JPH02145192A - Production of d-n-carbamoyl-alpha-amino acids - Google Patents
Production of d-n-carbamoyl-alpha-amino acidsInfo
- Publication number
- JPH02145192A JPH02145192A JP29962988A JP29962988A JPH02145192A JP H02145192 A JPH02145192 A JP H02145192A JP 29962988 A JP29962988 A JP 29962988A JP 29962988 A JP29962988 A JP 29962988A JP H02145192 A JPH02145192 A JP H02145192A
- Authority
- JP
- Japan
- Prior art keywords
- substituted
- carbamoyl
- amino acids
- group
- 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.)
- Pending
Links
- 238000004519 manufacturing process Methods 0.000 title claims description 9
- 150000001469 hydantoins Chemical class 0.000 claims abstract description 14
- 125000003710 aryl alkyl group Chemical group 0.000 claims abstract description 5
- 125000000217 alkyl group Chemical group 0.000 claims abstract description 3
- 125000002541 furyl group Chemical group 0.000 claims abstract description 3
- 125000002883 imidazolyl group Chemical group 0.000 claims abstract description 3
- 125000001041 indolyl group Chemical group 0.000 claims abstract description 3
- 125000004076 pyridyl group Chemical group 0.000 claims abstract description 3
- 125000000335 thiazolyl group Chemical group 0.000 claims abstract description 3
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 4
- 241001660259 Cereus <cactus> Species 0.000 claims description 3
- 125000000547 substituted alkyl group Chemical group 0.000 claims description 2
- 239000000126 substance Substances 0.000 claims 2
- 241000193755 Bacillus cereus Species 0.000 abstract description 11
- 244000005700 microbiome Species 0.000 abstract description 11
- 238000006243 chemical reaction Methods 0.000 abstract description 6
- 239000000758 substrate Substances 0.000 abstract description 4
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 abstract description 2
- VMAQYKGITHDWKL-UHFFFAOYSA-N 5-methylimidazolidine-2,4-dione Chemical compound CC1NC(=O)NC1=O VMAQYKGITHDWKL-UHFFFAOYSA-N 0.000 abstract description 2
- 102000016938 Catalase Human genes 0.000 abstract description 2
- 108010053835 Catalase Proteins 0.000 abstract description 2
- 239000003456 ion exchange resin Substances 0.000 abstract description 2
- 229920003303 ion-exchange polymer Polymers 0.000 abstract description 2
- 239000002689 soil Substances 0.000 abstract description 2
- 238000010186 staining Methods 0.000 abstract description 2
- 239000007795 chemical reaction product Substances 0.000 abstract 2
- -1 (substituted) phenyl Chemical group 0.000 abstract 1
- 241000304886 Bacilli Species 0.000 abstract 1
- 239000002253 acid Substances 0.000 abstract 1
- 150000007513 acids Chemical class 0.000 abstract 1
- 230000000813 microbial effect Effects 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 6
- 238000000034 method Methods 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 230000007062 hydrolysis Effects 0.000 description 3
- 238000006460 hydrolysis reaction Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 2
- 229910002651 NO3 Inorganic materials 0.000 description 2
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 2
- 150000001371 alpha-amino acids Chemical class 0.000 description 2
