JPH0198495A - Production of l-alpha-aminoadipic acid using microorganism - Google Patents
Production of l-alpha-aminoadipic acid using microorganismInfo
- Publication number
- JPH0198495A JPH0198495A JP25505587A JP25505587A JPH0198495A JP H0198495 A JPH0198495 A JP H0198495A JP 25505587 A JP25505587 A JP 25505587A JP 25505587 A JP25505587 A JP 25505587A JP H0198495 A JPH0198495 A JP H0198495A
- Authority
- JP
- Japan
- Prior art keywords
- acid
- aminoadipic acid
- aminoadipic
- microorganism
- alpha
- 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.)
- Granted
Links
- OYIFNHCXNCRBQI-BYPYZUCNSA-N L-2-aminoadipic acid Chemical compound OC(=O)[C@@H](N)CCCC(O)=O OYIFNHCXNCRBQI-BYPYZUCNSA-N 0.000 title claims abstract description 17
- 244000005700 microbiome Species 0.000 title claims abstract description 13
- 238000004519 manufacturing process Methods 0.000 title claims description 6
- 241000588986 Alcaligenes Species 0.000 claims abstract description 9
- 241000589516 Pseudomonas Species 0.000 claims abstract description 7
- OYIFNHCXNCRBQI-UHFFFAOYSA-N 2-aminoadipic acid Chemical compound OC(=O)C(N)CCCC(O)=O OYIFNHCXNCRBQI-UHFFFAOYSA-N 0.000 claims description 13
- HXEACLLIILLPRG-UHFFFAOYSA-N pipecolic acid Chemical compound OC(=O)C1CCCCN1 HXEACLLIILLPRG-UHFFFAOYSA-N 0.000 claims description 12
- HXEACLLIILLPRG-RXMQYKEDSA-N l-pipecolic acid Natural products OC(=O)[C@H]1CCCCN1 HXEACLLIILLPRG-RXMQYKEDSA-N 0.000 claims description 9
- HXEACLLIILLPRG-YFKPBYRVSA-N L-pipecolic acid Chemical compound [O-]C(=O)[C@@H]1CCCC[NH2+]1 HXEACLLIILLPRG-YFKPBYRVSA-N 0.000 abstract description 5
- 102000004196 processed proteins & peptides Human genes 0.000 abstract description 3
- 108090000765 processed proteins & peptides Proteins 0.000 abstract description 3
- 150000001875 compounds Chemical class 0.000 abstract 2
- 241000186809 Kurthia Species 0.000 abstract 1
- 238000012258 culturing Methods 0.000 abstract 1
- 239000007788 liquid Substances 0.000 abstract 1
- 239000000203 mixture Substances 0.000 abstract 1
- 241000894006 Bacteria Species 0.000 description 12
- 239000002609 medium Substances 0.000 description 7
- 239000002253 acid Substances 0.000 description 6
- 239000006916 nutrient agar Substances 0.000 description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 5
- 239000008103 glucose Substances 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 239000012228 culture supernatant Substances 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 241000193830 Bacillus <bacterium> Species 0.000 description 3
- 102000016938 Catalase Human genes 0.000 description 3
- 108010053835 Catalase Proteins 0.000 description 3
- 241000233866 Fungi Species 0.000 description 3
- 102000004316 Oxidoreductases Human genes 0.000 description 3
- 108090000854 Oxidoreductases Proteins 0.000 description 3
- 241001148470 aerobic bacillus Species 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000011888 foil Substances 0.000 description 3
- 210000004209 hair Anatomy 0.000 description 3
- 230000001766 physiological effect Effects 0.000 description 3
- 239000000049 pigment Substances 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 238000000108 ultra-filtration Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000012262 fermentative production Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- ZICZZIRIRHGROF-UHFFFAOYSA-N 1-$l^{1}-oxidanyl-2,2,4,5,5-pentamethylimidazole Chemical compound CC1=NC(C)(C)N([O])C1(C)C ZICZZIRIRHGROF-UHFFFAOYSA-N 0.000 description 1
- 229930186147 Cephalosporin Natural products 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 101000852543 Homo sapiens Importin-4 Proteins 0.000 description 1
