JPH04316496A - Production of (r)-(+)-halolactic acid - Google Patents
Production of (r)-(+)-halolactic acidInfo
- Publication number
- JPH04316496A JPH04316496A JP17156191A JP17156191A JPH04316496A JP H04316496 A JPH04316496 A JP H04316496A JP 17156191 A JP17156191 A JP 17156191A JP 17156191 A JP17156191 A JP 17156191A JP H04316496 A JPH04316496 A JP H04316496A
- Authority
- JP
- Japan
- Prior art keywords
- acid
- halolactic
- halolactic acid
- genus
- chlorolactic
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000002253 acid Substances 0.000 title claims abstract description 43
- 238000004519 manufacturing process Methods 0.000 title claims description 5
- 244000005700 microbiome Species 0.000 claims abstract description 21
- 241000222120 Candida <Saccharomycetales> Species 0.000 claims abstract description 9
- 241000589516 Pseudomonas Species 0.000 claims abstract description 8
- 241000186063 Arthrobacter Species 0.000 claims abstract description 4
- 241000235070 Saccharomyces Species 0.000 claims abstract description 4
- 241000588722 Escherichia Species 0.000 claims abstract description 3
- 241000311088 Schwanniomyces Species 0.000 claims abstract description 3
- 238000000034 method Methods 0.000 claims description 16
- OSLCJYYQMKPZHU-UHFFFAOYSA-N 3-chlorolactic acid Chemical group ClCC(O)C(O)=O OSLCJYYQMKPZHU-UHFFFAOYSA-N 0.000 claims description 14
- OSLCJYYQMKPZHU-REOHCLBHSA-N (2r)-3-chloro-2-hydroxypropanoic acid Chemical group ClC[C@H](O)C(O)=O OSLCJYYQMKPZHU-REOHCLBHSA-N 0.000 claims description 5
- 239000000758 substrate Substances 0.000 claims 2
- UKORLVILOUWVTJ-REOHCLBHSA-N (2r)-3-bromo-2-hydroxypropanoic acid Chemical group BrC[C@H](O)C(O)=O UKORLVILOUWVTJ-REOHCLBHSA-N 0.000 claims 1
- UKORLVILOUWVTJ-UHFFFAOYSA-N 3-bromo-2-hydroxypropanoic acid Chemical group BrCC(O)C(O)=O UKORLVILOUWVTJ-UHFFFAOYSA-N 0.000 claims 1
- 239000000126 substance Substances 0.000 abstract description 5
- 239000002994 raw material Substances 0.000 abstract description 4
- 239000013543 active substance Substances 0.000 abstract description 3
- 239000003814 drug Substances 0.000 abstract description 3
- 239000000203 mixture Substances 0.000 abstract description 3
- 150000001875 compounds Chemical class 0.000 abstract 1
- 230000001580 bacterial effect Effects 0.000 description 15
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 9
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 238000006243 chemical reaction Methods 0.000 description 9
- 239000000047 product Substances 0.000 description 8
- 239000013078 crystal Substances 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 230000003287 optical effect Effects 0.000 description 6
- 239000004310 lactic acid Substances 0.000 description 5
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- 235000002639 sodium chloride Nutrition 0.000 description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 239000001888 Peptone Substances 0.000 description 3
- 108010080698 Peptones Proteins 0.000 description 3
- 241000235648 Pichia Species 0.000 description 3
- 229940041514 candida albicans extract Drugs 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 235000013372 meat Nutrition 0.000 description 3
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 3
- BOPGDPNILDQYTO-NNYOXOHSSA-N nicotinamide-adenine dinucleotide Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-N 0.000 description 3
- 235000019319 peptone Nutrition 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 239000004094 surface-active agent Substances 0.000 description 3
- 239000012138 yeast extract Substances 0.000 description 3
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- 241000186074 Arthrobacter globiformis Species 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-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
- 241000178951 Endomyces Species 0.000 description 2
- 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 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- 241001645955 Pseudomonas chlororaphis subsp. aureofaciens Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 241000228389 Sporidiobolus Species 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 241000223230 Trichosporon Species 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 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
- 230000015572 biosynthetic process Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 229910000365 copper sulfate Inorganic materials 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
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- 235000019341 magnesium sulphate Nutrition 0.000 description 2
- 229940099596 manganese sulfate Drugs 0.000 description 2
- 239000011702 manganese sulphate Substances 0.000 description 2
