合物”在本文中是指能够被微生物代谢并且含有至少一个碳原子、两个氢原子和一个氧原子的任何碳源。本发明的碳水化合物优选选自葡萄糖、果糖、蔗糖、甘露糖、壳二糖、纤维二糖、海藻糖、半乳糖醇、甘露糖醇、山梨糖醇、半乳糖胺、N-乙酰-D-半乳糖胺、N-乙酰葡糖胺、N-乙酰胞壁酸、乳糖、半乳糖、山梨糖、麦芽糖、N,N'-二乙酰基壳二糖、抗坏血酸、β-葡萄糖苷。在本发明更优选的实施方案中，碳源选自葡萄糖、果糖、甘露糖、纤维二糖、蔗糖及其任意组合。
本领域技术人员可以容易地确定在根据本发明的方法中生长微生物所需的培养条件。特别地，熟知的是，细菌可以在包含20℃至55℃之间的温度发酵，优选在25℃至40℃之间。更特别地，大肠杆菌可以在包含约30℃至约37℃的温度培养。
该培养过程可以以分批过程、补料分批过程或连续过程进行，也可以在需氧、微需氧或厌氧条件下进行。
根据本发明方法的一个具体实施方案，编码PTS碳水化合物利用***的功能基因是异源的(重组微生物)或相对于经遗传修饰的微生物是天然的(野生型微生物)。
“基因”在本文中是指编码特定蛋白质(即多肽)的核酸分子或多核苷酸，或在某些情况下，编码功能性或结构性RNA分子。在本发明的上下文中，本文提及的基因编码蛋白质，例如酶、外排***或摄取转运蛋白。根据本发明的基因是内源基因或外源基因。
如本文所用的术语“重组微生物”或“经遗传修饰的微生物”是指在自然界中未发现并且在遗传上不同于天然存在的等价微生物的细菌、酵母或真菌。根据本发明，术语“修饰”表示在微生物中引入或诱导的任何遗传变化。可以通过引入新的遗传元件，增加或减弱内源或外源基因的表达或缺失内源遗传元件的来修饰微生物。此外，可以通过在特定选择压力下组合定向诱变和进化来迫使新代谢途径的发展和进化来修饰微生物(参见，例如，WO2004076659)。
在本发明的上下文中，术语“外源基因”(或者，“异源基因”或“转基因”)是指非天然存在于微生物中的基因。它可以是人工的，也可以源自另一种微生物。
应进一步理解，在本发明的上下文中，如果编码感兴趣蛋白质的外源基因在特定微生物中表达，则该基因的合成版本优选通过用编码相同氨基酸的所述微生物的优选密码子置换非优选密码子或较不优选的密码子的方法来构建。本领域确实熟知的是，密码子使用在微生物物种之间变化，其可能影响感兴趣蛋白质的重组表达水平。为了克服这个问题，已经开发了密码子优化方法，并且在Graf等(2000)、Deml等(2001)、或Davis&Olsen(2011)中对其进行了广泛的描述。已经开发了几种用于密码子优化确定的软件，例如软件(Lifetechnologies)或OptimumGeneTM软件(GenScript)。换句话说，编码感兴趣蛋白质的外源基因优选经密码子优化以在特定微生物中表达。
根据本发明方法的另一个实施方案中，经遗传修饰的微生物包含编码双功能ADP依赖性激酶-Pi-依赖性焦磷酸化酶蛋白的天然基因，其表达被减弱或缺失，所述蛋白调节磷酸烯醇式丙酮酸合酶(PPS)的表达。其他领域中，与未修饰的微生物相比，在所述经遗传修饰的微生物中，编码双功能ADP依赖性激酶-Pi-依赖性焦磷酸化酶蛋白的天然基因的表达减弱或缺失。优选地，在本发明的微生物中，缺失编码双功能ADP依赖性激酶-Pi-依赖性焦磷酸化酶蛋白的天然基因。
“天然基因”或“内源基因”在本文中是指所述基因天然存在于微生物中。
在本发明的上下文中，如果微生物经遗传修饰以“调节”一种或多种内源基因的表达水平，则在本文中表示与其自然表达水平相比，所述基因的表达水平被上调、下调(即减弱)、或者甚至完全消除。因此，这种调节理论上导致基因产物活性的增强，或者可选地，导致内源基因产物的活性较低或无活性。
还可以通过将突变引入内源基因编码序列以修饰基因产物来修饰内源基因活性和/或表达水平。还可以进行内源基因的缺失以完全抑制其在微生物内的表达。调节内源基因表达的另一种方法是将其启动子(即野生型启动子)与更强或更弱的启动子进行交换，以上调或下调该基因的表达水平。适用于此目的的启动子可以是同源的或异源的，并且是本领域熟知的。选择合适的启动子来调节内源基因的表达在本领域技术人员的能力范围内。
根据本发明的另一个实施方案，微生物选自表达功能性PTS糖***的微生物。优选地，微生物选自以下：肠杆菌科(Enterobacteriaceae)、梭菌科(Clostridiaceae)、芽孢杆菌科(Bacillaceae)、链霉菌科(Streptomycetaceae)、异常球菌科(Deinococcaceae)、亚硝化单胞菌科(Nitrosomonadaceae)、弧菌科(Vibrionaceae)、假单胞菌科(Pseudomonadaceae)、棒杆菌科(Corynebacteriaceae)、酵母科(Saccharomycetaceae)和酵母。更优选地，微生物是柠檬酸杆菌属(Citrobacter)、棒杆菌属(Corynebacterium)、异常球菌属(Deinococcus)、埃希氏菌属(Escherichia)、泛菌属(Pantoea)、克雷伯氏菌属(Klebsiella,)、亚硝化单胞菌属(Nitrosomonas)、光杆状菌属(Photorhabdus)、发光杆菌属(Photobacterium)、假单胞菌属(Pseudomonas)、沙门氏菌属(Salmonella)、沙雷氏菌属(Serratia)、志贺氏菌属(Shigella)和耶尔森氏菌属(Yersinia)的物种。甚至更优选地，微生物选自大肠杆菌(Escherichia coli)、肺炎克雷伯氏菌(Klebsiella pneumoniae)、产酸克雷伯氏菌(Klebisella oxytoca)、铜绿假单胞菌(Pseudomonas aeruginosa)、荧光假单胞菌(Pseudomonas fluorescens)、鼠伤寒沙门氏菌(Salmonalla typhimurium)、肠沙门氏菌(Salmonella enterica)、粘质沙雷氏菌(Serratia marcescens)、菠萝泛菌(Pantoeaananatis)、谷氨酸棒杆菌(Corynebacterium glutamicum)、耐放射异常球菌(Deinococcusradiodurans)、热解糖好热厌氧杆菌(Thermoanaerobacterium thermosaccharolyticum)、楔状梭菌(Clostridium sphenoides)和酿酒酵母(Saccharomyces cerevisiae)。
特别地，实施例显示了修饰的大肠杆菌菌株，但是这些修饰可以容易地在相同家族的其他微生物上进行。
大肠杆菌属于肠杆菌科家族，其包含革兰氏阴性、杆状、非孢子形成的成员，并且通常长度为1-5μm。大多数成员使用鞭毛移动，但少数属不运动。该家族的许多成员是在人类和其他动物的肠中发现的肠道菌群的正常部分，而其他成员在水或土壤中发现，或者是各种不同动物和植物的寄生虫。大肠杆菌是最重要的模式生物之一，但肠杆菌科的其他重要成员也包括克雷伯氏菌特别是肺炎克雷伯菌，以及沙门氏菌。
根据本发明方法的另一个实施方案，缺失了SEQ ID NO：1的基因ppsR(可以称为“ΔppsR”)，其编码SEQ ID NO：2的双功能ADP依赖性激酶-Pi-依赖性焦磷酸化酶DUF299蛋白。
如本文所用，术语“缺失”是指完全抑制基因的表达。这种表达的抑制可以是基因表达的抑制，基因表达所必需的全部或部分启动子区域的缺失，或基因编码区的缺失。缺失的基因可以被有助于根据本发明的菌株的鉴定、分离和纯化的选择标记基因置换。例如，可以通过同源重组技术来实现基因表达的抑制(Datsenko和Wanner，2000)。
在另一个实施方案中，可以减弱编码双功能ADP依赖性激酶-Pi-依赖性焦磷酸化酶DUF299蛋白的基因ppsR的表达。
如本文所用，术语“减弱”是指部分抑制基因的表达。这种表达的减弱可以是用较弱的天然或合成启动子与野生型启动子交换，或者是使用减少ppsR基因表达的试剂，包括反义RNA或干扰RNA(iRNA)，更具体是小干扰RNA(siRNAs)或短发夹RNA(shRNA)。例如，可以通过同源重组技术实现启动子交换(Datsenko和Wanner，2000)。
可以使用本领域技术人员已知的适于抑制蛋白质(尤其是这种蛋白质)的表达或功能的任何其他方法。
本发明的方法可用于大量生产甲硫氨酸或其羟基类似物形式。因此，本发明的方法允许增加甲硫氨酸或其羟基类似物形式的生产。
更具体地，本发明的方法允许改善甲硫氨酸或其衍生物的生产。
术语“改善的甲硫氨酸生产”是指与其亲本菌株即ppsR基因缺失或减弱前的微生物相比，甲硫氨酸的生产力提高和/或甲硫氨酸滴度增加和/或甲硫氨酸/碳源产量增加和/或甲硫氨酸纯度增加。通过本领域技术人员已知的标准分析方法，特别是HPLC，可以明确地记录培养液中微生物产生的甲硫氨酸。在专利申请WO2016034536、WO2014029592和WO2012091479中公开了一些增加甲硫氨酸生产的遗传修饰微生物，用于生产甲硫氨酸的大肠杆菌菌株的实例，以及专利申请WO2008080900和WO2012098042，用于生产甲硫氨酸的棒杆菌菌株的实例。所有这些公开内容都通过引用并入本文。
优选地，产生本发明的甲硫氨酸的微生物是大肠杆菌菌株，并且其至少包含：
-增强选自以下的至少一种基因的表达：SEQ ID NO：3的基因metA或如专利申请WO2005108561、US2010041108或WO2008127240中所公开的编码对甲硫氨酸或其衍生物具有降低的反馈抑制的酶的突变基因、SEQ ID NO：5的metH、SEQ ID NO：7-9-11-13-15的cysPUWAM、SEQ ID NO：17-19-21的cysJIH、SEQ ID NO：23-25-27的gcvTHP、SEQ ID NO：29中的metF、SEQ ID NO：31的serA、SEQ ID NO：33的serB、SEQ ID NO：35的serC、SEQ ID NO：37的cysE、SEQ ID NO：39的thrA或如专利申请WO2005111202中所述的编码对苏氨酸具有降低的反馈敏感性的酶的突变基因、SEQ ID NO：41的ptsG、SEQ ID NO：43和45中的ygaZH或其在专利申请WO2016034536中公开的同源基因、和SEQ ID NO：47的pyc，以及
-减弱选自以下的至少一种基因的表达：SEQ ID NO：49的metJ、SEQ ID NO：51的pykA、SEQ ID NO：53的pykF、SEQ ID NO：55的purU、SEQ ID NO：57的metE、SEQ ID NO：59的dgsA和SEQ ID NO：61的yncA。
表1：根据本发明的酶和基因(n/a：不适用)
如上所述，糖被转运到细菌细胞中并被磷酸烯醇式丙酮酸：糖磷酸转移酶***(PTS)磷酸化。高浓度的磷酸化糖，特别是磷酸化葡萄糖对细胞有毒性，因此PTS***受到高度调节。这与***复杂的事实相结合，使得操纵***非常困难。然而，如下所述，本发明人出乎意料地产生了经遗传修饰的微生物，其包含编码PTS碳水化合物利用***的功能基因，同时缺少至少一种调节PPS表达的蛋白质。
在第二方面，本发明因此涉及经遗传修饰的微生物，其用于增强从碳水化合物作为碳源的甲硫氨酸或其羟基类似物形式的生产，所述经遗传修饰的微生物包含编码PTS碳水化合物利用***的功能基因，并且具有降低的调节磷酸烯醇式丙酮酸合酶(PPS)表达的双功能ADP依赖性激酶-Pi-依赖性焦磷酸化酶蛋白表达。
该经遗传修饰的微生物具有与本发明方法中使用的微生物相同的遗传特征。特别地，在该微生物中，编码双功能ADP依赖性激酶-Pi-依赖性焦磷酸化酶DUF229的基因ppsR缺失或减弱。更优选地，在本发明的微生物中缺失基因ppsR。
因此，它可以用于根据本发明的发酵方法中以增加甲硫氨酸或其羟基类似物形式的产生。
优选地，所述微生物可以用于根据本发明的发酵方法中以增加甲硫氨酸的产生。
实施例
实施例1：菌株构建的方法
在下面给出的实施例中，使用本领域熟知的方法构建含有复制载体和/或各种染色体缺失的大肠杆菌菌株，并在大肠杆菌中使用Datsenko&Wanner(2000)描述的同源重组的方法进行取代。以相同的方式，使用质粒或载体在重组微生物中表达或过表达一种或几种基因是本领域技术人员熟知的。合适的大肠杆菌表达载体的实例包括pTrc、pACYC184npBR322、pUC18、pUC19、pKC30、pRep4、pHS1、pHS2、pPLc236等。
以下实施例中使用了几种方案。在专利申请EP 2532751中充分描述了本发明中使用的方案1(通过同源重组的染色体修饰，筛选重组体)，方案2(噬菌体P1的转导)和方案3(抗生素盒切除，必要时除去抗性基因)。抗生素抗性盒可以在pKD3、pKD4、pKD13或含有由FRT位点包围的另一抗生素抗性基因的任何其他质粒上扩增。用本领域技术人员能够设计的合适的寡核苷酸来进行PCR分析验证染色体修饰。
实施例2：构建菌株1和2
构建菌株1
用于构建菌株1的完整方案在专利申请号WO2012055798：菌株N°10中提供。
构建菌株2
使用同源重组策略(根据方案1和3)失活由ppsR基因编码的PEP合酶调节蛋白PSRP。DppsR的寡核苷酸：SEQ ID N°63和64用于PCR扩增抗性盒。保留的菌株命名为MG1655DppsR::Km或MG1655DppsR::Gt。最后，通过P1噬菌体转导(根据方案2)将DppsR::Km或MG1655DppsR::Gt缺失转移到菌株1中。
实施例3：摇瓶培养物和产量
在小锥形瓶中评估甲硫氨酸生产菌株。将5.5mL预培养物在30℃在混合培养基(10％LB培养基(Sigma 25％)，含有2.5g.L-1葡萄糖和90％基本培养基PC1(，如WO2012055798中所述)中生长21小时。将其接种到50mL PC1培养基中，培养至OD600为0.2。培养温度为37℃2小时、42℃2小时、37℃直至培养结束。当培养物达到OD600为5至7时，在OPA/Fmoc衍生化和GCMS-甲硅烷基化后通过HPLC定量甲硫氨酸(Met)和高羊毛氨酸(Homolanthionine,HLA)，并通过具有折射检测的HPLC定量葡萄糖。
对于所有培养物，必要时加入抗生素，浓度为50mg.L-1的卡那霉素和壮观霉素，浓度为30mg.L-1的氯霉素，浓度为10mg.L-1的庆大霉素。
表2：上述菌株的产量(g产物/g消耗的糖)
＝：与对照菌株无差异，+：产量高于对照菌株(110％-120％)，
++：产量高于对照菌株(120％-150％)，+++：产量高于对照菌株(>150％)
菌株2的产量高于相应的对照菌株1。
结论
如上述实施例所证明的，编码双功能ADP依赖性激酶-Pi-依赖性焦磷酸化酶DUF299蛋白的ppsR的缺失使得甲硫氨酸的生产增加。
BIBLIOGRAPHY
-Altschul S,Gish W,Miller W,Myers E,Lipman DJ(1990).J.Mol.Biol；215(3):403–410
-Anderson,Proc.Natl.Acad.Sci.USA.,1946,32:120-128.
-Burnell JN(2010)BMC Biochemistry 11(1)
-Canovas JL,Kornberg HL(1965).Biochim Biophys Acta 96；169-72.
-Cooper RA,Kornberg HL(1965).Biochim Biophys Acta 104(2)；618-20.
-Datsenko KAεt Wanner BL,(2000),Proc Natl Acad Sci U S A.,97:6640-6645
-Davis JJεt Olsen GJ.(2011).Mol.Biol.Evol.；28(1):211-221
-Deml L,Bojak A,Steck S,Graf M,Wild J,Schirmbeck R,Wolf H,Wagner R.(2011).J.Virol.,75(22):10991-11001.
-Gabor E,Kosfeld A,Staab A,Kremling A,Jahreis K.(2011).Eur.J.Cell Biol.,90(9):711-720
-Graf M,Bojak A,Deml L,Bieler K,Wolf H,Wagner R.(2000).J.Virol.；74(22):10/22-10826
-Gosset G,Yong-Xiao J,Berry A(1996).Journal of IndustrialMicrobiology 17:47–52.
-Kim P,Laivenieks M,Vieille C,Gregory Zeikus J.(2004).Applied andEnvironmentalMicrobiology 70(2):1238-1241.
-Kornberg HL,Malcovati M(1973).FEBS Lett 32(2)；257-9.
-Kotrba P,Inui M,Yukawa H(2001).J.BioscienceεtBioeng.92(6):502-517
-Meza E,Becker J,Bolivar F,Gosset G,Wittmann C(2012).Microbial CellFactories 11:127.
-Miller,1992；“A Short Course in Bacterial Genetics:A LaboratoryManual and Handbook forEscherichia coli and Related Bacteria”,Cold SpringHarbor Laboratory Press,Cold SpringHarbor,New York.
-Patnaik R,Roof WD,Young RF,Liao JC.(1992).Journal of Bacteriology174(23):7527-7532.
-Pittard J,Wallace BJ.(1966).Journal of Bacteriology 91(4):1494-1508.
-Postma,P.W.,Roseman,S.(1976).Biochim Biophys Acta 457(3-4)；213-57.
-Sambrook and Russell,(2001),Molecular Cloning:3rd edition,ColdSpring Harbor Laboratory Press,NY,Vol 1,2,3
-Schaefer U,Boos W,Takors R,Weuster-Botz D,(1999).Anal.Biochem.270:88-96.
-Tatarko M,Romeo T(2001).Current Microbiology 43:26–32.
序列表
<110> 赢创德固赛有限公司
<120> 通过含有编码糖磷酸转移酶***(PTS)的基因的微生物发酵生产甲硫氨酸或其羟基类似物形式的方法
<130> B372757D36346
<150> IB2016/001123
<151> 2016-07-08
<160> 64
<170> PatentIn version 3.5
<210> 1
<211> 834
<212> DNA
<213> Escherichia coli
<400> 1
atggataatg ctgttgatcg ccacgttttt tatatttctg atggtacggc aataactgcg 60
gaggtattag gacacgcagt aatgtcacaa tttcccgtca ctatcagcag catcacgctg 120
ccgtttgtcg aaaatgagag ccgtgcacgg gcagtgaagg atcagattga cgcaatttat 180
caccagacag gcgtgcgccc gctggtcttc tactccatcg tgttgccgga gattcgcgcc 240
atcatcttgc aaagtgaagg cttttgccag gatatcgttc aggcgctggt tgccccgcta 300
caacaagaga tgaaactgga tccaacgccg attgctcatc gtacccatgg ccttaaccct 360
aataatctca ataaatatga tgcgcgcatt gcggcgattg attacaccct cgcccacgat 420
gacggcattt cgttgcgcaa tctggaccag gctcaggtga tcctgctcgg tgtttctcgc 480
tgtggtaaaa cccccaccag tctgtatctg gcaatgcagt ttggtatccg cgcggcaaac 540
taccccttta ttgccgacga tatggataat ctggtgctac ccgcgtcgct caaaccgctt 600
cagcataaat tgttcggcct gactatcgac ccggaacgtc tggcggcgat tcgcgaggaa 660
cgtcgggaga acagtcgcta tgcctcgctt cgtcagtgca ggatggaagt cgcggaagtg 720
gaagccttgt accgtaaaaa tcagatcccg tggattaaca gtaccaatta ttcggtagaa 780
gagattgcca ccaagatcct cgatatcatg ggccttagtc gccgaatgta ctag 834
<210> 2
<211> 277
<212> PRT
<213> Escherichia coli
<400> 2
Met Asp Asn Ala Val Asp Arg His Val Phe Tyr Ile Ser Asp Gly Thr
1 5 10 15
Ala Ile Thr Ala Glu Val Leu Gly His Ala Val Met Ser Gln Phe Pro
20 25 30
Val Thr Ile Ser Ser Ile Thr Leu Pro Phe Val Glu Asn Glu Ser Arg
35 40 45
Ala Arg Ala Val Lys Asp Gln Ile Asp Ala Ile Tyr His Gln Thr Gly
50 55 60
Val Arg Pro Leu Val Phe Tyr Ser Ile Val Leu Pro Glu Ile Arg Ala
65 70 75 80
Ile Ile Leu Gln Ser Glu Gly Phe Cys Gln Asp Ile Val Gln Ala Leu
85 90 95
Val Ala Pro Leu Gln Gln Glu Met Lys Leu Asp Pro Thr Pro Ile Ala
100 105 110
His Arg Thr His Gly Leu Asn Pro Asn Asn Leu Asn Lys Tyr Asp Ala
115 120 125
Arg Ile Ala Ala Ile Asp Tyr Thr Leu Ala His Asp Asp Gly Ile Ser
130 135 140
Leu Arg Asn Leu Asp Gln Ala Gln Val Ile Leu Leu Gly Val Ser Arg
145 150 155 160
Cys Gly Lys Thr Pro Thr Ser Leu Tyr Leu Ala Met Gln Phe Gly Ile
165 170 175
Arg Ala Ala Asn Tyr Pro Phe Ile Ala Asp Asp Met Asp Asn Leu Val
180 185 190
Leu Pro Ala Ser Leu Lys Pro Leu Gln His Lys Leu Phe Gly Leu Thr
195 200 205
Ile Asp Pro Glu Arg Leu Ala Ala Ile Arg Glu Glu Arg Arg Glu Asn
210 215 220
Ser Arg Tyr Ala Ser Leu Arg Gln Cys Arg Met Glu Val Ala Glu Val
225 230 235 240
Glu Ala Leu Tyr Arg Lys Asn Gln Ile Pro Trp Ile Asn Ser Thr Asn
245 250 255
Tyr Ser Val Glu Glu Ile Ala Thr Lys Ile Leu Asp Ile Met Gly Leu
260 265 270
Ser Arg Arg Met Tyr
275
<210> 3
<211> 930
<212> DNA
<213> Escherichia coli
<400> 3
atgccgattc gtgtgccgga cgagctaccc gccgtcaatt tcttgcgtga agaaaacgtc 60
tttgtgatga caacttctcg tgcgtctggt caggaaattc gtccacttaa ggttctgatc 120
cttaacctga tgccgaagaa gattgaaact gaaaatcagt ttctgcgcct gctttcaaac 180
tcacctttgc aggtcgatat tcagctgttg cgcatcgatt cccgtgaatc gcgcaacacg 240
cccgcagagc atctgaacaa cttctactgt aactttgaag atattcagga tcagaacttt 300
gacggtttga ttgtaactgg tgcgccgctg ggcctggtgg agtttaatga tgtcgcttac 360
tggccgcaga tcaaacaggt gctggagtgg tcgaaagatc acgtcacctc gacgctgttt 420
gtctgctggg cggtacaggc cgcgctcaat atcctctacg gcattcctaa gcaaactcgc 480
accgaaaaac tctctggcgt ttacgagcat catattctcc atcctcatgc gcttctgacg 540
cgtggctttg atgattcatt cctggcaccg cattcgcgct atgctgactt tccggcagcg 600
ttgattcgtg attacaccga tctggaaatt ctggcagaga cggaagaagg ggatgcatat 660
ctgtttgcca gtaaagataa gcgcattgcc tttgtgacgg gccatcccga atatgatgcg 720
caaacgctgg cgcaggaatt tttccgcgat gtggaagccg gactagaccc ggatgtaccg 780
tataactatt tcccgcacaa tgatccgcaa aatacaccgc gagcgagctg gcgtagtcac 840
ggtaatttac tgtttaccaa ctggctcaac tattacgtct accagatcac gccatacgat 900
ctacggcaca tgaatccaac gctggattaa 930
<210> 4
<211> 309
<212> PRT
<213> Escherichia coli
<400> 4
Met Pro Ile Arg Val Pro Asp Glu Leu Pro Ala Val Asn Phe Leu Arg
1 5 10 15
Glu Glu Asn Val Phe Val Met Thr Thr Ser Arg Ala Ser Gly Gln Glu
20 25 30
Ile Arg Pro Leu Lys Val Leu Ile Leu Asn Leu Met Pro Lys Lys Ile
35 40 45
Glu Thr Glu Asn Gln Phe Leu Arg Leu Leu Ser Asn Ser Pro Leu Gln
50 55 60
Val Asp Ile Gln Leu Leu Arg Ile Asp Ser Arg Glu Ser Arg Asn Thr
65 70 75 80
Pro Ala Glu His Leu Asn Asn Phe Tyr Cys Asn Phe Glu Asp Ile Gln
85 90 95
Asp Gln Asn Phe Asp Gly Leu Ile Val Thr Gly Ala Pro Leu Gly Leu
100 105 110
Val Glu Phe Asn Asp Val Ala Tyr Trp Pro Gln Ile Lys Gln Val Leu
115 120 125
Glu Trp Ser Lys Asp His Val Thr Ser Thr Leu Phe Val Cys Trp Ala
130 135 140
Val Gln Ala Ala Leu Asn Ile Leu Tyr Gly Ile Pro Lys Gln Thr Arg
145 150 155 160
Thr Glu Lys Leu Ser Gly Val Tyr Glu His His Ile Leu His Pro His
165 170 175
Ala Leu Leu Thr Arg Gly Phe Asp Asp Ser Phe Leu Ala Pro His Ser
180 185 190
Arg Tyr Ala Asp Phe Pro Ala Ala Leu Ile Arg Asp Tyr Thr Asp Leu
195 200 205
Glu Ile Leu Ala Glu Thr Glu Glu Gly Asp Ala Tyr Leu Phe Ala Ser
210 215 220
Lys Asp Lys Arg Ile Ala Phe Val Thr Gly His Pro Glu Tyr Asp Ala
225 230 235 240
Gln Thr Leu Ala Gln Glu Phe Phe Arg Asp Val Glu Ala Gly Leu Asp
245 250 255
Pro Asp Val Pro Tyr Asn Tyr Phe Pro His Asn Asp Pro Gln Asn Thr
260 265 270
Pro Arg Ala Ser Trp Arg Ser His Gly Asn Leu Leu Phe Thr Asn Trp
275 280 285
Leu Asn Tyr Tyr Val Tyr Gln Ile Thr Pro Tyr Asp Leu Arg His Met
290 295 300
Asn Pro Thr Leu Asp
305
<210> 5
<211> 3684
<212> DNA
<213> Escherichia coli
<400> 5
gtgagcagca aagtggaaca actgcgtgcg cagttaaatg aacgtattct ggtgctggac 60
ggcggtatgg gcaccatgat ccagagttat cgactgaacg aagccgattt tcgtggtgaa 120
cgctttgccg actggccatg cgacctcaaa ggcaacaacg acctgctggt actcagtaaa 180
ccggaagtga tcgccgctat ccacaacgcc tactttgaag cgggcgcgga tatcatcgaa 240
accaacacct tcaactccac gaccattgcg atggcggatt accagatgga atccctgtcg 300
gcggaaatca actttgcggc ggcgaaactg gcgcgagctt gtgctgacga gtggaccgcg 360
cgcacgccag agaaaccgcg ctacgttgcc ggtgttctcg gcccgaccaa ccgcacggcg 420
tctatttctc cggacgtcaa cgatccggca tttcgtaata tcacttttga cgggctggtg 480
gcggcttatc gagagtccac caaagcgctg gtggaaggtg gcgcggatct gatcctgatt 540
gaaaccgttt tcgacaccct taacgccaaa gcggcggtat ttgcggtgaa aacggagttt 600
gaagcgctgg gcgttgagct gccgattatg atctccggca ccatcaccga cgcctccggg 660
cgcacgctct ccgggcagac caccgaagca ttttacaact cattgcgcca cgccgaagct 720
ctgacctttg gcctgaactg tgcgctgggg cccgatgaac tgcgccagta cgtgcaggag 780
ctgtcacgga ttgcggaatg ctacgtcacc gcgcacccga acgccgggct acccaacgcc 840
tttggtgagt acgatctcga cgccgacacg atggcaaaac agatacgtga atgggcgcaa 900
gcgggttttc tcaatatcgt cggcggctgc tgtggcacca cgccacaaca tattgcagcg 960
atgagtcgtg cagtagaagg attagcgccg cgcaaactgc cggaaattcc cgtagcctgc 1020
cgtttgtccg gcctggagcc gctgaacatt ggcgaagata gcctgtttgt gaacgtgggt 1080
gaacgcacca acgtcaccgg ttccgctaag ttcaagcgcc tgatcaaaga agagaaatac 1140
agcgaggcgc tggatgtcgc gcgtcaacag gtggaaaacg gcgcgcagat tatcgatatc 1200
aacatggatg aagggatgct cgatgccgaa gcggcgatgg tgcgttttct caatctgatt 1260
gccggtgaac cggatatcgc tcgcgtgccg attatgatcg actcctcaaa atgggacgtc 1320
attgaaaaag gtctgaagtg tatccagggc aaaggcattg ttaactctat ctcgatgaaa 1380
gagggcgtcg atgcctttat ccatcacgcg aaattgttgc gtcgctacgg tgcggcagtg 1440
gtggtaatgg cctttgacga acagggacag gccgatactc gcgcacggaa aatcgagatt 1500
tgccgtcggg cgtacaaaat cctcaccgaa gaggttggct tcccgccaga agatatcatc 1560
ttcgacccaa acatcttcgc ggtcgcaact ggcattgaag agcacaacaa ctacgcgcag 1620
gactttatcg gcgcgtgtga agacatcaaa cgcgaactgc cgcacgcgct gatttccggc 1680
ggcgtatcta acgtttcttt ctcgttccgt ggcaacgatc cggtgcgcga agccattcac 1740
gcagtgttcc tctactacgc tattcgcaat ggcatggata tggggatcgt caacgccggg 1800
caactggcga tttacgacga cctacccgct gaactgcgcg acgcggtgga agatgtgatt 1860
cttaatcgtc gcgacgatgg caccgagcgt ttactggagc ttgccgagaa atatcgcggc 1920
agcaaaaccg acgacaccgc caacgcccag caggcggagt ggcgctcgtg ggaagtgaat 1980
aaacgtctgg aatactcgct ggtcaaaggc attaccgagt ttatcgagca ggataccgaa 2040
gaagcccgcc agcaggctac gcgcccgatt gaagtgattg aaggcccgtt gatggacggc 2100
atgaatgtgg tcggcgacct gtttggcgaa gggaaaatgt tcctgccaca ggtggtcaaa 2160
tcggcgcgcg tcatgaaaca ggcggtggcc tacctcgaac cgtttattga agccagcaaa 2220
gagcagggca aaaccaacgg caagatggtg atcgccaccg tgaagggcga cgtccacgac 2280
atcggtaaaa atatcgttgg tgtggtgctg caatgtaaca actacgaaat tgtcgatctc 2340
ggcgttatgg tgcctgcgga aaaaattctc cgtaccgcta aagaagtgaa tgctgatctg 2400
attggccttt cggggcttat cacgccgtcg ctggacgaga tggttaacgt ggcgaaagag 2460
atggagcgtc agggcttcac tattccgtta ctgattggcg gcgcgacgac ctcaaaagcg 2520
cacacggcgg tgaaaatcga gcagaactac agcggcccga cggtgtatgt gcagaatgcc 2580
tcgcgtaccg ttggtgtggt ggcggcgctg ctttccgata cccagcgtga tgattttgtc 2640
gctcgtaccc gcaaggagta cgaaaccgta cgtattcagc acgggcgcaa gaaaccgcgc 2700
acaccaccgg tcacgctgga agcggcgcgc gataacgatt tcgcttttga ctggcaggct 2760
tacacgccgc cggtggcgca ccgtctcggc gtgcaggaag tcgaagccag catcgaaacg 2820
ctgcgtaatt acatcgactg gacaccgttc tttatgacct ggtcgctggc cgggaagtat 2880
ccgcgcattc tggaagatga agtggtgggc gttgaggcgc agcggctgtt taaagacgcc 2940
aacgacatgc tggataaatt aagcgccgag aaaacgctga atccgcgtgg cgtggtgggc 3000
ctgttcccgg caaaccgtgt gggcgatgac attgaaatct accgtgacga aacgcgtacc 3060
catgtgatca acgtcagcca ccatctgcgt caacagaccg aaaaaacagg cttcgctaac 3120
tactgtctcg ctgacttcgt tgcgccgaag ctttctggta aagcagatta catcggcgca 3180
tttgccgtga ctggcgggct ggaagaggac gcactggctg atgcctttga agcgcagcac 3240
gatgattaca acaaaatcat ggtgaaagcg cttgccgacc gtttagccga agcctttgcg 3300
gagtatctcc atgagcgtgt gcgtaaagtc tactggggct atgcgccgaa cgagaacctc 3360
agcaacgaag agctgatccg cgaaaactac cagggcatcc gtccggcacc gggctatccg 3420
gcctgcccgg aacatacgga aaaagccacc atctgggagc tgctggaagt ggaaaaacac 3480
actggcatga aactcacaga atctttcgcc atgtggcccg gtgcatcggt ttcgggttgg 3540
tacttcagcc acccggacag caagtactac gctgtagcac aaattcagcg cgatcaggtt 3600
gaagattatg cccgccgtaa aggtatgagc gttaccgaag ttgagcgctg gctggcaccg 3660
aatctggggt atgacgcgga ctga 3684
<210> 6
<211> 1227
<212> PRT
<213> Escherichia coli
<400> 6
Met Ser Ser Lys Val Glu Gln Leu Arg Ala Gln Leu Asn Glu Arg Ile
1 5 10 15
Leu Val Leu Asp Gly Gly Met Gly Thr Met Ile Gln Ser Tyr Arg Leu
20 25 30
Asn Glu Ala Asp Phe Arg Gly Glu Arg Phe Ala Asp Trp Pro Cys Asp
35 40 45
Leu Lys Gly Asn Asn Asp Leu Leu Val Leu Ser Lys Pro Glu Val Ile
50 55 60
Ala Ala Ile His Asn Ala Tyr Phe Glu Ala Gly Ala Asp Ile Ile Glu
65 70 75 80
Thr Asn Thr Phe Asn Ser Thr Thr Ile Ala Met Ala Asp Tyr Gln Met
85 90 95
Glu Ser Leu Ser Ala Glu Ile Asn Phe Ala Ala Ala Lys Leu Ala Arg
100 105 110
Ala Cys Ala Asp Glu Trp Thr Ala Arg Thr Pro Glu Lys Pro Arg Tyr
115 120 125
Val Ala Gly Val Leu Gly Pro Thr Asn Arg Thr Ala Ser Ile Ser Pro
130 135 140
Asp Val Asn Asp Pro Ala Phe Arg Asn Ile Thr Phe Asp Gly Leu Val
145 150 155 160
Ala Ala Tyr Arg Glu Ser Thr Lys Ala Leu Val Glu Gly Gly Ala Asp
165 170 175
Leu Ile Leu Ile Glu Thr Val Phe Asp Thr Leu Asn Ala Lys Ala Ala
180 185 190
Val Phe Ala Val Lys Thr Glu Phe Glu Ala Leu Gly Val Glu Leu Pro
195 200 205
Ile Met Ile Ser Gly Thr Ile Thr Asp Ala Ser Gly Arg Thr Leu Ser
210 215 220
Gly Gln Thr Thr Glu Ala Phe Tyr Asn Ser Leu Arg His Ala Glu Ala
225 230 235 240
Leu Thr Phe Gly Leu Asn Cys Ala Leu Gly Pro Asp Glu Leu Arg Gln
245 250 255
Tyr Val Gln Glu Leu Ser Arg Ile Ala Glu Cys Tyr Val Thr Ala His
260 265 270
Pro Asn Ala Gly Leu Pro Asn Ala Phe Gly Glu Tyr Asp Leu Asp Ala
275 280 285
Asp Thr Met Ala Lys Gln Ile Arg Glu Trp Ala Gln Ala Gly Phe Leu
290 295 300
Asn Ile Val Gly Gly Cys Cys Gly Thr Thr Pro Gln His Ile Ala Ala
305 310 315 320
Met Ser Arg Ala Val Glu Gly Leu Ala Pro Arg Lys Leu Pro Glu Ile
325 330 335
Pro Val Ala Cys Arg Leu Ser Gly Leu Glu Pro Leu Asn Ile Gly Glu
340 345 350
Asp Ser Leu Phe Val Asn Val Gly Glu Arg Thr Asn Val Thr Gly Ser
355 360 365
Ala Lys Phe Lys Arg Leu Ile Lys Glu Glu Lys Tyr Ser Glu Ala Leu
370 375 380
Asp Val Ala Arg Gln Gln Val Glu Asn Gly Ala Gln Ile Ile Asp Ile
385 390 395 400
Asn Met Asp Glu Gly Met Leu Asp Ala Glu Ala Ala Met Val Arg Phe
405 410 415
Leu Asn Leu Ile Ala Gly Glu Pro Asp Ile Ala Arg Val Pro Ile Met
420 425 430
Ile Asp Ser Ser Lys Trp Asp Val Ile Glu Lys Gly Leu Lys Cys Ile
435 440 445
Gln Gly Lys Gly Ile Val Asn Ser Ile Ser Met Lys Glu Gly Val Asp
450 455 460
Ala Phe Ile His His Ala Lys Leu Leu Arg Arg Tyr Gly Ala Ala Val
465 470 475 480
Val Val Met Ala Phe Asp Glu Gln Gly Gln Ala Asp Thr Arg Ala Arg
485 490 495
Lys Ile Glu Ile Cys Arg Arg Ala Tyr Lys Ile Leu Thr Glu Glu Val
500 505 510
Gly Phe Pro Pro Glu Asp Ile Ile Phe Asp Pro Asn Ile Phe Ala Val
515 520 525
Ala Thr Gly Ile Glu Glu His Asn Asn Tyr Ala Gln Asp Phe Ile Gly
530 535 540
Ala Cys Glu Asp Ile Lys Arg Glu Leu Pro His Ala Leu Ile Ser Gly