- 102100036341 Importin-4 Human genes 0.000 description 1
- ZOKXTWBITQBERF-UHFFFAOYSA-N Molybdenum Chemical compound [Mo] ZOKXTWBITQBERF-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 241000589774 Pseudomonas sp. Species 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000003782 beta lactam antibiotic agent Substances 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 229940124587 cephalosporin Drugs 0.000 description 1
- 150000001780 cephalosporins Chemical class 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229910017052 cobalt Inorganic materials 0.000 description 1
- 239000010941 cobalt Substances 0.000 description 1
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical class [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 229910052750 molybdenum Inorganic materials 0.000 description 1
- 239000011733 molybdenum Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 239000000813 peptide hormone Substances 0.000 description 1
- 235000021317 phosphate Nutrition 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000003910 polypeptide antibiotic agent Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 229910021654 trace metal Inorganic materials 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- 239000002132 β-lactam antibiotic Substances 0.000 description 1
- 229940124586 β-lactam antibiotics Drugs 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】 ン酸を製造する方法に関する。[Detailed description of the invention] The present invention relates to a method for producing phosphoric acid.
Ｌ−α−アミノアジピン酸は、非蛋白性のアミノ酸であ
る。このＬ−α−アミノアジピン酸は、ペプチド性抗生
物質やペプチドホルモンなどの生理活性ペプチドの末端
修飾剤として用いることができる。また、ペニシリンや
セファロスポリンに代表されるβ−ラクラム系抗生物質
の発酵生産において前駆体として利用できる。L-α-aminoadipic acid is a non-protein amino acid. This L-α-aminoadipic acid can be used as a terminal modifying agent for physiologically active peptides such as peptide antibiotics and peptide hormones. It can also be used as a precursor in the fermentative production of β-laclam antibiotics, such as penicillin and cephalosporin.
しかしなから、Ｌ−α−アミノアジピン酸を簡便に製造
する方法は未だ確立されていない、トウモロコシ種子等
中に少量存在するし一α−アミノアジピン醸を抽出する
方法などが知られているが、このような方法は、はん雑
であり、かつ原料の供給にも難点があるので、大量生産
には適さない。However, a method for easily producing L-α-aminoadipic acid has not yet been established; methods such as extracting L-α-aminoadipic acid, which exists in small amounts in corn seeds, etc., are known. Such a method is complicated and has difficulties in supplying raw materials, so it is not suitable for mass production.
発明が解決しようとする問題点〕
本発明者は、このような従来法に代わるＬ−α−ミノア
ジピン酸の製造について検討した結果。[Problems to be Solved by the Invention] The present inventor has investigated the production of L-α-minoadipic acid as an alternative to such conventional methods.
純粋なＬ−ピペコリン酸、或は、ＤＬ−ピペコリン酸中
に含まれるＬ−ピペコリン酸が、何種類かの微生物によ
りし一α−アミノアジピン酸に変換されることを見い出
した。It has been found that pure L-pipecolic acid or L-pipecolic acid contained in DL-pipecolic acid is converted to mono-α-aminoadipic acid by several types of microorganisms.
本発明は、この知見に基き完成されたものである。すな
わち、本発明は、アルカリゲネス属、シュウドモナス属
及びクルシア属より選ばれ、Ｌ−ビベコリン酸をＬ−α
−アミノアジピン酸に；変換する能力を有する微生物を
Ｌ−ピペコリン酸含有培地で培養し、培養物からし一α
−アミノアジピン酸を採取することを特徴とする微生物
によるし一α−アミノアジピン酸の製造法を提供するも
のである。The present invention was completed based on this knowledge. That is, the present invention provides L-bibecolic acid selected from the genus Alcaligenes, Pseudomonas, and Crucia.
- to aminoadipic acid; a microorganism having the ability to convert it into L-pipecolic acid is cultured in a medium containing L-pipecolic acid, and the culture is
The present invention provides a method for producing monoα-aminoadipic acid using a microorganism, which is characterized by collecting aminoadipic acid.