- 235000007079 manganese sulphate Nutrition 0.000 description 2
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 2
- 238000002844 melting Methods 0.000 description 2
- 230000008018 melting Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 238000006722 reduction reaction Methods 0.000 description 2
- 230000008929 regeneration Effects 0.000 description 2
- 238000011069 regeneration method Methods 0.000 description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 2
- 229910000368 zinc sulfate Inorganic materials 0.000 description 2
- 229960001763 zinc sulfate Drugs 0.000 description 2
- OSLCJYYQMKPZHU-UWTATZPHSA-N (2s)-3-chloro-2-hydroxypropanoic acid Chemical compound ClC[C@@H](O)C(O)=O OSLCJYYQMKPZHU-UWTATZPHSA-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
- 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
- DSCUFZSWIGAJKM-UHFFFAOYSA-N 2-chloro-2-hydroxypropanoic acid Chemical compound CC(O)(Cl)C(O)=O DSCUFZSWIGAJKM-UHFFFAOYSA-N 0.000 description 1
- 108010052386 2-haloacid dehalogenase Proteins 0.000 description 1
- RCWFNCXDFSOVLG-UHFFFAOYSA-N 3-chloro-2-oxopropanoic acid Chemical compound OC(=O)C(=O)CCl RCWFNCXDFSOVLG-UHFFFAOYSA-N 0.000 description 1
- 241000588986 Alcaligenes Species 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 241000908817 Aphanoascus Species 0.000 description 1
- 241000185996 Arthrobacter citreus Species 0.000 description 1
- 241000223651 Aureobasidium Species 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 241000222290 Cladosporium Species 0.000 description 1
- 241000186216 Corynebacterium Species 0.000 description 1
- 241001527609 Cryptococcus Species 0.000 description 1
- 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 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 239000005696 Diammonium phosphate Substances 0.000 description 1
- 101000925662 Enterobacteria phage PRD1 Endolysin Proteins 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241001136487 Eurotium Species 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 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
- 241000159512 Geotrichum Species 0.000 description 1
- 241000896533 Gliocladium Species 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 102000003855 L-lactate dehydrogenase Human genes 0.000 description 1
- 108700023483 L-lactate dehydrogenases Proteins 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- AFCARXCZXQIEQB-UHFFFAOYSA-N N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CCNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 AFCARXCZXQIEQB-UHFFFAOYSA-N 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 241000589774 Pseudomonas sp. Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241000877399 Saccharomyces chevalieri Species 0.000 description 1
- 241001123650 Schwanniomyces occidentalis Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 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
- 150000007513 acids Chemical class 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 238000004458 analytical method Methods 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
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 229960002713 calcium chloride Drugs 0.000 description 1
- 235000011148 calcium chloride Nutrition 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 235000013877 carbamide Nutrition 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 229960000355 copper sulfate Drugs 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 238000003795 desorption Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 235000019425 dextrin Nutrition 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
- 229910000388 diammonium phosphate Inorganic materials 0.000 description 1
- 235000019838 diammonium phosphate Nutrition 0.000 description 1
- 229940079919 digestives enzyme preparation Drugs 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 235000019797 dipotassium phosphate Nutrition 0.000 description 1
- 238000000921 elemental analysis Methods 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 229940023064 escherichia coli Drugs 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 229960001781 ferrous sulfate Drugs 0.000 description 1
- 239000011790 ferrous sulphate Substances 0.000 description 1
- 235000003891 ferrous sulphate Nutrition 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 235000001727 glucose Nutrition 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 239000003471 mutagenic agent Substances 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000012429 reaction media Substances 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 235000010344 sodium nitrate Nutrition 0.000 description 1
- 239000004317 sodium nitrate Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【０００１】0001