545 550 555 560
Gly Val Ser Asn Val Ser Phe Ser Phe Arg Gly Asn Asp Pro Val Arg
565 570 575
Glu Ala Ile His Ala Val Phe Leu Tyr Tyr Ala Ile Arg Asn Gly Met
580 585 590
Asp Met Gly Ile Val Asn Ala Gly Gln Leu Ala Ile Tyr Asp Asp Leu
595 600 605
Pro Ala Glu Leu Arg Asp Ala Val Glu Asp Val Ile Leu Asn Arg Arg
610 615 620
Asp Asp Gly Thr Glu Arg Leu Leu Glu Leu Ala Glu Lys Tyr Arg Gly
625 630 635 640
Ser Lys Thr Asp Asp Thr Ala Asn Ala Gln Gln Ala Glu Trp Arg Ser
645 650 655
Trp Glu Val Asn Lys Arg Leu Glu Tyr Ser Leu Val Lys Gly Ile Thr
660 665 670
Glu Phe Ile Glu Gln Asp Thr Glu Glu Ala Arg Gln Gln Ala Thr Arg
675 680 685
Pro Ile Glu Val Ile Glu Gly Pro Leu Met Asp Gly Met Asn Val Val
690 695 700
Gly Asp Leu Phe Gly Glu Gly Lys Met Phe Leu Pro Gln Val Val Lys
705 710 715 720
Ser Ala Arg Val Met Lys Gln Ala Val Ala Tyr Leu Glu Pro Phe Ile
725 730 735
Glu Ala Ser Lys Glu Gln Gly Lys Thr Asn Gly Lys Met Val Ile Ala
740 745 750
Thr Val Lys Gly Asp Val His Asp Ile Gly Lys Asn Ile Val Gly Val
755 760 765
Val Leu Gln Cys Asn Asn Tyr Glu Ile Val Asp Leu Gly Val Met Val
770 775 780
Pro Ala Glu Lys Ile Leu Arg Thr Ala Lys Glu Val Asn Ala Asp Leu
785 790 795 800
Ile Gly Leu Ser Gly Leu Ile Thr Pro Ser Leu Asp Glu Met Val Asn
805 810 815
Val Ala Lys Glu Met Glu Arg Gln Gly Phe Thr Ile Pro Leu Leu Ile
820 825 830
Gly Gly Ala Thr Thr Ser Lys Ala His Thr Ala Val Lys Ile Glu Gln
835 840 845
Asn Tyr Ser Gly Pro Thr Val Tyr Val Gln Asn Ala Ser Arg Thr Val
850 855 860
Gly Val Val Ala Ala Leu Leu Ser Asp Thr Gln Arg Asp Asp Phe Val
865 870 875 880
Ala Arg Thr Arg Lys Glu Tyr Glu Thr Val Arg Ile Gln His Gly Arg
885 890 895
Lys Lys Pro Arg Thr Pro Pro Val Thr Leu Glu Ala Ala Arg Asp Asn
900 905 910
Asp Phe Ala Phe Asp Trp Gln Ala Tyr Thr Pro Pro Val Ala His Arg
915 920 925
Leu Gly Val Gln Glu Val Glu Ala Ser Ile Glu Thr Leu Arg Asn Tyr
930 935 940
Ile Asp Trp Thr Pro Phe Phe Met Thr Trp Ser Leu Ala Gly Lys Tyr
945 950 955 960
Pro Arg Ile Leu Glu Asp Glu Val Val Gly Val Glu Ala Gln Arg Leu
965 970 975
Phe Lys Asp Ala Asn Asp Met Leu Asp Lys Leu Ser Ala Glu Lys Thr
980 985 990
Leu Asn Pro Arg Gly Val Val Gly Leu Phe Pro Ala Asn Arg Val Gly
995 1000 1005
Asp Asp Ile Glu Ile Tyr Arg Asp Glu Thr Arg Thr His Val Ile
1010 1015 1020
Asn Val Ser His His Leu Arg Gln Gln Thr Glu Lys Thr Gly Phe
1025 1030 1035
Ala Asn Tyr Cys Leu Ala Asp Phe Val Ala Pro Lys Leu Ser Gly
1040 1045 1050
Lys Ala Asp Tyr Ile Gly Ala Phe Ala Val Thr Gly Gly Leu Glu
1055 1060 1065
Glu Asp Ala Leu Ala Asp Ala Phe Glu Ala Gln His Asp Asp Tyr
1070 1075 1080
Asn Lys Ile Met Val Lys Ala Leu Ala Asp Arg Leu Ala Glu Ala
1085 1090 1095
Phe Ala Glu Tyr Leu His Glu Arg Val Arg Lys Val Tyr Trp Gly
1100 1105 1110
Tyr Ala Pro Asn Glu Asn Leu Ser Asn Glu Glu Leu Ile Arg Glu
1115 1120 1125
Asn Tyr Gln Gly Ile Arg Pro Ala Pro Gly Tyr Pro Ala Cys Pro
1130 1135 1140
Glu His Thr Glu Lys Ala Thr Ile Trp Glu Leu Leu Glu Val Glu
1145 1150 1155
Lys His Thr Gly Met Lys Leu Thr Glu Ser Phe Ala Met Trp Pro
1160 1165 1170
Gly Ala Ser Val Ser Gly Trp Tyr Phe Ser His Pro Asp Ser Lys
1175 1180 1185
Tyr Tyr Ala Val Ala Gln Ile Gln Arg Asp Gln Val Glu Asp Tyr
1190 1195 1200
Ala Arg Arg Lys Gly Met Ser Val Thr Glu Val Glu Arg Trp Leu
1205 1210 1215
Ala Pro Asn Leu Gly Tyr Asp Ala Asp
1220 1225
<210> 7
<211> 1017
<212> DNA
<213> Escherichia coli
<400> 7
atggccgtta acttactgaa aaagaactca ctcgcgctgg tcgcttctct gctgctggcg 60
ggccatgtac aggcaacgga actgctgaac agttcttatg acgtctcccg cgagctgttt 120
gccgccctga atccgccgtt tgagcaacaa tgggcaaaag ataacggcgg cgacaaactg 180
acgataaaac aatctcatgc cgggtcatca aaacaggcgc tggcgatttt acagggctta 240
aaagccgacg ttgtcactta taaccaggtg accgacgtac aaatcctgca cgataaaggc 300
aagctgatcc cggccgactg gcagtcgcgc ctgccgaata atagctcgcc gttctactcc 360
accatgggct tcctggtgcg taagggtaac ccgaagaata tccacgattg gaacgacctg 420
gtgcgctccg acgtgaagct gattttcccg aacccgaaaa cgtcgggtaa cgcgcgttat 480
acctatctgg cggcatgggg cgcagcggat aaagctgacg gtggtgacaa aggcaaaacc 540
gaacagttta tgacccagtt cctgaaaaac gttgaagtgt tcgatactgg cggtcgtggc 600
gcgaccacca cttttgccga gcgcggcctg ggcgatgtgc tgattagctt cgaatcggaa 660
gtgaacaaca tccgtaaaca gtatgaagcg cagggctttg aagtggtgat tccgaaaacc 720
aacattctgg cggaattccc ggtggcgtgg gttgataaaa acgtgcaggc caacggtacg 780
gaaaaagccg ccaaagccta tctgaactgg ctctatagcc cgcaggcgca aaccatcatc 840
accgactatt actaccgcgt gaataacccg gaggtgatgg acaaactgaa agacaaattc 900
ccgcagaccg agctgttccg cgtggaagac aaatttggct cctggccgga agtgatgaaa 960
acccacttca ccagcggcgg cgagttagac aagctgttag cggcggggcg taactga 1017
<210> 8
<211> 338
<212> PRT
<213> Escherichia coli
<400> 8
Met Ala Val Asn Leu Leu Lys Lys Asn Ser Leu Ala Leu Val Ala Ser
1 5 10 15
Leu Leu Leu Ala Gly His Val Gln Ala Thr Glu Leu Leu Asn Ser Ser
20 25 30
Tyr Asp Val Ser Arg Glu Leu Phe Ala Ala Leu Asn Pro Pro Phe Glu
35 40 45
Gln Gln Trp Ala Lys Asp Asn Gly Gly Asp Lys Leu Thr Ile Lys Gln
50 55 60
Ser His Ala Gly Ser Ser Lys Gln Ala Leu Ala Ile Leu Gln Gly Leu
65 70 75 80
Lys Ala Asp Val Val Thr Tyr Asn Gln Val Thr Asp Val Gln Ile Leu
85 90 95
His Asp Lys Gly Lys Leu Ile Pro Ala Asp Trp Gln Ser Arg Leu Pro
100 105 110
Asn Asn Ser Ser Pro Phe Tyr Ser Thr Met Gly Phe Leu Val Arg Lys
115 120 125
Gly Asn Pro Lys Asn Ile His Asp Trp Asn Asp Leu Val Arg Ser Asp
130 135 140
Val Lys Leu Ile Phe Pro Asn Pro Lys Thr Ser Gly Asn Ala Arg Tyr
145 150 155 160
Thr Tyr Leu Ala Ala Trp Gly Ala Ala Asp Lys Ala Asp Gly Gly Asp
165 170 175
Lys Gly Lys Thr Glu Gln Phe Met Thr Gln Phe Leu Lys Asn Val Glu
180 185 190
Val Phe Asp Thr Gly Gly Arg Gly Ala Thr Thr Thr Phe Ala Glu Arg
195 200 205
Gly Leu Gly Asp Val Leu Ile Ser Phe Glu Ser Glu Val Asn Asn Ile
210 215 220
Arg Lys Gln Tyr Glu Ala Gln Gly Phe Glu Val Val Ile Pro Lys Thr
225 230 235 240
Asn Ile Leu Ala Glu Phe Pro Val Ala Trp Val Asp Lys Asn Val Gln
245 250 255
Ala Asn Gly Thr Glu Lys Ala Ala Lys Ala Tyr Leu Asn Trp Leu Tyr
260 265 270
Ser Pro Gln Ala Gln Thr Ile Ile Thr Asp Tyr Tyr Tyr Arg Val Asn
275 280 285
Asn Pro Glu Val Met Asp Lys Leu Lys Asp Lys Phe Pro Gln Thr Glu
290 295 300
Leu Phe Arg Val Glu Asp Lys Phe Gly Ser Trp Pro Glu Val Met Lys
305 310 315 320
Thr His Phe Thr Ser Gly Gly Glu Leu Asp Lys Leu Leu Ala Ala Gly
325 330 335
Arg Asn
<210> 9
<211> 834
<212> DNA
<213> Escherichia coli
<400> 9
atgtttgctg tctcctccag acgcgtgctg ccgggcttta ccttaagcct cggcaccagt 60
ctgctgtttg tgtgcctgat tttgctgctg ccgctctccg cgctggtgat gcaactggcc 120
cagatgagct gggcgcagta ctgggaggtg atcaccaacc cgcaggtggt cgcggcctac 180
aaagtaacgc tgctgtcggc gtttgtggca tcgattttta acggcgtttt cggtctgctg 240
atggcgtgga tcctaacccg ctatcgcttc ccaggccgca cgctgcttga tgcgctgatg 300
gatttaccct ttgcgctgcc aacggctgtc gccggtttaa cgctggcctc gctcttttcc 360
gtaaacggtt tttacggtga atggctggcg aagtttgata tcaaagtcac ctatacatgg 420
ctggggattg cggtggctat ggcctttacc agcattccgt ttgtggtgcg taccgtgcag 480
ccggtgctgg aagagttagg cccggaatat gaagaagcgg cggaaacgct tggtgcaacg 540
cgctggcaga gtttctgcaa agtggtgctg ccggagcttt ctccggcgct ggtggcgggc 600
gtggcgctgt cgtttacccg tagtcttggt gaatttggcg cggtgatttt tatcgccgga 660
aatatcgcgt ggaagacgga agtgacgtcg ctgatgattt ttgtgcgctt acaggagttt 720
gattacccgg cagcgagcgc gattgcttcg gtgatcctcg cggcatctct gctgctgctg 780
ttctcaatta acactctgca aagtcgcttt ggtcggcgtg tggtaggtca ttaa 834
<210> 10
<211> 277
<212> PRT
<213> Escherichia coli
<400> 10
Met Phe Ala Val Ser Ser Arg Arg Val Leu Pro Gly Phe Thr Leu Ser
1 5 10 15
Leu Gly Thr Ser Leu Leu Phe Val Cys Leu Ile Leu Leu Leu Pro Leu
20 25 30
Ser Ala Leu Val Met Gln Leu Ala Gln Met Ser Trp Ala Gln Tyr Trp
35 40 45
Glu Val Ile Thr Asn Pro Gln Val Val Ala Ala Tyr Lys Val Thr Leu
50 55 60
Leu Ser Ala Phe Val Ala Ser Ile Phe Asn Gly Val Phe Gly Leu Leu
65 70 75 80
Met Ala Trp Ile Leu Thr Arg Tyr Arg Phe Pro Gly Arg Thr Leu Leu
85 90 95
Asp Ala Leu Met Asp Leu Pro Phe Ala Leu Pro Thr Ala Val Ala Gly
100 105 110
Leu Thr Leu Ala Ser Leu Phe Ser Val Asn Gly Phe Tyr Gly Glu Trp
115 120 125
Leu Ala Lys Phe Asp Ile Lys Val Thr Tyr Thr Trp Leu Gly Ile Ala
130 135 140
Val Ala Met Ala Phe Thr Ser Ile Pro Phe Val Val Arg Thr Val Gln
145 150 155 160
Pro Val Leu Glu Glu Leu Gly Pro Glu Tyr Glu Glu Ala Ala Glu Thr
165 170 175
Leu Gly Ala Thr Arg Trp Gln Ser Phe Cys Lys Val Val Leu Pro Glu
180 185 190
Leu Ser Pro Ala Leu Val Ala Gly Val Ala Leu Ser Phe Thr Arg Ser
195 200 205
Leu Gly Glu Phe Gly Ala Val Ile Phe Ile Ala Gly Asn Ile Ala Trp
210 215 220
Lys Thr Glu Val Thr Ser Leu Met Ile Phe Val Arg Leu Gln Glu Phe
225 230 235 240
Asp Tyr Pro Ala Ala Ser Ala Ile Ala Ser Val Ile Leu Ala Ala Ser
245 250 255
Leu Leu Leu Leu Phe Ser Ile Asn Thr Leu Gln Ser Arg Phe Gly Arg
260 265 270
Arg Val Val Gly His
275
<210> 11
<211> 876
<212> DNA
<213> Escherichia coli
<400> 11
atggcggaag ttacccaatt gaagcgttat gacgcgcgcc cgattaactg gggcaaatgg 60
tttctgattg gcatcgggat gctggtttcg gcgttcatcc tgctggtgcc gatgatttac 120
atcttcgtgc aggcattcag caaggggctg atgccggttt tacagaatct ggccgatccg 180
gacatgctgc acgccatctg gctgacggtg atgatcgcgc tgattgccgt accggtaaac 240
ctggtgttcg gcattctgct ggcctggctg gtgacgcgct ttaacttccc tggacgccag 300
ttactgctga cgctactgga cattccgttt gccgtatcgc cggtggttgc cggtctggtg 360
tatttgctgt tctacggctc taacggcccg ctcggcggtt ggctcgacga gcataacctg 420
caaattatgt tctcctggcc gggaatggtg ctggtcacca tcttcgtgac gtgtccgttt 480
gtggtgcgcg aactggtgcc ggtgatgtta agccagggca gccaggaaga cgaagcggcg 540
attttgcttg gcgcgtccgg ctggcagatg ttccgtcgcg tcacattacc gaacatccgc 600
tgggcgctgc tttatggcgt ggtgttgacc aacgcccgcg caattggcga gtttggcgcg 660
gtgtcggtgg tttccggctc gattcgcggc gaaaccctgt cgctgccgtt acagattgaa 720
ttgctggagc aggactacaa caccgtcggc tcctttaccg ctgcggcgct gttaacgctg 780
atggcgatta tcaccctgtt tttaaaaagt atgttgcagt ggcgcctgga gaatcaggaa 840
aaacgcgcac agcaggagga acatcatgag cattga 876
<210> 12
<211> 291
<212> PRT
<213> Escherichia coli
<400> 12
Met Ala Glu Val Thr Gln Leu Lys Arg Tyr Asp Ala Arg Pro Ile Asn
1 5 10 15
Trp Gly Lys Trp Phe Leu Ile Gly Ile Gly Met Leu Val Ser Ala Phe
20 25 30
Ile Leu Leu Val Pro Met Ile Tyr Ile Phe Val Gln Ala Phe Ser Lys
35 40 45
Gly Leu Met Pro Val Leu Gln Asn Leu Ala Asp Pro Asp Met Leu His
50 55 60
Ala Ile Trp Leu Thr Val Met Ile Ala Leu Ile Ala Val Pro Val Asn
65 70 75 80
Leu Val Phe Gly Ile Leu Leu Ala Trp Leu Val Thr Arg Phe Asn Phe
85 90 95
Pro Gly Arg Gln Leu Leu Leu Thr Leu Leu Asp Ile Pro Phe Ala Val
100 105 110
Ser Pro Val Val Ala Gly Leu Val Tyr Leu Leu Phe Tyr Gly Ser Asn
115 120 125
Gly Pro Leu Gly Gly Trp Leu Asp Glu His Asn Leu Gln Ile Met Phe
130 135 140
Ser Trp Pro Gly Met Val Leu Val Thr Ile Phe Val Thr Cys Pro Phe
145 150 155 160
Val Val Arg Glu Leu Val Pro Val Met Leu Ser Gln Gly Ser Gln Glu
165 170 175
Asp Glu Ala Ala Ile Leu Leu Gly Ala Ser Gly Trp Gln Met Phe Arg
180 185 190
Arg Val Thr Leu Pro Asn Ile Arg Trp Ala Leu Leu Tyr Gly Val Val
195 200 205
Leu Thr Asn Ala Arg Ala Ile Gly Glu Phe Gly Ala Val Ser Val Val
210 215 220
Ser Gly Ser Ile Arg Gly Glu Thr Leu Ser Leu Pro Leu Gln Ile Glu
225 230 235 240
Leu Leu Glu Gln Asp Tyr Asn Thr Val Gly Ser Phe Thr Ala Ala Ala
245 250 255
Leu Leu Thr Leu Met Ala Ile Ile Thr Leu Phe Leu Lys Ser Met Leu
260 265 270
Gln Trp Arg Leu Glu Asn Gln Glu Lys Arg Ala Gln Gln Glu Glu His
275 280 285
His Glu His
290
<210> 13
<211> 1098
<212> DNA
<213> Escherichia coli
<400> 13
atgagcattg agattgccaa tattaagaag tcgtttggtc gcacccaggt gctgaacgat 60
atctcactgg atattccttc aggtcagatg gtcgcgttgc tggggccgtc cggttccggg 120
aaaaccacgc tgctgcgcat tatcgccggg ctggagcatc aaaccagcgg gcatattcgc 180
ttccacggca ccgacgtgag ccgcctgcac gcacgtgatc gtaaagtcgg tttcgtgttc 240
cagcattacg cgctgttccg ccatatgacg gtgttcgaca atatcgcttt tggcctgacg 300
gtgctgccgc gtcgcgagcg cccgaatgcc gcagccatca aagcgaaagt gacaaaattg 360
ctggaaatgg tccagcttgc ccatctggcg gatcgttatc cggcgcagct ttccggcggc 420
cagaaacagc gcgtggcgct ggcgcgcgcg ctggctgtgg aaccgcaaat tctgctgctt 480
gatgaaccgt ttggcgcgct ggatgcgcag gtgcgtaaag agctgcgtcg ctggctgcgt 540
caactccatg aagaactaaa attcaccagc gtttttgtga cccacgatca ggaagaagcg 600
accgaagtag ctgatcgtgt agttgtgatg agccagggca atattgaaca ggctgacgcg 660
ccggatcagg tatggcgcga accggcgacc cgttttgtgc tcgaatttat gggcgaagtg 720
aaccgcctgc agggaaccat tcgcggcggg cagttccatg ttggcgcgca tcgctggccg 780
ctgggctaca cacctgcgta tcaggggccg gtggatctct tcctgcgccc ttgggaagtg 840
gatatcagcc gccgtaccag cctcgattcg ccgctgccgg tacaggtact ggaagccagc 900
ccgaaaggtc actacaccca attagtggtg cagccgctgg ggtggtacaa cgaaccgctg 960
acggtcgtga tgcatggcga cgatgccccg cagcgtggcg agcgtttatt cgttggtctg 1020
caacatgcgc ggctgtataa cggcgacgag cgtatcgaaa cccgcgatga ggaacttgct 1080
ctcgcacaaa gcgcctga 1098
<210> 14
<211> 365
<212> PRT
<213> Escherichia coli
<400> 14
Met Ser Ile Glu Ile Ala Asn Ile Lys Lys Ser Phe Gly Arg Thr Gln
1 5 10 15
Val Leu Asn Asp Ile Ser Leu Asp Ile Pro Ser Gly Gln Met Val Ala
20 25 30
Leu Leu Gly Pro Ser Gly Ser Gly Lys Thr Thr Leu Leu Arg Ile Ile
35 40 45
Ala Gly Leu Glu His Gln Thr Ser Gly His Ile Arg Phe His Gly Thr
50 55 60
Asp Val Ser Arg Leu His Ala Arg Asp Arg Lys Val Gly Phe Val Phe
65 70 75 80
Gln His Tyr Ala Leu Phe Arg His Met Thr Val Phe Asp Asn Ile Ala
85 90 95
Phe Gly Leu Thr Val Leu Pro Arg Arg Glu Arg Pro Asn Ala Ala Ala
100 105 110
Ile Lys Ala Lys Val Thr Lys Leu Leu Glu Met Val Gln Leu Ala His
115 120 125
Leu Ala Asp Arg Tyr Pro Ala Gln Leu Ser Gly Gly Gln Lys Gln Arg
130 135 140
Val Ala Leu Ala Arg Ala Leu Ala Val Glu Pro Gln Ile Leu Leu Leu
145 150 155 160
Asp Glu Pro Phe Gly Ala Leu Asp Ala Gln Val Arg Lys Glu Leu Arg
165 170 175
Arg Trp Leu Arg Gln Leu His Glu Glu Leu Lys Phe Thr Ser Val Phe
180 185 190
Val Thr His Asp Gln Glu Glu Ala Thr Glu Val Ala Asp Arg Val Val
195 200 205
Val Met Ser Gln Gly Asn Ile Glu Gln Ala Asp Ala Pro Asp Gln Val
210 215 220
Trp Arg Glu Pro Ala Thr Arg Phe Val Leu Glu Phe Met Gly Glu Val
225 230 235 240
Asn Arg Leu Gln Gly Thr Ile Arg Gly Gly Gln Phe His Val Gly Ala
245 250 255
His Arg Trp Pro Leu Gly Tyr Thr Pro Ala Tyr Gln Gly Pro Val Asp
260 265 270
Leu Phe Leu Arg Pro Trp Glu Val Asp Ile Ser Arg Arg Thr Ser Leu
275 280 285
Asp Ser Pro Leu Pro Val Gln Val Leu Glu Ala Ser Pro Lys Gly His
290 295 300
Tyr Thr Gln Leu Val Val Gln Pro Leu Gly Trp Tyr Asn Glu Pro Leu
305 310 315 320
Thr Val Val Met His Gly Asp Asp Ala Pro Gln Arg Gly Glu Arg Leu
325 330 335
Phe Val Gly Leu Gln His Ala Arg Leu Tyr Asn Gly Asp Glu Arg Ile
340 345 350
Glu Thr Arg Asp Glu Glu Leu Ala Leu Ala Gln Ser Ala
355 360 365
<210> 15
<211> 912
<212> DNA
<213> Escherichia coli
<400> 15
gtgagtacat tagaacaaac aataggcaat acgcctctgg tgaagttgca gcgaatgggg 60
ccggataacg gcagtgaagt gtggttaaaa ctggaaggca ataacccggc aggttcggtg 120
aaagatcgtg cggcactttc gatgatcgtc gaggcggaaa agcgcgggga aattaaaccg 180
ggtgatgtct taatcgaagc caccagtggt aacaccggca ttgcgctggc aatgattgcc 240
gcgctgaaag gctatcgcat gaaattgctg atgcccgaca acatgagcca ggaacgccgt 300
gcggcgatgc gtgcttatgg tgcggaactg attcttgtca ccaaagagca gggcatggaa 360
ggtgcgcgcg atctggcgct ggagatggcg aatcgtggcg aaggaaagct gctcgatcag 420
ttcaataatc ccgataaccc ttatgcgcat tacaccacca ctgggccgga aatctggcag 480
caaaccggcg ggcgcatcac tcattttgtc tccagcatgg ggacgaccgg cactatcacc 540
ggcgtctcac gctttatgcg cgaacaatcc aaaccggtga ccattgtcgg cctgcaaccg 600
gaagagggca gcagcattcc cggcattcgc cgctggccta cggaatatct gccggggatt 660
ttcaacgctt ctctggtgga tgaggtgctg gatattcatc agcgcgatgc ggaaaacacc 720
atgcgcgaac tggcggtgcg ggaaggaata ttctgtggcg tcagctccgg cggcgcggtt 780
gccggagcac tgcgggtggc aaaagctaac cctgacgcgg tggtggtggc gatcatctgc 840
gatcgtggcg atcgctacct ttctaccggg gtgtttgggg aagagcattt tagccagggg 900
gcggggattt aa 912
<210> 16
<211> 303
<212> PRT
<213> Escherichia coli
<400> 16
Met Ser Thr Leu Glu Gln Thr Ile Gly Asn Thr Pro Leu Val Lys Leu
1 5 10 15
Gln Arg Met Gly Pro Asp Asn Gly Ser Glu Val Trp Leu Lys Leu Glu
20 25 30
Gly Asn Asn Pro Ala Gly Ser Val Lys Asp Arg Ala Ala Leu Ser Met
35 40 45
Ile Val Glu Ala Glu Lys Arg Gly Glu Ile Lys Pro Gly Asp Val Leu
50 55 60
Ile Glu Ala Thr Ser Gly Asn Thr Gly Ile Ala Leu Ala Met Ile Ala
65 70 75 80
Ala Leu Lys Gly Tyr Arg Met Lys Leu Leu Met Pro Asp Asn Met Ser
85 90 95
Gln Glu Arg Arg Ala Ala Met Arg Ala Tyr Gly Ala Glu Leu Ile Leu
100 105 110
Val Thr Lys Glu Gln Gly Met Glu Gly Ala Arg Asp Leu Ala Leu Glu
115 120 125
Met Ala Asn Arg Gly Glu Gly Lys Leu Leu Asp Gln Phe Asn Asn Pro
130 135 140
Asp Asn Pro Tyr Ala His Tyr Thr Thr Thr Gly Pro Glu Ile Trp Gln
145 150 155 160
Gln Thr Gly Gly Arg Ile Thr His Phe Val Ser Ser Met Gly Thr Thr
165 170 175
Gly Thr Ile Thr Gly Val Ser Arg Phe Met Arg Glu Gln Ser Lys Pro
180 185 190
Val Thr Ile Val Gly Leu Gln Pro Glu Glu Gly Ser Ser Ile Pro Gly
195 200 205
Ile Arg Arg Trp Pro Thr Glu Tyr Leu Pro Gly Ile Phe Asn Ala Ser
210 215 220
Leu Val Asp Glu Val Leu Asp Ile His Gln Arg Asp Ala Glu Asn Thr
225 230 235 240
Met Arg Glu Leu Ala Val Arg Glu Gly Ile Phe Cys Gly Val Ser Ser
245 250 255
Gly Gly Ala Val Ala Gly Ala Leu Arg Val Ala Lys Ala Asn Pro Asp
260 265 270
Ala Val Val Val Ala Ile Ile Cys Asp Arg Gly Asp Arg Tyr Leu Ser
275 280 285
Thr Gly Val Phe Gly Glu Glu His Phe Ser Gln Gly Ala Gly Ile
290 295 300
<210> 17
<211> 1800
<212> DNA
<213> Escherichia coli
<400> 17
atgacgacac aggtcccacc ttccgcgttg cttccgttga acccggagca actggcacgc 60
cttcaggcgg ccacgaccga tttaactccc acccagcttg cctgggtttc tggctatttc 120
tggggcgtac tcaatcagca gcctgctgcg cttgcagcga cgccagcgcc agccgcagaa 180
atgccgggta taactattat ctccgcctcg caaaccggca atgcgcgccg ggttgctgaa 240
gcattacgtg atgatttatt agcagcaaaa ctgaacgtta agctggtgaa cgcgggcgac 300
tataaattca aacaaatcgc cagcgaaaaa ctgctcatcg tagtgacgtc aacgcaaggg 360
gaaggggaac cgccggaaga agccgtcgcg ctgcataagt tcctgttctc caaaaaagcg 420
ccaaagctgg aaaacaccgc gtttgccgtg tttagcctcg gcgatagctc ttatgaattt 480
ttctgccagt ccgggaaaga tttcgacagc aagctggcgg aactgggtgg tgaacgcctg 540
ctcgaccgtg tcgatgccga tgttgaatac caggctgctg ccagcgagtg gcgcgcccgc 600
gtggttgatg cgcttaaatc gcgtgcgcct gtcgcggcac cttcgcaatc cgtcgctact 660
ggcgcggtaa atgaaatcca caccagcccg tacagcaaag acgcgccgct ggtggctagc 720
ctctctgtta accagaaaat taccgggcgt aactctgaaa aagacgttcg ccatatcgaa 780
attgacttag gtgactcggg catgcgttac cagccgggtg acgcgctggg cgtctggtat 840
cagaacgatc cggcactggt gaaagaactt gtcgaactgc tgtggctgaa aggcgatgaa 900
cctgtcaccg tcgagggcaa aacgttgcct ctgaacgaag cgctacagtg gcacttcgaa 960
ctgaccgtca acaccgccaa cattgttgag aattacgcca cgcttacccg cagtgaaaca 1020
ctgctgccgc tggtgggcga taaagcgaag ttacagcatt acgccgcgac gacgccgatt 1080
gttgacatgg tgcgtttctc cccggcacag cttgatgccg aagcgctaat taatctgctg 1140
cgcccgctga cgccgcgtct gtattccatc gcctcctcgc aggcggaagt cgagaacgaa 1200
gtacacgtca ccgttggtgt ggtgcgttac gacgtggaag gccgcgcccg tgccggtggt 1260
gcctccagct tcctcgctga ccgcgtggaa gaagagggcg aagtccgcgt atttatcgaa 1320
cataacgata acttccgcct gccagccaat ccagaaaccc cggtgattat gattggccca 1380
ggcaccggta ttgcgccgtt ccgcgccttt atgcagcaac gcgccgccga cgaagcgcca 1440
ggtaaaaact ggctgttctt tggtaatccg cactttacgg aagacttcct gtaccaggtg 1500
gagtggcagc gctacgtcaa agatggcgtg ctgacacgta tcgatcttgc ctggtcgcgc 1560
gatcaaaaag aaaaagttta cgtacaagac aaactgcgcg aacagggcgc ggagctgtgg 1620
cgctggatca atgatggtgc ccacatttat gtctgcggcg acgctaatcg catggcgaaa 1680
gacgttgagc aggcacttct ggaagtgatt gccgaatttg gtggcatgga caccgaagcg 1740
gcggatgaat ttttaagtga gctgcgcgta gagcgccgtt atcagcgaga tgtctactaa 1800
<210> 18
<211> 599
<212> PRT
<213> Escherichia coli
<400> 18
Met Thr Thr Gln Val Pro Pro Ser Ala Leu Leu Pro Leu Asn Pro Glu
1 5 10 15
Gln Leu Ala Arg Leu Gln Ala Ala Thr Thr Asp Leu Thr Pro Thr Gln
20 25 30
Leu Ala Trp Val Ser Gly Tyr Phe Trp Gly Val Leu Asn Gln Gln Pro
35 40 45
Ala Ala Leu Ala Ala Thr Pro Ala Pro Ala Ala Glu Met Pro Gly Ile
50 55 60
Thr Ile Ile Ser Ala Ser Gln Thr Gly Asn Ala Arg Arg Val Ala Glu
65 70 75 80
Ala Leu Arg Asp Asp Leu Leu Ala Ala Lys Leu Asn Val Lys Leu Val
85 90 95
Asn Ala Gly Asp Tyr Lys Phe Lys Gln Ile Ala Ser Glu Lys Leu Leu
100 105 110
Ile Val Val Thr Ser Thr Gln Gly Glu Gly Glu Pro Pro Glu Glu Ala
115 120 125
Val Ala Leu His Lys Phe Leu Phe Ser Lys Lys Ala Pro Lys Leu Glu
130 135 140
Asn Thr Ala Phe Ala Val Phe Ser Leu Gly Asp Ser Ser Tyr Glu Phe
145 150 155 160
Phe Cys Gln Ser Gly Lys Asp Phe Asp Ser Lys Leu Ala Glu Leu Gly
165 170 175
Gly Glu Arg Leu Leu Asp Arg Val Asp Ala Asp Val Glu Tyr Gln Ala
180 185 190
Ala Ala Ser Glu Trp Arg Ala Arg Val Val Asp Ala Leu Lys Ser Arg
195 200 205
Ala Pro Val Ala Ala Pro Ser Gln Ser Val Ala Thr Gly Ala Val Asn
210 215 220
Glu Ile His Thr Ser Pro Tyr Ser Lys Asp Ala Pro Leu Val Ala Ser
225 230 235 240
Leu Ser Val Asn Gln Lys Ile Thr Gly Arg Asn Ser Glu Lys Asp Val
245 250 255
Arg His Ile Glu Ile Asp Leu Gly Asp Ser Gly Met Arg Tyr Gln Pro
