JPH02190196A - Production of optically active substance using actinomycete belonging to genus nocardia - Google Patents
Production of optically active substance using actinomycete belonging to genus nocardiaInfo
- Publication number
- JPH02190196A JPH02190196A JP1163789A JP1163789A JPH02190196A JP H02190196 A JPH02190196 A JP H02190196A JP 1163789 A JP1163789 A JP 1163789A JP 1163789 A JP1163789 A JP 1163789A JP H02190196 A JPH02190196 A JP H02190196A
- Authority
- JP
- Japan
- Prior art keywords
- amino
- salt
- acyl
- methylphosphinobutyric acid
- acid
- 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
- 241000187654 Nocardia Species 0.000 title claims abstract description 9
- 239000013543 active substance Substances 0.000 title abstract description 3
- 241001446247 uncultured actinomycete Species 0.000 title abstract 2
- 238000004519 manufacturing process Methods 0.000 title description 4
- NIBLDNQQTIFBAI-BYPYZUCNSA-N CPCC[C@H](N)C(O)=O Chemical compound CPCC[C@H](N)C(O)=O NIBLDNQQTIFBAI-BYPYZUCNSA-N 0.000 claims abstract description 19
- 230000003287 optical effect Effects 0.000 claims abstract description 12
- 125000002252 acyl group Chemical group 0.000 claims abstract description 10
- 150000003839 salts Chemical class 0.000 claims abstract description 10
- 239000002253 acid Substances 0.000 claims abstract description 8
- 150000001875 compounds Chemical class 0.000 claims abstract description 7
- 241000186361 Actinobacteria <class> Species 0.000 claims abstract description 4
- 241000187681 Nocardia sp. Species 0.000 claims abstract description 4
- 239000012736 aqueous medium Substances 0.000 claims abstract 2
- 238000000034 method Methods 0.000 claims description 14
- 244000005700 microbiome Species 0.000 claims description 12
- 239000000126 substance Substances 0.000 claims description 5
- 241000894006 Bacteria Species 0.000 claims description 3
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 2
- 150000003863 ammonium salts Chemical class 0.000 claims description 2
- 159000000007 calcium salts Chemical class 0.000 claims description 2
- 229910052698 phosphorus Inorganic materials 0.000 claims description 2
- 239000011574 phosphorus Substances 0.000 claims description 2
- 125000000218 acetic acid group Chemical group C(C)(=O)* 0.000 claims 1
- 150000001413 amino acids Chemical class 0.000 claims 1
- XAEFZNCEHLXOMS-UHFFFAOYSA-M potassium benzoate Chemical compound [K+].[O-]C(=O)C1=CC=CC=C1 XAEFZNCEHLXOMS-UHFFFAOYSA-M 0.000 claims 1
- 159000000000 sodium salts Chemical class 0.000 claims 1
- 239000002609 medium Substances 0.000 abstract description 13
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract description 5
- 229910052799 carbon Inorganic materials 0.000 abstract description 5
- 230000000813 microbial effect Effects 0.000 abstract description 5
- 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 abstract description 4
- 239000008103 glucose Substances 0.000 abstract description 3
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 3
- 229910052700 potassium Inorganic materials 0.000 abstract description 3
- 229910052708 sodium Inorganic materials 0.000 abstract description 3
- 239000000284 extract Substances 0.000 abstract description 2
- 229910017053 inorganic salt Inorganic materials 0.000 abstract description 2
- 235000013372 meat Nutrition 0.000 abstract description 2
- 210000004748 cultured cell Anatomy 0.000 abstract 2
- 239000007864 aqueous solution Substances 0.000 abstract 1
- 239000002689 soil Substances 0.000 abstract 1