- 235000008206 alpha-amino acids Nutrition 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- QYASYXWODYQOFU-SNVBAGLBSA-N (5R)-5-(1H-indol-2-ylmethyl)imidazolidine-2,4-dione Chemical compound N1C(=CC2=CC=CC=C12)C[C@@H]1C(NC(N1)=O)=O QYASYXWODYQOFU-SNVBAGLBSA-N 0.000 description 1
- ZUYPRBNQYTZYIK-BRJRFNKRSA-N (5R)-5-butan-2-ylimidazolidine-2,4-dione Chemical compound C(C)(CC)[C@@H]1C(NC(N1)=O)=O ZUYPRBNQYTZYIK-BRJRFNKRSA-N 0.000 description 1
- WLRZLHCGXUHRIG-RXMQYKEDSA-N (5r)-5-(2-methylpropyl)imidazolidine-2,4-dione Chemical compound CC(C)C[C@H]1NC(=O)NC1=O WLRZLHCGXUHRIG-RXMQYKEDSA-N 0.000 description 1
- PBNUQCWZHRMSMS-SCSAIBSYSA-N (5r)-5-propan-2-ylimidazolidine-2,4-dione Chemical compound CC(C)[C@H]1NC(=O)NC1=O PBNUQCWZHRMSMS-SCSAIBSYSA-N 0.000 description 1
- DBOMTIHROGSFTI-UHFFFAOYSA-N 5-benzylimidazolidine-2,4-dione Chemical compound O=C1NC(=O)NC1CC1=CC=CC=C1 DBOMTIHROGSFTI-UHFFFAOYSA-N 0.000 description 1
- FZRCKLPSHGTOAU-UHFFFAOYSA-N 6-amino-1,4-dimethylcyclohexa-2,4-diene-1-carbaldehyde Chemical compound CC1=CC(N)C(C)(C=O)C=C1 FZRCKLPSHGTOAU-UHFFFAOYSA-N 0.000 description 1
- 108010082340 Arginine deiminase Proteins 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000193764 Brevibacillus brevis Species 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- NXQJDVBMMRCKQG-SSDOTTSWSA-N D-5-phenylhydantoin Chemical compound O=C1NC(=O)N[C@@H]1C1=CC=CC=C1 NXQJDVBMMRCKQG-SSDOTTSWSA-N 0.000 description 1
- 150000008574 D-amino acids Chemical class 0.000 description 1
- 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 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 241000193386 Lysinibacillus sphaericus Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 239000003905 agrochemical Substances 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 239000000908 ammonium hydroxide Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- XEXFVRMLYUDDJY-UHFFFAOYSA-N azane;hydrate;hydrochloride Chemical compound [NH4+].[NH4+].[OH-].[Cl-] XEXFVRMLYUDDJY-UHFFFAOYSA-N 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 125000003917 carbamoyl group Chemical group [H]N([H])C(*)=O 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 239000012089 stop solution Substances 0.000 description 1
- 239000010902 straw Substances 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明は、Ｄ−５−置換ヒダントイン類をＤ−Ｎ−カル
バモイル−α−アミノ酸類に変換する能力を有するバチ
ルス・セレウス（Ｂａｃｉｌｌｕｓ Ｃｅｒｅｕｓ）（
ＦＥＲＭ Ｐ−１０３８４）を用いることにより、農薬
・抗生物置等の原料として工業的に重要な’ｌ’！質で
あるＤ−アミノ酸類の原料である［）−Ｎ−カルバモイ
ル−α−ごアミノ酸類を極めて有利に製造する方法に関
するものである。DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to Bacillus Cereus (
By using FERM P-10384), 'l', which is industrially important as a raw material for agricultural chemicals, antibiotic equipment, etc. The present invention relates to a very advantageous method for producing [)-N-carbamoyl-α-amino acids, which are raw materials for D-amino acids.
（従来の技術及び発明が解決しようとする課題）従来、
微生物の作用で５−置換ヒダントイン類から０４−カル
バモイル〜 α−アミノ酸類を製造する方法は、特公昭
６２−３０７５８号等にバチルス・プレビス（Ｂａｃｉ
ｌｌｕｓ Ｂｒｅｖｉｓ）およびバチルス・スファエリ
カス（Ｂａｃｉｌｌｕｓ ５ｐｈａｅｒｉｃｕｓ）等の
報告例が知られているが、いずれも収率が低く、゛実用
的ではない。(Prior art and problems to be solved by the invention) Conventionally,
A method for producing 04-carbamoyl to α-amino acids from 5-substituted hydantoins by the action of microorganisms is described in Japanese Patent Publication No. 30758/1983, etc.