260 265 270
Gly Asp Ala Leu Gly Val Trp Tyr Gln Asn Asp Pro Ala Leu Val Lys
275 280 285
Glu Leu Val Glu Leu Leu Trp Leu Lys Gly Asp Glu Pro Val Thr Val
290 295 300
Glu Gly Lys Thr Leu Pro Leu Asn Glu Ala Leu Gln Trp His Phe Glu
305 310 315 320
Leu Thr Val Asn Thr Ala Asn Ile Val Glu Asn Tyr Ala Thr Leu Thr
325 330 335
Arg Ser Glu Thr Leu Leu Pro Leu Val Gly Asp Lys Ala Lys Leu Gln
340 345 350
His Tyr Ala Ala Thr Thr Pro Ile Val Asp Met Val Arg Phe Ser Pro
355 360 365
Ala Gln Leu Asp Ala Glu Ala Leu Ile Asn Leu Leu Arg Pro Leu Thr
370 375 380
Pro Arg Leu Tyr Ser Ile Ala Ser Ser Gln Ala Glu Val Glu Asn Glu
385 390 395 400
Val His Val Thr Val Gly Val Val Arg Tyr Asp Val Glu Gly Arg Ala
405 410 415
Arg Ala Gly Gly Ala Ser Ser Phe Leu Ala Asp Arg Val Glu Glu Glu
420 425 430
Gly Glu Val Arg Val Phe Ile Glu His Asn Asp Asn Phe Arg Leu Pro
435 440 445
Ala Asn Pro Glu Thr Pro Val Ile Met Ile Gly Pro Gly Thr Gly Ile
450 455 460
Ala Pro Phe Arg Ala Phe Met Gln Gln Arg Ala Ala Asp Glu Ala Pro
465 470 475 480
Gly Lys Asn Trp Leu Phe Phe Gly Asn Pro His Phe Thr Glu Asp Phe
485 490 495
Leu Tyr Gln Val Glu Trp Gln Arg Tyr Val Lys Asp Gly Val Leu Thr
500 505 510
Arg Ile Asp Leu Ala Trp Ser Arg Asp Gln Lys Glu Lys Val Tyr Val
515 520 525
Gln Asp Lys Leu Arg Glu Gln Gly Ala Glu Leu Trp Arg Trp Ile Asn
530 535 540
Asp Gly Ala His Ile Tyr Val Cys Gly Asp Ala Asn Arg Met Ala Lys
545 550 555 560
Asp Val Glu Gln Ala Leu Leu Glu Val Ile Ala Glu Phe Gly Gly Met
565 570 575
Asp Thr Glu Ala Ala Asp Glu Phe Leu Ser Glu Leu Arg Val Glu Arg
580 585 590
Arg Tyr Gln Arg Asp Val Tyr
595
<210> 19
<211> 1713
<212> DNA
<213> Escherichia coli
<400> 19
atgagcgaaa aacatccagg gcctttagtg gtcgaaggaa aactgacaga cgccgagcgc 60
atgaagcatg aaagcaacta cctgcgcggc accattgcgg aagatttaaa cgacggtctg 120
accggcggct ttaagggcga caacttcctg ctgattcgct tccacggcat gtatcagcag 180
gatgaccgcg acatccgcgc cgaacgtgct gaacagaagc tggagccgcg ccacgcgatg 240
ctgcttcgct gtcgtctgcc gggtggggtg attaccacta aacagtggca ggcgatcgac 300
aaatttgccg gtgaaaacac catctatggc agcattcgcc tgaccaaccg ccagacgttt 360
cagttccacg gcattctgaa aaagaacgtc aaaccggtgc accagatgct gcactcggtc 420
ggtcttgatg cgctggcgac agctaacgac atgaaccgta acgtactctg cacctcgaac 480
ccttacgagt cgcagctgca cgcggaagcg tacgagtggg cgaagaagat ttctgagcat 540
ctgttgcctc gtacccgcgc gtatgcggag atctggctcg accaggaaaa agtcgccact 600
actgatgaag aaccgatcct cggccagacc tacctgccgc gtaaattcaa aaccacggta 660
gtgatcccgc cacagaacga tatcgatctg cacgccaacg acatgaactt cgtggcgatc 720
gccgaaaacg gcaagctggt gggctttaac ctgttggtgg gcggtgggct ttccatcgaa 780
cacggcaaca agaaaaccta cgcccgcacc gcgagtgagt ttggctatct gccgctggag 840
catacgctgg cggtggccga agccgtcgtg acaactcagc gtgactgggg taaccgaacc 900
gatcgtaaaa atgccaaaac caaatacacg ctggagcgcg tgggggttga gacgtttaaa 960
gcggaagtgg agcgtcgcgc ggggatcaaa tttgaaccga tccgtccata tgagttcacc 1020
ggacgaggcg atcgtattgg ctgggttaag ggcattgatg ataactggca cctgacgctg 1080
tttatcgaaa atggtcgcat ccttgattat ccggcgcgtc cgctgaaaac cggcctgctg 1140
gagatcgcga agatccacaa aggcgatttc cgcattacgg cgaaccagaa tctgatcatc 1200
gccggtgtac cggaaagcga gaaagcgaag atcgagaaga tcgccaaaga gagcgggtta 1260
atgaatgccg tcacgccgca gcgtgaaaac tcgatggctt gcgtgtcatt cccgacttgc 1320
ccgctggcga tggcggaagc agagcgtttc ctgccgtctt ttatcgacaa catcgataat 1380
ttaatggcga aacatggtgt cagcgatgag catatcgtga tgcgtgtaac aggctgcccg 1440
aacggttgtg gtcgcgcgat gctggcggaa gtgggcctgg tgggtaaagc gccgggtcgc 1500
tacaacctgc atcttggcgg caaccgcatt gggacacgta tcccacggat gtataaagaa 1560
aacatcaccg agccggaaat cctggcgtcg cttgatgaac tgatagggcg ctgggcgaaa 1620
gagcgcgaag cgggtgaagg cttcggcgac tttacggtgc gtgcgggcat cattcgcccg 1680
gtgctcgatc cggcgcgtga tttgtgggat taa 1713
<210> 20
<211> 570
<212> PRT
<213> Escherichia coli
<400> 20
Met Ser Glu Lys His Pro Gly Pro Leu Val Val Glu Gly Lys Leu Thr
1 5 10 15
Asp Ala Glu Arg Met Lys His Glu Ser Asn Tyr Leu Arg Gly Thr Ile
20 25 30
Ala Glu Asp Leu Asn Asp Gly Leu Thr Gly Gly Phe Lys Gly Asp Asn
35 40 45
Phe Leu Leu Ile Arg Phe His Gly Met Tyr Gln Gln Asp Asp Arg Asp
50 55 60
Ile Arg Ala Glu Arg Ala Glu Gln Lys Leu Glu Pro Arg His Ala Met
65 70 75 80
Leu Leu Arg Cys Arg Leu Pro Gly Gly Val Ile Thr Thr Lys Gln Trp
85 90 95
Gln Ala Ile Asp Lys Phe Ala Gly Glu Asn Thr Ile Tyr Gly Ser Ile
100 105 110
Arg Leu Thr Asn Arg Gln Thr Phe Gln Phe His Gly Ile Leu Lys Lys
115 120 125
Asn Val Lys Pro Val His Gln Met Leu His Ser Val Gly Leu Asp Ala
130 135 140
Leu Ala Thr Ala Asn Asp Met Asn Arg Asn Val Leu Cys Thr Ser Asn
145 150 155 160
Pro Tyr Glu Ser Gln Leu His Ala Glu Ala Tyr Glu Trp Ala Lys Lys
165 170 175
Ile Ser Glu His Leu Leu Pro Arg Thr Arg Ala Tyr Ala Glu Ile Trp
180 185 190
Leu Asp Gln Glu Lys Val Ala Thr Thr Asp Glu Glu Pro Ile Leu Gly
195 200 205
Gln Thr Tyr Leu Pro Arg Lys Phe Lys Thr Thr Val Val Ile Pro Pro
210 215 220
Gln Asn Asp Ile Asp Leu His Ala Asn Asp Met Asn Phe Val Ala Ile
225 230 235 240
Ala Glu Asn Gly Lys Leu Val Gly Phe Asn Leu Leu Val Gly Gly Gly
245 250 255
Leu Ser Ile Glu His Gly Asn Lys Lys Thr Tyr Ala Arg Thr Ala Ser
260 265 270
Glu Phe Gly Tyr Leu Pro Leu Glu His Thr Leu Ala Val Ala Glu Ala
275 280 285
Val Val Thr Thr Gln Arg Asp Trp Gly Asn Arg Thr Asp Arg Lys Asn
290 295 300
Ala Lys Thr Lys Tyr Thr Leu Glu Arg Val Gly Val Glu Thr Phe Lys
305 310 315 320
Ala Glu Val Glu Arg Arg Ala Gly Ile Lys Phe Glu Pro Ile Arg Pro
325 330 335
Tyr Glu Phe Thr Gly Arg Gly Asp Arg Ile Gly Trp Val Lys Gly Ile
340 345 350
Asp Asp Asn Trp His Leu Thr Leu Phe Ile Glu Asn Gly Arg Ile Leu
355 360 365
Asp Tyr Pro Ala Arg Pro Leu Lys Thr Gly Leu Leu Glu Ile Ala Lys
370 375 380
Ile His Lys Gly Asp Phe Arg Ile Thr Ala Asn Gln Asn Leu Ile Ile
385 390 395 400
Ala Gly Val Pro Glu Ser Glu Lys Ala Lys Ile Glu Lys Ile Ala Lys
405 410 415
Glu Ser Gly Leu Met Asn Ala Val Thr Pro Gln Arg Glu Asn Ser Met
420 425 430
Ala Cys Val Ser Phe Pro Thr Cys Pro Leu Ala Met Ala Glu Ala Glu
435 440 445
Arg Phe Leu Pro Ser Phe Ile Asp Asn Ile Asp Asn Leu Met Ala Lys
450 455 460
His Gly Val Ser Asp Glu His Ile Val Met Arg Val Thr Gly Cys Pro
465 470 475 480
Asn Gly Cys Gly Arg Ala Met Leu Ala Glu Val Gly Leu Val Gly Lys
485 490 495
Ala Pro Gly Arg Tyr Asn Leu His Leu Gly Gly Asn Arg Ile Gly Thr
500 505 510
Arg Ile Pro Arg Met Tyr Lys Glu Asn Ile Thr Glu Pro Glu Ile Leu
515 520 525
Ala Ser Leu Asp Glu Leu Ile Gly Arg Trp Ala Lys Glu Arg Glu Ala
530 535 540
Gly Glu Gly Phe Gly Asp Phe Thr Val Arg Ala Gly Ile Ile Arg Pro
545 550 555 560
Val Leu Asp Pro Ala Arg Asp Leu Trp Asp
565 570
<210> 21
<211> 735
<212> DNA
<213> Escherichia coli
<400> 21
atgtccaaac tcgatctaaa cgccctgaac gaactgccga aggtagatcg cattctggcg 60
ctggcggaaa ctaacgccga actggaaaaa ctggacgctg aaggccgcgt agcctgggcg 120
ctggataatc tgcccggtga atatgtactt tcttccagct ttggcattca ggcggcggtg 180
agcctgcatc tggtgaatca aattcgcccg gatattccgg tgatcctcac cgatacgggt 240
tacttgttcc cggaaaccta ccgctttatt gacgagttaa cggacaaact caagctcaac 300
ctgaaagtgt accgtgctac cgaaagcgca gcctggcagg aagcacgcta cggaaaactg 360
tgggagcagg gcgttgaagg cattgaaaag tacaatgaca tcaacaaagt cgaaccgatg 420
aaccgggctc tgaaagaact gaatgcgcaa acctggtttg ctggcctgcg ccgtgaacaa 480
tccggcagtc gtgccaattt accggtgctg gcaattcagc gtggcgtatt taaagtgctg 540
ccgattatcg actgggataa ccgaactatt tatcagtacc tgcaaaaaca tggcctgaaa 600
tatcacccat tatgggatga aggatattta tcggtgggcg atacccatac aacccgtaaa 660
tgggaacccg gcatggcgga agaagaaacg cgtttctttg gcttaaaaag ggaatgtggg 720
ttacacgaag ggtaa 735
<210> 22
<211> 244
<212> PRT
<213> Escherichia coli
<400> 22
Met Ser Lys Leu Asp Leu Asn Ala Leu Asn Glu Leu Pro Lys Val Asp
1 5 10 15
Arg Ile Leu Ala Leu Ala Glu Thr Asn Ala Glu Leu Glu Lys Leu Asp
20 25 30
Ala Glu Gly Arg Val Ala Trp Ala Leu Asp Asn Leu Pro Gly Glu Tyr
35 40 45
Val Leu Ser Ser Ser Phe Gly Ile Gln Ala Ala Val Ser Leu His Leu
50 55 60
Val Asn Gln Ile Arg Pro Asp Ile Pro Val Ile Leu Thr Asp Thr Gly
65 70 75 80
Tyr Leu Phe Pro Glu Thr Tyr Arg Phe Ile Asp Glu Leu Thr Asp Lys
85 90 95
Leu Lys Leu Asn Leu Lys Val Tyr Arg Ala Thr Glu Ser Ala Ala Trp
100 105 110
Gln Glu Ala Arg Tyr Gly Lys Leu Trp Glu Gln Gly Val Glu Gly Ile
115 120 125
Glu Lys Tyr Asn Asp Ile Asn Lys Val Glu Pro Met Asn Arg Ala Leu
130 135 140
Lys Glu Leu Asn Ala Gln Thr Trp Phe Ala Gly Leu Arg Arg Glu Gln
145 150 155 160
Ser Gly Ser Arg Ala Asn Leu Pro Val Leu Ala Ile Gln Arg Gly Val
165 170 175
Phe Lys Val Leu Pro Ile Ile Asp Trp Asp Asn Arg Thr Ile Tyr Gln
180 185 190
Tyr Leu Gln Lys His Gly Leu Lys Tyr His Pro Leu Trp Asp Glu Gly
195 200 205
Tyr Leu Ser Val Gly Asp Thr His Thr Thr Arg Lys Trp Glu Pro Gly
210 215 220
Met Ala Glu Glu Glu Thr Arg Phe Phe Gly Leu Lys Arg Glu Cys Gly
225 230 235 240
Leu His Glu Gly
<210> 23
<211> 1095
<212> DNA
<213> Escherichia coli
<400> 23
atggcacaac agactccttt gtacgaacaa cacacgcttt gcggcgctcg catggtggat 60
ttccacggct ggatgatgcc gctgcattac ggttcgcaaa tcgacgaaca tcatgcggta 120
cgtaccgatg ccggaatgtt tgatgtgtca catatgacca tcgtcgatct tcgcggcagc 180
cgcacccggg agtttctgcg ttatctgctg gcgaacgatg tggcgaagct caccaaaagc 240
ggcaaagccc tttactcggg gatgttgaat gcctctggcg gtgtgataga tgacctcatc 300
gtctactact ttactgaaga tttcttccgc ctcgttgtta actccgccac ccgcgaaaaa 360
gacctctcct ggattaccca acacgctgaa cctttcggca tcgaaattac cgttcgtgat 420
gacctttcca tgattgccgt gcaagggccg aatgcgcagg caaaagctgc cacactgttt 480
aatgacgccc agcgtcaggc ggtggaaggg atgaaaccgt tctttggcgt gcaggcgggc 540
gatctgttta ttgccaccac tggttatacc ggtgaagcgg gctatgaaat tgcgctgccc 600
aatgaaaaag cggccgattt ctggcgtgcg ctggtggaag cgggtgttaa gccatgtggc 660
ttgggcgcgc gtgacacgct gcgtctggaa gcgggcatga atctttatgg tcaggagatg 720
gacgaaacca tctctccttt agccgccaac atgggctgga ccatcgcctg ggaaccggca 780
gatcgtgact ttatcggtcg tgaagccctg gaagtgcagc gtgagcatgg tacagaaaaa 840
ctggttggtc tggtgatgac cgaaaaaggc gtgctgcgta atgaactgcc ggtacgcttt 900
accgatgcgc agggcaacca gcatgaaggc attatcacca gcggtacttt ctccccgacg 960
ctgggttaca gcattgcgct ggcgcgcgtg ccggaaggta ttggcgaaac ggcgattgtg 1020
caaattcgca accgtgaaat gccggttaaa gtgacaaaac ctgtttttgt gcgtaacggc 1080
aaagccgtcg cgtga 1095
<210> 24
<211> 364
<212> PRT
<213> Escherichia coli
<400> 24
Met Ala Gln Gln Thr Pro Leu Tyr Glu Gln His Thr Leu Cys Gly Ala
1 5 10 15
Arg Met Val Asp Phe His Gly Trp Met Met Pro Leu His Tyr Gly Ser
20 25 30
Gln Ile Asp Glu His His Ala Val Arg Thr Asp Ala Gly Met Phe Asp
35 40 45
Val Ser His Met Thr Ile Val Asp Leu Arg Gly Ser Arg Thr Arg Glu
50 55 60
Phe Leu Arg Tyr Leu Leu Ala Asn Asp Val Ala Lys Leu Thr Lys Ser
65 70 75 80
Gly Lys Ala Leu Tyr Ser Gly Met Leu Asn Ala Ser Gly Gly Val Ile
85 90 95
Asp Asp Leu Ile Val Tyr Tyr Phe Thr Glu Asp Phe Phe Arg Leu Val
100 105 110
Val Asn Ser Ala Thr Arg Glu Lys Asp Leu Ser Trp Ile Thr Gln His
115 120 125
Ala Glu Pro Phe Gly Ile Glu Ile Thr Val Arg Asp Asp Leu Ser Met
130 135 140
Ile Ala Val Gln Gly Pro Asn Ala Gln Ala Lys Ala Ala Thr Leu Phe
145 150 155 160
Asn Asp Ala Gln Arg Gln Ala Val Glu Gly Met Lys Pro Phe Phe Gly
165 170 175
Val Gln Ala Gly Asp Leu Phe Ile Ala Thr Thr Gly Tyr Thr Gly Glu
180 185 190
Ala Gly Tyr Glu Ile Ala Leu Pro Asn Glu Lys Ala Ala Asp Phe Trp
195 200 205
Arg Ala Leu Val Glu Ala Gly Val Lys Pro Cys Gly Leu Gly Ala Arg
210 215 220
Asp Thr Leu Arg Leu Glu Ala Gly Met Asn Leu Tyr Gly Gln Glu Met
225 230 235 240
Asp Glu Thr Ile Ser Pro Leu Ala Ala Asn Met Gly Trp Thr Ile Ala
245 250 255
Trp Glu Pro Ala Asp Arg Asp Phe Ile Gly Arg Glu Ala Leu Glu Val
260 265 270
Gln Arg Glu His Gly Thr Glu Lys Leu Val Gly Leu Val Met Thr Glu
275 280 285
Lys Gly Val Leu Arg Asn Glu Leu Pro Val Arg Phe Thr Asp Ala Gln
290 295 300
Gly Asn Gln His Glu Gly Ile Ile Thr Ser Gly Thr Phe Ser Pro Thr
305 310 315 320
Leu Gly Tyr Ser Ile Ala Leu Ala Arg Val Pro Glu Gly Ile Gly Glu
325 330 335
Thr Ala Ile Val Gln Ile Arg Asn Arg Glu Met Pro Val Lys Val Thr
340 345 350
Lys Pro Val Phe Val Arg Asn Gly Lys Ala Val Ala
355 360
<210> 25
<211> 390
<212> DNA
<213> Escherichia coli
<400> 25
atgagcaacg taccagcaga actgaaatac agcaaagaac acgaatggct gcgtaaagaa 60
gccgacggca cttacaccgt tggtattacc gaacatgctc aggagctgtt aggcgatatg 120
gtgtttgttg acctgccgga agtgggcgca acggttagcg cgggcgatga ctgcgcggtt 180
gccgaatcgg taaaagcggc gtcagacatt tatgcgccag taagcggtga aatcgtggcg 240
gtaaacgacg cactgagcga ttccccggaa ctggtgaaca gcgaaccgta tgcaggcggc 300
tggatcttta aaatcaaagc cagcgatgaa agcgaactgg aatcactgct ggatgcgacc 360
gcatacgaag cattgttaga agacgagtaa 390
<210> 26
<211> 129
<212> PRT
<213> Escherichia coli
<400> 26
Met Ser Asn Val Pro Ala Glu Leu Lys Tyr Ser Lys Glu His Glu Trp
1 5 10 15
Leu Arg Lys Glu Ala Asp Gly Thr Tyr Thr Val Gly Ile Thr Glu His
20 25 30
Ala Gln Glu Leu Leu Gly Asp Met Val Phe Val Asp Leu Pro Glu Val
35 40 45
Gly Ala Thr Val Ser Ala Gly Asp Asp Cys Ala Val Ala Glu Ser Val
50 55 60
Lys Ala Ala Ser Asp Ile Tyr Ala Pro Val Ser Gly Glu Ile Val Ala
65 70 75 80
Val Asn Asp Ala Leu Ser Asp Ser Pro Glu Leu Val Asn Ser Glu Pro
85 90 95
Tyr Ala Gly Gly Trp Ile Phe Lys Ile Lys Ala Ser Asp Glu Ser Glu
100 105 110
Leu Glu Ser Leu Leu Asp Ala Thr Ala Tyr Glu Ala Leu Leu Glu Asp
115 120 125
Glu
<210> 27
<211> 2874
<212> DNA
<213> Escherichia coli
<400> 27
atgacacaga cgttaagcca gcttgaaaac agcggcgctt ttattgaacg ccatatcgga 60
ccggacgccg cgcaacagca agaaatgctg aatgccgttg gtgcacaatc gttaaacgcg 120
ctgaccggcc agattgtgcc gaaagatatt caacttgcga caccaccgca ggttggcgca 180
ccggcgaccg aatacgccgc actggcagaa ctcaaggcta ttgccagtcg caataaacgc 240
ttcacgtctt acatcggcat gggttacacc gccgtgcagc taccgccggt tatcctgcgt 300
aacatgctgg aaaatccggg ctggtatacc gcgtacactc cgtatcaacc tgaagtctcc 360
cagggccgcc ttgaagcact gctcaacttc cagcaggtaa cgctggattt gactggactg 420
gatatggcct ctgcttctct tctggacgag gccaccgctg ccgccgaagc aatggcgatg 480
gcgaaacgcg tcagcaaact gaaaaatgcc aaccgcttct tcgtggcttc cgatgtgcat 540
ccgcaaacgc tggatgtggt ccgtactcgt gccgaaacct ttggttttga agtgattgtc 600
gatgacgcgc aaaaagtgct cgaccatcag gacgtcttcg gcgtgctgtt acagcaggta 660
ggcactaccg gtgaaattca cgactacact gcgcttatta gcgaactgaa atcacgcaaa 720
attgtggtca gcgttgccgc cgatattatg gcgctggtgc tgttaactgc gccgggtaaa 780
cagggcgcgg atattgtttt tggttcggcg caacgcttcg gcgtgccgat gggctacggt 840
ggcccacacg cggcattctt tgcggcgaaa gatgaataca aacgctcaat gccgggccgt 900
attatcggtg tatcgaaaga tgcagctggc aataccgcgc tgcgcatggc gatgcagact 960
cgcgagcaac atatccgccg tgagaaagcg aactccaaca tttgtacttc ccaggtactg 1020
ctggcaaaca tcgccagcct gtatgccgtt tatcacggcc cggttggcct gaaacgtatc 1080
gctaaccgca ttcaccgtct gaccgatatc ctggcggcgg gcctgcaaca aaaaggtctg 1140
aaactgcgcc atgcgcacta tttcgacacc ttgtgtgtgg aagtggccga caaagcgggc 1200
gtactgacgc gtgccgaagc ggctgaaatc aacctgcgta gcgatattct gaacgcggtt 1260
gggatcaccc ttgatgaaac aaccacgcgt gaaaacgtaa tgcagctttt caacgtgctg 1320
ctgggcgata accacggcct ggacatcgac acgctggaca aagacgtggc tcacgacagc 1380
cgctctatcc agcctgcgat gctgcgcgac gacgaaatcc tcacccatcc ggtgtttaat 1440
cgctaccaca gcgaaaccga aatgatgcgc tatatgcact cgctggagcg taaagatctg 1500
gcgctgaatc aggcgatgat cccgctgggt tcctgcacca tgaaactgaa cgccgccgcc 1560
gagatgatcc caatcacctg gccggaattt gccgaactgc acccgttctg cccgccggag 1620
caggccgaag gttatcagca gatgattgcg cagctggctg actggctggt gaaactgacc 1680
ggttacgacg ccgtttgtat gcagccgaac tctggcgcac agggcgaata cgcgggcctg 1740
ctggcgattc gtcattatca tgaaagccgc aacgaagggc atcgcgatat ctgcctgatc 1800
ccggcttctg cgcacggaac taaccccgct tctgcacata tggcaggaat gcaggtggtg 1860
gttgtggcgt gtgataaaaa cggcaacatc gatctgactg atctgcgcgc gaaagcggaa 1920
caggcgggcg ataacctctc ctgtatcatg gtgacttatc cttctaccca cggcgtgtat 1980
gaagaaacga tccgtgaagt gtgtgaagtc gtgcatcagt tcggcggtca ggtttacctt 2040
gatggcgcga acatgaacgc ccaggttggc atcacctcgc cgggctttat tggtgcggac 2100
gtttcacacc ttaacctaca taaaactttc tgcattccgc acggcggtgg tggtccgggt 2160
atgggaccga tcggcgtgaa agcgcatttg gcaccgtttg taccgggtca tagcgtggtg 2220
caaatcgaag gcatgttaac ccgtcagggc gcggtttctg cggcaccgtt cggtagcgcc 2280
tctatcctgc caatcagctg gatgtacatc cgcatgatgg gcgcagaagg gctgaaaaaa 2340
gcaagccagg tggcaatcct caacgccaac tatattgcca gccgcctgca ggatgccttc 2400
ccggtgctgt ataccggtcg cgacggtcgc gtggcgcacg aatgtattct cgatattcgc 2460
ccgctgaaag aagaaaccgg catcagcgag ctggatattg ccaagcgcct gatcgactac 2520
ggtttccacg cgccgacgat gtcgttcccg gtggcgggta cgctgatggt tgaaccgact 2580
gaatctgaaa gcaaagtgga actggatcgc tttatcgacg cgatgctggc tatccgcgca 2640
gaaattgacc aggtgaaagc cggtgtctgg ccgctggaag ataacccgct ggtgaacgcg 2700
ccgcacattc agagcgaact ggtcgccgag tgggcgcatc cgtacagccg tgaagttgcg 2760
gtattcccgg caggtgtggc agacaaatac tggccgacag tgaaacgtct ggatgatgtt 2820
tacggcgacc gtaacctgtt ctgctcctgc gtaccgatta gcgaatacca gtaa 2874
<210> 28
<211> 957
<212> PRT
<213> Escherichia coli
<400> 28
Met Thr Gln Thr Leu Ser Gln Leu Glu Asn Ser Gly Ala Phe Ile Glu
1 5 10 15
Arg His Ile Gly Pro Asp Ala Ala Gln Gln Gln Glu Met Leu Asn Ala
20 25 30
Val Gly Ala Gln Ser Leu Asn Ala Leu Thr Gly Gln Ile Val Pro Lys
35 40 45
Asp Ile Gln Leu Ala Thr Pro Pro Gln Val Gly Ala Pro Ala Thr Glu
50 55 60
Tyr Ala Ala Leu Ala Glu Leu Lys Ala Ile Ala Ser Arg Asn Lys Arg
65 70 75 80
Phe Thr Ser Tyr Ile Gly Met Gly Tyr Thr Ala Val Gln Leu Pro Pro
85 90 95
Val Ile Leu Arg Asn Met Leu Glu Asn Pro Gly Trp Tyr Thr Ala Tyr
100 105 110
Thr Pro Tyr Gln Pro Glu Val Ser Gln Gly Arg Leu Glu Ala Leu Leu
115 120 125
Asn Phe Gln Gln Val Thr Leu Asp Leu Thr Gly Leu Asp Met Ala Ser
130 135 140
Ala Ser Leu Leu Asp Glu Ala Thr Ala Ala Ala Glu Ala Met Ala Met
145 150 155 160
Ala Lys Arg Val Ser Lys Leu Lys Asn Ala Asn Arg Phe Phe Val Ala
165 170 175
Ser Asp Val His Pro Gln Thr Leu Asp Val Val Arg Thr Arg Ala Glu
180 185 190
Thr Phe Gly Phe Glu Val Ile Val Asp Asp Ala Gln Lys Val Leu Asp
195 200 205
His Gln Asp Val Phe Gly Val Leu Leu Gln Gln Val Gly Thr Thr Gly
210 215 220
Glu Ile His Asp Tyr Thr Ala Leu Ile Ser Glu Leu Lys Ser Arg Lys
225 230 235 240
Ile Val Val Ser Val Ala Ala Asp Ile Met Ala Leu Val Leu Leu Thr
245 250 255
Ala Pro Gly Lys Gln Gly Ala Asp Ile Val Phe Gly Ser Ala Gln Arg
260 265 270
Phe Gly Val Pro Met Gly Tyr Gly Gly Pro His Ala Ala Phe Phe Ala
275 280 285
Ala Lys Asp Glu Tyr Lys Arg Ser Met Pro Gly Arg Ile Ile Gly Val
290 295 300
Ser Lys Asp Ala Ala Gly Asn Thr Ala Leu Arg Met Ala Met Gln Thr
305 310 315 320
Arg Glu Gln His Ile Arg Arg Glu Lys Ala Asn Ser Asn Ile Cys Thr
325 330 335
Ser Gln Val Leu Leu Ala Asn Ile Ala Ser Leu Tyr Ala Val Tyr His
340 345 350
Gly Pro Val Gly Leu Lys Arg Ile Ala Asn Arg Ile His Arg Leu Thr
355 360 365
Asp Ile Leu Ala Ala Gly Leu Gln Gln Lys Gly Leu Lys Leu Arg His
370 375 380
Ala His Tyr Phe Asp Thr Leu Cys Val Glu Val Ala Asp Lys Ala Gly
385 390 395 400
Val Leu Thr Arg Ala Glu Ala Ala Glu Ile Asn Leu Arg Ser Asp Ile
405 410 415
Leu Asn Ala Val Gly Ile Thr Leu Asp Glu Thr Thr Thr Arg Glu Asn
420 425 430
Val Met Gln Leu Phe Asn Val Leu Leu Gly Asp Asn His Gly Leu Asp
435 440 445
Ile Asp Thr Leu Asp Lys Asp Val Ala His Asp Ser Arg Ser Ile Gln
450 455 460
Pro Ala Met Leu Arg Asp Asp Glu Ile Leu Thr His Pro Val Phe Asn
465 470 475 480
Arg Tyr His Ser Glu Thr Glu Met Met Arg Tyr Met His Ser Leu Glu
485 490 495
Arg Lys Asp Leu Ala Leu Asn Gln Ala Met Ile Pro Leu Gly Ser Cys
500 505 510
Thr Met Lys Leu Asn Ala Ala Ala Glu Met Ile Pro Ile Thr Trp Pro
515 520 525
Glu Phe Ala Glu Leu His Pro Phe Cys Pro Pro Glu Gln Ala Glu Gly
530 535 540
Tyr Gln Gln Met Ile Ala Gln Leu Ala Asp Trp Leu Val Lys Leu Thr
545 550 555 560
Gly Tyr Asp Ala Val Cys Met Gln Pro Asn Ser Gly Ala Gln Gly Glu
565 570 575
Tyr Ala Gly Leu Leu Ala Ile Arg His Tyr His Glu Ser Arg Asn Glu
580 585 590
Gly His Arg Asp Ile Cys Leu Ile Pro Ala Ser Ala His Gly Thr Asn
595 600 605
Pro Ala Ser Ala His Met Ala Gly Met Gln Val Val Val Val Ala Cys
610 615 620
Asp Lys Asn Gly Asn Ile Asp Leu Thr Asp Leu Arg Ala Lys Ala Glu
625 630 635 640
Gln Ala Gly Asp Asn Leu Ser Cys Ile Met Val Thr Tyr Pro Ser Thr
645 650 655
His Gly Val Tyr Glu Glu Thr Ile Arg Glu Val Cys Glu Val Val His
660 665 670
Gln Phe Gly Gly Gln Val Tyr Leu Asp Gly Ala Asn Met Asn Ala Gln
675 680 685
Val Gly Ile Thr Ser Pro Gly Phe Ile Gly Ala Asp Val Ser His Leu
690 695 700
Asn Leu His Lys Thr Phe Cys Ile Pro His Gly Gly Gly Gly Pro Gly
705 710 715 720
Met Gly Pro Ile Gly Val Lys Ala His Leu Ala Pro Phe Val Pro Gly
725 730 735
His Ser Val Val Gln Ile Glu Gly Met Leu Thr Arg Gln Gly Ala Val
740 745 750
Ser Ala Ala Pro Phe Gly Ser Ala Ser Ile Leu Pro Ile Ser Trp Met
755 760 765
Tyr Ile Arg Met Met Gly Ala Glu Gly Leu Lys Lys Ala Ser Gln Val
770 775 780
Ala Ile Leu Asn Ala Asn Tyr Ile Ala Ser Arg Leu Gln Asp Ala Phe
785 790 795 800
Pro Val Leu Tyr Thr Gly Arg Asp Gly Arg Val Ala His Glu Cys Ile
805 810 815
Leu Asp Ile Arg Pro Leu Lys Glu Glu Thr Gly Ile Ser Glu Leu Asp
820 825 830
Ile Ala Lys Arg Leu Ile Asp Tyr Gly Phe His Ala Pro Thr Met Ser