- 238000009210 therapy by ultrasound Methods 0.000 abstract 1
- 108090000790 Enzymes Proteins 0.000 description 10
- 102000004190 Enzymes Human genes 0.000 description 10
- 229920001817 Agar Polymers 0.000 description 9
- 239000008272 agar Substances 0.000 description 9
- 238000006243 chemical reaction Methods 0.000 description 9
- 239000000047 product Substances 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 8
- -1 methylphosphinobutyric acid -2-amino-4-methylphosphinobutyric acid Chemical compound 0.000 description 6
- 239000000758 substrate Substances 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- NIBLDNQQTIFBAI-UHFFFAOYSA-N 2-amino-4-methylphosphanylbutanoic acid Chemical compound CPCCC(N)C(O)=O NIBLDNQQTIFBAI-UHFFFAOYSA-N 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 3
- 230000002363 herbicidal effect Effects 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical class [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- GXCLVBGFBYZDAG-UHFFFAOYSA-N N-[2-(1H-indol-3-yl)ethyl]-N-methylprop-2-en-1-amine Chemical compound CN(CCC1=CNC2=C1C=CC=C2)CC=C GXCLVBGFBYZDAG-UHFFFAOYSA-N 0.000 description 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 2
- 241000122973 Stenotrophomonas maltophilia Species 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 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 2
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 2
- 229960001230 asparagine Drugs 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 2
- 229910000366 copper(II) sulfate Inorganic materials 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000012156 elution solvent Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 239000004009 herbicide Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 239000011591 potassium Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 238000004809 thin layer chromatography Methods 0.000 description 2
- 238000011282 treatment Methods 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- GMKMEZVLHJARHF-UHFFFAOYSA-N (2R,6R)-form-2.6-Diaminoheptanedioic acid Natural products OC(=O)C(N)CCCC(N)C(O)=O GMKMEZVLHJARHF-UHFFFAOYSA-N 0.000 description 1
- 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 description 1
- TZRAICQNZRDZPX-UHFFFAOYSA-N 2-methylphosphanylbutanoic acid Chemical compound CCC(PC)C(O)=O TZRAICQNZRDZPX-UHFFFAOYSA-N 0.000 description 1
- RXQVPOHYECKXPQ-UHFFFAOYSA-N 4-methylphosphanylbutanoic acid Chemical compound CPCCCC(O)=O RXQVPOHYECKXPQ-UHFFFAOYSA-N 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- RFSUNEUAIZKAJO-VRPWFDPXSA-N D-Fructose Natural products OC[C@H]1OC(O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-VRPWFDPXSA-N 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 1
- YXHKONLOYHBTNS-UHFFFAOYSA-N Diazomethane Chemical compound C=[N+]=[N-] YXHKONLOYHBTNS-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- SRBFZHDQGSBBOR-HWQSCIPKSA-N L-arabinopyranose Chemical compound O[C@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-HWQSCIPKSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-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
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical class [Mg] FYYHWMGAXLPEAU-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
- MUPFEKGTMRGPLJ-RMMQSMQOSA-N Raffinose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 MUPFEKGTMRGPLJ-RMMQSMQOSA-N 0.000 description 1
- MUPFEKGTMRGPLJ-UHFFFAOYSA-N UNPD196149 Natural products OC1C(O)C(CO)OC1(CO)OC1C(O)C(O)C(O)C(COC2C(C(O)C(O)C(CO)O2)O)O1 MUPFEKGTMRGPLJ-UHFFFAOYSA-N 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