Although reported examples include Bacillus brevis and Bacillus sphaericus, both have low yields and are not practical.
そこで、本発明は［１−５−置換ヒダントイン類をり。Therefore, the present invention provides [1-5-substituted hydantoins].
Ｎ−カルバモイル−α、アミノ酸類に変換する能力を有
するバチルス・セレウス（Ｂａｃｉｌｌｕｓ Ｃｅｒｅ
ｕｓ）（ＦＥＲＭ Ｐ−１０３８４）を用いることによ
り、工業的に有利に製造する方法を提供するものである
。Bacillus cereus has the ability to convert N-carbamoyl-α into amino acids.
US) (FERM P-10384) provides an industrially advantageous manufacturing method.
〔課題を解決するための手段及び作用〕本発明者らは、
このような従来の製造法に対し、より効率のよい製造方
法を研究した結果、バチルス・セレウス（Ｂａｃｉｌｌ
ｕｓ Ｃｅｒｅｕｓ） （ＦＥＲＭ Ｐ−１０３８４）
にＤ−５−？１［換ヒダントイン類を［１−Ｎ−カルバ
モイル−α−アミノ酸類に効率よく、変換する能力を有
することを見出し、本発明を完成するに至った。[Means and effects for solving the problem] The present inventors,
As a result of research into a more efficient manufacturing method compared to the conventional manufacturing method, we discovered that Bacillus cereus (Bacillus cereus)
(FERM P-10384)
D-5-? The present inventors have discovered that the present invention has the ability to efficiently convert hydantoins into [1-N-carbamoyl-α-amino acids, and have completed the present invention.
すなわち、本発明は
一般式（１）
％式％
（式中、Ｒはアルキル基、置換アルキル基、アラルキル
基、置換アラルキル基、フェニル基、置換フェニル基、
フリル基、ピリジル基、チアゾリル基、イミダゾリル基
またはインドリル基を示す。That is, the present invention is based on the general formula (1) % formula % (wherein R is an alkyl group, a substituted alkyl group, an aralkyl group, a substituted aralkyl group, a phenyl group, a substituted phenyl group,
It represents a furyl group, a pyridyl group, a thiazolyl group, an imidazolyl group, or an indolyl group.
）で表されるり、５．置換ヒダントイン類にＤ−５−置
換ヒダントイン類をＤ−Ｎ−カルバモイル、α−アミノ
酸類に変換する能力を育するバチルス・セレウス（Ｂａ
ｃｉｌｌｕｓ Ｃｅｒｅｕｓ）（ＦＥＲＭ Ｐ−１０３
８４）を作用せしめてＤ−Ｎ−カルバモイル−α−アミ
ノ酸類に変換せしめることを特徴とする一瓜式（ＩＩ）
（式中、Ｒは一瓜式（１） と間し。）で表されるＤ−
Ｎ−カルバモイル−α−アミノ酸類の製造方法に関する
ものである。) or 5. Bacillus cereus (Ba.
cillus Cereus) (FERM P-103
84) to convert it into D-N-carbamoyl-α-amino acids. D-
The present invention relates to a method for producing N-carbamoyl-α-amino acids.
本発明で使用する微生物は、土壌から採取・分離された
細菌で、以下に示す菌学的性状の所見よりバチルス・セ
レウス（Ｂａｃｉｌｌｕｓ Ｃｅｒｅｕｓ）（ＦＥＲＭ
Ｐ−１０３８４） と同定した。The microorganisms used in the present invention are bacteria collected and isolated from soil, and based on the findings of the mycological properties shown below, they are Bacillus Cereus (FERM).
P-10384).
