JPH0479895A - Production of d-amino acid - Google Patents
Production of d-amino acidInfo
- Publication number
- JPH0479895A JPH0479895A JP19167790A JP19167790A JPH0479895A JP H0479895 A JPH0479895 A JP H0479895A JP 19167790 A JP19167790 A JP 19167790A JP 19167790 A JP19167790 A JP 19167790A JP H0479895 A JPH0479895 A JP H0479895A
- Authority
- JP
- Japan
- Prior art keywords
- amino acid
- racemic
- compound
- nitrile
- microorganisms
- 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 5
- -1 benzonitrile compound Chemical class 0.000 claims abstract description 17
- 244000005700 microbiome Species 0.000 claims abstract description 14
- 150000003839 salts Chemical class 0.000 claims abstract description 9
- 150000002825 nitriles Chemical class 0.000 claims abstract description 7
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims abstract description 7
- 230000003301 hydrolyzing effect Effects 0.000 claims abstract description 5
- 150000008574 D-amino acids Chemical class 0.000 claims description 11
- 239000000126 substance Substances 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 2
- 241000186073 Arthrobacter sp. Species 0.000 claims 1
- 239000002994 raw material Substances 0.000 abstract description 10
- 230000003287 optical effect Effects 0.000 abstract description 6
- 241000186063 Arthrobacter Species 0.000 abstract description 5
- JFDZBHWFFUWGJE-UHFFFAOYSA-N benzenecarbonitrile Natural products N#CC1=CC=CC=C1 JFDZBHWFFUWGJE-UHFFFAOYSA-N 0.000 abstract description 5
- 150000001413 amino acids Chemical class 0.000 abstract description 3
- 239000003814 drug Substances 0.000 abstract description 2
- 229940079593 drug Drugs 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 19
- 230000001580 bacterial effect Effects 0.000 description 17
- 238000000034 method Methods 0.000 description 16
- ZGUNAGUHMKGQNY-SSDOTTSWSA-N D-alpha-phenylglycine Chemical compound OC(=O)[C@H](N)C1=CC=CC=C1 ZGUNAGUHMKGQNY-SSDOTTSWSA-N 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 230000000813 microbial effect Effects 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- ZOCYQVNGROEVLU-UHFFFAOYSA-N Isopentadecylic acid Natural products CC(C)CCCCCCCCCCCC(O)=O ZOCYQVNGROEVLU-UHFFFAOYSA-N 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- FVSKHRXBFJPNKK-UHFFFAOYSA-N propionitrile Chemical compound CCC#N FVSKHRXBFJPNKK-UHFFFAOYSA-N 0.000 description 3
- XKLJLHAPJBUBNL-UHFFFAOYSA-N 12-methyltetradecanoic acid Chemical compound CCC(C)CCCCCCCCCCC(O)=O XKLJLHAPJBUBNL-UHFFFAOYSA-N 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 2
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 238000005191 phase separation Methods 0.000 description 2
- SUSQOBVLVYHIEX-UHFFFAOYSA-N phenylacetonitrile Chemical compound N#CCC1=CC=CC=C1 SUSQOBVLVYHIEX-UHFFFAOYSA-N 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- FTNJQNQLEGKTGD-UHFFFAOYSA-N 1,3-benzodioxole Chemical compound C1=CC=C2OCOC2=C1 FTNJQNQLEGKTGD-UHFFFAOYSA-N 0.000 description 1
- ZONJATNKKGGVSU-UHFFFAOYSA-N 14-methylpentadecanoic acid Chemical compound CC(C)CCCCCCCCCCCCC(O)=O ZONJATNKKGGVSU-UHFFFAOYSA-N 0.000 description 1
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 description 1
- NLHHRLWOUZZQLW-UHFFFAOYSA-N Acrylonitrile Chemical compound C=CC#N NLHHRLWOUZZQLW-UHFFFAOYSA-N 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 239000004254 Ammonium phosphate Substances 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 1
- ZOXJGFHDIHLPTG-UHFFFAOYSA-N Boron Chemical compound [B] ZOXJGFHDIHLPTG-UHFFFAOYSA-N 0.000 description 1
- FXUKWLSZZHVEJD-UHFFFAOYSA-N C16:0-14-methyl Natural products CCC(C)CCCCCCCCCCCCC(O)=O FXUKWLSZZHVEJD-UHFFFAOYSA-N 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 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