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- PNNNRSAQSRJVSB-BXKVDMCESA-N aldehydo-L-rhamnose Chemical compound C[C@H](O)[C@H](O)[C@@H](O)[C@@H](O)C=O PNNNRSAQSRJVSB-BXKVDMCESA-N 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 235000011114 ammonium hydroxide 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
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 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
- 125000003435 aroyl group Chemical group 0.000 description 1
- 125000003236 benzoyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C(*)=O 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 125000004063 butyryl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 210000002318 cardia Anatomy 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
- 210000002421 cell wall Anatomy 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 238000003381 deacetylation reaction Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 125000002485 formyl group Chemical group [H]C(*)=O 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- JEGUKCSWCFPDGT-UHFFFAOYSA-N h2o hydrate Chemical compound O.O JEGUKCSWCFPDGT-UHFFFAOYSA-N 0.000 description 1
- 125000005843 halogen group Chemical group 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 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
- 235000010355 mannitol Nutrition 0.000 description 1
- GMKMEZVLHJARHF-SYDPRGILSA-N meso-2,6-diaminopimelic acid Chemical compound [O-]C(=O)[C@@H]([NH3+])CCC[C@@H]([NH3+])C([O-])=O GMKMEZVLHJARHF-SYDPRGILSA-N 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 239000006877 oatmeal agar Substances 0.000 description 1
- 125000001477 organic nitrogen group Chemical group 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 125000001501 propionyl group Chemical group O=C([*])C([H])([H])C([H])([H])[H] 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose 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[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 235000013547 stew Nutrition 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 230000001131 transforming effect Effects 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
- 238000002525 ultrasonication Methods 0.000 description 1
- 125000003774 valeryl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Abstract
Description
（産業上の利用分野）
本発明は、安価な化学的合成法で得られたＮ−アシル−
Ｄ、 Ｌ−２−アミノ−４−メチルホスフィノ酪酸また
はその塩に微生物の培養菌体またはその培養処理物を作
用させて酵素的にＬ−２−アミノ−４−メチルホスフィ
ノ酪酸を製造する方法に関するものである。
なお、上記の含燐化合物は除草剤の活性成分として有用
である。また、そのＬ体は０１体の約２倍の活性を有す
る。
（従来の技術）
従来、Ｎ−アシル−Ｄ、Ｌ−２−アミノ−４−メチルホ
スフィノ酪酸又はその塩の化学的合成法（特開昭４８９
１０１９及び特開昭５２−１３９７２７ ）は知られて
いるが、この場合はラセミ体、すなわち光学異性体であ
るＤ一体とＬ一体の混合物として得られので、活性本体
であるし一体以外に余分なり一体も得られることになり
、経済的に有効な方法とはいえない。
一方、酵素化学的な検討はほとんどなされていないが、
シュードモナス・マルトフイリアＢＮ−２３３（Ｐｓｅ
ｕｄｏｍｏｎａｓ ｍａｌｔｏｐｈｉｌｉａ ＢＮ−２
３３）の培養菌体を用いてＮ−アシル−Ｄ、Ｌ−２−ア
ミノ−４−メチルホスフィノｍＭからＬ−２−アミノ−
４−メチルホスフィノ酪酸が生じることが報告されてい
る（特開昭５５４７６３０ ）。
しかしながら、この方法は微生物の属が異なる点に於て
本発明の光学分割法とは全く異なる。
（発明が解決しようとする問題点）
従って本発明は、本発明の微生物の培養菌体またはその
処理物を用いて、温和な条件下で一般式(Industrial Application Field) The present invention provides N-acyl-
D. Enzymatic production of L-2-amino-4-methylphosphinobutyric acid by reacting cultured microorganisms or cultured products thereof with L-2-amino-4-methylphosphinobutyric acid or its salt. It is about the method. Note that the above-mentioned phosphorus-containing compounds are useful as active ingredients of herbicides. Furthermore, the L form has approximately twice the activity of the 01 form. (Prior art) Conventionally, a method for chemically synthesizing N-acyl-D, L-2-amino-4-methylphosphinobutyric acid or a salt thereof (Japanese Unexamined Patent Application Publication No. 489/1989)