835 840 845
Phe Pro Val Ala Gly Thr Leu Met Val Glu Pro Thr Glu Ser Glu Ser
850 855 860
Lys Val Glu Leu Asp Arg Phe Ile Asp Ala Met Leu Ala Ile Arg Ala
865 870 875 880
Glu Ile Asp Gln Val Lys Ala Gly Val Trp Pro Leu Glu Asp Asn Pro
885 890 895
Leu Val Asn Ala Pro His Ile Gln Ser Glu Leu Val Ala Glu Trp Ala
900 905 910
His Pro Tyr Ser Arg Glu Val Ala Val Phe Pro Ala Gly Val Ala Asp
915 920 925
Lys Tyr Trp Pro Thr Val Lys Arg Leu Asp Asp Val Tyr Gly Asp Arg
930 935 940
Asn Leu Phe Cys Ser Cys Val Pro Ile Ser Glu Tyr Gln
945 950 955
<210> 29
<211> 891
<212> DNA
<213> Escherichia coli
<400> 29
atgagctttt ttcacgccag ccagcgggat gccctgaatc agagcctggc agaagtccag 60
gggcagatta acgtttcgtt cgagtttttc ccgccgcgta ccagtgaaat ggagcagacc 120
ctgtggaact ccatcgatcg ccttagcagc ctgaaaccga agtttgtatc ggtgacctat 180
ggcgcgaact ccggcgagcg cgaccgtacg cacagcatta ttaaaggcat taaagatcgc 240
actggtctgg aagcggcacc gcatcttact tgcattgatg cgacgcccga cgagctgcgc 300
accattgcac gcgactactg gaataacggt attcgtcata tcgtggcgct gcgtggcgat 360
ctgccgccgg gaagtggtaa gccagaaatg tatgcttctg acctggtgac gctgttaaaa 420
gaagtggcag atttcgatat ctccgtggcg gcgtatccgg aagttcaccc ggaagcaaaa 480
agcgctcagg cggatttgct taatctgaaa cgcaaagtgg atgccggagc caaccgcgcg 540
attactcagt tcttcttcga tgtcgaaagc tacctgcgtt ttcgtgaccg ctgtgtatcg 600
gcgggcattg atgtggaaat tattccggga attttgccgg tatctaactt taaacaggcg 660
aagaaatttg ccgatatgac caacgtgcgt attccggcgt ggatggcgca aatgttcgac 720
ggtctggatg atgatgccga aacccgcaaa ctggttggcg cgaatattgc catggatatg 780
gtgaagattt taagccgtga aggagtgaaa gatttccact tctatacgct taaccgtgct 840
gaaatgagtt acgcgatttg ccatacgctg ggggttcgac ctggtttata a 891
<210> 30
<211> 296
<212> PRT
<213> Escherichia coli
<400> 30
Met Ser Phe Phe His Ala Ser Gln Arg Asp Ala Leu Asn Gln Ser Leu
1 5 10 15
Ala Glu Val Gln Gly Gln Ile Asn Val Ser Phe Glu Phe Phe Pro Pro
20 25 30
Arg Thr Ser Glu Met Glu Gln Thr Leu Trp Asn Ser Ile Asp Arg Leu
35 40 45
Ser Ser Leu Lys Pro Lys Phe Val Ser Val Thr Tyr Gly Ala Asn Ser
50 55 60
Gly Glu Arg Asp Arg Thr His Ser Ile Ile Lys Gly Ile Lys Asp Arg
65 70 75 80
Thr Gly Leu Glu Ala Ala Pro His Leu Thr Cys Ile Asp Ala Thr Pro
85 90 95
Asp Glu Leu Arg Thr Ile Ala Arg Asp Tyr Trp Asn Asn Gly Ile Arg
100 105 110
His Ile Val Ala Leu Arg Gly Asp Leu Pro Pro Gly Ser Gly Lys Pro
115 120 125
Glu Met Tyr Ala Ser Asp Leu Val Thr Leu Leu Lys Glu Val Ala Asp
130 135 140
Phe Asp Ile Ser Val Ala Ala Tyr Pro Glu Val His Pro Glu Ala Lys
145 150 155 160
Ser Ala Gln Ala Asp Leu Leu Asn Leu Lys Arg Lys Val Asp Ala Gly
165 170 175
Ala Asn Arg Ala Ile Thr Gln Phe Phe Phe Asp Val Glu Ser Tyr Leu
180 185 190
Arg Phe Arg Asp Arg Cys Val Ser Ala Gly Ile Asp Val Glu Ile Ile
195 200 205
Pro Gly Ile Leu Pro Val Ser Asn Phe Lys Gln Ala Lys Lys Phe Ala
210 215 220
Asp Met Thr Asn Val Arg Ile Pro Ala Trp Met Ala Gln Met Phe Asp
225 230 235 240
Gly Leu Asp Asp Asp Ala Glu Thr Arg Lys Leu Val Gly Ala Asn Ile
245 250 255
Ala Met Asp Met Val Lys Ile Leu Ser Arg Glu Gly Val Lys Asp Phe
260 265 270
His Phe Tyr Thr Leu Asn Arg Ala Glu Met Ser Tyr Ala Ile Cys His
275 280 285
Thr Leu Gly Val Arg Pro Gly Leu
290 295
<210> 31
<211> 1233
<212> DNA
<213> Escherichia coli
<400> 31
atggcaaagg tatcgctgga gaaagacaag attaagtttc tgctggtaga aggcgtgcac 60
caaaaggcgc tggaaagcct tcgtgcagct ggttacacca acatcgaatt tcacaaaggc 120
gcgctggatg atgaacaatt aaaagaatcc atccgcgatg cccacttcat cggcctgcga 180
tcccgtaccc atctgactga agacgtgatc aacgccgcag aaaaactggt cgctattggc 240
tgtttctgta tcggaacaaa ccaggttgat ctggatgcgg cggcaaagcg cgggatcccg 300
gtatttaacg caccgttctc aaatacgcgc tctgttgcgg agctggtgat tggcgaactg 360
ctgctgctat tgcgcggcgt gccggaagcc aatgctaaag cgcaccgtgg cgtgtggaac 420
aaactggcgg cgggttcttt tgaagcgcgc ggcaaaaagc tgggtatcat cggctacggt 480
catattggta cgcaattggg cattctggct gaatcgctgg gaatgtatgt ttacttttat 540
gatattgaaa ataaactgcc gctgggcaac gccactcagg tacagcatct ttctgacctg 600
ctgaatatga gcgatgtggt gagtctgcat gtaccagaga atccgtccac caaaaatatg 660
atgggcgcga aagaaatttc actaatgaag cccggctcgc tgctgattaa tgcttcgcgc 720
ggtactgtgg tggatattcc ggcgctgtgt gatgcgctgg cgagcaaaca tctggcgggg 780
gcggcaatcg acgtattccc gacggaaccg gcgaccaata gcgatccatt tacctctccg 840
ctgtgtgaat tcgacaacgt ccttctgacg ccacacattg gcggttcgac tcaggaagcg 900
caggagaata tcggcctgga agttgcgggt aaattgatca agtattctga caatggctca 960
acgctctctg cggtgaactt cccggaagtc tcgctgccac tgcacggtgg gcgtcgtctg 1020
atgcacatcc acgaaaaccg tccgggcgtg ctaactgcgc tgaacaaaat cttcgccgag 1080
cagggcgtca acatcgccgc gcaatatctg caaacttccg cccagatggg ttatgtggtt 1140
attgatattg aagccgacga agacgttgcc gaaaaagcgc tgcaggcaat gaaagctatt 1200
ccgggtacca ttcgcgcccg tctgctgtac taa 1233
<210> 32
<211> 410
<212> PRT
<213> Escherichia coli
<400> 32
Met Ala Lys Val Ser Leu Glu Lys Asp Lys Ile Lys Phe Leu Leu Val
1 5 10 15
Glu Gly Val His Gln Lys Ala Leu Glu Ser Leu Arg Ala Ala Gly Tyr
20 25 30
Thr Asn Ile Glu Phe His Lys Gly Ala Leu Asp Asp Glu Gln Leu Lys
35 40 45
Glu Ser Ile Arg Asp Ala His Phe Ile Gly Leu Arg Ser Arg Thr His
50 55 60
Leu Thr Glu Asp Val Ile Asn Ala Ala Glu Lys Leu Val Ala Ile Gly
65 70 75 80
Cys Phe Cys Ile Gly Thr Asn Gln Val Asp Leu Asp Ala Ala Ala Lys
85 90 95
Arg Gly Ile Pro Val Phe Asn Ala Pro Phe Ser Asn Thr Arg Ser Val
100 105 110
Ala Glu Leu Val Ile Gly Glu Leu Leu Leu Leu Leu Arg Gly Val Pro
115 120 125
Glu Ala Asn Ala Lys Ala His Arg Gly Val Trp Asn Lys Leu Ala Ala
130 135 140
Gly Ser Phe Glu Ala Arg Gly Lys Lys Leu Gly Ile Ile Gly Tyr Gly
145 150 155 160
His Ile Gly Thr Gln Leu Gly Ile Leu Ala Glu Ser Leu Gly Met Tyr
165 170 175
Val Tyr Phe Tyr Asp Ile Glu Asn Lys Leu Pro Leu Gly Asn Ala Thr
180 185 190
Gln Val Gln His Leu Ser Asp Leu Leu Asn Met Ser Asp Val Val Ser
195 200 205
Leu His Val Pro Glu Asn Pro Ser Thr Lys Asn Met Met Gly Ala Lys
210 215 220
Glu Ile Ser Leu Met Lys Pro Gly Ser Leu Leu Ile Asn Ala Ser Arg
225 230 235 240
Gly Thr Val Val Asp Ile Pro Ala Leu Cys Asp Ala Leu Ala Ser Lys
245 250 255
His Leu Ala Gly Ala Ala Ile Asp Val Phe Pro Thr Glu Pro Ala Thr
260 265 270
Asn Ser Asp Pro Phe Thr Ser Pro Leu Cys Glu Phe Asp Asn Val Leu
275 280 285
Leu Thr Pro His Ile Gly Gly Ser Thr Gln Glu Ala Gln Glu Asn Ile
290 295 300
Gly Leu Glu Val Ala Gly Lys Leu Ile Lys Tyr Ser Asp Asn Gly Ser
305 310 315 320
Thr Leu Ser Ala Val Asn Phe Pro Glu Val Ser Leu Pro Leu His Gly
325 330 335
Gly Arg Arg Leu Met His Ile His Glu Asn Arg Pro Gly Val Leu Thr
340 345 350
Ala Leu Asn Lys Ile Phe Ala Glu Gln Gly Val Asn Ile Ala Ala Gln
355 360 365
Tyr Leu Gln Thr Ser Ala Gln Met Gly Tyr Val Val Ile Asp Ile Glu
370 375 380
Ala Asp Glu Asp Val Ala Glu Lys Ala Leu Gln Ala Met Lys Ala Ile
385 390 395 400
Pro Gly Thr Ile Arg Ala Arg Leu Leu Tyr
405 410
<210> 33
<211> 969
<212> DNA
<213> Escherichia coli
<400> 33
atgcctaaca ttacctggtg cgacctgcct gaagatgtct ctttatggcc gggtctgcct 60
ctttcattaa gtggtgatga agtgatgcca ctggattacc acgcaggtcg tagcggctgg 120
ctgctgtatg gtcgtgggct ggataaacaa cgtctgaccc aataccagag caaactgggt 180
gcggcgatgg tgattgttgc cgcctggtgc gtggaagatt atcaggtgat tcgtctggca 240
ggttcactca ccgcacgggc tacacgcctg gcccacgaag cgcagctgga tgtcgccccg 300
ctggggaaaa tcccgcacct gcgcacgccg ggtttgctgg tgatggatat ggactccacc 360
gccatccaga ttgaatgtat tgatgaaatt gccaaactgg ccggaacggg cgagatggtg 420
gcggaagtaa ccgaacgggc gatgcgcggc gaactcgatt ttaccgccag cctgcgcagc 480
cgtgtggcga cgctgaaagg cgctgacgcc aatattctgc aacaggtgcg tgaaaatctg 540
ccgctgatgc caggcttaac gcaactggtg ctcaagctgg aaacgctggg ctggaaagtg 600
gcgattgcct ccggcggctt tactttcttt gctgaatacc tgcgcgacaa gctgcgcctg 660
accgccgtgg tagccaatga actggagatc atggacggta aatttaccgg caatgtgatc 720
ggcgacatcg tagacgcgca gtacaaagcg aaaactctga ctcgcctcgc gcaggagtat 780
gaaatcccgc tggcgcagac cgtggcgatt ggcgatggag ccaatgacct gccgatgatc 840
aaagcggcag ggctggggat tgcctaccat gccaagccaa aagtgaatga aaaggcggaa 900
gtcaccatcc gtcacgctga cctgatgggg gtattctgca tcctctcagg cagcctgaat 960
cagaagtaa 969
<210> 34
<211> 322
<212> PRT
<213> Escherichia coli
<400> 34
Met Pro Asn Ile Thr Trp Cys Asp Leu Pro Glu Asp Val Ser Leu Trp
1 5 10 15
Pro Gly Leu Pro Leu Ser Leu Ser Gly Asp Glu Val Met Pro Leu Asp
20 25 30
Tyr His Ala Gly Arg Ser Gly Trp Leu Leu Tyr Gly Arg Gly Leu Asp
35 40 45
Lys Gln Arg Leu Thr Gln Tyr Gln Ser Lys Leu Gly Ala Ala Met Val
50 55 60
Ile Val Ala Ala Trp Cys Val Glu Asp Tyr Gln Val Ile Arg Leu Ala
65 70 75 80
Gly Ser Leu Thr Ala Arg Ala Thr Arg Leu Ala His Glu Ala Gln Leu
85 90 95
Asp Val Ala Pro Leu Gly Lys Ile Pro His Leu Arg Thr Pro Gly Leu
100 105 110
Leu Val Met Asp Met Asp Ser Thr Ala Ile Gln Ile Glu Cys Ile Asp
115 120 125
Glu Ile Ala Lys Leu Ala Gly Thr Gly Glu Met Val Ala Glu Val Thr
130 135 140
Glu Arg Ala Met Arg Gly Glu Leu Asp Phe Thr Ala Ser Leu Arg Ser
145 150 155 160
Arg Val Ala Thr Leu Lys Gly Ala Asp Ala Asn Ile Leu Gln Gln Val
165 170 175
Arg Glu Asn Leu Pro Leu Met Pro Gly Leu Thr Gln Leu Val Leu Lys
180 185 190
Leu Glu Thr Leu Gly Trp Lys Val Ala Ile Ala Ser Gly Gly Phe Thr
195 200 205
Phe Phe Ala Glu Tyr Leu Arg Asp Lys Leu Arg Leu Thr Ala Val Val
210 215 220
Ala Asn Glu Leu Glu Ile Met Asp Gly Lys Phe Thr Gly Asn Val Ile
225 230 235 240
Gly Asp Ile Val Asp Ala Gln Tyr Lys Ala Lys Thr Leu Thr Arg Leu
245 250 255
Ala Gln Glu Tyr Glu Ile Pro Leu Ala Gln Thr Val Ala Ile Gly Asp
260 265 270
Gly Ala Asn Asp Leu Pro Met Ile Lys Ala Ala Gly Leu Gly Ile Ala
275 280 285
Tyr His Ala Lys Pro Lys Val Asn Glu Lys Ala Glu Val Thr Ile Arg
290 295 300
His Ala Asp Leu Met Gly Val Phe Cys Ile Leu Ser Gly Ser Leu Asn
305 310 315 320
Gln Lys
<210> 35
<211> 1089
<212> DNA
<213> Escherichia coli
<400> 35
atggctcaaa tcttcaattt tagttctggt ccggcaatgc taccggcaga ggtgcttaaa 60
caggctcaac aggaactgcg cgactggaac ggtcttggta cgtcggtgat ggaagtgagt 120
caccgtggca aagagttcat tcaggttgca gaggaagccg agaaggattt tcgcgatctt 180
cttaatgtcc cctccaacta caaggtatta ttctgccatg gcggtggtcg cggtcagttt 240
gctgcggtac cgctgaatat tctcggtgat aaaaccaccg cagattatgt tgatgccggt 300
tactgggcgg caagtgccat taaagaagcg aaaaaatact gcacgcctaa tgtctttgac 360
gccaaagtga ctgttgatgg tctgcgcgcg gttaagccaa tgcgtgaatg gcaactctct 420
gataatgctg cttatatgca ttattgcccg aatgaaacca tcgatggtat cgccatcgac 480
gaaacgccag acttcggcgc agatgtggtg gtcgccgctg acttctcttc aaccattctt 540
tcccgtccga ttgacgtcag ccgttatggt gtaatttacg ctggcgcgca gaaaaatatc 600
ggcccggctg gcctgacaat cgtcatcgtt cgtgaagatt tgctgggcaa agcgaatatc 660
gcgtgtccgt cgattctgga ttattccatc ctcaacgata acggctccat gtttaacacg 720
ccgccgacat ttgcctggta tctatctggt ctggtcttta aatggctgaa agcgaacggc 780
ggtgtagctg aaatggataa aatcaatcag caaaaagcag aactgctata tggggtgatt 840
gataacagcg atttctaccg caatgacgtg gcgaaagcta accgttcgcg gatgaacgtg 900
ccgttccagt tggcggacag tgcgcttgac aaattgttcc ttgaagagtc ttttgctgct 960
ggccttcatg cactgaaagg tcaccgtgtg gtcggcggaa tgcgcgcttc tatttataac 1020
gccatgccgc tggaaggcgt taaagcgctg acagacttca tggttgagtt cgaacgccgt 1080
cacggttaa 1089
<210> 36
<211> 362
<212> PRT
<213> Escherichia coli
<400> 36
Met Ala Gln Ile Phe Asn Phe Ser Ser Gly Pro Ala Met Leu Pro Ala
1 5 10 15
Glu Val Leu Lys Gln Ala Gln Gln Glu Leu Arg Asp Trp Asn Gly Leu
20 25 30
Gly Thr Ser Val Met Glu Val Ser His Arg Gly Lys Glu Phe Ile Gln
35 40 45
Val Ala Glu Glu Ala Glu Lys Asp Phe Arg Asp Leu Leu Asn Val Pro
50 55 60
Ser Asn Tyr Lys Val Leu Phe Cys His Gly Gly Gly Arg Gly Gln Phe
65 70 75 80
Ala Ala Val Pro Leu Asn Ile Leu Gly Asp Lys Thr Thr Ala Asp Tyr
85 90 95
Val Asp Ala Gly Tyr Trp Ala Ala Ser Ala Ile Lys Glu Ala Lys Lys
100 105 110
Tyr Cys Thr Pro Asn Val Phe Asp Ala Lys Val Thr Val Asp Gly Leu
115 120 125
Arg Ala Val Lys Pro Met Arg Glu Trp Gln Leu Ser Asp Asn Ala Ala
130 135 140
Tyr Met His Tyr Cys Pro Asn Glu Thr Ile Asp Gly Ile Ala Ile Asp
145 150 155 160
Glu Thr Pro Asp Phe Gly Ala Asp Val Val Val Ala Ala Asp Phe Ser
165 170 175
Ser Thr Ile Leu Ser Arg Pro Ile Asp Val Ser Arg Tyr Gly Val Ile
180 185 190
Tyr Ala Gly Ala Gln Lys Asn Ile Gly Pro Ala Gly Leu Thr Ile Val
195 200 205
Ile Val Arg Glu Asp Leu Leu Gly Lys Ala Asn Ile Ala Cys Pro Ser
210 215 220
Ile Leu Asp Tyr Ser Ile Leu Asn Asp Asn Gly Ser Met Phe Asn Thr
225 230 235 240
Pro Pro Thr Phe Ala Trp Tyr Leu Ser Gly Leu Val Phe Lys Trp Leu
245 250 255
Lys Ala Asn Gly Gly Val Ala Glu Met Asp Lys Ile Asn Gln Gln Lys
260 265 270
Ala Glu Leu Leu Tyr Gly Val Ile Asp Asn Ser Asp Phe Tyr Arg Asn
275 280 285
Asp Val Ala Lys Ala Asn Arg Ser Arg Met Asn Val Pro Phe Gln Leu
290 295 300
Ala Asp Ser Ala Leu Asp Lys Leu Phe Leu Glu Glu Ser Phe Ala Ala
305 310 315 320
Gly Leu His Ala Leu Lys Gly His Arg Val Val Gly Gly Met Arg Ala
325 330 335
Ser Ile Tyr Asn Ala Met Pro Leu Glu Gly Val Lys Ala Leu Thr Asp
340 345 350
Phe Met Val Glu Phe Glu Arg Arg His Gly
355 360
<210> 37
<211> 822
<212> DNA
<213> Escherichia coli
<400> 37
atgtcgtgtg aagaactgga aattgtctgg aacaatatta aagccgaagc cagaacgctg 60
gcggactgtg agccaatgct ggccagtttt taccacgcga cgctactcaa gcacgaaaac 120
cttggcagtg cactgagcta catgctggcg aacaagctgt catcgccaat tatgcctgct 180
attgctatcc gtgaagtggt ggaagaagcc tacgccgctg acccggaaat gatcgcctct 240
gcggcctgtg atattcaggc ggtgcgtacc cgcgacccgg cagtcgataa atactcaacc 300
ccgttgttat acctgaaggg ttttcatgcc ttgcaggcct atcgcatcgg tcactggttg 360
tggaatcagg ggcgtcgcgc actggcaatc tttctgcaaa accaggtttc tgtgacgttc 420
caggtcgata ttcacccggc agcaaaaatt ggtcgcggta tcatgcttga ccacgcgaca 480
ggcatcgtcg ttggtgaaac ggcggtgatt gaaaacgacg tatcgattct gcaatctgtg 540
acgcttggcg gtacgggtaa atctggtggt gaccgtcacc cgaaaattcg tgaaggtgtg 600
atgattggcg cgggcgcgaa aatcctcggc aatattgaag ttgggcgcgg cgcgaagatt 660
ggcgcaggtt ccgtggtgct gcaaccggtg ccgccgcata ccaccgccgc tggcgttccg 720
gctcgtattg tcggtaaacc agacagcgat aagccatcaa tggatatgga ccagcatttc 780
aacggtatta accatacatt tgagtatggg gatgggatct aa 822
<210> 38
<211> 273
<212> PRT
<213> Escherichia coli
<400> 38
Met Ser Cys Glu Glu Leu Glu Ile Val Trp Asn Asn Ile Lys Ala Glu
1 5 10 15
Ala Arg Thr Leu Ala Asp Cys Glu Pro Met Leu Ala Ser Phe Tyr His
20 25 30
Ala Thr Leu Leu Lys His Glu Asn Leu Gly Ser Ala Leu Ser Tyr Met
35 40 45
Leu Ala Asn Lys Leu Ser Ser Pro Ile Met Pro Ala Ile Ala Ile Arg
50 55 60
Glu Val Val Glu Glu Ala Tyr Ala Ala Asp Pro Glu Met Ile Ala Ser
65 70 75 80
Ala Ala Cys Asp Ile Gln Ala Val Arg Thr Arg Asp Pro Ala Val Asp
85 90 95
Lys Tyr Ser Thr Pro Leu Leu Tyr Leu Lys Gly Phe His Ala Leu Gln
100 105 110
Ala Tyr Arg Ile Gly His Trp Leu Trp Asn Gln Gly Arg Arg Ala Leu
115 120 125
Ala Ile Phe Leu Gln Asn Gln Val Ser Val Thr Phe Gln Val Asp Ile
130 135 140
His Pro Ala Ala Lys Ile Gly Arg Gly Ile Met Leu Asp His Ala Thr
145 150 155 160
Gly Ile Val Val Gly Glu Thr Ala Val Ile Glu Asn Asp Val Ser Ile
165 170 175
Leu Gln Ser Val Thr Leu Gly Gly Thr Gly Lys Ser Gly Gly Asp Arg
180 185 190
His Pro Lys Ile Arg Glu Gly Val Met Ile Gly Ala Gly Ala Lys Ile
195 200 205
Leu Gly Asn Ile Glu Val Gly Arg Gly Ala Lys Ile Gly Ala Gly Ser
210 215 220
Val Val Leu Gln Pro Val Pro Pro His Thr Thr Ala Ala Gly Val Pro
225 230 235 240
Ala Arg Ile Val Gly Lys Pro Asp Ser Asp Lys Pro Ser Met Asp Met
245 250 255
Asp Gln His Phe Asn Gly Ile Asn His Thr Phe Glu Tyr Gly Asp Gly
260 265 270
Ile
<210> 39
<211> 2463
<212> DNA
<213> Escherichia coli
<400> 39
atgcgagtgt tgaagttcgg cggtacatca gtggcaaatg cagaacgttt tctgcgtgtt 60
gccgatattc tggaaagcaa tgccaggcag gggcaggtgg ccaccgtcct ctctgccccc 120
gccaaaatca ccaaccacct ggtggcgatg attgaaaaaa ccattagcgg ccaggatgct 180
ttacccaata tcagcgatgc cgaacgtatt tttgccgaac ttttgacggg actcgccgcc 240
gcccagccgg ggttcccgct ggcgcaattg aaaactttcg tcgatcagga atttgcccaa 300
ataaaacatg tcctgcatgg cattagtttg ttggggcagt gcccggatag catcaacgct 360
gcgctgattt gccgtggcga gaaaatgtcg atcgccatta tggccggcgt attagaagcg 420
cgcggtcaca acgttactgt tatcgatccg gtcgaaaaac tgctggcagt ggggcattac 480
ctcgaatcta ccgtcgatat tgctgagtcc acccgccgta ttgcggcaag ccgcattccg 540
gctgatcaca tggtgctgat ggcaggtttc accgccggta atgaaaaagg cgaactggtg 600
gtgcttggac gcaacggttc cgactactct gctgcggtgc tggctgcctg tttacgcgcc 660
gattgttgcg agatttggac ggacgttgac ggggtctata cctgcgaccc gcgtcaggtg 720
cccgatgcga ggttgttgaa gtcgatgtcc taccaggaag cgatggagct ttcctacttc 780
ggcgctaaag ttcttcaccc ccgcaccatt acccccatcg cccagttcca gatcccttgc 840
ctgattaaaa ataccggaaa tcctcaagca ccaggtacgc tcattggtgc cagccgtgat 900
gaagacgaat taccggtcaa gggcatttcc aatctgaata acatggcaat gttcagcgtt 960
tctggtccgg ggatgaaagg gatggtcggc atggcggcgc gcgtctttgc agcgatgtca 1020
cgcgcccgta tttccgtggt gctgattacg caatcatctt ccgaatacag catcagtttc 1080
tgcgttccac aaagcgactg tgtgcgagct gaacgggcaa tgcaggaaga gttctacctg 1140
gaactgaaag aaggcttact ggagccgctg gcagtgacgg aacggctggc cattatctcg 1200
gtggtaggtg atggtatgcg caccttgcgt gggatctcgg cgaaattctt tgccgcactg 1260
gcccgcgcca atatcaacat tgtcgccatt gctcagggat cttctgaacg ctcaatctct 1320
gtcgtggtaa ataacgatga tgcgaccact ggcgtgcgcg ttactcatca gatgctgttc 1380
aataccgatc aggttatcga agtgtttgtg attggcgtcg gtggcgttgg cggtgcgctg 1440
ctggagcaac tgaagcgtca gcaaagctgg ctgaagaata aacatatcga cttacgtgtc 1500
tgcggtgttg ccaactcgaa ggctctgctc accaatgtac atggccttaa tctggaaaac 1560
tggcaggaag aactggcgca agccaaagag ccgtttaatc tcgggcgctt aattcgcctc 1620
gtgaaagaat atcatctgct gaacccggtc attgttgact gcacttccag ccaggcagtg 1680
gcggatcaat atgccgactt cctgcgcgaa ggtttccacg ttgtcacgcc gaacaaaaag 1740
gccaacacct cgtcgatgga ttactaccat cagttgcgtt atgcggcgga aaaatcgcgg 1800
cgtaaattcc tctatgacac caacgttggg gctggattac cggttattga gaacctgcaa 1860
aatctgctca atgcaggtga tgaattgatg aagttctccg gcattctttc tggttcgctt 1920
tcttatatct tcggcaagtt agacgaaggc atgagtttct ccgaggcgac cacgctggcg 1980
cgggaaatgg gttataccga accggacccg cgagatgatc tttctggtat ggatgtggcg 2040
cgtaaactat tgattctcgc tcgtgaaacg ggacgtgaac tggagctggc ggatattgaa 2100
attgaacctg tgctgcccgc agagtttaac gccgagggtg atgttgccgc ttttatggcg 2160
aatctgtcac aactcgacga tctctttgcc gcgcgcgtgg cgaaggcccg tgatgaagga 2220
aaagttttgc gctatgttgg caatattgat gaagatggcg tctgccgcgt gaagattgcc 2280
gaagtggatg gtaatgatcc gctgttcaaa gtgaaaaatg gcgaaaacgc cctggccttc 2340
tatagccact attatcagcc gctgccgttg gtactgcgcg gatatggtgc gggcaatgac 2400
gttacagctg ccggtgtctt tgctgatctg ctacgtaccc tctcatggaa gttaggagtc 2460
tga 2463
<210> 40
<211> 820
<212> PRT
<213> Escherichia coli
<400> 40
Met Arg Val Leu Lys Phe Gly Gly Thr Ser Val Ala Asn Ala Glu Arg
1 5 10 15
Phe Leu Arg Val Ala Asp Ile Leu Glu Ser Asn Ala Arg Gln Gly Gln
20 25 30
Val Ala Thr Val Leu Ser Ala Pro Ala Lys Ile Thr Asn His Leu Val
35 40 45
Ala Met Ile Glu Lys Thr Ile Ser Gly Gln Asp Ala Leu Pro Asn Ile
50 55 60
Ser Asp Ala Glu Arg Ile Phe Ala Glu Leu Leu Thr Gly Leu Ala Ala
65 70 75 80
Ala Gln Pro Gly Phe Pro Leu Ala Gln Leu Lys Thr Phe Val Asp Gln
85 90 95
Glu Phe Ala Gln Ile Lys His Val Leu His Gly Ile Ser Leu Leu Gly
100 105 110
Gln Cys Pro Asp Ser Ile Asn Ala Ala Leu Ile Cys Arg Gly Glu Lys
115 120 125
Met Ser Ile Ala Ile Met Ala Gly Val Leu Glu Ala Arg Gly His Asn
130 135 140
Val Thr Val Ile Asp Pro Val Glu Lys Leu Leu Ala Val Gly His Tyr
145 150 155 160
Leu Glu Ser Thr Val Asp Ile Ala Glu Ser Thr Arg Arg Ile Ala Ala
165 170 175
Ser Arg Ile Pro Ala Asp His Met Val Leu Met Ala Gly Phe Thr Ala
180 185 190
Gly Asn Glu Lys Gly Glu Leu Val Val Leu Gly Arg Asn Gly Ser Asp
195 200 205
Tyr Ser Ala Ala Val Leu Ala Ala Cys Leu Arg Ala Asp Cys Cys Glu
210 215 220
Ile Trp Thr Asp Val Asp Gly Val Tyr Thr Cys Asp Pro Arg Gln Val
225 230 235 240
Pro Asp Ala Arg Leu Leu Lys Ser Met Ser Tyr Gln Glu Ala Met Glu
245 250 255
Leu Ser Tyr Phe Gly Ala Lys Val Leu His Pro Arg Thr Ile Thr Pro
260 265 270
Ile Ala Gln Phe Gln Ile Pro Cys Leu Ile Lys Asn Thr Gly Asn Pro
275 280 285
Gln Ala Pro Gly Thr Leu Ile Gly Ala Ser Arg Asp Glu Asp Glu Leu
290 295 300
Pro Val Lys Gly Ile Ser Asn Leu Asn Asn Met Ala Met Phe Ser Val
305 310 315 320
Ser Gly Pro Gly Met Lys Gly Met Val Gly Met Ala Ala Arg Val Phe
325 330 335
Ala Ala Met Ser Arg Ala Arg Ile Ser Val Val Leu Ile Thr Gln Ser
340 345 350
Ser Ser Glu Tyr Ser Ile Ser Phe Cys Val Pro Gln Ser Asp Cys Val
355 360 365
Arg Ala Glu Arg Ala Met Gln Glu Glu Phe Tyr Leu Glu Leu Lys Glu
370 375 380
Gly Leu Leu Glu Pro Leu Ala Val Thr Glu Arg Leu Ala Ile Ile Ser
385 390 395 400
Val Val Gly Asp Gly Met Arg Thr Leu Arg Gly Ile Ser Ala Lys Phe
405 410 415
Phe Ala Ala Leu Ala Arg Ala Asn Ile Asn Ile Val Ala Ile Ala Gln
420 425 430
Gly Ser Ser Glu Arg Ser Ile Ser Val Val Val Asn Asn Asp Asp Ala
435 440 445
Thr Thr Gly Val Arg Val Thr His Gln Met Leu Phe Asn Thr Asp Gln
450 455 460
Val Ile Glu Val Phe Val Ile Gly Val Gly Gly Val Gly Gly Ala Leu
465 470 475 480
Leu Glu Gln Leu Lys Arg Gln Gln Ser Trp Leu Lys Asn Lys His Ile
485 490 495
Asp Leu Arg Val Cys Gly Val Ala Asn Ser Lys Ala Leu Leu Thr Asn
500 505 510