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 229910021578 Iron(III) chloride Inorganic materials 0.000 description 1
- ZGUNAGUHMKGQNY-ZETCQYMHSA-N L-alpha-phenylglycine zwitterion Chemical compound OC(=O)[C@@H](N)C1=CC=CC=C1 ZGUNAGUHMKGQNY-ZETCQYMHSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 229910021380 Manganese Chloride Inorganic materials 0.000 description 1
- GLFNIEUTAYBVOC-UHFFFAOYSA-L Manganese chloride Chemical compound Cl[Mn]Cl GLFNIEUTAYBVOC-UHFFFAOYSA-L 0.000 description 1
- GYCMBHHDWRMZGG-UHFFFAOYSA-N Methylacrylonitrile Chemical compound CC(=C)C#N GYCMBHHDWRMZGG-UHFFFAOYSA-N 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 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
- 230000003213 activating effect Effects 0.000 description 1
- UAMZETBJZRERCQ-UHFFFAOYSA-N alpha-aminopropionitrile Chemical compound CC(N)C#N UAMZETBJZRERCQ-UHFFFAOYSA-N 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 229910000148 ammonium phosphate Inorganic materials 0.000 description 1
- 235000019289 ammonium phosphates Nutrition 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 229910052796 boron Inorganic materials 0.000 description 1
- KVNRLNFWIYMESJ-UHFFFAOYSA-N butyronitrile Chemical compound CCCC#N KVNRLNFWIYMESJ-UHFFFAOYSA-N 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 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
- NKKMVIVFRUYPLQ-NSCUHMNNSA-N crotononitrile Chemical compound C\C=C\C#N NKKMVIVFRUYPLQ-NSCUHMNNSA-N 0.000 description 1
- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 239000011790 ferrous sulphate Substances 0.000 description 1
- 235000003891 ferrous sulphate Nutrition 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000007952 growth promoter Substances 0.000 description 1
- WJRBRSLFGCUECM-UHFFFAOYSA-N hydantoin Chemical compound O=C1CNC(=O)N1 WJRBRSLFGCUECM-UHFFFAOYSA-N 0.000 description 1
- 229940091173 hydantoin Drugs 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
- LRDFRRGEGBBSRN-UHFFFAOYSA-N isobutyronitrile Chemical compound CC(C)C#N LRDFRRGEGBBSRN-UHFFFAOYSA-N 0.000 description 1
- WOFDVDFSGLBFAC-UHFFFAOYSA-N lactonitrile Chemical compound CC(O)C#N WOFDVDFSGLBFAC-UHFFFAOYSA-N 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 239000011565 manganese chloride Substances 0.000 description 1
- 235000002867 manganese chloride Nutrition 0.000 description 1
- 229940099607 manganese chloride Drugs 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 150000002897 organic nitrogen compounds Chemical class 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 229910000160 potassium phosphate Inorganic materials 0.000 description 1
- 239000008057 potassium phosphate buffer Substances 0.000 description 1
- 235000011009 potassium phosphates Nutrition 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000004317 sodium nitrate Substances 0.000 description 1
- 235000010344 sodium nitrate Nutrition 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 235000011008 sodium phosphates Nutrition 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- IAHFWCOBPZCAEA-UHFFFAOYSA-N succinonitrile Chemical compound N#CCCC#N IAHFWCOBPZCAEA-UHFFFAOYSA-N 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】
り粟上圓肌■公団
本発明は、微生物を利用してラセミ体の２−アミノベン
ゾニトリル化合物あるいはその塩から相当するＤ−アミ
ノ酸またはその塩を製造する方法に関する。Ｄ−アミノ
酸中、特にＤ−フェニルグリシンはアンピシリン等の抗
生物質の中間体として用いられており、医薬品原料とし
て有用である。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing a corresponding D-amino acid or a salt thereof from a racemic 2-aminobenzonitrile compound or a salt thereof using microorganisms. Among D-amino acids, D-phenylglycine in particular is used as an intermediate for antibiotics such as ampicillin, and is useful as a raw material for pharmaceuticals.