し−ピペコリン酸をＬ−α−アミノアジピン酸に変換す
る能力を有する微生物として、アルカリゲネス属、シュ
ウドモナス属、クルシア属より選ばれ、代表菌として本
発明者らが自然界より分離したアルカリゲネス、［Ｎａ
３０９Ｂ１菌、シュードモナス属Ｎα５２０Ｂ菌、クル
シア属Ｎａ１１３Ｂ菌を例示できる。Microorganisms that have the ability to convert L-pipecolic acid into L-α-aminoadipic acid are selected from the genera Alcaligenes, Pseudomonas, and Crucia, and representative microorganisms include Alcaligenes, [Na
309B1 bacteria, Pseudomonas sp. Nα520B bacterium, and Crucia sp. Na113B bacterium are examples.
これらの微生物の菌学的性質は以下に示すとおりである
。The mycological properties of these microorganisms are as shown below.
（１）アルカリゲネス属Ｎα３０９Ｂｌ菌（ａ）形態
・直径約１μ膣、長さ２μｍ程度の桿菌（栄養寒天上で
生育した菌体） ′
・ダラム染色：陰性
・運動性であり、周ペン毛を有する
・芽胞形成能：無し
くｂ）栄養寒天培地上ぎの生育状態
・生育良好
・生育状態は拡布状
・箔層の***は偏平状
・光沢は純光
・色素生産性：無し
くｃ）生理学的性質
・好気性菌
・カタラーゼ：生産性有り
・オキシダーゼ：生産性有り
・グルコースより酸を生成しない
・Ｏ−Ｆテスト：グルコースを分解しない以上の特徴は
従来知られているアルカリゲネス属（Ａｌｃａｌｉｇｅ
ｎｅｓ属）の特徴と一致している。(1) Alcaligenes Nα309Bl bacterium (a) Morphology: Bacillus with a diameter of approximately 1 μm and a length of approximately 2 μm (bacteria grown on nutrient agar) ′ - Durham staining: negative, motile, and has peripen hairs・Spore forming ability: None b) Growth condition on nutrient agar medium ・Good growth ・Growth condition is spread-like ・Protuberance of foil layer is flat ・Gloss is pure light ・Pigment productivity: None c) Physiological properties・Aerobic bacteria ・Catalase: Productive ・Oxidase: Productive ・Does not produce acid from glucose ・O-F test: Does not degrade glucose
The characteristics match those of the genus nes.
本菌はアルカリゲネス属Ｎα３０９旧と命名され、ＦＥ
ＲＭ Ｐ−９３１４として寄託されている。This bacterium was named Alcaligenes Nα309 old, and FE
It has been deposited as RM P-9314.
（２）シュードモナス５２０Ｂ菌
（ａ）形態
・直径約１μｍ、長さ約２μｍ程度の桿菌（栄養寒天上
での生育した菌体）
・ダラム染色：陰性
・運動性であり、１本の極ベン毛を有する・芽胞形成能
：無し
くｂ）栄養寒天培地上での生育状態
・生育良好
・生育状態は拡布状
・箔層の***は偏平状
・光沢は純光
・色素生産性：無し
くｃ）生理学的性質
・好気性菌
・カタラーゼ：生産性有り
・オキシダーゼ：生産性有り
・グルコースより酸を生成する
・Ｏ−Ｆテスト：酸化的
以上の特徴は従来公知のシュードモナス３（Ｐｓｅｕｄ
ｏｍｏｎａｓ属）の特徴と一致している。(2) Pseudomonas 520B bacterium (a) Morphology - Bacillus with a diameter of about 1 μm and a length of about 2 μm (bacteria grown on nutrient agar) - Durham staining: negative, motile, with one polar Ben hair・Spore forming ability: No b) Growth condition on nutrient agar medium ・Good growth ・Growth condition is spread-like ・Protuberance of foil layer is flat ・Gloss is pure light ・Pigment productivity: No c) Physiological properties・Aerobic bacteria・Catalase: Productive・Oxidase: Productive・Produces acid from glucose・O-F test: Characteristics other than oxidative
omonas genus).