Val His Gly Leu Asn Leu Glu Asn Trp Gln Glu Glu Leu Ala Gln Ala
515 520 525
Lys Glu Pro Phe Asn Leu Gly Arg Leu Ile Arg Leu Val Lys Glu Tyr
530 535 540
His Leu Leu Asn Pro Val Ile Val Asp Cys Thr Ser Ser Gln Ala Val
545 550 555 560
Ala Asp Gln Tyr Ala Asp Phe Leu Arg Glu Gly Phe His Val Val Thr
565 570 575
Pro Asn Lys Lys Ala Asn Thr Ser Ser Met Asp Tyr Tyr His Gln Leu
580 585 590
Arg Tyr Ala Ala Glu Lys Ser Arg Arg Lys Phe Leu Tyr Asp Thr Asn
595 600 605
Val Gly Ala Gly Leu Pro Val Ile Glu Asn Leu Gln Asn Leu Leu Asn
610 615 620
Ala Gly Asp Glu Leu Met Lys Phe Ser Gly Ile Leu Ser Gly Ser Leu
625 630 635 640
Ser Tyr Ile Phe Gly Lys Leu Asp Glu Gly Met Ser Phe Ser Glu Ala
645 650 655
Thr Thr Leu Ala Arg Glu Met Gly Tyr Thr Glu Pro Asp Pro Arg Asp
660 665 670
Asp Leu Ser Gly Met Asp Val Ala Arg Lys Leu Leu Ile Leu Ala Arg
675 680 685
Glu Thr Gly Arg Glu Leu Glu Leu Ala Asp Ile Glu Ile Glu Pro Val
690 695 700
Leu Pro Ala Glu Phe Asn Ala Glu Gly Asp Val Ala Ala Phe Met Ala
705 710 715 720
Asn Leu Ser Gln Leu Asp Asp Leu Phe Ala Ala Arg Val Ala Lys Ala
725 730 735
Arg Asp Glu Gly Lys Val Leu Arg Tyr Val Gly Asn Ile Asp Glu Asp
740 745 750
Gly Val Cys Arg Val Lys Ile Ala Glu Val Asp Gly Asn Asp Pro Leu
755 760 765
Phe Lys Val Lys Asn Gly Glu Asn Ala Leu Ala Phe Tyr Ser His Tyr
770 775 780
Tyr Gln Pro Leu Pro Leu Val Leu Arg Gly Tyr Gly Ala Gly Asn Asp
785 790 795 800
Val Thr Ala Ala Gly Val Phe Ala Asp Leu Leu Arg Thr Leu Ser Trp
805 810 815
Lys Leu Gly Val
820
<210> 41
<211> 1434
<212> DNA
<213> Escherichia coli
<400> 41
atgtttaaga atgcatttgc taacctgcaa aaggtcggta aatcgctgat gctgccggta 60
tccgtactgc ctatcgcagg tattctgctg ggcgtcggtt ccgcgaattt cagctggctg 120
cccgccgttg tatcgcatgt tatggcagaa gcaggcggtt ccgtctttgc aaacatgcca 180
ctgatttttg cgatcggtgt cgccctcggc tttaccaata acgatggcgt atccgcgctg 240
gccgcagttg ttgcctatgg catcatggtt aaaaccatgg ccgtggttgc gccactggta 300
ctgcatttac ctgctgaaga aatcgcctct aaacacctgg cggatactgg cgtactcgga 360
gggattatct ccggtgcgat cgcagcgtac atgtttaacc gtttctaccg tattaagctg 420
cctgagtatc ttggcttctt tgccggtaaa cgctttgtgc cgatcatttc tggcctggct 480
gccatcttta ctggcgttgt gctgtccttc atttggccgc cgattggttc tgcaatccag 540
accttctctc agtgggctgc ttaccagaac ccggtagttg cgtttggcat ttacggtttc 600
atcgaacgtt gcctggtacc gtttggtctg caccacatct ggaacgtacc tttccagatg 660
cagattggtg aatacaccaa cgcagcaggt caggttttcc acggcgacat tccgcgttat 720
atggcgggtg acccgactgc gggtaaactg tctggtggct tcctgttcaa aatgtacggt 780
ctgccagctg ccgcaattgc tatctggcac tctgctaaac cagaaaaccg cgcgaaagtg 840
ggcggtatta tgatctccgc ggcgctgacc tcgttcctga ccggtatcac cgagccgatc 900
gagttctcct tcatgttcgt tgcgccgatc ctgtacatca tccacgcgat tctggcaggc 960
ctggcattcc caatctgtat tcttctgggg atgcgtgacg gtacgtcgtt ctcgcacggt 1020
ctgatcgact tcatcgttct gtctggtaac agcagcaaac tgtggctgtt cccgatcgtc 1080
ggtatcggtt atgcgattgt ttactacacc atcttccgcg tgctgattaa agcactggat 1140
ctgaaaacgc cgggtcgtga agacgcgact gaagatgcaa aagcgacagg taccagcgaa 1200
atggcaccgg ctctggttgc tgcatttggt ggtaaagaaa acattactaa cctcgacgca 1260
tgtattaccc gtctgcgcgt cagcgttgct gatgtgtcta aagtggatca ggccggcctg 1320
aagaaactgg gcgcagcggg cgtagtggtt gctggttctg gtgttcaggc gattttcggt 1380
actaaatccg ataacctgaa aaccgagatg gatgagtaca tccgtaacca ctaa 1434
<210> 42
<211> 477
<212> PRT
<213> Escherichia coli
<400> 42
Met Phe Lys Asn Ala Phe Ala Asn Leu Gln Lys Val Gly Lys Ser Leu
1 5 10 15
Met Leu Pro Val Ser Val Leu Pro Ile Ala Gly Ile Leu Leu Gly Val
20 25 30
Gly Ser Ala Asn Phe Ser Trp Leu Pro Ala Val Val Ser His Val Met
35 40 45
Ala Glu Ala Gly Gly Ser Val Phe Ala Asn Met Pro Leu Ile Phe Ala
50 55 60
Ile Gly Val Ala Leu Gly Phe Thr Asn Asn Asp Gly Val Ser Ala Leu
65 70 75 80
Ala Ala Val Val Ala Tyr Gly Ile Met Val Lys Thr Met Ala Val Val
85 90 95
Ala Pro Leu Val Leu His Leu Pro Ala Glu Glu Ile Ala Ser Lys His
100 105 110
Leu Ala Asp Thr Gly Val Leu Gly Gly Ile Ile Ser Gly Ala Ile Ala
115 120 125
Ala Tyr Met Phe Asn Arg Phe Tyr Arg Ile Lys Leu Pro Glu Tyr Leu
130 135 140
Gly Phe Phe Ala Gly Lys Arg Phe Val Pro Ile Ile Ser Gly Leu Ala
145 150 155 160
Ala Ile Phe Thr Gly Val Val Leu Ser Phe Ile Trp Pro Pro Ile Gly
165 170 175
Ser Ala Ile Gln Thr Phe Ser Gln Trp Ala Ala Tyr Gln Asn Pro Val
180 185 190
Val Ala Phe Gly Ile Tyr Gly Phe Ile Glu Arg Cys Leu Val Pro Phe
195 200 205
Gly Leu His His Ile Trp Asn Val Pro Phe Gln Met Gln Ile Gly Glu
210 215 220
Tyr Thr Asn Ala Ala Gly Gln Val Phe His Gly Asp Ile Pro Arg Tyr
225 230 235 240
Met Ala Gly Asp Pro Thr Ala Gly Lys Leu Ser Gly Gly Phe Leu Phe
245 250 255
Lys Met Tyr Gly Leu Pro Ala Ala Ala Ile Ala Ile Trp His Ser Ala
260 265 270
Lys Pro Glu Asn Arg Ala Lys Val Gly Gly Ile Met Ile Ser Ala Ala
275 280 285
Leu Thr Ser Phe Leu Thr Gly Ile Thr Glu Pro Ile Glu Phe Ser Phe
290 295 300
Met Phe Val Ala Pro Ile Leu Tyr Ile Ile His Ala Ile Leu Ala Gly
305 310 315 320
Leu Ala Phe Pro Ile Cys Ile Leu Leu Gly Met Arg Asp Gly Thr Ser
325 330 335
Phe Ser His Gly Leu Ile Asp Phe Ile Val Leu Ser Gly Asn Ser Ser
340 345 350
Lys Leu Trp Leu Phe Pro Ile Val Gly Ile Gly Tyr Ala Ile Val Tyr
355 360 365
Tyr Thr Ile Phe Arg Val Leu Ile Lys Ala Leu Asp Leu Lys Thr Pro
370 375 380
Gly Arg Glu Asp Ala Thr Glu Asp Ala Lys Ala Thr Gly Thr Ser Glu
385 390 395 400
Met Ala Pro Ala Leu Val Ala Ala Phe Gly Gly Lys Glu Asn Ile Thr
405 410 415
Asn Leu Asp Ala Cys Ile Thr Arg Leu Arg Val Ser Val Ala Asp Val
420 425 430
Ser Lys Val Asp Gln Ala Gly Leu Lys Lys Leu Gly Ala Ala Gly Val
435 440 445
Val Val Ala Gly Ser Gly Val Gln Ala Ile Phe Gly Thr Lys Ser Asp
450 455 460
Asn Leu Lys Thr Glu Met Asp Glu Tyr Ile Arg Asn His
465 470 475
<210> 43
<211> 738
<212> DNA
<213> Escherichia coli
<400> 43
atggaaagcc ctactccaca gcctgctcct ggttcggcga ccttcatgga aggatgcaaa 60
gacagtttac cgattgttat tagttatatt ccggtggcct ttgcgttcgg tctgaatgcg 120
acccgtctgg gattctctcc tctcgaaagc gtttttttct cctgcatcat ttatgcaggc 180
gcgagccagt tcgtcattac cgcgatgctg gcagccggga gtagtttgtg gattgctgca 240
ctgaccgtca tggcaatgga tgttcgccat gtgttgtatg gcccgtcact gcgtagccgt 300
attattcagc gtctgcaaaa atcgaaaacc gccctgtggg cgtttggcct gacggatgag 360
gtttttgccg ccgcaaccgc aaaactggta cgcaataatc gccgctggag cgagaactgg 420
atgatcggca ttgccttcag ttcatggtca tcgtgggtat ttggtacggt aataggggca 480
ttctccggca gcggcttgct gcaaggttat cccgccgttg aagctgcatt aggttttatg 540
cttccggcac tctttatgag tttcctgctc gcctctttcc agcgcaaaca atctctttgc 600
gttaccgcag cgttagttgg tgcccttgca ggcgtaacgc tattttctat tcccgtcgcc 660
attctggcag gcattgtctg tggctgcctc actgcgttaa tccaggcatt ctggcaagga 720
gcgcccgatg agctatga 738
<210> 44
<211> 245
<212> PRT
<213> Escherichia coli
<400> 44
Met Glu Ser Pro Thr Pro Gln Pro Ala Pro Gly Ser Ala Thr Phe Met
1 5 10 15
Glu Gly Cys Lys Asp Ser Leu Pro Ile Val Ile Ser Tyr Ile Pro Val
20 25 30
Ala Phe Ala Phe Gly Leu Asn Ala Thr Arg Leu Gly Phe Ser Pro Leu
35 40 45
Glu Ser Val Phe Phe Ser Cys Ile Ile Tyr Ala Gly Ala Ser Gln Phe
50 55 60
Val Ile Thr Ala Met Leu Ala Ala Gly Ser Ser Leu Trp Ile Ala Ala
65 70 75 80
Leu Thr Val Met Ala Met Asp Val Arg His Val Leu Tyr Gly Pro Ser
85 90 95
Leu Arg Ser Arg Ile Ile Gln Arg Leu Gln Lys Ser Lys Thr Ala Leu
100 105 110
Trp Ala Phe Gly Leu Thr Asp Glu Val Phe Ala Ala Ala Thr Ala Lys
115 120 125
Leu Val Arg Asn Asn Arg Arg Trp Ser Glu Asn Trp Met Ile Gly Ile
130 135 140
Ala Phe Ser Ser Trp Ser Ser Trp Val Phe Gly Thr Val Ile Gly Ala
145 150 155 160
Phe Ser Gly Ser Gly Leu Leu Gln Gly Tyr Pro Ala Val Glu Ala Ala
165 170 175
Leu Gly Phe Met Leu Pro Ala Leu Phe Met Ser Phe Leu Leu Ala Ser
180 185 190
Phe Gln Arg Lys Gln Ser Leu Cys Val Thr Ala Ala Leu Val Gly Ala
195 200 205
Leu Ala Gly Val Thr Leu Phe Ser Ile Pro Val Ala Ile Leu Ala Gly
210 215 220
Ile Val Cys Gly Cys Leu Thr Ala Leu Ile Gln Ala Phe Trp Gln Gly
225 230 235 240
Ala Pro Asp Glu Leu
245
<210> 45
<211> 336
<212> DNA
<213> Escherichia coli
<400> 45
atgagctatg aggttctgct gcttgggtta ctagttggcg tggcgaatta ttgcttccgc 60
tatttgccgc tgcgcctgcg tgtgggtaat gcccgcccaa ccaaacgtgg cgcggtaggt 120
attttgctcg acaccattgg catcgcctcg atatgcgctc tgctggttgt ctctaccgca 180
ccagaagtga tgcacgatac acgccgtttc gtgcccacgc tggtcggctt cgcggtactg 240
ggtgccagtt tctataaaac acgcagcatt atcatcccaa cactgcttag tgcgctggcc 300
tatgggctcg cctggaaagt gatggcgatt atataa 336
<210> 46
<211> 111
<212> PRT
<213> Escherichia coli
<400> 46
Met Ser Tyr Glu Val Leu Leu Leu Gly Leu Leu Val Gly Val Ala Asn
1 5 10 15
Tyr Cys Phe Arg Tyr Leu Pro Leu Arg Leu Arg Val Gly Asn Ala Arg
20 25 30
Pro Thr Lys Arg Gly Ala Val Gly Ile Leu Leu Asp Thr Ile Gly Ile
35 40 45
Ala Ser Ile Cys Ala Leu Leu Val Val Ser Thr Ala Pro Glu Val Met
50 55 60
His Asp Thr Arg Arg Phe Val Pro Thr Leu Val Gly Phe Ala Val Leu
65 70 75 80
Gly Ala Ser Phe Tyr Lys Thr Arg Ser Ile Ile Ile Pro Thr Leu Leu
85 90 95
Ser Ala Leu Ala Tyr Gly Leu Ala Trp Lys Val Met Ala Ile Ile
100 105 110
<210> 47
<211> 3114
<212> DNA
<213> Rhizobium etli
<400> 47
atgcgccagc ttggcaacaa ggtcgcagcg cgcaacctgg cgatctcggt cggcgtaccg 60
gtcgtgccgg cgaccgagcc actgccggac gatatggccg aagtggcgaa gatggcggcg 120
gcgatcggct atcccgtcat gctgaaggca tcctggggcg gcggcggtcg cggcatgcgc 180
gtcattcgtt ccgaggccga cctcgccaag gaagtgacgg aagccaagcg cgaggcgatg 240
gcggccttcg gcaaggacga ggtctatctc gaaaaactgg tcgagcgcgc ccgccacgtc 300
gaaagccaga tcctcggcga cacccacggc aatgtcgtgc atctcttcga gcgcgactgt 360
tccgttcagc gccgcaatca gaaggtcgtc gagcgcgcgc ccgcacccta tctttcggaa 420
gcgcagcgcc aggaactcgc cgcctattcg ctgaagatcg caggggcgac caactatatc 480
ggcgccggca ccgtcgaata tctgatggat gccgataccg gcaaatttta cttcatcgaa 540
gtcaatccgc gcatccaggt cgagcacacg gtgaccgaag tcgtcaccgg catcgatatc 600
gtcaaggcgc agatccacat cctggacggc gccgcgatcg gcacgccgca atccggcgtg 660
ccgaaccagg aagacatccg tctcaacggt cacgccctgc agtgccgcgt gacgacggaa 720
gatccggagc acaacttcat tccggattac ggccgcatca ccgcctatcg ctcggcttcc 780
ggcttcggca tccggcttga cggcggcacc tcttattccg gcgccatcat cacccgctat 840
tacgatccgc tgctcgtcaa ggtcacggcc tgggcgccga acccgctgga agccatttcc 900
cgcatggacc gggcgctgcg cgaattccgc atccgtggcg tcgccaccaa cctgaccttc 960
ctcgaagcga tcatcggcca tccgaaattc cgcgacaaca gctacaccac ccgcttcatc 1020
gacacgacgc cggagctctt ccagcaggtc aagcgccagg accgcgcgac gaagcttctg 1080
acctatctcg ccgacgtcac cgtcaatggc catcccgagg ccaaggacag gccgaagccc 1140
ctcgagaatg ccgccaggcc ggtggtgccc tatgccaatg gcaacggggt gaaggacggc 1200
accaagcagc tgctcgatac gctcggcccg aaaaaattcg gcgaatggat gcgcaatgag 1260
aagcgcgtgc ttctgaccga caccacgatg cgcgacggcc accagtcgct gctcgcaacc 1320
cgcatgcgta cctatgacat cgccaggatc gccggcacct attcgcatgc gctgccgaac 1380
ctcttgtcgc tcgaatgctg gggcggcgcc accttcgacg tctcgatgcg cttcctcacc 1440
gaagatccgt gggagcggct ggcgctgatc cgagaggggg cgccgaacct gctcctgcag 1500
atgctgctgc gcggcgccaa tggcgtcggt tacaccaact atcccgacaa tgtcgtcaaa 1560
tacttcgtcc gccaggcggc caaaggcggc atcgatctct tccgcgtctt cgactgcctg 1620
aactgggtcg agaatatgcg ggtgtcgatg gatgcgattg ccgaggagaa caagctctgc 1680
gaggcggcga tctgctacac cggcgatatc ctcaattccg cccgcccgaa atacgacttg 1740
aaatattaca ccaaccttgc cgtcgagctt gagaaggccg gcgcccatat cattgcggtc 1800
aaggatatgg cgggccttct gaagccggct gctgccaagg ttctgttcaa ggcgctgcgt 1860
gaagcaaccg gcctgccgat ccatttccac acgcatgaca cctcgggcat tgcggcggca 1920
acggttcttg ccgccgtcga agccggtgtc gatgccgtcg atgcggcgat ggatgcgctc 1980
tccggcaaca cctcgcaacc ctgtctcggc tcgatcgtcg aggcgctctc cggctccgag 2040
cgcgatcccg gcctcgatcc ggcatggatc cgccgcatct ccttctattg ggaagcggtg 2100
cgcaaccagt atgccgcctt cgaaagcgac ctcaagggac cggcatcgga agtctatctg 2160
catgaaatgc cgggcggcca gttcaccaac ctcaaggagc aggcccgctc gctggggctg 2220
gaaacccgct ggcaccaggt ggcgcaggcc tatgccgacg ccaaccagat gttcggcgat 2280
atcgtcaagg tgacgccatc ctccaaggtc gtcggcgaca tggcgctgat gatggtctcc 2340
caggacctga ccgtcgccga tgtcgtcagc cccgaccgcg aagtctcctt cccggaatcg 2400
gtcgtctcga tgctgaaggg cgatctcggc cagcctccgt ctggatggcc ggaagcgctg 2460
cagaagaaag cattgaaggg cgaaaagccc tatacggtgc gccccggctc gctgctcaag 2520
gaagccgatc tcgatgcgga acgcaaagtc atcgagaaga agcttgagcg cgaggtcagc 2580
gacttcgaat tcgcttccta tctgatgtat ccgaaggtct tcaccgactt tgcgcttgcc 2640
tccgatacct acggtccggt ttcggtgctg ccgacgcccg cctattttta cgggttggcg 2700
gacggcgagg agctgttcgc cgacatcgag aagggcaaga cgctcgtcat cgtcaatcag 2760
gcggtgagcg ccaccgacag ccagggcatg gtcactgtct tcttcgagct caacggccag 2820
ccgcgccgta tcaaggtgcc cgatcgggcc cacggggcga cgggagccgc cgtgcgccgc 2880
aaggccgaac ccggcaatgc cgcccatgtc ggtgcgccga tgccgggcgt catcagccgt 2940
gtctttgtct cttcaggcca ggccgtcaat gccggcgacg tgctcgtctc catcgaggcc 3000
atgaagatgg aaaccgcgat ccatgcggaa aaggacggca ccattgccga agtgctggtc 3060
aaggccggcg atcagatcga tgccaaggac ctgctggcgg tttacggcgg atga 3114
<210> 48
<211> 1037
<212> PRT
<213> Rhizobium etli
<400> 48
Met Arg Gln Leu Gly Asn Lys Val Ala Ala Arg Asn Leu Ala Ile Ser
1 5 10 15
Val Gly Val Pro Val Val Pro Ala Thr Glu Pro Leu Pro Asp Asp Met
20 25 30
Ala Glu Val Ala Lys Met Ala Ala Ala Ile Gly Tyr Pro Val Met Leu
35 40 45
Lys Ala Ser Trp Gly Gly Gly Gly Arg Gly Met Arg Val Ile Arg Ser
50 55 60
Glu Ala Asp Leu Ala Lys Glu Val Thr Glu Ala Lys Arg Glu Ala Met
65 70 75 80
Ala Ala Phe Gly Lys Asp Glu Val Tyr Leu Glu Lys Leu Val Glu Arg
85 90 95
Ala Arg His Val Glu Ser Gln Ile Leu Gly Asp Thr His Gly Asn Val
100 105 110
Val His Leu Phe Glu Arg Asp Cys Ser Val Gln Arg Arg Asn Gln Lys
115 120 125
Val Val Glu Arg Ala Pro Ala Pro Tyr Leu Ser Glu Ala Gln Arg Gln
130 135 140
Glu Leu Ala Ala Tyr Ser Leu Lys Ile Ala Gly Ala Thr Asn Tyr Ile
145 150 155 160
Gly Ala Gly Thr Val Glu Tyr Leu Met Asp Ala Asp Thr Gly Lys Phe
165 170 175
Tyr Phe Ile Glu Val Asn Pro Arg Ile Gln Val Glu His Thr Val Thr
180 185 190
Glu Val Val Thr Gly Ile Asp Ile Val Lys Ala Gln Ile His Ile Leu
195 200 205
Asp Gly Ala Ala Ile Gly Thr Pro Gln Ser Gly Val Pro Asn Gln Glu
210 215 220
Asp Ile Arg Leu Asn Gly His Ala Leu Gln Cys Arg Val Thr Thr Glu
225 230 235 240
Asp Pro Glu His Asn Phe Ile Pro Asp Tyr Gly Arg Ile Thr Ala Tyr
245 250 255
Arg Ser Ala Ser Gly Phe Gly Ile Arg Leu Asp Gly Gly Thr Ser Tyr
260 265 270
Ser Gly Ala Ile Ile Thr Arg Tyr Tyr Asp Pro Leu Leu Val Lys Val
275 280 285
Thr Ala Trp Ala Pro Asn Pro Leu Glu Ala Ile Ser Arg Met Asp Arg
290 295 300
Ala Leu Arg Glu Phe Arg Ile Arg Gly Val Ala Thr Asn Leu Thr Phe
305 310 315 320
Leu Glu Ala Ile Ile Gly His Pro Lys Phe Arg Asp Asn Ser Tyr Thr
325 330 335
Thr Arg Phe Ile Asp Thr Thr Pro Glu Leu Phe Gln Gln Val Lys Arg
340 345 350
Gln Asp Arg Ala Thr Lys Leu Leu Thr Tyr Leu Ala Asp Val Thr Val
355 360 365
Asn Gly His Pro Glu Ala Lys Asp Arg Pro Lys Pro Leu Glu Asn Ala
370 375 380
Ala Arg Pro Val Val Pro Tyr Ala Asn Gly Asn Gly Val Lys Asp Gly
385 390 395 400
Thr Lys Gln Leu Leu Asp Thr Leu Gly Pro Lys Lys Phe Gly Glu Trp
405 410 415
Met Arg Asn Glu Lys Arg Val Leu Leu Thr Asp Thr Thr Met Arg Asp
420 425 430
Gly His Gln Ser Leu Leu Ala Thr Arg Met Arg Thr Tyr Asp Ile Ala
435 440 445
Arg Ile Ala Gly Thr Tyr Ser His Ala Leu Pro Asn Leu Leu Ser Leu
450 455 460
Glu Cys Trp Gly Gly Ala Thr Phe Asp Val Ser Met Arg Phe Leu Thr
465 470 475 480
Glu Asp Pro Trp Glu Arg Leu Ala Leu Ile Arg Glu Gly Ala Pro Asn
485 490 495
Leu Leu Leu Gln Met Leu Leu Arg Gly Ala Asn Gly Val Gly Tyr Thr
500 505 510
Asn Tyr Pro Asp Asn Val Val Lys Tyr Phe Val Arg Gln Ala Ala Lys
515 520 525
Gly Gly Ile Asp Leu Phe Arg Val Phe Asp Cys Leu Asn Trp Val Glu
530 535 540
Asn Met Arg Val Ser Met Asp Ala Ile Ala Glu Glu Asn Lys Leu Cys
545 550 555 560
Glu Ala Ala Ile Cys Tyr Thr Gly Asp Ile Leu Asn Ser Ala Arg Pro
565 570 575
Lys Tyr Asp Leu Lys Tyr Tyr Thr Asn Leu Ala Val Glu Leu Glu Lys
580 585 590
Ala Gly Ala His Ile Ile Ala Val Lys Asp Met Ala Gly Leu Leu Lys
595 600 605
Pro Ala Ala Ala Lys Val Leu Phe Lys Ala Leu Arg Glu Ala Thr Gly
610 615 620
Leu Pro Ile His Phe His Thr His Asp Thr Ser Gly Ile Ala Ala Ala
625 630 635 640
Thr Val Leu Ala Ala Val Glu Ala Gly Val Asp Ala Val Asp Ala Ala
645 650 655
Met Asp Ala Leu Ser Gly Asn Thr Ser Gln Pro Cys Leu Gly Ser Ile
660 665 670
Val Glu Ala Leu Ser Gly Ser Glu Arg Asp Pro Gly Leu Asp Pro Ala
675 680 685
Trp Ile Arg Arg Ile Ser Phe Tyr Trp Glu Ala Val Arg Asn Gln Tyr
690 695 700
Ala Ala Phe Glu Ser Asp Leu Lys Gly Pro Ala Ser Glu Val Tyr Leu
705 710 715 720
His Glu Met Pro Gly Gly Gln Phe Thr Asn Leu Lys Glu Gln Ala Arg
725 730 735
Ser Leu Gly Leu Glu Thr Arg Trp His Gln Val Ala Gln Ala Tyr Ala
740 745 750
Asp Ala Asn Gln Met Phe Gly Asp Ile Val Lys Val Thr Pro Ser Ser
755 760 765
Lys Val Val Gly Asp Met Ala Leu Met Met Val Ser Gln Asp Leu Thr
770 775 780
Val Ala Asp Val Val Ser Pro Asp Arg Glu Val Ser Phe Pro Glu Ser
785 790 795 800
Val Val Ser Met Leu Lys Gly Asp Leu Gly Gln Pro Pro Ser Gly Trp
805 810 815
Pro Glu Ala Leu Gln Lys Lys Ala Leu Lys Gly Glu Lys Pro Tyr Thr
820 825 830
Val Arg Pro Gly Ser Leu Leu Lys Glu Ala Asp Leu Asp Ala Glu Arg
835 840 845
Lys Val Ile Glu Lys Lys Leu Glu Arg Glu Val Ser Asp Phe Glu Phe
850 855 860
Ala Ser Tyr Leu Met Tyr Pro Lys Val Phe Thr Asp Phe Ala Leu Ala
865 870 875 880
Ser Asp Thr Tyr Gly Pro Val Ser Val Leu Pro Thr Pro Ala Tyr Phe
885 890 895
Tyr Gly Leu Ala Asp Gly Glu Glu Leu Phe Ala Asp Ile Glu Lys Gly
900 905 910
Lys Thr Leu Val Ile Val Asn Gln Ala Val Ser Ala Thr Asp Ser Gln
915 920 925
Gly Met Val Thr Val Phe Phe Glu Leu Asn Gly Gln Pro Arg Arg Ile
930 935 940
Lys Val Pro Asp Arg Ala His Gly Ala Thr Gly Ala Ala Val Arg Arg
945 950 955 960
Lys Ala Glu Pro Gly Asn Ala Ala His Val Gly Ala Pro Met Pro Gly
965 970 975
Val Ile Ser Arg Val Phe Val Ser Ser Gly Gln Ala Val Asn Ala Gly
980 985 990
Asp Val Leu Val Ser Ile Glu Ala Met Lys Met Glu Thr Ala Ile His
995 1000 1005
Ala Glu Lys Asp Gly Thr Ile Ala Glu Val Leu Val Lys Ala Gly
1010 1015 1020
Asp Gln Ile Asp Ala Lys Asp Leu Leu Ala Val Tyr Gly Gly
1025 1030 1035
<210> 49
<211> 318
<212> DNA
<213> Escherichia coli
<400> 49
atggctgaat ggagcggcga atatatcagc ccatacgctg agcacggcaa gaagagtgaa 60
caagtcaaaa agattacggt ttccattcct cttaaggtgt taaaaatcct caccgatgaa 120
cgcacgcgtc gtcaggtgaa caacctgcgt cacgctacca acagcgagct gctgtgcgaa 180
gcgtttctgc atgcctttac cgggcaacct ttgccggatg atgccgatct gcgtaaagag 240
cgcagcgacg aaatcccgga agcggcaaaa gagatcatgc gtgagatggg gattaacccg 300
gagacgtggg aatactaa 318
<210> 50
<211> 105
<212> PRT
<213> Escherichia coli
<400> 50
Met Ala Glu Trp Ser Gly Glu Tyr Ile Ser Pro Tyr Ala Glu His Gly
1 5 10 15
Lys Lys Ser Glu Gln Val Lys Lys Ile Thr Val Ser Ile Pro Leu Lys
20 25 30
Val Leu Lys Ile Leu Thr Asp Glu Arg Thr Arg Arg Gln Val Asn Asn
35 40 45
Leu Arg His Ala Thr Asn Ser Glu Leu Leu Cys Glu Ala Phe Leu His
50 55 60
Ala Phe Thr Gly Gln Pro Leu Pro Asp Asp Ala Asp Leu Arg Lys Glu
65 70 75 80
Arg Ser Asp Glu Ile Pro Glu Ala Ala Lys Glu Ile Met Arg Glu Met
85 90 95
Gly Ile Asn Pro Glu Thr Trp Glu Tyr
100 105
<210> 51
<211> 1443
<212> DNA
<213> Escherichia coli
<400> 51
atgtccagaa ggcttcgcag aacaaaaatc gttaccacgt taggcccagc aacagatcgc 60
gataataatc ttgaaaaagt tatcgcggcg ggtgccaacg ttgtacgtat gaacttttct 120
cacggctcgc ctgaagatca caaaatgcgc gcggataaag ttcgtgagat tgccgcaaaa 180
ctggggcgtc atgtggctat tctgggtgac ctccaggggc ccaaaatccg tgtatccacc 240
tttaaagaag gcaaagtttt cctcaatatt ggggataaat tcctgctcga cgccaacctg 300
ggtaaaggtg aaggcgacaa agaaaaagtc ggtatcgact acaaaggcct gcctgctgac 360
gtcgtgcctg gtgacatcct gctgctggac gatggtcgcg tccagttaaa agtactggaa 420
gttcagggca tgaaagtgtt caccgaagtc accgtcggtg gtcccctctc caacaataaa 480
ggtatcaaca aacttggcgg cggtttgtcg gctgaagcgc tgaccgaaaa agacaaagca 540
gacattaaga ctgcggcgtt gattggcgta gattacctgg ctgtctcctt cccacgctgt 600
ggcgaagatc tgaactatgc ccgtcgcctg gcacgcgatg caggatgtga tgcgaaaatt 660
gttgccaagg ttgaacgtgc ggaagccgtt tgcagccagg atgcaatgga tgacatcatc 720
ctcgcctctg acgtggtaat ggttgcacgt ggcgacctcg gtgtggaaat tggcgacccg 780
gaactggtcg gcattcagaa agcgttgatc cgtcgtgcgc gtcagctaaa ccgagcggta 840
atcacggcga cccagatgat ggagtcaatg attactaacc cgatgccgac gcgtgcagaa 900
gtcatggacg tagcaaacgc cgttctggat ggtactgacg ctgtgatgct gtctgcagaa 960
actgccgctg ggcagtatcc gtcagaaacc gttgcagcca tggcgcgcgt ttgcctgggt 1020
gcggaaaaaa tcccgagcat caacgtttct aaacaccgtc tggacgttca gttcgacaat 1080
gtggaagaag ctattgccat gtcagcaatg tacgcagcta accacctgaa aggcgttacg 1140
gcgatcatca ccatgaccga atcgggtcgt accgcgctga tgacctcccg tatcagctct 1200
ggtctgccaa ttttcgccat gtcgcgccat gaacgtacgc tgaacctgac tgctctctat 1260
cgtggcgtta cgccggtgca ctttgatagc gctaatgacg gcgtagcagc tgccagcgaa 1320
gcggttaatc tgctgcgcga taaaggttac ttgatgtctg gtgacctggt gattgtcacc 1380
cagggcgacg tgatgagtac cgtgggttct actaatacca cgcgtatttt aacggtagag 1440
taa 1443
<210> 52
<211> 480
<212> PRT
<213> Escherichia coli
<400> 52
Met Ser Arg Arg Leu Arg Arg Thr Lys Ile Val Thr Thr Leu Gly Pro
1 5 10 15
Ala Thr Asp Arg Asp Asn Asn Leu Glu Lys Val Ile Ala Ala Gly Ala