1019 and JP-A-52-139727) are known, but in this case, it is obtained as a racemate, that is, a mixture of optical isomers D and L, so it is the active substance and there is no excess in addition to the monomer. This is not an economically effective method. On the other hand, although little enzymatic chemistry has been investigated,
Pseudomonas maltophilia BN-233 (Pse
udomonas maltophilia BN-2
33) was used to prepare L-2-amino-D, L-2-amino-4-methylphosphino from mM.
It has been reported that 4-methylphosphinobutyric acid is produced (JP-A-5547630). However, this method is completely different from the optical resolution method of the present invention in that the genus of microorganisms is different. (Problems to be Solved by the Invention) Therefore, the present invention aims to solve the following problems by using the cultured microorganisms of the present invention or their processed products, under mild conditions.
【式中、Ｒは非
置換又はハロゲン置換アシル基を表す。】で表されるＮ
−アシル−Ｄ、Ｌ−２−アミノ−４−メチルホスフィノ
酪酸の光学分割を行う新規な方法を提供しようとするも
のである。
（問題点を解決するための手段）
前記の目的は、ノカルデイア（Ｎｏｃａｒｄｉａ）属に
属する放線菌から選ばれたＮ−アシル−Ｄ、Ｌ−２−ア
ミノ−４−メチルホスフィノ酪酸をＬ−２−アミノ−４
−メチルホスフィノ酪酸に変換する能力を有する微生物
の菌体またはその処理物の存在下でＮ−アシル−Ｄ、Ｌ
−２−アミノ−４−メチルホスフィノ酪酸をＬ−２−ア
ミノ−４−メチルホスフィノ＃酸とＮ−アシル−Ｄ−２
アミノ−４−メチルホスフィノ酪酸との混合物に変換す
ることを特徴とする光学分割法により達成される。
（具体的な説明）
Ｄ、Ｌ−２−アミノ−４−メチルホスフィノ酪酸は除草
剤、特に多年生雑草及び雑潅木防除用除草剤として有用
である。しかしながら、本物質の除草作用の研究から除
草活性の本体はＬ−２−アミノ−４−メチルホスフィノ
酪酸に起因することが判明している。
本発明者らは、前記目的を達成するために鋭意研究を重
ねた結果、Ｎ−アシル−Ｄ、Ｌ−２−アミノ−４メチル
ホスフイノ酪酸のうち、Ｌ一体だけを選択的に脱アセチ
ル化して、Ｌ−２−アミノ−４−メチルホスフィノ酪酸
に変換することの出来る微生物を見いだし本発明を完成
するに至った。
本発明に使用する微生物としては、ノカルデイア（Ｎｏ
ｃａｒｄ ｉａ）属から選ばれた微生物のうち、Ｎ−ア
シル−Ｄ、Ｌ−２−アミノ−４−メチルホスフィノ酪酸
のＬ一体だけを選択的に脱アセチル化して、Ｌ−２−ア
ミノ−４−メチルホスフィノ酪酸に変換することの出来
る微生物であればよい。この様な微生物は一般に入手ま
たは購入が容易である保存株から選択することが出来る
し、または自然界から分離することが出来る。
なお、これらの菌株に変異を生じさせて一層生産性の高
い菌株を得ることもできる。また、これら菌株の細胞中
に存在する酵素の生産に関与する遺伝子を切り出し、こ
れを適切なベクター例えばプラスミドに挿入し、このベ
クターを用いて適当な宿主、例えばエシェリッヒア・コ
リ
（Ｕｓｃｈｅｒｉｃｈｉａ ｃｏｌｉ）や酵母のごとき
異種宿主または同種宿主を形質転換することにより、本
発明の酵素生産株を人為的に創製することもできる。
本発明に用いるノカルデイア属放線菌の例としては、埼
玉県白岡町の土壌より分離したノカルディア・エスピー
ＮＣＢ ２６ （Ｎｏｃａｒｄｉａ ｓｐ、 ＮＣＢ
２６）株が挙げられる。本菌株の菌学的性状を示すと
、次の通りである。
（１）形態的特徴
若い細胞は菌糸状に生育し、 分岐が観察される。菌糸
は培養の進行に従って桿菌あるいは長桿菌状に分断する
。気菌糸は、イースト・麦芽寒天で良好に着生する。菌
糸の直径は約０．６〜０．８μである。
（２）細胞壁組成
ジアミノピメリン酸 メソ型
糖組成 アラビノース、ガラクトース（３）各培地に
おける生育状態（２５°Ｃ１２１日間観察）■シュクロ
ース・硝酸塩寒天培地
生育は中程度であり、コロニーの色は白色である。
■グルコース・アスパラギン寒天培地
生育は中程度であり、コロニーの色は白色ないし淡いク
リーム色である。
■グリセリン・アスパラギン寒天培地
生育は中程度であり、コロニーの色は白色である。
■スターチ寒天培地
生育は貧弱であり、コロニーの色は白色である。
■チロシン寒天培地
生育は貧弱であり、コロニーの色は白色である。
■栄養寒天培地
生育は中程度であり、コロニーの色は淡黄色である。
■イースト・麦芽寒天培地
生育は豊富であり、コロニーの色は淡黄色ないし橙色で
ある。
■オートミール寒天培地
生育は貧弱であり、コロニーの色は白色である。
（４）生理的性質
■生育温度範囲：１０〜４０°Ｃ０最適、２５〜３０”
Ｃ。
■生育ｐ＋１範囲：５．０〜１０．０゜最適、７．０〜
８．ｏ０■ゼラチンの液化：陽性
■スターチの加水分解：陽性
■脱脂牛乳の凝固、ペプトン化：ともに陰性■メラニン
様色素の生成：陰性
（５）各種炭素源の同化性（プリドハム・ゴドリーブ寒
天培地上）
Ｌ−アラビノース 士
Ｄ−キシロース ＋
Ｄ−グルコース ＋
Ｄ−フラクトース ＋
シュクロース ＋
イノシトール ＋
Ｌ−ラムノース
ラフィノース
Ｄ−マンニット 士
ＮＣ８２６株の菌学的性質を、バーシーズ・マニュアル
・オブ・システマティク・バクテリオロジーの記載にも
とづいて検索した結果、本菌株はノカルデイア属に属す
る一菌株と認められる。本菌株は、工業技術院微生物工
業技術研究所に微生物受託番号微工研菌寄第１０４６０
号として寄託されている。なお、本発明に使用出来る微
生物株は上記の例に限定されるものではない。
本発明に用いる微生物の培養は、通常、振とう培養ある
いは通気撹はん深部培養等の好気的条件下で行う。培養
温度は１０〜３７°Ｃ１培養ｐｌ＋は６〜９で、１〜６
日間培養する。
培地には、使用菌が資化し得る炭素源、窒素源、無機塩
及び微量有機栄養源が含まれる。
即ち、炭素源としては、グルコース、デンプン加水分解
液、糖蜜などの炭水化物等も使用できる。
窒素源としては、アンモニア、硫酸アンモニウム、塩化
アンモニウム等の各種の無機及び有機のアンモニウム塩
類または肉エキス、酵母エキス、コーン・スチーブ・リ
カー、カゼイン加水分解物等の天然有機窒素源も使用可
能である。
無機塩としては、マグネシウム、鉄、マンガン、カリウ
ム、ナトリウム等の塩が適時用いられる。
また、目的変換酵素活性を誘導あるいは酵素活性を高め