【産業上の利用分野】本発明は、（Ｒ）−（＋）−３−
ハロ乳酸の製造法に関する。（Ｒ）−（＋）−３−ハロ
乳酸は医薬品等の光学活性な生理活性物質の合成原料と
して有用な物質である。[Industrial Application Field] The present invention relates to (R)-(+)-3-
This invention relates to a method for producing halo-lactic acid. (R)-(+)-3-halolactic acid is a substance useful as a raw material for the synthesis of optically active physiologically active substances such as pharmaceuticals.
【０００２】0002
【従来の技術】光学活性な（Ｒ）−（＋）−３−クロロ
乳酸の生化学的な調製法に関しては、乳酸脱水素酵素を
用いて、クロロピルビン酸の不斉還元法による製法が知
られている（Ｊｏｕｒｎａｌ ｏｆ Ａｍｅｒｉｃ
ａｎ Ｃｈｅｍｉｃａｌ Ｓｏｃｉｅｔｙ，１０４
巻，４４５２頁、１９８２年）。この方法では、反応時
に補酵素としてＮＡＤＨを必要とするので何らかの方法
でＮＡＤＨを連続的に供給する必要があり、ＮＡＤＨ再
生系と組み合せる方法がとられるが、この種のＮＡＤＨ
再生系を組み合せる酵素的還元反応は、工業的実施に困
難な問題が多い。第２の方法として、（±）−３−ハロ
−１，２−プロパンジオールの微生物的不斉酸化反応に
より光学活性な（Ｒ）−（＋）−３−ハロ乳酸を製造す
る方法が知られている（特公開昭５９−２２０１９３号
：特公開昭６１−２６８１９７号：Ｊｏｕｒｎａｌ
ｏｆ Ｆｅｒｍｅｎｔａｔｉｏｎ Ｔｅｃｈｎｏｌ
ｏｇｙ，６４巻，２５１−２５４頁，１９８６年）。微
生物としてゲオトリカム属、トリコスポロン属、エンド
ミセス属、ハンゼヌラ属、キャンジタ属、スポリジオボ
ルス属、ロデロミセス属の微生物が用いられている。こ
の方法では収率が低い難点がある。第３の方法として、
（±）−３−ハロ乳酸に微生物を作用させて（Ｒ）−（
＋）−３−ハロ乳酸をえる方法（特公開昭６１−２６８
１９７号）が知られている。微生物としては、キャンジ
ダ、クリプトコッカス、エンドミセス、ハンゼヌラ、ロ
デロミセス、ピチヤ、スポリジオボルス、トリコスポロ
ン、アルカリゲネス、バチルス、コリネバクテリウム、
アファノアスカス、オーレオバシディウム、バクジェラ
、バクジア、ビソシラミス、フロリディウム、クラドス
ポリウム、コニオケティデウム、ユーロティウムおよび
グリオクラディウムの各属に属する微生物が使用されて
いる。この方法では、収率のよい場合は光学純度が低く
、光学純度の高い場合は収量が低い難点がある。第４の
方法として、２，３−ジハロプロピオン酸に２−ハロ酸
デハロゲナーゼを作用させる方法（（特公開平２−１１
３８９０）が知られている。この方法では、原料である
２，３−ジハロプロピオン酸のコストに難点がある。[Prior Art] Regarding the biochemical preparation of optically active (R)-(+)-3-chlorolactic acid, there is a known method for producing optically active (R)-(+)-3-chlorolactic acid by asymmetric reduction of chloropyruvic acid using lactate dehydrogenase. Journal of America
a Chemical Society, 104
Vol. 4452, 1982). This method requires NADH as a coenzyme during the reaction, so it is necessary to continuously supply NADH by some means, and a method is used in which it is combined with an NADH regeneration system.
Enzymatic reduction reactions that combine regeneration systems have many problems that are difficult to implement industrially. The second method is known to produce optically active (R)-(+)-3-halolactic acid by a microbial asymmetric oxidation reaction of (±)-3-halo-1,2-propanediol. (Special Publication No. 59-220193: Publication No. 61-268197: Journal
of Fermentation Technology
ogy, vol. 64, pp. 251-254, 1986). Microorganisms of the genus Geotrichum, Trichosporon, Endomyces, Hansenula, Candida, Sporidiobolus, and Rhoderomyces are used as the microorganisms. This method has the disadvantage of low yield. As a third method,
(±)-3-Halolactic acid is treated with (R)-(
+) Method for obtaining -3-halolactic acid (Japanese Patent Publication No. 61-268
No. 197) is known. Microorganisms include Candida, Cryptococcus, Endomyces, Hansenula, Rhoderomyces, Pichia, Sporidiobolus, Trichosporon, Alcaligenes, Bacillus, Corynebacterium,
Microorganisms belonging to the genera Aphanoascus, Aureobasidium, Bacgela, Bacdia, Bysocilamis, Floridium, Cladosporium, Coniocetideum, Eurotium and Gliocladium have been used. This method has the disadvantage that when the yield is good, the optical purity is low, and when the optical purity is high, the yield is low. The fourth method is a method in which 2-halo acid dehalogenase is allowed to act on 2,3-dihalopropionic acid ((Japanese Patent Publication No. 2011-11)
3890) is known. This method has a drawback in terms of the cost of the raw material, 2,3-dihalopropionic acid.
【０００３】0003
【発明が解決しようとする課題】上に述べたように従来
の技術が工業的実施になお難点をもっているため、より
効率的な（Ｒ）−（＋）−３−ハロ乳酸の製造法を確立
するために研究を重ねた結果、本発明を完成するに至っ
た。[Problems to be Solved by the Invention] As mentioned above, the conventional technology still has difficulties in industrial implementation, so a more efficient method for producing (R)-(+)-3-halolactic acid was established. As a result of repeated research, we have completed the present invention.
【０００４】0004
【課題を解決するための手段】本発明によれば、シュー
ドモナス（Ｐｓｅｕｄｏｍｏｎａｓ）属、アースロバク
ター（Ａｒｔｈｒｏｂａｃｔｅｒ）属、エッセリヒア（
Ｅｓｃｈｅｒｉｃｈｉａ）属、サッカロミセス（Ｓａｃ
ｃｈａｒｏｍｙｃｅｓ）属、シュワニオミセス（Ｓｃｈ
ｗａｎｎｉｏｍｙｃｅｓ）属、あるいはトルロプシス（
Ｔｏｒｕｌｏｐｓｉｓ）属に属し、（±）−３−ハロ乳
酸から（Ｒ）−（＋）−３−ハロ乳酸を生成する能力を
有する微生物の菌体、培養液またはそれらの処理物を（
±）−３−ハロ乳酸に接触させることにより、（Ｒ）−
（＋）−３−ハロ乳酸を生成させ、これを採取すること
により（Ｒ）−（＋）−３−ハロ乳酸を製造することが
できる。[Means for Solving the Problems] According to the present invention, the genus Pseudomonas, the genus Arthrobacter, and the genus Esserichia (
Genus Escherichia, Saccharomyces
charomyces), Schwaniomyces (Sch
wanniomyces) or Torulopsis (
The cells, culture solution, or processed products of microorganisms belonging to the genus Torulopsis and having the ability to produce (R)-(+)-3-halolactic acid from (±)-3-halolactic acid (
±)-By contacting with 3-halolactic acid, (R)-
(R)-(+)-3-halolactic acid can be produced by producing (+)-3-halolactic acid and collecting it.