20 25 30
Asn Val Val Arg Met Asn Phe Ser His Gly Ser Pro Glu Asp His Lys
35 40 45
Met Arg Ala Asp Lys Val Arg Glu Ile Ala Ala Lys Leu Gly Arg His
50 55 60
Val Ala Ile Leu Gly Asp Leu Gln Gly Pro Lys Ile Arg Val Ser Thr
65 70 75 80
Phe Lys Glu Gly Lys Val Phe Leu Asn Ile Gly Asp Lys Phe Leu Leu
85 90 95
Asp Ala Asn Leu Gly Lys Gly Glu Gly Asp Lys Glu Lys Val Gly Ile
100 105 110
Asp Tyr Lys Gly Leu Pro Ala Asp Val Val Pro Gly Asp Ile Leu Leu
115 120 125
Leu Asp Asp Gly Arg Val Gln Leu Lys Val Leu Glu Val Gln Gly Met
130 135 140
Lys Val Phe Thr Glu Val Thr Val Gly Gly Pro Leu Ser Asn Asn Lys
145 150 155 160
Gly Ile Asn Lys Leu Gly Gly Gly Leu Ser Ala Glu Ala Leu Thr Glu
165 170 175
Lys Asp Lys Ala Asp Ile Lys Thr Ala Ala Leu Ile Gly Val Asp Tyr
180 185 190
Leu Ala Val Ser Phe Pro Arg Cys Gly Glu Asp Leu Asn Tyr Ala Arg
195 200 205
Arg Leu Ala Arg Asp Ala Gly Cys Asp Ala Lys Ile Val Ala Lys Val
210 215 220
Glu Arg Ala Glu Ala Val Cys Ser Gln Asp Ala Met Asp Asp Ile Ile
225 230 235 240
Leu Ala Ser Asp Val Val Met Val Ala Arg Gly Asp Leu Gly Val Glu
245 250 255
Ile Gly Asp Pro Glu Leu Val Gly Ile Gln Lys Ala Leu Ile Arg Arg
260 265 270
Ala Arg Gln Leu Asn Arg Ala Val Ile Thr Ala Thr Gln Met Met Glu
275 280 285
Ser Met Ile Thr Asn Pro Met Pro Thr Arg Ala Glu Val Met Asp Val
290 295 300
Ala Asn Ala Val Leu Asp Gly Thr Asp Ala Val Met Leu Ser Ala Glu
305 310 315 320
Thr Ala Ala Gly Gln Tyr Pro Ser Glu Thr Val Ala Ala Met Ala Arg
325 330 335
Val Cys Leu Gly Ala Glu Lys Ile Pro Ser Ile Asn Val Ser Lys His
340 345 350
Arg Leu Asp Val Gln Phe Asp Asn Val Glu Glu Ala Ile Ala Met Ser
355 360 365
Ala Met Tyr Ala Ala Asn His Leu Lys Gly Val Thr Ala Ile Ile Thr
370 375 380
Met Thr Glu Ser Gly Arg Thr Ala Leu Met Thr Ser Arg Ile Ser Ser
385 390 395 400
Gly Leu Pro Ile Phe Ala Met Ser Arg His Glu Arg Thr Leu Asn Leu
405 410 415
Thr Ala Leu Tyr Arg Gly Val Thr Pro Val His Phe Asp Ser Ala Asn
420 425 430
Asp Gly Val Ala Ala Ala Ser Glu Ala Val Asn Leu Leu Arg Asp Lys
435 440 445
Gly Tyr Leu Met Ser Gly Asp Leu Val Ile Val Thr Gln Gly Asp Val
450 455 460
Met Ser Thr Val Gly Ser Thr Asn Thr Thr Arg Ile Leu Thr Val Glu
465 470 475 480
<210> 53
<211> 1413
<212> DNA
<213> Escherichia coli
<400> 53
atgaaaaaga ccaaaattgt ttgcaccatc ggaccgaaaa ccgaatctga agagatgtta 60
gctaaaatgc tggacgctgg catgaacgtt atgcgtctga acttctctca tggtgactat 120
gcagaacacg gtcagcgcat tcagaatctg cgcaacgtga tgagcaaaac tggtaaaacc 180
gccgctatcc tgcttgatac caaaggtccg gaaatccgca ccatgaaact ggaaggcggt 240
aacgacgttt ctctgaaagc tggtcagacc tttactttca ccactgataa atctgttatc 300
ggcaacagcg aaatggttgc ggtaacgtat gaaggtttca ctactgacct gtctgttggc 360
aacaccgtac tggttgacga tggtctgatc ggtatggaag ttaccgccat tgaaggtaac 420
aaagttatct gtaaagtgct gaacaacggt gacctgggcg aaaacaaagg tgtgaacctg 480
cctggcgttt ccattgctct gccagcactg gctgaaaaag acaaacagga cctgatcttt 540
ggttgcgaac aaggcgtaga ctttgttgct gcttccttta ttcgtaagcg ttctgacgtt 600
atcgaaatcc gtgagcacct gaaagcgcac ggcggcgaaa acatccacat catctccaaa 660
atcgaaaacc aggaaggcct caacaacttc gacgaaatcc tcgaagcctc tgacggcatc 720
atggttgcgc gtggcgacct gggtgtagaa atcccggtag aagaagttat cttcgcccag 780
aagatgatga tcgaaaaatg tatccgtgca cgtaaagtcg ttatcactgc gacccagatg 840
ctggattcca tgatcaaaaa cccacgcccg actcgcgcag aagccggtga cgttgcaaac 900
gccatcctcg acggtactga cgcagtgatg ctgtctggtg aatccgcaaa aggtaaatac 960
ccgctggaag cggtttctat catggcgacc atctgcgaac gtaccgaccg cgtgatgaac 1020
agccgtctcg agttcaacaa tgacaaccgt aaactgcgca ttaccgaagc ggtatgccgt 1080
ggtgccgttg aaactgctga aaaactggat gctccgctga tcgtggttgc tactcagggc 1140
ggtaaatctg ctcgcgcagt acgtaaatac ttcccggatg ccaccatcct ggcactgacc 1200
accaacgaaa aaacggctca tcagttggta ctgagcaaag gcgttgtgcc gcagcttgtt 1260
aaagagatca cttctactga tgatttctac cgtctgggta aagaactggc tctgcagagc 1320
ggtctggcac acaaaggtga cgttgtagtt atggtttctg gtgcactggt accgagcggc 1380
actactaaca ccgcatctgt tcacgtcctg taa 1413
<210> 54
<211> 470
<212> PRT
<213> Escherichia coli
<400> 54
Met Lys Lys Thr Lys Ile Val Cys Thr Ile Gly Pro Lys Thr Glu Ser
1 5 10 15
Glu Glu Met Leu Ala Lys Met Leu Asp Ala Gly Met Asn Val Met Arg
20 25 30
Leu Asn Phe Ser His Gly Asp Tyr Ala Glu His Gly Gln Arg Ile Gln
35 40 45
Asn Leu Arg Asn Val Met Ser Lys Thr Gly Lys Thr Ala Ala Ile Leu
50 55 60
Leu Asp Thr Lys Gly Pro Glu Ile Arg Thr Met Lys Leu Glu Gly Gly
65 70 75 80
Asn Asp Val Ser Leu Lys Ala Gly Gln Thr Phe Thr Phe Thr Thr Asp
85 90 95
Lys Ser Val Ile Gly Asn Ser Glu Met Val Ala Val Thr Tyr Glu Gly
100 105 110
Phe Thr Thr Asp Leu Ser Val Gly Asn Thr Val Leu Val Asp Asp Gly
115 120 125
Leu Ile Gly Met Glu Val Thr Ala Ile Glu Gly Asn Lys Val Ile Cys
130 135 140
Lys Val Leu Asn Asn Gly Asp Leu Gly Glu Asn Lys Gly Val Asn Leu
145 150 155 160
Pro Gly Val Ser Ile Ala Leu Pro Ala Leu Ala Glu Lys Asp Lys Gln
165 170 175
Asp Leu Ile Phe Gly Cys Glu Gln Gly Val Asp Phe Val Ala Ala Ser
180 185 190
Phe Ile Arg Lys Arg Ser Asp Val Ile Glu Ile Arg Glu His Leu Lys
195 200 205
Ala His Gly Gly Glu Asn Ile His Ile Ile Ser Lys Ile Glu Asn Gln
210 215 220
Glu Gly Leu Asn Asn Phe Asp Glu Ile Leu Glu Ala Ser Asp Gly Ile
225 230 235 240
Met Val Ala Arg Gly Asp Leu Gly Val Glu Ile Pro Val Glu Glu Val
245 250 255
Ile Phe Ala Gln Lys Met Met Ile Glu Lys Cys Ile Arg Ala Arg Lys
260 265 270
Val Val Ile Thr Ala Thr Gln Met Leu Asp Ser Met Ile Lys Asn Pro
275 280 285
Arg Pro Thr Arg Ala Glu Ala Gly Asp Val Ala Asn Ala Ile Leu Asp
290 295 300
Gly Thr Asp Ala Val Met Leu Ser Gly Glu Ser Ala Lys Gly Lys Tyr
305 310 315 320
Pro Leu Glu Ala Val Ser Ile Met Ala Thr Ile Cys Glu Arg Thr Asp
325 330 335
Arg Val Met Asn Ser Arg Leu Glu Phe Asn Asn Asp Asn Arg Lys Leu
340 345 350
Arg Ile Thr Glu Ala Val Cys Arg Gly Ala Val Glu Thr Ala Glu Lys
355 360 365
Leu Asp Ala Pro Leu Ile Val Val Ala Thr Gln Gly Gly Lys Ser Ala
370 375 380
Arg Ala Val Arg Lys Tyr Phe Pro Asp Ala Thr Ile Leu Ala Leu Thr
385 390 395 400
Thr Asn Glu Lys Thr Ala His Gln Leu Val Leu Ser Lys Gly Val Val
405 410 415
Pro Gln Leu Val Lys Glu Ile Thr Ser Thr Asp Asp Phe Tyr Arg Leu
420 425 430
Gly Lys Glu Leu Ala Leu Gln Ser Gly Leu Ala His Lys Gly Asp Val
435 440 445
Val Val Met Val Ser Gly Ala Leu Val Pro Ser Gly Thr Thr Asn Thr
450 455 460
Ala Ser Val His Val Leu
465 470
<210> 55
<211> 843
<212> DNA
<213> Escherichia coli
<400> 55
atgcattcac tccaacgtaa agttctgcgt actatttgtc cggaccaaaa aggtctgatc 60
gcacgtatta ccaatatttg ctacaagcac gagttaaata tcgtacagaa caatgaattt 120
gttgatcacc gtaccgggcg cttttttatg cgcacggaac tggaagggat ttttaatgat 180
tccaccctgc tggcggatct cgatagcgca ttgccagaag gctccgtgcg tgagctgaat 240
cctgccggtc gtcgccggat agtgattctg gtcactaaag aagcgcattg ccttggcgat 300
ttgttgatga aagccaatta cggcggcctg gatgtcgaaa tcgcggcagt tattggtaac 360
cacgatactt tacgttctct ggttgagcgt tttgatattc cgtttgagct ggtaagccat 420
gaagggttaa cccgcaacga gcacgatcaa aagatggcgg atgccattga tgcttatcaa 480
cctgactacg tggtgctggc gaagtatatg cgggtattaa cgccggaatt tgtggcacgc 540
ttcccgaata agatcatcaa tattcaccat tcattcctgc cagcgtttat tggcgcacgt 600
ccttatcacc aggcctatga acgtggtgtg aagattattg gcgcaaccgc tcactatgtg 660
aatgacaatc tggacgaagg cccaatcatc atgcaggacg ttattcatgt cgatcatacc 720
tacacagctg aagatatgat gcgcgcaggt cgtgacgtcg agaaaaacgt cttaagtcgt 780
gcactataca aagtactggc acagcgcgtc tttgtttacg gtaatcgaac gattattctt 840
taa 843
<210> 56
<211> 280
<212> PRT
<213> Escherichia coli
<400> 56
Met His Ser Leu Gln Arg Lys Val Leu Arg Thr Ile Cys Pro Asp Gln
1 5 10 15
Lys Gly Leu Ile Ala Arg Ile Thr Asn Ile Cys Tyr Lys His Glu Leu
20 25 30
Asn Ile Val Gln Asn Asn Glu Phe Val Asp His Arg Thr Gly Arg Phe
35 40 45
Phe Met Arg Thr Glu Leu Glu Gly Ile Phe Asn Asp Ser Thr Leu Leu
50 55 60
Ala Asp Leu Asp Ser Ala Leu Pro Glu Gly Ser Val Arg Glu Leu Asn
65 70 75 80
Pro Ala Gly Arg Arg Arg Ile Val Ile Leu Val Thr Lys Glu Ala His
85 90 95
Cys Leu Gly Asp Leu Leu Met Lys Ala Asn Tyr Gly Gly Leu Asp Val
100 105 110
Glu Ile Ala Ala Val Ile Gly Asn His Asp Thr Leu Arg Ser Leu Val
115 120 125
Glu Arg Phe Asp Ile Pro Phe Glu Leu Val Ser His Glu Gly Leu Thr
130 135 140
Arg Asn Glu His Asp Gln Lys Met Ala Asp Ala Ile Asp Ala Tyr Gln
145 150 155 160
Pro Asp Tyr Val Val Leu Ala Lys Tyr Met Arg Val Leu Thr Pro Glu
165 170 175
Phe Val Ala Arg Phe Pro Asn Lys Ile Ile Asn Ile His His Ser Phe
180 185 190
Leu Pro Ala Phe Ile Gly Ala Arg Pro Tyr His Gln Ala Tyr Glu Arg
195 200 205
Gly Val Lys Ile Ile Gly Ala Thr Ala His Tyr Val Asn Asp Asn Leu
210 215 220
Asp Glu Gly Pro Ile Ile Met Gln Asp Val Ile His Val Asp His Thr
225 230 235 240
Tyr Thr Ala Glu Asp Met Met Arg Ala Gly Arg Asp Val Glu Lys Asn
245 250 255
Val Leu Ser Arg Ala Leu Tyr Lys Val Leu Ala Gln Arg Val Phe Val
260 265 270
Tyr Gly Asn Arg Thr Ile Ile Leu
275 280
<210> 57
<211> 2262
<212> DNA
<213> Escherichia coli
<400> 57
atgacaatat tgaatcacac cctcggtttc cctcgcgttg gcctgcgtcg cgagctgaaa 60
aaagcgcaag aaagttattg ggcggggaac tccacgcgtg aagaactgct ggcggtaggg 120
cgtgaattgc gtgctcgtca ctgggatcaa caaaagcaag cgggtatcga cctgctgccg 180
gtgggcgatt ttgcctggta cgatcatgta ctgaccacca gtctgctgct gggtaacgtt 240
ccggcgcgtc atcagaacaa agatggttcg gtagatatcg acaccctgtt ccgtattggt 300
cgtggacgtg cgccgactgg cgaacctgcg gcggcagcgg aaatgaccaa atggtttaac 360
accaactatc actacatggt gccggagttc gttaaaggcc aacagttcaa actgacctgg 420
acgcagctgc tggacgaagt ggacgaggcg ctggcgctgg gccacaaggt gaaacctgtg 480
ctgctggggc cggttacctg gctgtggctg gggaaagtga aaggtgaaca atttgaccgc 540
ctgagcctgc tgaacgacat tctgccggtt tatcagcaag tgctggcaga actggcgaaa 600
cgcggcatcg agtgggtaca gattgatgaa cccgcgctgg tactggaact accacaggcg 660
tggctggacg catacaaacc cgcttacgac gcgctccagg gacaggtgaa actgctgctg 720
accacctatt ttgaaggcgt aacgccaaat ctcgacacga ttactgcgct gcctgttcag 780
ggtctgcatg ttgacctcgt acatggtaaa gatgacgttg ctgaactgca caagcgcctg 840
ccttctgact ggttgctgtc tgcgggtctg atcaatggtc gtaacgtctg gcgcgccgat 900
cttaccgaga aatatgcgca aattaaggac attgtcggca aacgtgattt gtgggtggca 960
tcttcctgct cgttgctgca cagccccatc gacctgagcg tggaaacgcg tcttgatgca 1020
gaagtgaaaa gctggtttgc cttcgcccta caaaaatgcc atgaactggc actgctgcgc 1080
gatgcgctga acagtggtga cacggcagct ctggcagagt ggagcgcccc gattcaggca 1140
cgtcgtcact ctacccgcgt acataatccg gcggtagaaa agcgtctggc ggcgatcacc 1200
gcccaggaca gccagcgtgc gaatgtctat gaagtgcgtg ctgaagccca gcgtgcgcgt 1260
tttaaactgc cagcgtggcc gaccaccacg attggttcct tcccgcaaac cacggaaatt 1320
cgtaccctgc gtctggattt caaaaagggc aatctcgacg ccaacaacta ccgcacgggc 1380
attgcggaac atatcaagca ggccattgtt gagcaggaac gtttgggact ggatgtgctg 1440
gtacatggcg aggccgagcg taatgacatg gtggaatact ttggcgagca cctcgacgga 1500
tttgtcttta cgcaaaacgg ttgggtacag agctacggtt cccgctgcgt gaagccaccg 1560
attgtcattg gtgacattag ccgcccggca ccgattaccg tggagtgggc gaagtatgcg 1620
caatcgctga ccgacaaacc ggtgaaaggg atgctgacgg ggccggtgac catactctgc 1680
tggtcgttcc cgcgtgaaga tgtcagccgt gaaaccatcg ccaaacagat tgcgctggcg 1740
ctgcgtgatg aagtggccga tctggaagcc gctggaattg gcatcatcca gattgacgaa 1800
ccggcgctgc gcgaaggttt accgctgcgt cgtagcgact gggatgcgta tctccagtgg 1860
ggcgtagagg ccttccgtat caacgccgcc gtggcgaaag atgacacaca aatccacact 1920
cacatgtgtt attgcgagtt caacgacatc atggattcga ttgcggcgct ggacgcagac 1980
gtcatcacca tcgaaacctc gcgttccgac atggagttgc tggagtcgtt tgaagagttt 2040
gattatccaa atgaaatcgg tcctggcgtc tatgacattc actcgccaaa cgtaccgagc 2100
gtggaatgga ttgaagcctt gctgaagaaa gcggcaaaac gcattccggc agagcgcctg 2160
tgggtcaacc cggactgtgg cctgaaaacg cgcggctggc cagaaacccg cgcggcactg 2220
gcgaacatgg tgcaggcggc gcagaacttg cgtcgggggt aa 2262
<210> 58
<211> 753
<212> PRT
<213> Escherichia coli
<400> 58
Met Thr Ile Leu Asn His Thr Leu Gly Phe Pro Arg Val Gly Leu Arg
1 5 10 15
Arg Glu Leu Lys Lys Ala Gln Glu Ser Tyr Trp Ala Gly Asn Ser Thr
20 25 30
Arg Glu Glu Leu Leu Ala Val Gly Arg Glu Leu Arg Ala Arg His Trp
35 40 45
Asp Gln Gln Lys Gln Ala Gly Ile Asp Leu Leu Pro Val Gly Asp Phe
50 55 60
Ala Trp Tyr Asp His Val Leu Thr Thr Ser Leu Leu Leu Gly Asn Val
65 70 75 80
Pro Ala Arg His Gln Asn Lys Asp Gly Ser Val Asp Ile Asp Thr Leu
85 90 95
Phe Arg Ile Gly Arg Gly Arg Ala Pro Thr Gly Glu Pro Ala Ala Ala
100 105 110
Ala Glu Met Thr Lys Trp Phe Asn Thr Asn Tyr His Tyr Met Val Pro
115 120 125
Glu Phe Val Lys Gly Gln Gln Phe Lys Leu Thr Trp Thr Gln Leu Leu
130 135 140
Asp Glu Val Asp Glu Ala Leu Ala Leu Gly His Lys Val Lys Pro Val
145 150 155 160
Leu Leu Gly Pro Val Thr Trp Leu Trp Leu Gly Lys Val Lys Gly Glu
165 170 175
Gln Phe Asp Arg Leu Ser Leu Leu Asn Asp Ile Leu Pro Val Tyr Gln
180 185 190
Gln Val Leu Ala Glu Leu Ala Lys Arg Gly Ile Glu Trp Val Gln Ile
195 200 205
Asp Glu Pro Ala Leu Val Leu Glu Leu Pro Gln Ala Trp Leu Asp Ala
210 215 220
Tyr Lys Pro Ala Tyr Asp Ala Leu Gln Gly Gln Val Lys Leu Leu Leu
225 230 235 240
Thr Thr Tyr Phe Glu Gly Val Thr Pro Asn Leu Asp Thr Ile Thr Ala
245 250 255
Leu Pro Val Gln Gly Leu His Val Asp Leu Val His Gly Lys Asp Asp
260 265 270
Val Ala Glu Leu His Lys Arg Leu Pro Ser Asp Trp Leu Leu Ser Ala
275 280 285
Gly Leu Ile Asn Gly Arg Asn Val Trp Arg Ala Asp Leu Thr Glu Lys
290 295 300
Tyr Ala Gln Ile Lys Asp Ile Val Gly Lys Arg Asp Leu Trp Val Ala
305 310 315 320
Ser Ser Cys Ser Leu Leu His Ser Pro Ile Asp Leu Ser Val Glu Thr
325 330 335
Arg Leu Asp Ala Glu Val Lys Ser Trp Phe Ala Phe Ala Leu Gln Lys
340 345 350
Cys His Glu Leu Ala Leu Leu Arg Asp Ala Leu Asn Ser Gly Asp Thr
355 360 365
Ala Ala Leu Ala Glu Trp Ser Ala Pro Ile Gln Ala Arg Arg His Ser
370 375 380
Thr Arg Val His Asn Pro Ala Val Glu Lys Arg Leu Ala Ala Ile Thr
385 390 395 400
Ala Gln Asp Ser Gln Arg Ala Asn Val Tyr Glu Val Arg Ala Glu Ala
405 410 415
Gln Arg Ala Arg Phe Lys Leu Pro Ala Trp Pro Thr Thr Thr Ile Gly
420 425 430
Ser Phe Pro Gln Thr Thr Glu Ile Arg Thr Leu Arg Leu Asp Phe Lys
435 440 445
Lys Gly Asn Leu Asp Ala Asn Asn Tyr Arg Thr Gly Ile Ala Glu His
450 455 460
Ile Lys Gln Ala Ile Val Glu Gln Glu Arg Leu Gly Leu Asp Val Leu
465 470 475 480
Val His Gly Glu Ala Glu Arg Asn Asp Met Val Glu Tyr Phe Gly Glu
485 490 495
His Leu Asp Gly Phe Val Phe Thr Gln Asn Gly Trp Val Gln Ser Tyr
500 505 510
Gly Ser Arg Cys Val Lys Pro Pro Ile Val Ile Gly Asp Ile Ser Arg
515 520 525
Pro Ala Pro Ile Thr Val Glu Trp Ala Lys Tyr Ala Gln Ser Leu Thr
530 535 540
Asp Lys Pro Val Lys Gly Met Leu Thr Gly Pro Val Thr Ile Leu Cys
545 550 555 560
Trp Ser Phe Pro Arg Glu Asp Val Ser Arg Glu Thr Ile Ala Lys Gln
565 570 575
Ile Ala Leu Ala Leu Arg Asp Glu Val Ala Asp Leu Glu Ala Ala Gly
580 585 590
Ile Gly Ile Ile Gln Ile Asp Glu Pro Ala Leu Arg Glu Gly Leu Pro
595 600 605
Leu Arg Arg Ser Asp Trp Asp Ala Tyr Leu Gln Trp Gly Val Glu Ala
610 615 620
Phe Arg Ile Asn Ala Ala Val Ala Lys Asp Asp Thr Gln Ile His Thr
625 630 635 640
His Met Cys Tyr Cys Glu Phe Asn Asp Ile Met Asp Ser Ile Ala Ala
645 650 655
Leu Asp Ala Asp Val Ile Thr Ile Glu Thr Ser Arg Ser Asp Met Glu
660 665 670
Leu Leu Glu Ser Phe Glu Glu Phe Asp Tyr Pro Asn Glu Ile Gly Pro
675 680 685
Gly Val Tyr Asp Ile His Ser Pro Asn Val Pro Ser Val Glu Trp Ile
690 695 700
Glu Ala Leu Leu Lys Lys Ala Ala Lys Arg Ile Pro Ala Glu Arg Leu
705 710 715 720
Trp Val Asn Pro Asp Cys Gly Leu Lys Thr Arg Gly Trp Pro Glu Thr
725 730 735
Arg Ala Ala Leu Ala Asn Met Val Gln Ala Ala Gln Asn Leu Arg Arg
740 745 750
Gly
<210> 59
<211> 1221
<212> DNA
<213> Escherichia coli
<400> 59
gtggttgctg aaaaccagcc tgggcacatt gatcaaataa agcagaccaa cgcgggcgcg 60
gtttatcgcc tgattgatca gcttggtcca gtctcgcgta tcgatctttc ccgtctggcg 120
caactggctc ctgccagtat cactaaaatt gtccgtgaga tgctcgaagc acacctggtg 180
caagagctgg aaatcaaaga agcggggaac cgtggccgtc cggcggtggg gctggtggtt 240
gaaactgaag cctggcacta tctttctctg cgcattagtc gcggggagat tttccttgct 300
ctgcgcgatc tgagcagcaa actggtggtg gaagagtcgc aggaactggc gttaaaagat 360
gacttgccat tgctggatcg tattatttcc catatcgatc agttttttat ccgccaccag 420
aaaaaacttg agcgtctaac ttcgattgcc ataaccttgc cgggaattat tgatacggaa 480
aatggtattg tacatcgcat gccgttctac gaggatgtaa aagagatgcc gctcggcgag 540
gcgctggagc agcataccgg cgttccggtt tatattcagc atgatatcag cgcatggacg 600
atggcagagg ccttgtttgg tgcctcacgc ggggcgcgcg atgtgattca ggtggttatc 660
gatcacaacg tgggggcggg cgtcattacc gatggtcatc tgctacacgc aggcagcagt 720
agtctcgtgg aaataggcca cacacaggtc gacccgtatg ggaaacgctg ttattgcggg 780
aatcacggct gcctcgaaac catcgccagc gtggacagta ttcttgagct ggcacagctg 840
cgtcttaatc aatccatgag ctcgatgtta catggacaac cgttaaccgt ggactcattg 900
tgtcaggcgg cattgcgcgg cgatctactg gcaaaagaca tcattaccgg ggtgggcgcg 960
catgtcgggc gcattcttgc catcatggtg aatttattta acccacaaaa aatactgatt 1020
ggctcaccgt taagtaaagc ggcagatatc ctcttcccgg tcatctcaga cagcatccgt 1080
cagcaggccc ttcctgcgta tagtcagcac atcagcgttg agagtactca gttttctaac 1140
cagggcacga tggcaggcgc tgcactggta aaagacgcga tgtataacgg ttctttgttg 1200
attcgtctgt tgcagggtta a 1221
<210> 60
<211> 406
<212> PRT
<213> Escherichia coli
<400> 60
Met Val Ala Glu Asn Gln Pro Gly His Ile Asp Gln Ile Lys Gln Thr
1 5 10 15
Asn Ala Gly Ala Val Tyr Arg Leu Ile Asp Gln Leu Gly Pro Val Ser
20 25 30
Arg Ile Asp Leu Ser Arg Leu Ala Gln Leu Ala Pro Ala Ser Ile Thr
35 40 45
Lys Ile Val Arg Glu Met Leu Glu Ala His Leu Val Gln Glu Leu Glu
50 55 60
Ile Lys Glu Ala Gly Asn Arg Gly Arg Pro Ala Val Gly Leu Val Val
65 70 75 80
Glu Thr Glu Ala Trp His Tyr Leu Ser Leu Arg Ile Ser Arg Gly Glu
85 90 95
Ile Phe Leu Ala Leu Arg Asp Leu Ser Ser Lys Leu Val Val Glu Glu
100 105 110
Ser Gln Glu Leu Ala Leu Lys Asp Asp Leu Pro Leu Leu Asp Arg Ile
115 120 125
Ile Ser His Ile Asp Gln Phe Phe Ile Arg His Gln Lys Lys Leu Glu
130 135 140
Arg Leu Thr Ser Ile Ala Ile Thr Leu Pro Gly Ile Ile Asp Thr Glu
145 150 155 160
Asn Gly Ile Val His Arg Met Pro Phe Tyr Glu Asp Val Lys Glu Met
165 170 175
Pro Leu Gly Glu Ala Leu Glu Gln His Thr Gly Val Pro Val Tyr Ile
180 185 190
Gln His Asp Ile Ser Ala Trp Thr Met Ala Glu Ala Leu Phe Gly Ala
195 200 205
Ser Arg Gly Ala Arg Asp Val Ile Gln Val Val Ile Asp His Asn Val
210 215 220
Gly Ala Gly Val Ile Thr Asp Gly His Leu Leu His Ala Gly Ser Ser
225 230 235 240
Ser Leu Val Glu Ile Gly His Thr Gln Val Asp Pro Tyr Gly Lys Arg
245 250 255
Cys Tyr Cys Gly Asn His Gly Cys Leu Glu Thr Ile Ala Ser Val Asp
260 265 270
Ser Ile Leu Glu Leu Ala Gln Leu Arg Leu Asn Gln Ser Met Ser Ser
275 280 285
Met Leu His Gly Gln Pro Leu Thr Val Asp Ser Leu Cys Gln Ala Ala
290 295 300
Leu Arg Gly Asp Leu Leu Ala Lys Asp Ile Ile Thr Gly Val Gly Ala
305 310 315 320
His Val Gly Arg Ile Leu Ala Ile Met Val Asn Leu Phe Asn Pro Gln
325 330 335
Lys Ile Leu Ile Gly Ser Pro Leu Ser Lys Ala Ala Asp Ile Leu Phe
340 345 350
Pro Val Ile Ser Asp Ser Ile Arg Gln Gln Ala Leu Pro Ala Tyr Ser
355 360 365
Gln His Ile Ser Val Glu Ser Thr Gln Phe Ser Asn Gln Gly Thr Met
370 375 380
Ala Gly Ala Ala Leu Val Lys Asp Ala Met Tyr Asn Gly Ser Leu Leu
385 390 395 400
Ile Arg Leu Leu Gln Gly
405
<210> 61
<211> 519
<212> DNA
<213> Escherichia coli
<400> 61
atgtccatcc gttttgcccg caaagccgac tgtgctgcca ttgcggaaat ttataaccac 60
gccgtgttgt atacggcggc tatctggaat gaccaaacgg tggatgctga taaccgcatt 120
gcctggtttg aagcgcggac tttagcaggt tatccagtgc tggtgagcga ggaaaacggc 180
gtagtgacgg gatatgcctc gtttggcgac tggcgtagtt tcgatggttt tcgccatacc 240
gtggaacatt cggtttatgt ccatcccgat catcagggca aaggtctggg gcgtaaattg 300
ttaagccgat tgattgatga agcgcgggat tgcgggaagc atgtcatggt cgccgggatc 360
gaatcgcaaa atcaggcctc gctgcatctc caccagtcgc tgggatttgt cgtcaccgcg 420
caaatgccgc aggtaggcac taaatttggt cgttggctgg atctgacatt tatgcagttg 480
caactcgacg agcgcactga accggacgcg attggatga 519
<210> 62
<211> 172
<212> PRT
<213> Escherichia coli
<400> 62
Met Ser Ile Arg Phe Ala Arg Lys Ala Asp Cys Ala Ala Ile Ala Glu
1 5 10 15
Ile Tyr Asn His Ala Val Leu Tyr Thr Ala Ala Ile Trp Asn Asp Gln
20 25 30
Thr Val Asp Ala Asp Asn Arg Ile Ala Trp Phe Glu Ala Arg Thr Leu
35 40 45
Ala Gly Tyr Pro Val Leu Val Ser Glu Glu Asn Gly Val Val Thr Gly
50 55 60
Tyr Ala Ser Phe Gly Asp Trp Arg Ser Phe Asp Gly Phe Arg His Thr
65 70 75 80
Val Glu His Ser Val Tyr Val His Pro Asp His Gln Gly Lys Gly Leu
85 90 95
Gly Arg Lys Leu Leu Ser Arg Leu Ile Asp Glu Ala Arg Asp Cys Gly
100 105 110
Lys His Val Met Val Ala Gly Ile Glu Ser Gln Asn Gln Ala Ser Leu
115 120 125
His Leu His Gln Ser Leu Gly Phe Val Val Thr Ala Gln Met Pro Gln
130 135 140
Val Gly Thr Lys Phe Gly Arg Trp Leu Asp Leu Thr Phe Met Gln Leu
145 150 155 160
Gln Leu Asp Glu Arg Thr Glu Pro Asp Ala Ile Gly
165 170
<210> 63
<211> 100
<212> DNA
<213> artificial
<220>
<223> oligo DppsR
<400> 63
cgttctacgc ttctgtgcgt ttttaattta tgctttcata gaattatgtc tgcatcacgg 60
gaagaacaaa atgattccgg ggatccgtcg acctgcagtt 100
<210> 64
<211> 100
<212> DNA
<213> artificial
<220>
<223> oligo DppsR
<400> 64
gtcacacctc gccgggtggt tagcatgata acaaaaaaat aagctaatgc actagttctc 60
tagtacattc ggcgactaag tgtaggctgg agctgcttcg 100