るために、培養初期あるいは培養途中に本酵素の基質と
なる一般式（■）により表される化合物あるいは本酵素
の基質となる一般式（１）により表される化合物の構造
類似体等を微生物の生育を妨げない程度添加し培養する
ことも出来る。
上記の方法で得られた培養物またはその処理物が本発明
の光学分割に用いられるのであるが、ここでいう培養処
理物とは、例えば培養物から集菌洗浄された菌体、菌体
の超音波処理、ゴーリン破砕やその他の処理で光学分割
活性を保持した酵素を抽出精製したものや、菌体や抽出
精製酵素に更に固形化処理を施したもの等を示す。
また本酵素の基質となる一般式（１）により表される化
合物中の置換基Ｒであるアシル基は、本微生物酵素で分
解されてＬ−２−アミノ−４−メチルホスフィノ酪酸を
生成し得るアシル基であればどんな基であってもよいが
、通常工業的に用いられるアシル基はアルカノイル基及
びアロイル基で、その中でも、ホルミル、アセチル、プ
ロピオニル、ノルマルブチリル、イソブチリル、ノルマ
ルペンタノイルおよびイソペンタノイルの様な炭素数１
〜５のアルカノイル基及びベンゾイル基等が例示される
。これらの基の水素原子がハロゲン原子と置換されてい
ることは本酵素反応を妨げない限り何等制限されるもの
ではない。
本発明の方法に用いられる基質は、上記のＮ−アシル−
Ｄ、Ｌ−２−アミノ−４−メチルホスフィノ酪酸だけで
なく、その塩（例えばナトリウム塩、カリウム塩、アン
モニウム塩またはカルシウム塩）も包含する。
これらの基質を、特に濃度に制限はないが、通常０．５
〜１０％で使用する。 反応温度は１０〜４０°Ｃ１好
ましくは２０〜３７°Ｃである。 反応ｐＨは４〜１０
、好ましくは６〜９の範囲で１〜４日間反応する。
反応液からＬ−２−アミノ−４−メチルホスフィノ酪酸
とＮ−アシル−Ｄ−２−アミノ−４−メチルホスフィノ
酪酸を分離するには、例えば濃縮、等電点沈澱等による
直接晶析法や、イオン交換樹脂処理等の公知の方法によ
り行うことが出来る。
生成したＬ−２−アミノ−４−メチルホスフィノ酪酸の
定性と定量は薄層クロマトグラフィー（ＴＬＣ）、高速
液体クロマトグラフィー（ＨＰＬＣ）または／およびバ
イオアッセイによる方法を用いることにより出来る。
また、光学的異性体は、旋光度分析、光学異性体分離カ
ラムを用いることにより判別することが出来る。
なお、未反応のＮ−７’シル−Ｄ−２−アミノ−４−メ
チルホスフィノ酪酸は常法により化学的にラセミ化し、
再び上述の反応に供することが出来る。
（実施例）
以下、本発明を実施例に基づいて詳細に説明するが、本
発明はこれに限定されるものではない。
参考例
Ｎ−アセチル−Ｄ、Ｌ−２−アミノ−４−メチルホスフ
ィノ酪酸の合成
り、Ｌ−２−アミノ−４−メチルホスフィノ酪酸４ｇ（
０，０２２モル）を、室温で水と酢酸（１χ１重量比）
の混合溶液１６０ｇに溶解後、撹拌しながら無水酢酸３
２０ｇを加えた。
２時間後、発熱が起こり８０°Ｃに温度が上昇したので
、水−水浴で２０°Ｃに冷却した。冷却後、更に２０°
Ｃで２時間攪拌を行った。
反応生成物を、減圧下′ａ縮した後、残渣を高速液体ク
ロマトグラフィーで分析したところ、Ｄ、Ｌ２−アミノ
−４−メチルホスフィノ酪酸のＮ−アセチル化物への転
化率は１００％であった。
また残渣をジアゾメタンでメチル化し、ガスクロマトグ
ラフィー−質量分析にて分析したところ、メチル化物は
Ｎ−アセチル−Ｄ、Ｌ−２−アミノ−４−メチルホスフ
ィノ酪酸メチルエステルであり、収率１００％であった
。
実施例
５００ｍ１容三角フラスコに、酵母エキス０．４χ、麦
芽エキス１．０χ、グルコース０．４χ（ｐＨ７，３）
の組成からなる滅菌培地１００ ｍｌを加え、これにノ
カルディア・エスピーＮＣＢ ２６ のスラントから
１白金耳を植菌し、２８°Ｃで９６時間１分間１５０回
転で旋回振とう培養を行った。
培養液を遠心分離（８０００ｒｐｍ、 ２０ｎ＋ｉｎ、
） シ、菌体を得た。得られた菌体５００ｍｇと濃アン
モニア−水でｐｌ＋ ８．０に調整したＮ−アセチル−
Ｄ、Ｌ−２−アミノ−４−メチルホスフィノ酪酸１００
ｍｇを０．１Ｍ）リス・塩酸緩衝液（ｐＨ８，０） １
０ｍ１に懸濁した。
この懸濁液を１００ｍ１容三角フラスコに加え、１分間
２００回転で旋回振とうしつつ、２８℃で７２時間反応
を行った。
反応終了後遠心分離により除菌し、上澄を光学異性体分
離カラム（ＭＣＩ ＧＥＬ 三菱化成製、キラルパッ
ク引１ ダイセル化学工業製）を用いて分析した。この
際に、生成産物の分析は以下の条件で行った。
分析条件：カラム ＭＣＩ ＧＥＬ ＣＲ５ｌ０Ｗ（Ｄ
ＬＡＡ） ４．６ｍｍＸ５０ｍｍ （三菱化成製）、溶
出溶媒 ２．ＯｍＭ ＣｕＳＯ４、流量 １ｍｌ／ｌｌ
１ｉｎ、、温度 ３０°Ｃ，ＵＶ２５４ｎｍで検出。
また未反応基質（残存している基ｉｔ）の分析について
は下記の条件で行った。
分析条件：カラム ＣＨＩＲＡＬＰＡＫ Ｗｔｌ ４．
６ｍ１ＩＸ２５０慴預（ダイセル化学工業製）、溶出溶
媒 ０．２５Ｍ ＣｕＳＯ４、流量 １．５ｍ１１０＋
ｉｎ、、温度 ３０“Ｃ，、ＵＶ２２０ｎｍで検出。
その結果、生成産物はＬ−２−アミノ−４−メチルホス
フィノ酪酸であった。
Ｎ−アセチル−Ｌ−２−アミノ−４−メチルホスフィノ
酪酸からＬ−２−アミノ−４−メチルホスフィノ酪酸へ
の変換率６０％。
Ｎ−アセチル−Ｄ、Ｌ−２−アミノ−４−メチルホスフ
ィノ酪酸のＬ体だけの脱アセチル化反応（加水分解反応
）の選択率１００％。
（発明の効果）
本発明によれば、簡単な工程で、かつ温和な条件でＮ−
アシル−Ｄ、 Ｌ−２−アミノ−４−メチルホスフィノ
酪酸から光学活性体であるＬ−２−アミノ−４−メチル
ホスフィノ酪酸が選択的に生成でき、その工業的価値は
極めて大である。[In the formula, R represents an unsubstituted or halogen-substituted acyl group. ] N
The present invention aims to provide a novel method for optical resolution of -acyl-D, L-2-amino-4-methylphosphinobutyric acid. (Means for Solving the Problems) The above object is to convert N-acyl-D, L-2-amino-4-methylphosphinobutyric acid selected from actinomycetes belonging to the genus Nocardia into L-2 -Amino-4
-N-acyl-D,L in the presence of microbial cells or their processed products that have the ability to convert into methylphosphinobutyric acid
-2-amino-4-methylphosphinobutyric acid and L-2-amino-4-methylphosphinobutyric acid and N-acyl-D-2