本菌はシュードモナス５２０Ｂ菌と命名され、ＦＥＲＭ
Ｐ−Ｎａ９３１３として寄託されている。This bacterium was named Pseudomonas 520B, and FERM
It has been deposited as P-Na9313.
（３）クルシア属ＮＱ１１３Ｂ菌
（ａ）形態
・直径約１μｍ、長さ２μｍ程度の桿菌（栄養寒天培地
上で生育した菌体）
・ダラム染色：陰性
・運動性であり、１本の周ペン毛を有する・芽胞形成能
：無し
くｂ）栄養寒天培地上での生育状態
・生育良好
・生育状態は拡布状
・箔層の***は偏平状
・光沢は純光
・色素生産性：無し
くｃ）生理学的性質
・好気性菌
・カタラーゼ：生産性有り
・オキシダーゼ：生産性有り
・グルコースから酸を生成しない
・Ｏ−Ｆテスト：グルコースを分解しない以上の特徴は
従来公知のクルシア属（Ｋｕｒｔｈｉａ属）の特徴と一
致する。(3) Crucia sp. NQ113B bacteria (a) Morphology - Bacillus with a diameter of approximately 1 μm and a length of approximately 2 μm (bacteria grown on a nutrient agar medium) - Durham staining: negative, motile, with one pericysmal hair・Spore forming ability: No b) Growth condition on nutrient agar medium ・Good growth ・Growth condition is spread-like ・Protuberance of foil layer is flat ・Gloss is pure light ・Pigment productivity: No c) Physiological properties, aerobic bacteria, catalase: productive, oxidase: productive, does not produce acid from glucose, O-F test: does not decompose glucose. Match the features.
木菌はクルシア属Ｎα゛１１３Ｂ菌と命名され、ＦＥＲ
Ｎ Ｐ−Ｎα９３１２として寄託されている。The wood fungus is named Crucia genus Nα゛113B fungus, and FER
It has been deposited as NP-Nα9312.
本発明における具体的な培養は、し−ピペコリン酸、或
は、ＤＬ−ピペコリン酸の１〜２０％水溶液に、（ＮＩ
＋４）２Ｓｏ、などの窒素源、Ｋ、）ＩＰＯ４やＫＨ２
ＰＯ，などリン酸塩、各種微量金属塩を加え培地とした
もので前記の微生物を培養する。この際の培地組成には
、特に限定はない。また培養条件にも制約はないが、Ｐ
ｌｌは４〜９、望ましくは７、温度は、２０℃〜５０℃
、望ましくは３０℃、培養方法は、振とう培養ないし通
気培養が望ましい。The specific culture in the present invention is carried out in a 1 to 20% aqueous solution of ni-pipecolic acid or DL-pipecolic acid.
+4) Nitrogen sources such as 2So, K,) IPO4 and KH2
The above-mentioned microorganisms are cultured in a medium containing phosphates such as PO, and various trace metal salts. There is no particular limitation on the medium composition at this time. There are also no restrictions on the culture conditions, but P
ll is 4 to 9, preferably 7, temperature is 20°C to 50°C
, preferably at 30° C., and shaking culture or aeration culture is preferred as the culture method.
培養の経過と共に、培地中のし一ピペコリン酸がＬ−α
−アミノアジピン酸に変換され、培養液中に蓄積される
。すべてのし−ピペコリン酸がＬ−α−アミノアジピン
酸に変換されるのに要する時間は、Ｌ−ピペコリン酸或
はＤＬ−ピペコリン酸の初期濃度、用いる微生物及び培
養条件による。As the culture progresses, the monopipecolic acid in the medium becomes L-α.
- Converted to aminoadipic acid and accumulated in the culture medium. The time required for all of the pipecolic acid to be converted to L-α-aminoadipic acid depends on the initial concentration of L-pipecolic acid or DL-pipecolic acid, the microorganism used, and the culture conditions.