従来勿伎歪
従来、Ｄ−フェニルグリシン等のＤ−アミノ酸を製造す
る方法としては、スルフォン酸などを分割剤として物理
化学的に晶出させる方法（特開昭５１−９８２３７号公
報） 、５−１換ヒダントインに微生物の菌体を作用さ
せる方法（特開昭５５−４５１９５号公報）等が知られ
ている。しかしこれらの方法は数段の反応を経ることか
ら工程が複雑となる欠点を有している。Traditionally, as a method for producing D-amino acids such as D-phenylglycine, there is a method of physicochemically crystallizing using sulfonic acid or the like as a resolving agent (Japanese Patent Application Laid-open No. 51-98237), 5- A method is known in which monoconverted hydantoin is made to act on microbial cells (Japanese Unexamined Patent Publication No. 55-45195). However, these methods have the disadvantage that the steps are complicated because they involve several stages of reaction.
しよ゛と る 占
本発明は、上記の欠点を解決することを目的としてなさ
れたものであって、ラセミ体の２−アミノベンゾニトリ
ル化合物から直接相当するＤ−アミノ酸を製造する方法
を提供しようとするものである。The present invention has been made to solve the above-mentioned drawbacks, and provides a method for directly producing the corresponding D-amino acid from a racemic 2-aminobenzonitrile compound. That is.
１ ”るための
本発明は、アルスロバククー属（Ａｒｔｈｒｏｂａｃｔ
ｅγｓｐ、 ）に属するニトリル加水分解能を有する微
生物をラセミ体の２−アミノベンゾニトリル化合物に作
用させると相当するＤ−アミノ酸を選択的に産生ずると
いう知見に基いてなされたものである。1” The present invention is directed to the use of Arthrobacterium
This invention was based on the knowledge that when a microorganism having the ability to hydrolyze nitrile belonging to the group eγsp, ) is allowed to act on a racemic 2-aminobenzonitrile compound, the corresponding D-amino acid is selectively produced.
本発明の構成上の特徴は、アルスロバクタ−属に属する
群から選択されるニトリル加水分解能を有する微生物を
、ラセミ体の２−アミノベンゾニトリル化合物に作用さ
せて相当するＤ−アミノ酸を選択的に生産させることに
ある。The structural feature of the present invention is to selectively produce the corresponding D-amino acid by allowing a microorganism having a nitrile hydrolyzing ability selected from the group belonging to the genus Arthrobacter to act on a racemic 2-aminobenzonitrile compound. It's about letting people know.
本発明における２−アミノベンゾニトリル化合物は、次
の一般式（１）で表される化合物あるいはその塩がある
。The 2-aminobenzonitrile compound in the present invention includes a compound represented by the following general formula (1) or a salt thereof.
ＮＨ。N.H.
（式中、Ｒはフェニル基、置換フェニル基を意味する。(In the formula, R means a phenyl group or a substituted phenyl group.
）
置換フェニル基の代表的なものとして４−ヒドロキシル
置換フェニル基を挙げることができる。また、塩として
は、塩酸塩、硝酸塩等の塩を挙げることができる。) A typical substituted phenyl group includes a 4-hydroxyl substituted phenyl group. Further, examples of the salt include salts such as hydrochloride and nitrate.