This is achieved by an optical resolution method characterized by conversion into a mixture with amino-4-methylphosphinobutyric acid. (Specific Description) D,L-2-amino-4-methylphosphinobutyric acid is useful as a herbicide, particularly for controlling perennial weeds and shrubs. However, studies on the herbicidal activity of this substance have revealed that the main herbicidal activity is due to L-2-amino-4-methylphosphinobutyric acid. As a result of extensive research in order to achieve the above object, the present inventors selectively deacetylated only L of N-acyl-D, L-2-amino-4 methylphosphinobutyric acid, A microorganism capable of converting L-2-amino-4-methylphosphinobutyric acid was discovered and the present invention was completed. The microorganisms used in the present invention include Nocardia (No.
Among microorganisms selected from the genus Cardia), N-acyl-D, L-2-amino-4-methylphosphinobutyric acid is selectively deacetylated to produce L-2-amino-4. - Any microorganism that can convert it into methylphosphinobutyric acid may be used. Such microorganisms can be selected from stock stocks that are generally available or easily purchased, or can be isolated from nature. In addition, it is also possible to obtain strains with even higher productivity by causing mutations in these strains. In addition, genes involved in the production of enzymes present in the cells of these strains are excised, inserted into an appropriate vector such as a plasmid, and this vector is used to infect a suitable host such as Escherichia coli or yeast. The enzyme-producing strain of the present invention can also be artificially created by transforming a heterologous host or a homologous host such as the following. As an example of the actinomycetes of the genus Nocardia used in the present invention, Nocardia sp.
26) Stocks are mentioned. The mycological properties of this strain are as follows. (1) Morphological characteristics Young cells grow like hyphae and branching is observed. As the culture progresses, the hyphae divide into bacilli or long bacilli. Aerial mycelium grows well on yeast and malt agar. The diameter of the hyphae is approximately 0.6-0.8μ. (2) Cell wall composition Diaminopimelic acid Meso sugar composition Arabinose, galactose (3) Growth status in each medium (observed at 25°C for 121 days) ■Sucrose/nitrate agar medium Growth is moderate, and the colony color is white. . ■Glucose-asparagine agar medium Growth is moderate, and colonies are white to pale cream in color. ■Glycerin-asparagine agar medium Growth is moderate and colonies are white in color. ■Growth on starch agar medium is poor, and colonies are white in color. ■Growth on tyrosine agar medium is poor, and colonies are white in color. ■Nutritional agar medium Growth is moderate and colonies are pale yellow in color. ■Yeast/malt agar medium Growth is abundant, and colonies are pale yellow to orange in color. ■Growth on oatmeal agar medium is poor, and colonies are white in color. (4) Physiological properties ■ Growth temperature range: 10-40°C0 optimum, 25-30"
C. ■Growth p+1 range: 5.0~10.0° optimal, 7.0~
8. o0 ■ Liquefaction of gelatin: Positive ■ Hydrolysis of starch: Positive ■ Coagulation and peptonization of skimmed milk: Both negative ■ Production of melanin-like pigments: Negative (5) Assimilation of various carbon sources (on Pridham-Godelive agar medium) L-arabinose D-xylose + D-glucose + D-fructose + sucrose + inositol + L-rhamnose raffinose D-mannitol The mycological properties of NC826 strain were determined according to the Bersey's Manual of Systematic Bacteriology. As a result of a search based on the description, this strain was recognized as a strain belonging to the genus Nocardia. This strain was submitted to the Institute of Microbial Technology, Agency of Industrial Science and Technology, with microbial accession number 10460.