蓄積されたし一α−アミノアジピン酸の培養液からの回
収は、通常の任意の方法を用いることができるが、等重
点沈殿法が最も簡便である。すなわち、培養液から遠′
心分雌などにより菌体を除き、培養液上清を得る。この
培養液上清から蛋白質等の高分子性物質を限外濾過によ
り除く。次いで、得られた濾液のＰＨを、し−α−アミ
ノアジピン酸の等電点である３、２に塩酸等で調整し、
−晩、低温下（４℃程度）に放置するとＬ−α−アミノ
アジピン酸が沈殿する。この沈殿したし一α−アミノア
ジピン酸を濾取し、冷水で洗浄した後、乾燥すると、
Ｌ−α−アミノアジピン酸の結晶が得られる。Although any conventional method can be used to recover the accumulated mono-α-aminoadipic acid from the culture solution, the isocenter precipitation method is the simplest. In other words, it is far from the culture medium.
Remove the bacterial cells using a centrifuge or the like to obtain a culture supernatant. Polymer substances such as proteins are removed from this culture supernatant by ultrafiltration. Next, the pH of the obtained filtrate was adjusted to 3.2, which is the isoelectric point of α-aminoadipic acid, with hydrochloric acid, etc.
- L-α-aminoadipic acid precipitates when left overnight at a low temperature (about 4°C). This precipitated mono-α-aminoadipic acid is filtered, washed with cold water, and dried.
Crystals of L-α-aminoadipic acid are obtained.
以上述べたように、本発明は、比較的安価でかつ大量に
入手できるＤＬ−ピペコリン酸から簡便にＬ−α−アミ
ノアジピン酸を製造することを可能にする。As described above, the present invention makes it possible to easily produce L-α-aminoadipic acid from DL-pipecolic acid, which is relatively inexpensive and available in large quantities.
次に、本発明を実施例により更に詳細に説明する。Next, the present invention will be explained in more detail with reference to Examples.
ＯＬ−ピペコリン酸部、Ｋ１１２Ｐ０．０．３％、Ｋ、
ＨＰＱ４０．３％、（Ｎ）１．）、５Ｏ４０，２％、
Ｍｇ５Ｏ，・７８．ＯＯ，０３％、 ＮａＣ１０００１
％、 ＣａＣ１，・２ＨｔＯＯ，０１％、及び微量のマ
ンガン、亜鉛、銅、鉄、モリブデン、コバルトの塩を含
むＰｌｌ７，０の液体培地を滅菌し、これＩＲ’Ｖ）；
″Ｌ／、／Ｊリゲネス属Ｎａ３０９Ｂ１菌（ＦＥＲＭ
Ｐ−９３１４）、シュードモナス属５２０Ｂ菌（ＦＥＲ
Ｍ Ｐ−９３１３）、クルシア屈魔１１３Ｂ菌（ＦＥＲ
河Ｐ−’１３１２）をそれぞれ接種し、３日間３０℃で
振どう培養した。遠心分離により菌体を培養液から除去
し、培養液上清を得た。この培養液上清を分画分子量１
０，０００の限外濾過膜（アミコン社製、商品名ダイア
フローメンブレンＰＭＩＯ）を用いて限外濾過を行ない
高分子性物質を除いた。得られた濾液のｐＨを５規定濃
度の塩酸により３．２に調整し庫内温度４℃の冷蔵庫内
に１晩放置した。沈殿したＬ−α−アミノアジピン酸を
濾過により集め冷水で数回洗浄した後、乾燥した。いず
れの菌を用いた場合でも、得られたし一α−アミノアジ
ピン酸は、アミノ酸分析システム（島津製作所製、商品
名ＬＣ−６Ａアミノ酸分析システム）において１ピーク
を与え、そのピークの保持時間は、標準品のし一α−ア
ミノアジピン酸の保持時間と一致していた。また、得ら
れたＬ−α−アミノアジピン酸の性状、融点１元素分析
値、比施光度は標準品のし一α−アミノアジピン酸と一
致していた。それぞＡ１’ＧＷ”Ｌ用いた場合の収率を
次表に示す。OL-pipecolic acid part, K112P0.0.3%, K,
HPQ40.3%, (N)1. ), 5O40.2%,
Mg5O, 78. OO, 03%, NaC10001
%, CaC1,·2HtOO,01%, and trace amounts of salts of manganese, zinc, copper, iron, molybdenum, and cobalt.