本発明に用いられる微生物としては、アルスロバクタ−
属に属する微生物で、ニトリル加水分解能を有するもの
が用いられ、その例としては土壌から採取されたＡｒｔ
ｈｒｏｂａｃｔｅｒ ｓｐ、 ＰＣ−３（寄託番号微工
研条寄１５６０）を挙げることができる。このＡｒＬｒ
ｏｂａｃｔｅｒ ｓｐ、 ＰＣ−３の菌学的性質を以
下に示す。As the microorganisms used in the present invention, Arthrobacter
Microorganisms belonging to the genus genus that have the ability to hydrolyze nitrile are used; examples include Art.
hrobacter sp, PC-3 (deposit number: KAIKEN JOKYO 1560). This ArLr
The mycological properties of PC-3 are shown below.
ダラム染色性 十
胞 子
運 動 性
形 状 桿菌、ヘン毛無
カタラーゼ
＋
オキシダーゼ
（＋）
叶テスト
細胞の化学分析
ミコール酸を含まない。細胞壁のジアミノ酸はリジンで
ある。脂肪酸組成は１２−メチルテトラデカン酸しｊｃ
＋ｓ＋ｏ；６９χ）、１４−メチルヘキサデカン酸（ａ
ｉｃ＋ｔ＋ｏ；２１χ）が主であり、少量の１３−メチ
ルテトラデカン酸（ｉｃ１ｓ＋ａ；約４χ）、１４−メ
チルペンタデカン酸（ｉｃｉｔｈ＋。；約４Ｋ）を含む
。Durham staining Decaspore Motility Shape Bacillus, no catalase + oxidase (+) Chemical analysis of leaf test cells Contains no mycolic acid. The diamino acid in the cell wall is lysine. The fatty acid composition is 12-methyltetradecanoic acid.
+s+o; 69χ), 14-methylhexadecanoic acid (a
ic+t+o; 21χ), and contains small amounts of 13-methyltetradecanoic acid (ic1s+a; about 4χ) and 14-methylpentadecanoic acid (icith+; about 4K).
本発明においては、上記したような２−アミノベンゾニ
トリル化合物を原料とし、これに上記微生物を作用させ
るものであるが、この方法としては、例えば、次の（ａ
）〜（Ｃ）のいずれかの方法を適用することが好ましい
。In the present invention, the above-mentioned 2-aminobenzonitrile compound is used as a raw material, and the above-mentioned microorganism is allowed to act on it.
) to (C) are preferably applied.
すなわち、（ａ）上記微生物を、誘導基質としてのニト
リル化合物、例えばプロピオニトリルを含む培地中で培
養して増殖して得られた菌体に、原料を接触させて反応
させる方法、（ｂ）＠生物を予め培養し、増殖して得ら
れた菌体を誘導基質としてのニトリル化合物に接触させ
た後、該菌体に原料を加えて反応させる方法、及び（Ｃ
）微生物を予め培養し、増殖して得られた菌体に原料を
直接接触させて反応させる方法を通用する。That is, (a) a method of culturing and multiplying the above-mentioned microorganism in a medium containing a nitrile compound such as propionitrile as an induction substrate, and causing the resulting bacterial cells to contact with a raw material to react; (b) @ A method of culturing an organism in advance, bringing the obtained microbial cells into contact with a nitrile compound as an induction substrate, and then adding a raw material to the microbial cells and causing a reaction, and (C
) A method is commonly used in which microorganisms are cultured in advance and the resulting microorganisms are brought into direct contact with the raw material to cause a reaction.
上記（ａ）及び（ｂ）の方法で用いる誘導基質としての
ニトリル化合物は、プロピオニトリルのほかに、アセト
ニトリル、ｎ−ブチロニトリル、トカプロニトリル、メ
タクリロニトリル、イソブチロニトリル、ゲルタロニト
リル、トリアクリロニトリル、クロトノニトリル、ラク
トニトリル、サクシノニトリル、アクリロニトリル、ベ
ンゾニトリル及びフェニルアセトニトリル等を例示し得
る。In addition to propionitrile, the nitrile compounds used in the methods (a) and (b) above include acetonitrile, n-butyronitrile, tocapronitrile, methacrylonitrile, isobutyronitrile, geltalonitrile, Examples include triacrylonitrile, crotononitrile, lactonitrile, succinonitrile, acrylonitrile, benzonitrile, and phenylacetonitrile.