Claims (12)

1.一种在发酵过程中通过转化碳源来生产甲硫氨酸或其羟基类似物形式的方法，包含以下步骤：
-在包含以碳水化合物作为碳源的适当培养基中培养经遗传修饰的微生物以生产甲硫氨酸或其羟基类似物形式，以及
-从培养基中回收甲硫氨酸或其羟基类似物形式，
其中所述经遗传修饰的微生物包含编码PTS碳水化合物利用***的功能基因，并且
其中在所述经遗传修饰的微生物中，调节磷酸烯醇式丙酮酸合酶(PPS)表达的双功能ADP依赖性激酶-Pi-依赖性焦磷酸化酶蛋白的表达降低。
2.权利要求1的方法，其中所述编码PTS碳水化合物利用***的功能基因对于经遗传修饰的微生物是异源的。
3.权利要求1的方法，其中所述编码PTS碳水化合物利用***的功能基因对于经遗传修饰的微生物是天然的。
4.权利要求1至3任一项的方法，其中编码所述调节磷酸烯醇式丙酮酸合酶(PPS)表达的双功能ADP依赖性激酶-Pi-依赖性焦磷酸化酶蛋白的天然基因在所述经遗传修饰的微生物中减弱或缺失。
5.权利要求1至4任一项的方法，其中所述微生物选自：肠杆菌科(Enterobacteriaceae)、芽孢杆菌科(Bacillaceae)、梭菌科(Clostridiaceae)、链霉菌科(Streptomycetaceae)、棒杆菌科(Corynebacteriaceae)、酵母科(Saccharomyceteceae)和酵母。
6.权利要求5的方法，其中所述微生物选自：大肠杆菌(Escherichia coli)、肺炎克雷伯氏菌(Klebsiella pneumoniae)、热解糖好热厌氧杆菌(Thermoanaerobacteriumthermosaccharolyticum)、楔状梭菌(Clostridium sphenoides)或酿酒酵母(Saccharomyces cerevisiae)。
7.权利要求1至6任一项的方法，其中所述微生物是大肠杆菌。
8.权利要求7的方法，其中编码所述双功能ADP依赖性激酶-Pi-依赖性焦磷酸化酶DUF299蛋白的基因ppsR减弱或缺失。
9.权利要求1至8任一项的方法，其中所述微生物经遗传修饰以增强甲硫氨酸或其羟基类似物形式的生产。
10.一种经遗传修饰的微生物，其用于增强从碳水化合物作为碳源的甲硫氨酸或其羟基类似物形式的生产，其中所述经遗传修饰的微生物包含编码PTS碳水化合物利用***的功能基因，并且
其中在所述经遗传修饰的微生物中，调节磷酸烯醇式丙酮酸合酶(PPS)表达的双功能ADP依赖性激酶-Pi-依赖性焦磷酸化酶蛋白的表达降低。
11.权利要求10的经遗传修饰的微生物，其中所述微生物如权利要求2至8任一项所述进行修饰。
12.权利要求10或11的经遗传修饰的微生物，其中编码所述双功能ADP依赖性激酶-Pi-依赖性焦磷酸化酶DUF229的基因ppsR减弱或缺失。
CN201780040978.2A 2016-07-08 2017-07-06 通过含有编码糖磷酸转移酶***(pts)的基因的微生物发酵生产甲硫氨酸或其羟基类似物形式的方法 Pending CN109415417A (zh)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
IBPCT/IB2016/001123 2016-07-08
IB2016001123 2016-07-08
PCT/EP2017/067036 WO2018007565A1 (en) 2016-07-08 2017-07-06 Method for the fermentative production of methionine or its hydroxy analog form by microorganisms comprising genes coding sugar phosphotransferase system (pts)