【０００５】以下に本発明を詳細に説明する。本発明に
用いられる微生物は、シュードモナス属、アースロバク
ター属、エッセリヒア属、サッカロミセス属、シュワニ
オミセス属またはトルロプシス属に属し、（±）−３−
ハロ乳酸に作用して（−）−３−ハロ乳酸を消費変換し
て（Ｒ）−（＋）−３−ハロ乳酸を残留させ、結果とし
て（Ｒ）−（＋）−３−ハロ乳酸を生成する能力を有す
る微生物であればよい。具体的な例としては、シュード
モナス・オーレオファシエンス（Ｐｓｅｕｄｏｍｏｎａ
ｓ ａｕｒｅｏｆａｃｉｅｎｓ）ＡＴＣＣ １３９
８６、シュードモナス・デハロゲナンス（Ｐｓｅｕｄｏ
ｍｏｎａｓ ｄｅｈａｌｏｇｅｎａｎｓ）ＮＣＩＢ
９０６１、アースロバクター・シトレウス（Ａｒｔｈ
ｒｏｂａｃｔｅｒ ｃｉｔｒｅｕｓ）ＡＴＣＣ １
１６２４、アースロバクター・グロビホルミス（Ａｒｔ
ｈｒｏｂａｃｔｅｒ ｇｌｏｂｉｆｏｒｍｉｓ）ＡＴ
ＣＣ ８０１０、エッセリヒア・コリ（Ｅｓｃｈｅｒ
ｉｃｈｉａ ｃｏｌｉ）ＡＴＣＣ １１３０３、サ
ッカロミセス・シュバリエリ（Ｓａｃｃｈａｒａｍｙｃ
ｅｓ ｃｈｅｖａｌｉｅｒｉ）ＩＦＯ ０２２２、
シュワニオミセス・オキシデンタリス（Ｓｃｈｗａｎｎ
ｉｏｍｙｃｅｓ ｏｃｃｉｄｅｎｔａｌｉｓ）ＡＴＣ
Ｃ ２６０７４、トルロプシス・スフェリカ（Ｔｏｒ
ｕｌｏｐｓｉｓ ｓｐｈａｅｒｉｃａ）ＡＴＣＣ
８５４９などが挙げられる。The present invention will be explained in detail below. The microorganisms used in the present invention belong to the genus Pseudomonas, Arthrobacter, Esselichia, Saccharomyces, Schwaniomyces, or Torulopsis, and are (±)-3-
It acts on halo-lactic acid and consumes and converts (-)-3-halolactic acid to leave (R)-(+)-3-halolactic acid, resulting in (R)-(+)-3-halolactic acid. Any microorganism can be used as long as it has the ability to produce it. A specific example is Pseudomonas aureofaciens.
ATCC 139
86, Pseudomonas dehalogenans (Pseudo
monas dehalogenans) NCIB
9061, Arthlobacter citreus (Arth
robacter citreus) ATCC 1
1624, Arthrobacter globiformis (Art
hrobacter globiformis) AT
CC 8010, Escheria coli (Escher
ichia coli) ATCC 11303, Saccharomyces chevalieri (Saccharamyc
es chevalieri) IFO 0222,
Schwann
iomyces occidentalis) ATC
C 26074, Torulopsis spherica (Tor
ulopsis sphaerica) ATCC
Examples include 8549.
【０００６】これらの菌株を人工的変異方法、たとえば
紫外線照射、Ｘ線照射、変異誘起剤処理などによって変
異させた変異株も用いることができる。[0006] Mutant strains obtained by mutating these strains by artificial mutation methods such as ultraviolet irradiation, X-ray irradiation, mutagenic agent treatment, etc. can also be used.