上記（ａ）の方法では、誘導基質としてのニトリル化合
物のほかに、炭素源としてグルコース、シュクロース、
糖蜜、澱粉加水分解物のようなｔｌ！質、もしくは酢酸
等のごとき菌体増殖作用を有する物質を培地に添加し、
更に、塩化アンモニウム、硫酸アンモニウム、リン酸ア
ンモニウム、硝酸アンモニウム、尿素、アンモニア水、
硝酸ナトリウム、アミノ酸及びその他の資化性有機窒素
化合物のような窒素源、リン酸カリウム、リン酸ナトリ
ウム、硫酸マグネシウム、硫酸マンガン、硫酸第１鉄、
塩化第２鉄、塩化カルシウム、塩化マンガンのごとき無
機塩類、及びホウ素、銅、亜鉛などの塩、すなわち、い
わゆる微量元素、更には必要に応じてビタミン類、酵母
エキス、コーンステープリカーの如き成長促進物質を添
加した培地に、上記各微生物の種菌を接種し、好気的条
件下で培養して菌体を増殖させる。In the method (a) above, in addition to the nitrile compound as the inducing substrate, glucose, sucrose,
tl like molasses, starch hydrolyzate! Adding a substance that has a bacterial growth effect, such as acetic acid or acetic acid, to the medium,
Furthermore, ammonium chloride, ammonium sulfate, ammonium phosphate, ammonium nitrate, urea, aqueous ammonia,
Nitrogen sources such as sodium nitrate, amino acids and other assimilable organic nitrogen compounds, potassium phosphate, sodium phosphate, magnesium sulfate, manganese sulfate, ferrous sulfate,
Inorganic salts such as ferric chloride, calcium chloride, manganese chloride, salts such as boron, copper, zinc, ie so-called trace elements, and optionally growth promoters such as vitamins, yeast extract, corn staple liquor. Inoculum of each of the above-mentioned microorganisms is inoculated into a medium to which a substance has been added, and the cells are grown by culturing under aerobic conditions.
次に、このようにして得られた菌体培養物を原料と接触
させて反応させるが、この場合、この菌体培養物以外の
、この培養物から分離した菌体の懸濁液あるいは菌体の
破砕物、乾燥菌体、または分離精製されたニトリル加水
分解酵素などの菌体処理物、さらには常法に従って固定
化した菌体および菌体処理物を用いることもできる。本
発明ではこれらを微生物調製物という。Next, the bacterial cell culture obtained in this way is brought into contact with the raw material and reacted, but in this case, a suspension of bacterial cells or bacterial cells isolated from this culture other than this bacterial cell culture is used. It is also possible to use crushed products, dried bacterial cells, or treated bacterial cells such as isolated and purified nitrile hydrolase, as well as bacterial cells fixed according to conventional methods and treated bacterial cells. In the present invention, these are referred to as microbial preparations.
この反応は、ｐＨ４〜１３、温度２０〜７０°Ｃの範囲
で１〜１５日間行う。また、反応中に菌体増殖に用いた
上記炭素源、窒素源、その他の成分を適宜添加して菌体
濃度や菌体のニトリル加水分解能を維持し、かつ高める
ことができる。また、各種有ｊａ溶媒を添加してもよい
。This reaction is carried out at a pH of 4 to 13 and a temperature of 20 to 70°C for 1 to 15 days. In addition, the above-mentioned carbon source, nitrogen source, and other components used for cell proliferation can be appropriately added during the reaction to maintain and increase the cell concentration and the nitrile hydrolyzing ability of the cell. Additionally, various types of solvents may be added.
上記反応により生成した光学活性な有機酸は、相分離、
濾過、抽出、カラムクロマトグラフィー等の公知の手段
を適用して分離、採取する。The optically active organic acid produced by the above reaction undergoes phase separation,
Separation and collection are performed by applying known means such as filtration, extraction, and column chromatography.