Publications (1)

Publication Number Publication Date
CN109415417A true CN109415417A (zh) 2019-03-01

Family

ID=56741149

Family Applications (2)

Application Number Title Priority Date Filing Date
CN201780042466.XA Active CN109415418B (zh) 2016-07-08 2017-07-06 通过包含编码糖磷酸转移酶***(pts)的基因的微生物发酵产生感兴趣的分子的方法
CN201780040978.2A Pending CN109415417A (zh) 2016-07-08 2017-07-06 通过含有编码糖磷酸转移酶***(pts)的基因的微生物发酵生产甲硫氨酸或其羟基类似物形式的方法

Family Applications Before (1)

Application Number Title Priority Date Filing Date
CN201780042466.XA Active CN109415418B (zh) 2016-07-08 2017-07-06 通过包含编码糖磷酸转移酶***(pts)的基因的微生物发酵产生感兴趣的分子的方法

Country Status (11)

Country Link
US (2) US20190276859A1 (zh)
EP (2) EP3481850A1 (zh)
JP (1) JP7321089B2 (zh)
KR (2) KR102430878B1 (zh)
CN (2) CN109415418B (zh)
AU (1) AU2017291975B2 (zh)
BR (2) BR112019000132A2 (zh)
CA (1) CA3028871A1 (zh)
MX (1) MX2019000075A (zh)
RU (1) RU2760290C1 (zh)
WO (2) WO2018007560A1 (zh)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20240023684A (ko) 2016-09-13 2024-02-22 알레간 인코포레이티드 안정화된 비단백질 클로스트리듐 독소 조성물
CN112384516A (zh) 2018-07-10 2021-02-19 日本农药株式会社 具有任选被卤代的亚烷基二氧基的苯并咪唑化合物或其盐类、以及含有这些化合物的农业园艺用杀虫剂及其使用方法
CN109468254B (zh) * 2018-11-13 2021-03-26 江南大学 一种提高l-丙氨酸生产效率的方法及其应用
JPWO2020230719A1 (zh) * 2019-05-10 2020-11-19
KR102558672B1 (ko) * 2022-05-17 2023-07-25 주식회사 엑티브온 1,3-프로판디올 생산 변이 미생물 및 이를 이용한 1,3-프로판디올의 생산 방법

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002022829A2 (en) * 2000-09-13 2002-03-21 Degussa Ag Nucleotide sequences which code for the ppsa gene
DE10130192A1 (de) * 2000-09-30 2002-04-11 Degussa Verfahren zur fermentativen Herstellung von L-Aminosäuren unter Verwendung von Stämmen der Familie Enterobacteriaceae
CN1539015A (zh) * 2001-08-06 2004-10-20 德古萨股份公司 用遗传修饰的谷氨酸棒杆菌生产l-赖氨酸

Family Cites Families (37)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4032441B2 (ja) * 1995-08-30 2008-01-16 味の素株式会社 Ｌ−アミノ酸の製造方法
US6087140A (en) 1997-02-19 2000-07-11 Wisconsin Alumni Research Foundation Microbial production of 1,2-propanediol from sugar
JP4110641B2 (ja) 1998-11-17 2008-07-02 味の素株式会社 発酵法によるｌ−メチオニンの製造法
AU9635901A (en) * 2000-09-29 2002-04-08 Cargill Inc Isoprenoid production
AU2001293795A1 (en) 2000-09-30 2002-04-15 Degussa A.G. Fermentation process for the preparation of l-amino acids using strains of the family enterobacteriaceae
AU2003282687A1 (en) 2002-10-04 2004-05-04 Genencor International, Inc. Glucose transport mutants for production of biomaterial
ES2356526T3 (es) * 2002-10-04 2011-04-08 E.I. Du Pont De Nemours And Company Procedimiento para la producción biológica de 1,3-propanodiol con elevado rendimiento.
JP2006517796A (ja) 2003-02-18 2006-08-03 メタボリック エクスプローラー 代謝経路の生成または改変を可能とする進化した微生物の生産方法
FR2864967B1 (fr) * 2004-01-12 2006-05-19 Metabolic Explorer Sa Microorganisme evolue pour la production de 1,2-propanediol
WO2005111202A1 (en) 2004-05-12 2005-11-24 Metabolic Explorer Recombinant enzyme with altered feedback sensitivity
EP2400027A1 (en) 2006-10-31 2011-12-28 Metabolic Explorer Process for the biological production of 1,3-propanediol from glycerol with high yield
WO2008052596A1 (en) * 2006-10-31 2008-05-08 Metabolic Explorer Process for the biological production of n-butanol with high yield
US8252555B2 (en) 2006-12-29 2012-08-28 Evonik Degussa Ag Nucleic acid encoding a cobalamin-dependent methionine synthase polypeptide
WO2008116852A1 (en) 2007-03-23 2008-10-02 Metabolic Explorer New micro-organisms for the production of 1,2-propanediol obtained by a combination of evolution and rational design.
DK2126101T3 (en) * 2007-03-23 2015-09-28 Metabolic Explorer Sa MICROORGANISMS AND PROCESSES FOR PREPARING 1,2-propane diol, and acetol
WO2008116848A1 (en) * 2007-03-23 2008-10-02 Metabolic Explorer Metabolically engineered microorganism useful for the production of 1,2-propanediol
EP2808382B1 (en) 2007-04-11 2016-03-16 CJ CheilJedang Corporation Compositions and methods of producing methionine
ES2549003T3 (es) 2008-12-31 2015-10-22 Metabolic Explorer Método para la preparación de dioles
EP2233562A1 (en) 2009-03-24 2010-09-29 Metabolic Explorer Method for producing high amount of glycolic acid by fermentation
EP2248904A1 (en) 2009-05-05 2010-11-10 Metabolic Explorer Continuous culture for 1,3-propanediol production using high glycerine concentration
EP2582828B1 (en) 2010-06-15 2015-07-15 Metabolic Explorer Use of inducible promoters in the production of glycolic acid
TWI500768B (zh) 2010-07-05 2015-09-21 Metabolic Explorer Sa 由蔗糖製備１，３－丙二醇之方法
CN103189517A (zh) 2010-08-27 2013-07-03 代谢探索者公司 用经修饰的微生物改善乙醇酸发酵产生
AR083468A1 (es) 2010-10-25 2013-02-27 Metabolic Explorer Sa Aumento de la disponibilidad de nadph para la produccion de metionina
AR083799A1 (es) 2010-11-10 2013-03-20 Metabolic Explorer Sa Microorganismos para la produccion de 1,3-propanodiol usando concentracion de glicerina alta
JP6368488B2 (ja) 2010-12-29 2018-08-01 シージェイ チェイルジェダン コーポレーション Ｌ−メチオニン及び関連産物の生産方法
EP2479279A1 (de) 2011-01-20 2012-07-25 Evonik Degussa GmbH Verfahren zur fermentativen Herstellung schwefelhaltiger Aminosäuren
EP2532751A1 (en) 2011-06-10 2012-12-12 Metabolic Explorer Use of inducible promoters in the fermentative production of 1,2-propanediol
EP2721167A1 (en) 2011-06-15 2014-04-23 B.R.A.I.N. Biotechnology Research And Information Network AG New means and methods for producing propanediol
FI125136B (en) 2011-10-04 2015-06-15 Teknologian Tutkimuskeskus Vtt Oy Eukaryotic cells and a method for preparing glycolic acid
ES2689754T3 (es) 2012-08-20 2018-11-15 Evonik Degussa Gmbh Procedimiento para la preparación fermentativa de L-aminoácidos utilizando cepas mejoradas de la familia Enterobacteriaceae
FI20135333L (fi) 2013-04-05 2014-10-06 Teknologian Tutkimuskeskus Vtt Oy Hapon / happojen ja alkoholin tuotto sokereista käyttämällä hiivaa
US9363091B2 (en) * 2013-12-23 2016-06-07 Louroe Electronics Power over Ethernet devices, systems and methods
CN106559997B (zh) 2014-05-12 2021-04-09 代谢探索者公司 基于nadph依赖型丙酮醇还原酶和改进的nadph供给用于生产1,2-丙二醇的新微生物和方法
WO2015177800A2 (en) * 2014-05-22 2015-11-26 Yeda Research And Development Co. Ltd. Recombinant microorganisms capable of carbon fixation
JP2017525379A (ja) 2014-09-01 2017-09-07 メタボリック エクスプローラー メチオニン排出を改善した発酵によるメチオニン産生の方法及び微生物
WO2016050959A2 (en) 2014-10-03 2016-04-07 Metabolic Explorer Mutant glutamate dehydrogenase for the conversion of homoserine into 4-hydroxy-2-ketobutyrate

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002022829A2 (en) * 2000-09-13 2002-03-21 Degussa Ag Nucleotide sequences which code for the ppsa gene
DE10130192A1 (de) * 2000-09-30 2002-04-11 Degussa Verfahren zur fermentativen Herstellung von L-Aminosäuren unter Verwendung von Stämmen der Familie Enterobacteriaceae
CN1539015A (zh) * 2001-08-06 2004-10-20 德古萨股份公司 用遗传修饰的谷氨酸棒杆菌生产l-赖氨酸

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
ELISABETH GABOR: "The phosphoenolpyruvate-dependent glucose-phosphotransferase system from Escherichia coli K-12 as the center of a network regulating carbohydrate flux in the cell", 《EUR J CELL BIOL》 *
JIM N BURNELL: "Cloning and characterization of Escherichia coli DUF299: a bifunctional ADP-dependent kinase--Pi-dependent pyrophosphorylase from bacteria", 《BMC BIOCHEM》 *
NIV ANTONOVSKY等: "Sugar synthesis from CO2 in Escherichia coli", 《CELL》 *

Also Published As

Publication number Publication date
MX2019000075A (es) 2019-05-02
WO2018007560A1 (en) 2018-01-11
RU2760290C1 (ru) 2021-11-23
CN109415418A (zh) 2019-03-01
CN109415418B (zh) 2024-02-09
US10961499B2 (en) 2021-03-30
KR102430878B1 (ko) 2022-08-09
EP3481851A1 (en) 2019-05-15
AU2017291975A1 (en) 2019-01-24
BR112019000132A2 (pt) 2019-04-16
KR20190026851A (ko) 2019-03-13
US20190276859A1 (en) 2019-09-12
AU2017291975B2 (en) 2021-06-03
JP7321089B2 (ja) 2023-08-04
EP3481851B1 (en) 2021-11-24
EP3481850A1 (en) 2019-05-15
KR20190025969A (ko) 2019-03-12
WO2018007565A1 (en) 2018-01-11
US20190276794A1 (en) 2019-09-12
JP2019520081A (ja) 2019-07-18
BR112019000360A2 (pt) 2019-04-24
CA3028871A1 (en) 2018-01-11

Similar Documents

Publication Publication Date Title
CN109415417A (zh) 通过含有编码糖磷酸转移酶***(pts)的基因的微生物发酵生产甲硫氨酸或其羟基类似物形式的方法
Tapia V et al. Optimization of lipid production by the oleaginous yeast Lipomyces starkeyi by random mutagenesis coupled to cerulenin screening
RU2549689C2 (ru) Микроорганизм с повышенной продукцией l-аминокислот и способ получения l-аминокислот с его применением
WO2017150304A1 (ja) エルゴチオネインの発酵生産
Feng et al. Halorubrum alkaliphilum sp. nov., a novel haloalkaliphile isolated from a soda lake in Xinjiang, China
CN107690482A (zh) 用于2,4‑二羟基丁酸的优化生产的经修饰的微生物
Liu et al. Enhancement of pyruvate productivity in Torulopsis glabrata: increase of NAD+ availability
JPH10507082A (ja) 混合微生物培養物を用いて炭水化物から１，３−プロパンジオールを製造する方法
Lindner et al. NCgl2620 encodes a class II polyphosphate kinase in Corynebacterium glutamicum
CN105385646A (zh) 5-羟色氨酸的前体定向生物合成
Zamboni et al. The phosphoenolpyruvate carboxykinase also catalyzes C3 carboxylation at the interface of glycolysis and the TCA cycle of Bacillus subtilis
Borgos et al. Mapping global effects of the anti-sigma factor MucA in Pseudomonas fluorescens SBW25 through genome-scale metabolic modeling
CN105112436A (zh) 一种己二酸的全生物合成方法
CN109937257A (zh) 生产目标物质的方法
CN109890960A (zh) 用于生产醛的方法
CN104540935A (zh) 代谢途径产物的生成方法
Pérez-García et al. From brown seaweed to a sustainable microbial feedstock for the production of riboflavin
Patil et al. Anaerobium acetethylicum gen. nov., sp. nov., a strictly anaerobic, gluconate-fermenting bacterium isolated from a methanogenic bioreactor
CN105377878B (zh) 新的RhtB蛋白变体和使用其产生O-磷酸丝氨酸的方法
Bao et al. Isolation of the stable strain Labrys sp. BK-8 for L (+)-tartaric acid production
Liu et al. RETRACTED ARTICLE: Comparative analysis of the Corynebacterium glutamicum transcriptome in response to changes in dissolved oxygen levels
Schulze et al. Investigation of exopolysaccharide formation and its impact on anaerobic succinate production with Vibrio natriegens
CN106701649A (zh) 生产l‑谷氨酰胺的菌株和生产l‑谷氨酰胺的方法
Mukti et al. Medium development and production of carotenoids and exopolysaccharides by the extremophile Rhodothermus marinus DSM16675 in glucose-based defined media
David et al. Cellular physiology controls photoautotrophic production of 1, 2‐propanediol from pools of CO2 and glycogen

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
CB02 Change of applicant information
CB02 Change of applicant information

Address after: Essen, Germany

Applicant after: Evonik Operations Ltd.

Address before: Essen, Germany

Applicant before: EVONIK DEGUSSA GmbH

WD01 Invention patent application deemed withdrawn after publication
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20190301