Ａ＝形態
■細胞の形状、大きさ＝１．２〜Ｌ、５ Ｘ３．Ｏ〜５
．０μ桿菌
無二あり
成：あり、楕円形
性：あり
■運動性の有
■胞 子 の 形
■ダラム染色
Ｂ：生理学的性質
■生 育 の 範 囲：温度４５°Ｃまで生育■無 機
窒 素 源：硝酸塩 十
■カ タ ラ − ゼ： ＋
■酸素に対する態度 ：好気性
■ｖｐ テ ス ト：
＠糖類からの酸ニゲルコース −
二Ｌ−アラビノース
：キシロース
：アンニトール
■グルコー久からのガス：
■し シ チ す − ゼ：
■スターチの加水分Ｍ ：
■グルタチンの加水分解：
■カゼインの加水分解 ：
■クエン酸の資化性：＋
■プロピオン酸の資化性：
■チロシンの分Ｍ：＋
■硝酸塩の利用：＋
［相］インドールの精製；
■フェニルアラニンジアミナーゼ：−
■アルギニンジヒドロラーゼ：十
Ｄ−５−ｆ換ヒダントイン類に本発明のバチルス・セレ
ウス（Ｂａｃｉｌｌｕｓ Ｃｅｒｅｕｓ）（ＦＥＲＭ
Ｐ−１０３８４）を作用せしめる方法は、本微生物の菌
体または菌体の処理物を水溶液中で接触せしめる方法で
ある。A = Morphology ■Cell shape, size = 1.2-L, 5 X3. O~5
．． 0μ Bacillus Unique formation: Yes, ellipticity: Yes ■Motile ■Spore shape ■Durham staining B: physiological properties ■Growth range: Grows up to 45°C ■Inorganic nitrogen Source: Nitrate Catalase: + ■Attitude towards oxygen: Aerobic ■vp Test: @ Acid Nigelcose from sugars - L-arabinose: Xylose: Annitol ■ Gas from glucose: ■ Hydrolysis of starch M: ■Hydrolysis of glutatine: ■Hydrolysis of casein: ■Assimilation of citric acid: + ■Assimilation of propionic acid: ■Tyrosine M: + ■ Utilization of nitrate: + [Phase] Purification of indole; ■ Phenylalanine diaminase: - ■ Arginine dihydrolase: Bacillus Cereus (FERM) of the present invention to 10D-5-f-converted hydantoins.
P-10384) is a method in which cells of the present microorganism or a treated product of the cells are brought into contact with each other in an aqueous solution.
本微生物の培養に用いられる培地は、通常、責化しうる
炭素源、窒素源および微生物の生育に必要な無機塩栄養
素を含有させる、通常の培地である。The medium used for culturing the present microorganism is a conventional medium that usually contains a carbon source, a nitrogen source, and inorganic salt nutrients necessary for the growth of the microorganism.
培養条件は、好気的条件下にて、ｐＨ・４〜９、温度２
５〜４５’Ｃの適当な箱面に制御しつつ行えば望ましい
。The culture conditions are aerobic, pH 4-9, temperature 2.
It is desirable to control the temperature to a suitable box surface of 5 to 45'C.
本発明で用いられる微生物は、自然界に存在する野性法
からＤ−５−置換ヒダントイン類をＤ−Ｎ−カルハモイ
ル−α−アミノ酸類に変換する能力の有無を調べること
によって分離、選択されたものである。The microorganisms used in the present invention were isolated and selected by examining whether they have the ability to convert D-5-substituted hydantoins into D-N-carhamoyl-α-amino acids using wild methods that exist in nature. be.
この０．５．置換ヒダントイン類をＤ−Ｎ−カルバモイ
ル４α−アミノ酸類への変換する能力のｔ★定方法とし
ては、例えば、次のような方法が用いられる。This 0.5. For example, the following method is used to determine the ability to convert substituted hydantoins into DN-carbamoyl 4α-amino acids.