次に、前記（ｂ）の方法では、上記（ａ）の方法におけ
る菌体の培養増殖時に誘導基質としてのニトリル化合物
を加えずに、菌体の増殖後に当該二ｌ−ＩＪル化合物を
加えて該菌体微生物のニトリル加水分解能を活性化した
後、原料を反応させて相当するＤアミノ酸、例えばＤ−
フェニルグリシン、Ｄ−ヒドロキシフェニルグリシン等
を生産させる。Next, in the method (b) above, the nitrile compound as an inducing substrate is not added during the culture and growth of the bacterial cells in the method (a) above, but the nitrile compound is added after the bacterial cells have grown. After activating the nitrile hydrolyzing ability of the microorganism, the raw materials are reacted to form the corresponding D-amino acid, for example, D-
Phenylglycine, D-hydroxyphenylglycine, etc. are produced.
また、前記（Ｃ）の方法は、上記（ｂ）の方法における
菌体の増殖後に直ちに原料を加えて反応させて同様に相
当するＤ−アミノ酸を生産させるものである。In addition, in the method (C), the raw materials are added immediately after the bacterial cells grow in the method (b), and the reaction is caused to produce the corresponding D-amino acid.
なお、前記（ｂ）及び（Ｃ）のいずれの方法においても
、培養条件、反応条件及び生成した有機酸の分離、採取
には、前記（ａ）の方法におけるものを適用し得［実施
例〕
実施例１
シュークｔｌｌ−ス１χ、ＮａｚＨＰＯａ ’ １２Ｈ
ｚ。０．２５Ｘ、ＫＨｚＰＯ４０，２ｘ、Ｍｇ５Ｏ４Ｈ
７Ｈｚｏ Ｏ，０５χ、ＦｅＳＯ４・７１１ｚｏ Ｏ，
０３χＣａＣＥ ｚ ’ ２Ｈｚｏ Ｏ，００６χ酵母
エキス０．１χ及びプロピオニトリル０，５χを含みｐ
Ｈ７，２に調整した培地１００ｍにアルスロバクタ−（
Ａｒｔｈｒｏｂａｃｔｅｒ）ＰＣ−３株を１白金耳植菌
し、３０°Ｃで９６時間培養した。得られた培養液を１
００ＯＯＧで１０分間遠心分離して、得られた菌体を０
．１Ｍリン酸カリウム緩衝液中に光学的濃度（０，Ｄ）
が７５．６となるように懸濁した。上記菌液ｌ−を試験
管に入れ、この中に２−アミノヘンジニトリル５■を加
えて３０°Ｃ１６０分間毎分１５０往復の速度で振のを
行った。得られた反応液を遠心除菌後高速波体クロマト
グラフィーによってＤ−フェニルグリシンを単離した。In addition, in both methods (b) and (C), the culture conditions, reaction conditions, and separation and collection of the produced organic acid can be the same as those in method (a) above [Example] Example 1 Shuktll-su1χ, NazHPOa' 12H
z. 0.25X, KHzPO40,2x, Mg5O4H
7Hzo O, 05χ, FeSO4・711zo O,
03χCaCE z' 2Hzo O, 006χ Contains 0.1χ of yeast extract and 0.5χ of propionitrile p
Arthrobacter (
One platinum loop of Arthrobacter) PC-3 strain was inoculated and cultured at 30°C for 96 hours. 1 of the obtained culture solution
Centrifuge for 10 minutes at 00OOG, and the resulting bacterial cells are
．． Optical density (0,D) in 1M potassium phosphate buffer
It was suspended so that the value was 75.6. The above bacterial solution 1- was placed in a test tube, 5 μm of 2-aminohendinitrile was added thereto, and the mixture was shaken at 30° C. for 160 minutes at a speed of 150 reciprocations per minute. The resulting reaction solution was centrifuged to remove bacteria, and then D-phenylglycine was isolated by high-speed wave chromatography.