【０００７】本発明で用いられる微生物の培養において
は通常の微生物の培養法が一般に用いられる。培地とし
ては、微生物が資化可能な炭素源、窒素源、無機物およ
び微生物の生育に必要な物質をほどよく含有する培地で
あれば、合成培地、天然培地のいずれでも使用可能であ
る。炭素源としては、グルコース、でん粉、デキストリ
ン、マンノース、フラクトース、シュクロース、ラクト
ース、キシロース、アラビノース、マニトール、糖蜜な
どが単独または組み合わせて用いられる。さらに微生物
の資化能によっては、炭化水素、アルコール類、有機酸
なども用いられる。窒素源としては、塩化アンモニウム
、硫酸アンモニウム、硝酸アンモニウム、硝酸ナトリウ
ム、尿素、ペプトン、肉エキス、酵母エキス、乾燥酵母
、コーン・スチープ・リカー、大豆粉、カザミノ酸など
が単独あるいは組み合わせて用いられる。そのほか、必
要に応じて、食塩、塩化カリウム、硫酸マグネシウム、
炭酸カルシウム、リン酸二水素カリウム、リン酸水素二
カリウム、硫酸第一鉄、塩化カルシウム、硫酸マンガン
、硫酸亜鉛、硫酸銅などの無機塩類を加えることができ
る。 培養は、振とう培養または通気攪拌培養などの
好気条件下、温度４〜４３℃、好ましくは２０〜３５℃
、ｐＨ４〜１０、好ましくは６．０〜９．５で行われ、
通常１〜１０日で終了する。培地のｐＨは水酸化ナトリ
ウムなどを用いて調整するのが好ましい。[0007] In culturing the microorganisms used in the present invention, conventional microorganism culturing methods are generally used. As the medium, either a synthetic medium or a natural medium can be used as long as it contains a suitable amount of carbon sources, nitrogen sources, inorganic substances that can be assimilated by microorganisms, and substances necessary for the growth of microorganisms. As the carbon source, glucose, starch, dextrin, mannose, fructose, sucrose, lactose, xylose, arabinose, mannitol, molasses, etc. are used alone or in combination. Furthermore, depending on the assimilation ability of microorganisms, hydrocarbons, alcohols, organic acids, etc. may also be used. As the nitrogen source, ammonium chloride, ammonium sulfate, ammonium nitrate, sodium nitrate, urea, peptone, meat extract, yeast extract, dried yeast, corn steep liquor, soybean flour, casamino acid, etc. are used alone or in combination. In addition, salt, potassium chloride, magnesium sulfate,
Inorganic salts such as calcium carbonate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, ferrous sulfate, calcium chloride, manganese sulfate, zinc sulfate, copper sulfate, etc. can be added. The culture is carried out under aerobic conditions such as shaking culture or aerated agitation culture at a temperature of 4 to 43°C, preferably 20 to 35°C.
, carried out at pH 4-10, preferably 6.0-9.5,
Usually completed in 1 to 10 days. The pH of the medium is preferably adjusted using sodium hydroxide or the like.
【０００８】使用微生物の（±）−３−ハロ乳酸との接
触は、このような微生物の生育培養中に行うこともでき
るし、また生育した培養に（±）−３−ハロ乳酸を加え
て反応させることもできる。さらに、培養から菌体を分
けてから、この菌体あるいは菌体処理物と（±）−３−
ハロ乳酸を適当な培地中、例えば緩衝液中で接触反応さ
せることもできる。培養液もしくは菌体の処理は、菌体
中の（−）−３−ハロ乳酸変換活性を損なうことなく、
酵素反応がより容易に進む方法を適宜選択しておこなわ
れる。具体的処理物としては、培養物の濃縮物、乾燥物
、界面活性剤および／または有機溶剤処理物、さらに培
養物を遠心分離してえられる菌体、菌体破砕物、菌体の
乾燥物、アセトン処理物、界面活性剤および／または有
機溶剤処理物、溶菌酵素処理物、固定化菌体あるいは菌
体からの抽出酵素標品などがあげられる。[0008] The microorganism used can be brought into contact with (±)-3-halolactic acid during the growth and culture of such microorganisms, or it can be carried out by adding (±)-3-halolactic acid to the grown culture. It can also be reacted. Furthermore, after separating the bacterial cells from the culture, the bacterial cells or the treated bacterial cells and (±)-3-
Halolactic acids can also be reacted in a suitable medium, for example in a buffer. The culture solution or bacterial cells can be treated without impairing the (-)-3-halolactic acid converting activity in the bacterial cells.
A method that facilitates the enzymatic reaction is appropriately selected and carried out. Specific processed products include culture concentrates, dried products, products treated with surfactants and/or organic solvents, as well as bacterial cells obtained by centrifuging the culture, crushed bacterial cells, and dried bacterial cells. Examples include acetone-treated products, surfactant and/or organic solvent-treated products, lytic enzyme-treated products, immobilized bacterial cells, and enzyme preparations extracted from bacterial cells.
【０００９】本発明で用いられる（±）−３−ハロ乳酸
は遊離もしくは塩酸塩のような塩の形で用いられる。
（±）−３−ハロ乳酸の濃度にはとくに制限はないが、
０．１〜１０％（重量対容量）が好ましい。（＋）−３
−ハロ乳酸を生成させる反応を実施するに際しては、微
生物またはその処理物と、（±）−３−ハロ乳酸を好適
条件下、温度１０〜６０℃、好ましくは２０〜４０℃、
ｐＨ４〜１０、好ましくは７．０〜９．５で反応させる
。反応は通常１〜１０日で終了する。反応中のｐＨは好
適ｐＨ（７．０〜９．５）に維持することが好ましい。
この為には反応媒体として緩衝液を用いたり、反応中ｐ
Ｈを水酸化ナトリウムなどを用いて調整することが好ま
しい。The (±)-3-halolactic acid used in the present invention is used in the form of free or salt such as hydrochloride. There is no particular limit to the concentration of (±)-3-halolactic acid, but
0.1-10% (weight to volume) is preferred. (+)-3
- When carrying out the reaction to produce halo-lactic acid, microorganisms or their processed material and (±)-3-halo-lactic acid are mixed under suitable conditions at a temperature of 10 to 60°C, preferably 20 to 40°C,
The reaction is carried out at a pH of 4 to 10, preferably 7.0 to 9.5. The reaction usually completes in 1 to 10 days. The pH during the reaction is preferably maintained at a suitable pH (7.0 to 9.5). For this purpose, a buffer solution may be used as the reaction medium, or p
It is preferable to adjust H using sodium hydroxide or the like.