検定微生物の培養液５ｍ２を採取し、遠心分離によって
集菌した後、この藁Ｎ菌体を同容積の殺菌した生理食塩
水で洗浄後、２Ｉｌ１１の０．５重量％Ｄ−イソプロピ
ルヒダントインの塩化アンモニウム−水酸化アンモニウ
ム緩衝液（０，１Ｍ濃度ｐｌ（９，０）基質液中に分散
させて、温度３５°Ｃ１２４時間反応させる。After collecting 5 m2 of the culture solution of the test microorganisms and collecting them by centrifugation, the straw N bacterial cells were washed with the same volume of sterilized physiological saline, and then 2Il11 of 0.5% by weight of D-isopropylhydantoin ammonium chloride was added. - Ammonium hydroxide buffer (0.1M concentration pl(9,0)) is dispersed in the substrate solution and reacted at a temperature of 35°C for 124 hours.
次いで、１２重量％トリクロル酢酸水溶液０．５ｍｌを
添加して反応を停止させる。Next, 0.5 ml of a 12% by weight aqueous trichloroacetic acid solution is added to stop the reaction.
反応停止液中に１０重量％パラジメチルアミノベンズア
ルデヒドの濃塩酸溶液０．５ｍ ｆｔを添加し、１００
００ｒｐ−で１０分間遠心分雄して上澄み液を得る。Add 0.5 m ft of a concentrated hydrochloric acid solution of 10% by weight paradimethylaminobenzaldehyde to the reaction stop solution, and
Centrifuge at 00 rpm for 10 minutes to obtain a supernatant.
このようにして、カルバモイル基の発色により、４２０
ｎｉにて、生成したＤ−Ｎ−カルバミル−バリンを比色
定量する。In this way, due to the color development of the carbamoyl group, 420
The produced DN-carbamyl-valine is colorimetrically determined at Ni.
上記のようにして、り−５−置換ヒダントイン類をＤ−
Ｎ−カルバモイル−α−アミノ酸類に変換する能力を有
すると認められた菌株について、さらに、生成したＤ−
Ｎ−カルバモイル−α−アミノ酸類を常法により単離・
精製し、旋光度を測定することにより検定した。 本発
明で用いられる微生物であるバチルス・セレウス（Ｂａ
ｃｉｌｌｕｓ Ｃｅｒｅｕｓ）（ＦＥＲＭ Ｐ−１０３
８４）は、前記の検定に合格したものである。As described above, the di-5-substituted hydantoins were converted to D-
For strains recognized to have the ability to convert into N-carbamoyl-α-amino acids, the produced D-
Isolate N-carbamoyl-α-amino acids by conventional methods.
It was purified and assayed by measuring the optical rotation. The microorganism used in the present invention, Bacillus cereus (Ba
cillus Cereus) (FERM P-103
84) passed the above test.
本発明に用いられる酵素反応基質とは、各種Ｄ−５−置
換ヒダントイン類で具体的に例示すると、Ｄ−５−メチ
ルヒダントイン、Ｄ−５−イソプロピルヒダントイン、
Ｄ−５−イソブチルヒダントイン、Ｄ−５−ｓｅｃブチ
ルヒダントイン、［）−５−メチルチオエチルヒダント
イン、Ｄ−５−フェニルヒダントイン、Ｄ−５−ベンジ
ルヒダントイン、Ｄ−５−インドリルメチルヒダントイ
ン等がある。The enzyme reaction substrate used in the present invention includes various D-5-substituted hydantoins, such as D-5-methylhydantoin, D-5-isopropylhydantoin,
Examples include D-5-isobutylhydantoin, D-5-sec butylhydantoin, [)-5-methylthioethylhydantoin, D-5-phenylhydantoin, D-5-benzylhydantoin, and D-5-indolylmethylhydantoin.
酵素反応における反応基質の濃度は、０．１〜１０重量
％の濃度まで用いることができる。The concentration of the reaction substrate in the enzyme reaction can range from 0.1 to 10% by weight.
反応温度は、使用する微生物の［＋−Ｎ−カルバモイル
−α−アミノ酸類への変換する能力を持つ酵素の至ｉｉ
！Ｉ’ｉ度が採用されるが、通常、２０〜６０°Ｃの範
囲にある。The reaction temperature is determined by the temperature of the microorganism used and the enzyme capable of converting into [+-N-carbamoyl-α-amino acids].