この結果、光学純度１００χｅｅのＤ−フェニルグリシ
ン１．６２■を得た。生産したフェニルグリシンの定量
はカラム充填剤として、イナートシルＯＤＳ （ガスク
ロ工業）を用いて行い、光学純度の測定はＣｈｉｒａｌ
ｐａｋ ＷＥ（ダイセル化学工業）を用いて行った。As a result, 1.62 .mu. of D-phenylglycine with an optical purity of 100 .chi.ee was obtained. The produced phenylglycine was quantitatively determined using Inertsil ODS (Gascro Industries) as a column packing material, and the optical purity was measured using Chiral.
pak WE (Daicel Chemical Industries) was used.
実施例２
実施例１と同様に培養、集菌した菌体を００３２．４に
なるように２８ ＮＨ，Ｃｊ！／Ｎ）１３バツフ ｙ
（ｐＨ１０，０）に懸濁した。上記菌液１雌に２−ア
ミノヘンジニトリル５ｍｇを加えて、６０°Ｃ１６０分
間、毎分１５０往復の速度で振盪を行った。得られた反
応液を実施例１と同様に分析したところ、光学純度９２
．７％ｅｅのＤ−フェニルグリシン１．９０■を得た。Example 2 Bacterial cells cultured and collected in the same manner as in Example 1 were adjusted to 28 NH, Cj! to give a value of 0032.4. /N) 13 batsufu y
(pH 10,0). 5 mg of 2-aminohendinitrile was added to the above bacterial solution 1, and the mixture was shaken at 60° C. for 160 minutes at a speed of 150 reciprocations per minute. When the obtained reaction solution was analyzed in the same manner as in Example 1, the optical purity was 92.
．． 1.90 ml of D-phenylglycine with 7% ee was obtained.
光皿■羞来
本発明によると、ラセミ体の２−アミノプロピオニトリ
ルから光学純度の高い相当するＤ−アミノ酸を高収率で
直接得ることができる。従って、本発明の方法は、医薬
原料として価値の高いＤ−アミノ酸、特にＤ−フェニル
グリシンを工業的有利に生産する方法である。According to the present invention, the corresponding D-amino acid with high optical purity can be directly obtained in high yield from racemic 2-aminopropionitrile. Therefore, the method of the present invention is an industrially advantageous method for producing D-amino acids, particularly D-phenylglycine, which are highly valuable as pharmaceutical raw materials.
Claims (1)
ミノベンゾニトリル化合物及び／又はその塩に、▲数式
、化学式、表等があります▼（ I ） （式中、Ｒはフェニル基または置換フェニル基を意味す
る。） アルスロバクター属（Ａｒｔｈｒｏｂａｃｔｅｒｓｐ．
）に属し、ニトリル加水分解活性を有する微生物または
その調製物を作用せしめることを特徴とするＤ−アミノ
酸の製造法。(1) The racemic 2-aminobenzonitrile compound and/or its salt represented by the following general formula (I) has ▲numerical formulas, chemical formulas, tables, etc.▼(I) (wherein, R is a phenyl group or a substituted phenyl group.) Arthrobacter sp.
1.) A method for producing D-amino acids, which comprises reacting with a microorganism having nitrile hydrolyzing activity or a preparation thereof.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP19167790A JPH0479895A (en) | 1990-07-19 | 1990-07-19 | Production of d-amino acid |
| US08/277,775 US5587303A (en) | 1988-03-08 | 1994-07-20 | Production process of L-amino acids with bacteria |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP19167790A JPH0479895A (en) | 1990-07-19 | 1990-07-19 | Production of d-amino acid |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH0479895A true JPH0479895A (en) | 1992-03-13 |
Family
ID=16278621
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP19167790A Pending JPH0479895A (en) | 1988-03-08 | 1990-07-19 | Production of d-amino acid |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0479895A (en) |
-
1990
- 1990-07-19 JP JP19167790A patent/JPH0479895A/en active Pending
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