It has been deposited as No. Note that the microorganism strains that can be used in the present invention are not limited to the above examples. The microorganisms used in the present invention are usually cultured under aerobic conditions such as shaking culture or submerged culture with aeration and agitation. Culture temperature is 10-37°C 1 culture pl+ is 6-9, 1-6
Incubate for days. The medium contains carbon sources, nitrogen sources, inorganic salts, and trace organic nutrient sources that can be assimilated by the bacteria used. That is, carbohydrates such as glucose, starch hydrolyzate, and molasses can also be used as carbon sources. As a nitrogen source, various inorganic and organic ammonium salts such as ammonia, ammonium sulfate, ammonium chloride, etc. or natural organic nitrogen sources such as meat extract, yeast extract, corn stew liquor, casein hydrolyzate, etc. can also be used. As the inorganic salt, salts of magnesium, iron, manganese, potassium, sodium, etc. are used as appropriate. In addition, in order to induce the target converting enzyme activity or increase the enzyme activity, a compound represented by the general formula (■) that serves as a substrate for this enzyme or a compound represented by the general formula (1) that serves as a substrate for this enzyme may be added at the beginning or during the culture. It is also possible to add and culture a structural analog of the compound represented by to the extent that it does not interfere with the growth of microorganisms. The culture obtained by the above method or the processed product thereof is used for the optical resolution of the present invention, and the culture processed product here refers to, for example, bacterial cells collected and washed from the culture, or bacterial cells collected and washed from the culture. These include enzymes that have been extracted and purified with optical resolution activity retained through ultrasonication, Gorlin disruption, and other treatments, and those that have been further solidified on bacterial cells and extracted and purified enzymes. Furthermore, the acyl group, which is the substituent R in the compound represented by general formula (1), which is the substrate of this enzyme, is decomposed by this microbial enzyme to produce L-2-amino-4-methylphosphinobutyric acid. Any acyl group may be used as long as it can be obtained, but the acyl groups usually used industrially are alkanoyl groups and aroyl groups, among which formyl, acetyl, propionyl, normal butyryl, isobutyryl, normal pentanoyl and 1 carbon number like isopentanoyl
-5 alkanoyl groups, benzoyl groups, etc. are exemplified. There are no restrictions on the substitution of hydrogen atoms in these groups with halogen atoms as long as this does not interfere with the enzyme reaction. The substrate used in the method of the present invention is the above-mentioned N-acyl-
It includes not only D,L-2-amino-4-methylphosphinobutyric acid, but also its salts, such as the sodium, potassium, ammonium or calcium salts. There is no particular limit to the concentration of these substrates, but the concentration is usually 0.5
Use at ~10%. The reaction temperature is 10-40°C, preferably 20-37°C. Reaction pH is 4-10
, preferably in the range of 6 to 9, for 1 to 4 days. To separate L-2-amino-4-methylphosphinobutyric acid and N-acyl-D-2-amino-4-methylphosphinobutyric acid from the reaction solution, for example, direct crystallization by concentration, isoelectric precipitation, etc. This can be carried out by a known method such as a method or treatment with an ion exchange resin. The produced L-2-amino-4-methylphosphinobutyric acid can be qualitatively and quantitatively determined by thin layer chromatography (TLC), high performance liquid chromatography (HPLC) and/or bioassay.
Further, optical isomers can be distinguished by optical rotation analysis and by using an optical isomer separation column. In addition, unreacted N-7' syl-D-2-amino-4-methylphosphinobutyric acid was chemically racemized by a conventional method,
It can be subjected to the above reaction again. (Examples) Hereinafter, the present invention will be described in detail based on Examples, but the present invention is not limited thereto. Reference Example Synthesis of N-acetyl-D, L-2-amino-4-methylphosphinobutyric acid, 4 g of L-2-amino-4-methylphosphinobutyric acid (
0,022 mol) in water and acetic acid (1x1 weight ratio) at room temperature.
After dissolving in 160 g of a mixed solution of
Added 20g. After 2 hours, an exotherm occurred and the temperature rose to 80°C, so it was cooled to 20°C in a water-water bath. After cooling, further 20°
Stirring was performed at C for 2 hours. After condensing the reaction product under reduced pressure, the residue was analyzed by high performance liquid chromatography, and the conversion rate of D, L2-amino-4-methylphosphinobutyric acid to N-acetylated product was 100%. Ta. Further, the residue was methylated with diazomethane and analyzed by gas chromatography-mass spectrometry, and the methylated product was N-acetyl-D, L-2-amino-4-methylphosphinobutyric acid methyl ester, yield 100%. Met. Example 500ml Erlenmeyer flask, yeast extract 0.4χ, malt extract 1.0χ, glucose 0.4χ (pH 7.3)
100 ml of a sterile medium having the following composition was added, one platinum loop of Nocardia sp. NCB 26 was inoculated from a slant, and cultured with orbital shaking at 150 rpm for 1 minute at 28° C. for 96 hours. Centrifuge the culture solution (8000 rpm, 20n+in,
) Bacterial cells were obtained. 500 mg of the obtained bacterial cells and N-acetyl-adjusted to pl+ 8.0 with concentrated ammonia water.