【００１０】反応はまた、微生物の培養中に（±）−３
−ハロ乳酸を培養液に添加すする方式によってもおこな
うことができる。この場合、（±）−３−ハロ乳酸の添
加量は、前記した菌体またはその処理物を用いた反応と
同様の濃度になるよう添加すればよい。さらに必要に応
じて微生物の増殖を阻害しない量の前記の界面活性剤を
培養液中に存在させることにより酵素反応を促進させる
ことができる。反応終了後、反応液中に生成（残存）し
た（＋）−３−ハロ乳酸は、公知の方法、例えばイオン
交換樹脂への吸着、脱離、濃縮、結晶化などの手段によ
り単離される。The reaction also occurs during the cultivation of microorganisms (±)-3
- It can also be carried out by adding halo-lactic acid to the culture solution. In this case, the amount of (±)-3-halolactic acid to be added may be the same concentration as in the reaction using the above-mentioned bacterial cells or a treated product thereof. Furthermore, if necessary, the enzymatic reaction can be promoted by making the above-mentioned surfactant present in the culture solution in an amount that does not inhibit the growth of microorganisms. After the reaction is completed, the (+)-3-halolactic acid produced (remaining) in the reaction solution is isolated by known methods such as adsorption to an ion exchange resin, desorption, concentration, and crystallization.
【００１１】[0011]
【実施例】以下に本発明の実施例を示す。実施例におけ
る高速液体クロマトグラフィーによる（Ｒ）−（＋）−
３−クロロ乳酸と、（Ｓ）−（−）−３−クロロ乳酸の
分別分析の条件は次のとおりである。
カラム：ＭＣＩ ＧＥＬ ＣＲＳ １０Ｗ、４．
６×５０ｍｍ（三菱化成製）、溶出液：マイクロモル硫
酸銅、流速：毎分１ｍｌ、サンプル量：１０マイクロリ
ッター、検出：ＵＶ（２５４ｎｍ）。
実施例１
グルコース２％、りん酸２アンモニウム１％、りん酸１
カリウム１％、肉エキス１％、ペプトン１％、酵母エキ
ス０．５％、塩化ナトリウム０．１％、（±）−３−ク
ロロ乳酸０．２％、ｐＨ７．０の組成の滅菌培地１０ｍ
ｌをふくむ太型試験管に表１に示した微生物を植菌し、
２６℃で４８時間振とう培養してから、培養液から遠心
分離により菌体を集め、５０ミリモルのりん酸緩衝液（
ｐＨ７．０）で洗浄後、菌体を再度遠心分離してから、
（±）−３−クロロ乳酸をふくむ５０ミリモルりん酸緩
衝液（ｐＨ７．０）に菌体をけん濁して１０ｍｌとする
。（±）−３−クロロ乳酸の濃度は最終０．２％とした
。この反応混合物を２６℃で４８時間振とうして反応さ
せた後、遠心上清に含まれる３−クロロ乳酸を高速液体
クロマトグラフィーにより分析した。その結果は表１に
示した如くで、菌株により異なるが５０〜８０％の３−
クロロ乳酸が残留し、この残留する（Ｒ）−（＋）−３
−クロロ乳酸の光学異性体過剰率（ｅ．ｅ．）は１００
〜３８％であった。収量は添加した（±）−３−クロロ
乳酸の重量に対する１００分率で示した。[Examples] Examples of the present invention are shown below. (R)-(+)- by high performance liquid chromatography in Examples
The conditions for the differential analysis of 3-chlorolactic acid and (S)-(-)-3-chlorolactic acid are as follows. Column: MCI GEL CRS 10W, 4.
6 x 50 mm (manufactured by Mitsubishi Kasei), eluent: micromolar copper sulfate, flow rate: 1 ml per minute, sample amount: 10 microliters, detection: UV (254 nm). Example 1 2% glucose, 1% diammonium phosphate, 1% phosphoric acid
10 m of sterile medium with the following composition: potassium 1%, meat extract 1%, peptone 1%, yeast extract 0.5%, sodium chloride 0.1%, (±)-3-chlorolactic acid 0.2%, pH 7.0
Inoculate the microorganisms shown in Table 1 into a large test tube containing
After culturing with shaking at 26°C for 48 hours, the bacterial cells were collected from the culture medium by centrifugation, and added to 50 mmol phosphate buffer (
After washing with pH 7.0), the bacterial cells were centrifuged again, and then
The bacterial cells are suspended in 50 mmol phosphate buffer (pH 7.0) containing (±)-3-chlorolactic acid to make 10 ml. The final concentration of (±)-3-chlorolactic acid was 0.2%. The reaction mixture was shaken and reacted at 26° C. for 48 hours, and then 3-chlorolactic acid contained in the centrifuged supernatant was analyzed by high performance liquid chromatography. The results are shown in Table 1, and although it varies depending on the strain, 50 to 80% of the 3-
Chlorolactic acid remains, and this residual (R)-(+)-3
-The optical isomer excess (ee) of chlorolactic acid is 100
It was ~38%. The yield was expressed as a percentage of the weight of (±)-3-chlorolactic acid added.