! I'i degrees are employed, but are usually in the range of 20-60°C.
反応中のｐＨは、使用する微生物のＤ−Ｎ−カルバモイ
ル、α−アミノ酸類への変換する能力を持つ酵素の至適
ｐＨが採用されるが、通常、ｐＨ・５〜９の範囲にある
。特に好ましくは、温度２０〜５０°Ｃ％ｐＨ・６〜１
０である。The pH during the reaction is the optimum pH of the microorganism used for the enzyme having the ability to convert into DN-carbamoyl and α-amino acids, and is usually in the range of pH 5 to 9. Particularly preferably, temperature: 20-50°C% pH: 6-1
It is 0.
前述したようなり−５−置換ヒダントイン類を不斉的に
変換して生成した［）−Ｎ−カルバモイル−α−アミノ
酸類の単離は、濃縮・中和・イオン交換樹脂処理等の公
知の方法を利用することにより、巨的物であるｎ−Ｎ−
カルバモイル−α−アミノ酸類を取得できる。[)-N-carbamoyl-α-amino acids produced by asymmetrically converting -5-substituted hydantoins as described above can be isolated by known methods such as concentration, neutralization, and ion exchange resin treatment. By using the gigantic n-N-
Carbamoyl-α-amino acids can be obtained.
本発明の実施においては、技術常識に従い適宜界面活性
剤を併用することができる。In carrying out the present invention, a surfactant may be appropriately used in combination according to common general knowledge.
〔実施例］
以下、実施例により本発明を具体的に説明するが、本発
明はこれらの例のみに限定されるものではない。[Examples] Hereinafter, the present invention will be specifically explained using Examples, but the present invention is not limited only to these Examples.
実施例１
表１に示した培地を２５０ｍ ｌ三角フラスコに２０！
ｌｌ入れ、１２０℃、１５分間殺菌し、ＤＬ−イソプロ
ピルヒダントインは、別に殺菌して混合した。これにブ
イヨン寒天培地で温度２８°Ｃ２１６時間培養したバチ
ルス・セレウス（Ｂａｃｉｌｌｕｓ Ｃｅｒｅｕｓ）（
ＦＥＲＭ Ｐ−１０３８４）を１白金耳接種し、温度２
８°Ｃ１２４時間培養した。 この培養液を遠心分離に
より面体を採取し、培養液と同量の殺菌された生理食塩
水にて１回洗浄し、面体を集めた。Example 1 20ml of the culture medium shown in Table 1 was placed in a 250ml Erlenmeyer flask.
The mixture was sterilized at 120° C. for 15 minutes, and DL-isopropylhydantoin was sterilized and mixed separately. To this, Bacillus Cereus (
FERM P-10384) was inoculated with one platinum loop, and the temperature was 2.
Cultured at 8°C for 124 hours. The facepieces were collected by centrifuging the culture solution, washed once with sterilized physiological saline in the same amount as the culture solution, and collected.
二の７体を表２に示すＤ−５−２換ヒダントイン類のい
ずれか一種壱Ｌｏｇ／ ｆ含む０．１ｊ塩化アンモニウ
ムー水酸化アンモニウム緩衝液（ｐＨ・９．０）・・・
終末５ｍｊ２・・・に１．＃！になるように添加し、温
度３６°Ｃ８２０時間反応した。生成するＤ−Ｎ−カル
バモイル−α−アミノ酸類は、前記の方法にて測定し、
また、これらのＤ−Ｎ−カルバモイル−α−アミノ酸類
を分離・精製し、旋光度の測定を行った結果、生成する
アミノ酸は、すべての場合り体であることを確認した。A 0.1j ammonium chloride-ammonium hydroxide buffer (pH 9.0) containing one Log/f of any one of the D-5-2-converted hydantoins shown in Table 2.