D, L-2-amino-4-methylphosphinobutyric acid 100
mg to 0.1M) Lis-HCl buffer (pH 8,0) 1
Suspended in 0ml. This suspension was added to a 100 ml Erlenmeyer flask, and the reaction was carried out at 28° C. for 72 hours while shaking at 200 revolutions per minute. After the reaction was completed, bacteria were removed by centrifugation, and the supernatant was analyzed using an optical isomer separation column (MCI GEL, manufactured by Mitsubishi Chemical Industries, Ltd., Chiral Pack 1, manufactured by Daicel Chemical Industries, Ltd.). At this time, the product was analyzed under the following conditions. Analysis conditions: Column MCI GEL CR5l0W (D
LAA) 4.6mmX50mm (manufactured by Mitsubishi Kasei), elution solvent 2. OmM CuSO4, flow rate 1ml/ll
1 inch, temperature 30°C, UV 254nm detection. Furthermore, analysis of unreacted substrate (remaining group it) was conducted under the following conditions. Analysis conditions: Column CHIRALPAK Wtl 4.
6m1IX250 (manufactured by Daicel Chemical Industries), elution solvent 0.25M CuSO4, flow rate 1.5m110+
in, temperature 30"C, UV 220nm detection. As a result, the product was L-2-amino-4-methylphosphinobutyric acid. N-acetyl-L-2-amino-4-methylphosphino Conversion rate of butyric acid to L-2-amino-4-methylphosphinobutyric acid: 60%. Deacetylation reaction of only the L form of N-acetyl-D, L-2-amino-4-methylphosphinobutyric acid (hydration 100% selectivity for decomposition reaction. (Effects of the invention) According to the present invention, N-
L-2-amino-4-methylphosphinobutyric acid, an optically active form, can be selectively produced from acyl-D and L-2-amino-4-methylphosphinobutyric acid, and its industrial value is extremely large. .
Claims (3)
チルホスフィノ酪酸またはその塩（ナトリウム塩、カリ
ウム塩、アンモニウム塩またはカルシウム塩）に、ノカ
ルディア（Ｎｏｃａｒｄｉａ）属に属する放線菌から選
ばれた前記化合物中のアシル基分解能を有する微生物の
培養菌体またはその処理物を水性媒体下に作用させ、Ｌ
−２−アミノ−４−メチルホスフィノ酪酸とＮ−アシル
−Ｄ−２−アミノ−４−メチルホスフィノ酪酸との混合
物、またはこれらの塩の混合物に変換して光学分割を行
い、Ｌ−２−アミノ−４−メチルホスフィノ酪酸及びＮ
−アシル−Ｄ−２−アミノ−４−メチルホスフィノ酪酸
を分別採取することを特徴とする含燐アミノ酸とその誘
導体の光学分割法。(1) General formula (I) ▲Mathematical formulas, chemical formulas, tables, etc.▼(I) [In the formula, R represents an unsubstituted or halogen-substituted acyl group. ] N-acyl-D,L-2-amino-4-methylphosphinobutyric acid or its salt (sodium salt, potassium salt, ammonium salt, or calcium salt) is added to the actinic acid belonging to the genus Nocardia. A cultured microorganism selected from bacteria having the ability to decompose the acyl group in the compound or a processed product thereof is allowed to act in an aqueous medium, and L
L-2 -amino-4-methylphosphinobutyric acid and N
-An optical resolution method for phosphorus-containing amino acids and their derivatives, which comprises fractionally collecting -acyl-D-2-amino-4-methylphosphinobutyric acid.
ＮＣＢ２６（Ｎｏｃａｒｄｉａ ｓｐ． ＮＣＢ２６）
株である請求項（１）に記載の方法。(2) Actinomycetes of the genus Nocardia are Nocardia sp. NCB26
The method according to claim (1), which is a strain.
）または（２）に記載の方法。(3) Claim (1) wherein R in general formula (I) is acetyl.
) or the method described in (2).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1163789A JPH02190196A (en) | 1989-01-20 | 1989-01-20 | Production of optically active substance using actinomycete belonging to genus nocardia |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1163789A JPH02190196A (en) | 1989-01-20 | 1989-01-20 | Production of optically active substance using actinomycete belonging to genus nocardia |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH02190196A true JPH02190196A (en) | 1990-07-26 |
Family
ID=11783460
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP1163789A Pending JPH02190196A (en) | 1989-01-20 | 1989-01-20 | Production of optically active substance using actinomycete belonging to genus nocardia |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH02190196A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992018513A1 (en) | 1991-04-16 | 1992-10-29 | Alkaloida Vegyészeti Gyár Rt. | New non-hygroscopic mono-ammonium salts |
-
1989
- 1989-01-20 JP JP1163789A patent/JPH02190196A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992018513A1 (en) | 1991-04-16 | 1992-10-29 | Alkaloida Vegyészeti Gyár Rt. | New non-hygroscopic mono-ammonium salts |
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