″L/,/J Ligenes genus Na309B1 (FERM
P-9314), Pseudomonas 520B (FER
M P-9313), Crucia 113B fungus (FER
Kawa P-'1312) was inoculated and cultured with shaking at 30°C for 3 days. The bacterial cells were removed from the culture solution by centrifugation to obtain a culture supernatant. This culture supernatant was collected with a molecular weight cutoff of 1
Ultrafiltration was performed using a 0,000 ultrafiltration membrane (manufactured by Amicon, trade name: Diaflow Membrane PMIO) to remove polymeric substances. The pH of the obtained filtrate was adjusted to 3.2 with 5N hydrochloric acid and left overnight in a refrigerator at an internal temperature of 4°C. The precipitated L-α-aminoadipic acid was collected by filtration, washed several times with cold water, and then dried. Regardless of which bacteria is used, the obtained monoα-aminoadipic acid gives one peak in an amino acid analysis system (manufactured by Shimadzu Corporation, trade name: LC-6A amino acid analysis system), and the retention time of that peak is The retention time was consistent with that of the standard 1-α-aminoadipic acid. Furthermore, the properties, melting point, single element analysis value, and specific light intensity of the obtained L-α-aminoadipic acid were consistent with those of the standard product, L-α-aminoadipic acid. The yields when using A1'GW"L are shown in the following table.
表−１
〔本発明の効果〕
本発明は、Ｌ−α−アミノアジピン酸の大規模製造を可
能にする。従って、本発明により、生理活性ペプチドの
改良や、β−ラクタム系抗生物質の発酵生産へのし一α
−アミノアジピン酸の応用が促進されると予想される。Table 1 [Effects of the present invention] The present invention enables large-scale production of L-α-aminoadipic acid. Therefore, the present invention provides an opportunity for improving bioactive peptides and for fermentative production of β-lactam antibiotics.
- It is expected that the application of aminoadipic acid will be promoted.
Claims (1)
ア属より選ばれ、Ｌ−ピペコリン酸をＬ−α−アミノア
ジピン酸に変換する能力を有する微生物をＬ−ピペコリ
ン酸含有培地で培養し培養物からＬ−α−アミノアジピ
ン酸を採取することを特徴とする微生物によるＬ−α−
アミノアジピン酸の製造法。(1) A microorganism selected from the genus Alcaligenes, Pseudomonas, and Crucia that has the ability to convert L-pipecolic acid to L-α-aminoadipic acid is cultured in a medium containing L-pipecolic acid, and L-α is extracted from the culture. -L-α- by a microorganism characterized by collecting aminoadipic acid
Method for producing aminoadipic acid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP25505587A JPH0198495A (en) | 1987-10-09 | 1987-10-09 | Production of l-alpha-aminoadipic acid using microorganism |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP25505587A JPH0198495A (en) | 1987-10-09 | 1987-10-09 | Production of l-alpha-aminoadipic acid using microorganism |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0198495A true JPH0198495A (en) | 1989-04-17 |
| JPH046355B2 JPH046355B2 (en) | 1992-02-05 |
Family
ID=17273521
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP25505587A Granted JPH0198495A (en) | 1987-10-09 | 1987-10-09 | Production of l-alpha-aminoadipic acid using microorganism |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0198495A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1996031616A1 (en) * | 1995-04-07 | 1996-10-10 | Mercian Corporation | Process for producing l-2-aminoadipic acid |
-
1987
- 1987-10-09 JP JP25505587A patent/JPH0198495A/en active Granted
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1996031616A1 (en) * | 1995-04-07 | 1996-10-10 | Mercian Corporation | Process for producing l-2-aminoadipic acid |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH046355B2 (en) | 1992-02-05 |
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