1 at the end 5mj2... #! The mixture was added so as to be reacted at a temperature of 36° C. for 820 hours. The produced DN-carbamoyl-α-amino acids are measured by the method described above,
In addition, as a result of separating and purifying these DN-carbamoyl-α-amino acids and measuring their optical rotations, it was confirmed that the produced amino acids were all in the form of a polymer.
結果は表２に示す。The results are shown in Table 2.
（以下、余白）
〔発明の効果］
本発明は、［１−５＝置換ヒダントイン類をＤ−Ｎ−カ
ルバモイル−α−アミノ酸類に変換する能力を有するバ
チルス・セレウス（Ｂａｃｉｌｌｕｓ Ｃｅｒｅｕｓ）
（ＦＥＲＭ Ｐ−１０３８４）を用いることにより、Ｄ
−５−置換ヒダントイン類から容易に高収率でＤ−Ｎ−
カルバモイル−α−アミノ酸類を取得できるので、Ｄ−
Ｎ−カルバモイル−α−アミノ酸類の製造に際し、極め
て有利な方ン去である。(Hereinafter, the margins) [Effects of the Invention] The present invention is directed to the use of Bacillus Cereus, which has the ability to convert [1-5=substituted hydantoins into D-N-carbamoyl-α-amino acids].
By using (FERM P-10384), D
D-N- can be easily obtained in high yield from -5-substituted hydantoins.
Since carbamoyl-α-amino acids can be obtained, D-
This is an extremely advantageous method for producing N-carbamoyl-α-amino acids.
特許出願人 三井東圧化学株式会社Patent applicant: Mitsui Toatsu Chemical Co., Ltd.
Claims (1)
基、置換アラルキル基、フェニル基、置換フェニル基、
フリル基、ピリジル基、チアゾリル基、イミダゾリル基
またはインドリル基を示す。 ）で表されるＤ−５−置換ヒダントイン類にＤ−５−置
換ヒダントイン類をＤ−Ｎ−カルバモイル−α−アミノ
酸類に変換する能力を有するバチルス・セレウス（Ｂａ
ｃｉｌｌｕｓ Ｃｅｒｅｕｓ）（ＦＥＲＭＰ−１０３８
４）を作用せしめてＤ−Ｎ−カルバモイル−α−アミノ
酸類に変換せしめることを特徴とする一般式（II） ▲数式、化学式、表等があります▼（II） （式中、Ｒは一般式（ I ）と同じ。）で表されるＤ−
Ｎ−カルバモイル−α−アミノ酸類の製造方法。[Claims] 1 General formula (I) ▲ Numerical formulas, chemical formulas, tables, etc. ▼ (I) (In the formula, R is an alkyl group, a substituted alkyl group, an aralkyl group, a substituted aralkyl group, a phenyl group, a substituted phenyl group) basis,
It represents a furyl group, a pyridyl group, a thiazolyl group, an imidazolyl group, or an indolyl group. ) has the ability to convert D-5-substituted hydantoins into D-N-carbamoyl-α-amino acids.
cillus Cereus) (FERMP-1038
General formula (II) characterized by converting 4) into D-N-carbamoyl-α-amino acids ▲There are mathematical formulas, chemical formulas, tables, etc.▼(II) (In the formula, R is the general formula (same as (I))
Method for producing N-carbamoyl-α-amino acids.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP29962988A JPH02145192A (en) | 1988-11-29 | 1988-11-29 | Production of d-n-carbamoyl-alpha-amino acids |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP29962988A JPH02145192A (en) | 1988-11-29 | 1988-11-29 | Production of d-n-carbamoyl-alpha-amino acids |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH02145192A true JPH02145192A (en) | 1990-06-04 |
Family
ID=17875074
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP29962988A Pending JPH02145192A (en) | 1988-11-29 | 1988-11-29 | Production of d-n-carbamoyl-alpha-amino acids |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH02145192A (en) |
-
1988
- 1988-11-29 JP JP29962988A patent/JPH02145192A/en active Pending
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