【表１】[Table 1]
【００１２】実施例２
（±）−３−クロロ乳酸３％、硫酸アンモニウム０．２
％、りん酸２カリ０．２％、硫酸マグネシウム０．０５
％、肉エキス０．５％、ペプトン１．０％、酵母エキス
０．５％、グリセロール２％、硫酸マンガン０．００１
％、硫酸亜鉛０．００１％の組成でｐＨを９．０に水酸
化ナトリウムで調整した滅菌培地１０ｍｌを含む太型試
験管に、シュードモナス属菌種Ｈ１−１（微工研菌寄第
１２１２８号）を植菌し、２６℃で４８時間振とう
培養したとき、培養液上清中に（＋）−３−クロロ乳酸
が１ｍｌあたり１６ｍｇ濃度に生成し、その光学異性体
過剰率（ｅ．ｅ．）は７８％であった。Example 2 (±)-3-chlorolactic acid 3%, ammonium sulfate 0.2
%, dipotassium phosphate 0.2%, magnesium sulfate 0.05
%, meat extract 0.5%, peptone 1.0%, yeast extract 0.5%, glycerol 2%, manganese sulfate 0.001
%, zinc sulfate 0.001% and the pH was adjusted to 9.0 with sodium hydroxide in a large test tube containing Pseudomonas sp. ) was inoculated and cultured with shaking at 26°C for 48 hours, (+)-3-chlorolactic acid was produced in the culture supernatant at a concentration of 16 mg per ml, and the optical isomer excess rate (e.e. ) was 78%.
【００１３】実施例３
シュードモナス・オーレオファシエンスＡＴＣＣ １
３９８６を用いて実施例１と同様に実施してえた反応液
１８００ｍｌから菌体を遠心分離により除いた上清を２
０分の１の液量まで減圧濃縮し、濃縮液のｐＨを硫酸で
ｐＨ２．５として２倍量の酢酸エチルで３回抽出した。
抽出液を合わせて減圧濃縮して冷室に放置し、析出した
結晶を濾別して褐色の結晶１．２ｇをえた。この結晶を
５ｍｌの酢酸エチルにとかし、シリカゲル（１．０×３
０ｃｍ）のカラムを通して吸着させた後、カラムにｎ−
ヘキサン５０ｍｌを流して洗った後、ｎ−ヘキサンと酢
酸エチルの混合液（容積比で９：１）で溶出し、（Ｒ）
−（＋）−３−クロロ乳酸の分画を集めて濃縮し、乾固
して１８０ｍｇの白色結晶をえた。この結晶は融点（８
９℃）、〔α〕２５Ｄ＋４．３３°（Ｃ＝１０、水）、
ＩＲ（Ｎｕｊｏｒ）ｃｍ−１：３４５０，１７２２、元
素分析値（Ｃ ２８．８４、Ｈ３．８５、Ｃｌ ２
８．３６）のデータから文献値と一致し、（Ｒ）−（＋
）−３−クロロ乳酸と同定された。Example 3 Pseudomonas aureofaciens ATCC 1
3986 in the same manner as in Example 1, and the supernatant after removing the bacterial cells by centrifugation was
It was concentrated under reduced pressure to 1/0 of the liquid volume, and the pH of the concentrated liquid was adjusted to pH 2.5 with sulfuric acid, and extracted three times with twice the amount of ethyl acetate. The combined extracts were concentrated under reduced pressure and left in a cold room, and the precipitated crystals were filtered off to obtain 1.2 g of brown crystals. The crystals were dissolved in 5 ml of ethyl acetate, and silica gel (1.0 x 3
After adsorption through a column of 0 cm), the column was filled with n-
After washing with 50 ml of hexane, elution was carried out with a mixture of n-hexane and ethyl acetate (9:1 by volume), and (R)
The fractions of -(+)-3-chlorolactic acid were collected, concentrated, and dried to give 180 mg of white crystals. This crystal has a melting point (8
9℃), [α]25D+4.33° (C=10, water),
IR (Nujor) cm-1: 3450, 1722, elemental analysis value (C 28.84, H 3.85, Cl 2
8.36) agrees with the literature value, and (R)−(+
)-3-chlorolactic acid.
【００１４】実施例４
実施例１において（±）−３−クロロ乳酸の代りに（±
）−３−ブロモ乳酸を用いる他は実施例１と同様に実施
した場合の結果は表２に示すとおりで、光学異性体過剰
率（ｅ．ｅ．）３７〜１００％の（＋）−３−ブロモ乳
酸が生成した。Example 4 In Example 1, (±)-3-chlorolactic acid was replaced with (±)-3-chlorolactic acid.
)-3-bromo lactic acid was used, and the results were shown in Table 2. (+)-3 with an optical isomer excess (ee) of 37 to 100% - Bromo lactic acid was produced.
【表２】[Table 2]
【００１５】実施例５
実施例３において（±）−３−クロロ乳酸の代りに（±
）−３−ブロモ乳酸を用いる他は実施例３と同様に実施
して（Ｒ）−（＋）−３−クロロ乳酸の結晶をえた。融
点７８℃、〔α〕２５Ｄ＋１．８°（Ｃ＝１０、水）、
光学異性体過剰率（ｅ．ｅ．）１００％であった。Example 5 In Example 3, (±)-3-chlorolactic acid was replaced with (±)-3-chlorolactic acid.
)-3-bromo lactic acid was used in the same manner as in Example 3 to obtain crystals of (R)-(+)-3-chlorolactic acid. Melting point 78°C, [α]25D+1.8° (C=10, water),
The optical isomer excess (ee) was 100%.
【００１６】[0016]
【発明の効果】本発明によれば（±）−３−ハロ乳酸か
ら、医薬品などの光学活性生理活性物の合成原料として
有用な（Ｒ）−（＋）−３−ハロ乳酸を効率よく製造す
ることができる。Effects of the Invention According to the present invention, (R)-(+)-3-halolactic acid, which is useful as a raw material for the synthesis of optically active physiologically active substances such as pharmaceuticals, can be efficiently produced from (±)-3-halolactic acid. can do.
Claims (3)
ａｓ）属、アースロバクター（Ａｒｔｈｒｏｂａｃｔｅ
ｒ）属、エッセリヒア（Ｅｓｃｈｅｒｉｃｈｉａ）属、
サッカロミセス（Ｓａｃｃｈａｒｏｍｙｃｅｓ）属、シ
ュワニオミセス（Ｓｃｈｗａｎｎｉｏｍｙｃｅｓ）属、
あるいはトルロプシス（Ｔｏｒｕｌｏｐｓｉｓ）属に属
し、（±）−３−ハロ乳酸から（Ｒ）−（＋）−３−ハ
ロ乳酸を生成する能力を有する微生物またはその処理物
を（±）−３−ハロ乳酸に接触反応させ、生成する（Ｒ
）−（＋）−３−ハロ乳酸を採取することを特徴とする
（Ｒ）−（＋）−３−ハロ乳酸の製造法。[Claim 1] Pseudomonas
as) genus, Arthrobacter
r) genus Escherichia,
Genus Saccharomyces, Genus Schwanniomyces,
Alternatively, a microorganism belonging to the genus Torulopsis and having the ability to produce (R)-(+)-3-halolactic acid from (±)-3-halolactic acid or a processed product thereof may be used to produce (±)-3-halolactic acid. to produce (R
A method for producing (R)-(+)-3-halolactic acid, which comprises collecting )-(+)-3-halolactic acid.
、生成物が（Ｒ）−（＋）−３−クロロ乳酸である請求
項１の製造法。2. The method of claim 1, wherein the substrate is (±)-3-chlorolactic acid and the product is (R)-(+)-3-chlorolactic acid.
、生成物が（Ｒ）−（＋）−３−ブロモ乳酸である請求
項１の製造法。3. The method of claim 1, wherein the substrate is (±)-3-bromo lactic acid and the product is (R)-(+)-3-bromo lactic acid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP17156191A JP2936551B2 (en) | 1991-04-15 | 1991-04-15 | Method for producing (R)-(+)-3-halolactic acid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP17156191A JP2936551B2 (en) | 1991-04-15 | 1991-04-15 | Method for producing (R)-(+)-3-halolactic acid |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH04316496A true JPH04316496A (en) | 1992-11-06 |
| JP2936551B2 JP2936551B2 (en) | 1999-08-23 |
Family
ID=15925423
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP17156191A Expired - Fee Related JP2936551B2 (en) | 1991-04-15 | 1991-04-15 | Method for producing (R)-(+)-3-halolactic acid |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2936551B2 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009050251A (en) * | 2007-05-14 | 2009-03-12 | Sumitomo Chemical Co Ltd | Method for producing lactic acid |
| WO2014017327A1 (en) * | 2012-07-23 | 2014-01-30 | 旭硝子株式会社 | Method for producing organic acid |
-
1991
- 1991-04-15 JP JP17156191A patent/JP2936551B2/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009050251A (en) * | 2007-05-14 | 2009-03-12 | Sumitomo Chemical Co Ltd | Method for producing lactic acid |
| WO2014017327A1 (en) * | 2012-07-23 | 2014-01-30 | 旭硝子株式会社 | Method for producing organic acid |
| JP5890905B2 (en) * | 2012-07-23 | 2016-03-22 | 旭硝子株式会社 | Method for producing organic acid |
| JPWO2014017327A1 (en) * | 2012-07-23 | 2016-07-11 | 旭硝子株式会社 | Method for producing organic acid |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2936551B2 (en) | 1999-08-23 |
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