CN104531577B - Arthrobacter nicotinovorans WYG001 and application thereof in preparation of N-BOC-L-homoserine lactone - Google Patents
Arthrobacter nicotinovorans WYG001 and application thereof in preparation of N-BOC-L-homoserine lactone Download PDFInfo
- Publication number
- CN104531577B CN104531577B CN201410800564.3A CN201410800564A CN104531577B CN 104531577 B CN104531577 B CN 104531577B CN 201410800564 A CN201410800564 A CN 201410800564A CN 104531577 B CN104531577 B CN 104531577B
- Authority
- CN
- China
- Prior art keywords
- boc
- wyg001
- application
- reaction
- homoserine
- 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.)
- Active
Links
- 241001147838 Paenarthrobacter nicotinovorans Species 0.000 title claims abstract description 9
- 238000002360 preparation method Methods 0.000 title claims abstract description 6
- IMWMFJMYEKHYKG-LURJTMIESA-N tert-butyl n-[(3s)-2-oxooxolan-3-yl]carbamate Chemical compound CC(C)(C)OC(=O)N[C@H]1CCOC1=O IMWMFJMYEKHYKG-LURJTMIESA-N 0.000 title abstract description 10
- 238000006243 chemical reaction Methods 0.000 claims abstract description 71
- 238000000034 method Methods 0.000 claims abstract description 24
- 239000000758 substrate Substances 0.000 claims abstract description 16
- 238000004108 freeze drying Methods 0.000 claims abstract description 6
- 239000003054 catalyst Substances 0.000 claims abstract description 5
- 239000012429 reaction media Substances 0.000 claims abstract description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 48
- 239000000243 solution Substances 0.000 claims description 33
- -1 N-BOC-DL- homoserine lactones Chemical class 0.000 claims description 30
- SNICXCGAKADSCV-JTQLQIEISA-N (-)-Nicotine Chemical compound CN1CCC[C@H]1C1=CC=CN=C1 SNICXCGAKADSCV-JTQLQIEISA-N 0.000 claims description 29
- 229960002715 nicotine Drugs 0.000 claims description 29
- SNICXCGAKADSCV-UHFFFAOYSA-N nicotine Natural products CN1CCCC1C1=CC=CN=C1 SNICXCGAKADSCV-UHFFFAOYSA-N 0.000 claims description 29
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 22
- 238000003756 stirring Methods 0.000 claims description 19
- 239000008363 phosphate buffer Substances 0.000 claims description 17
- 239000006228 supernatant Substances 0.000 claims description 16
- 239000001963 growth medium Substances 0.000 claims description 15
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 14
- 239000002904 solvent Substances 0.000 claims description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 239000012530 fluid Substances 0.000 claims description 12
- 238000005292 vacuum distillation Methods 0.000 claims description 12
- 239000002609 medium Substances 0.000 claims description 11
- 238000011218 seed culture Methods 0.000 claims description 11
- 239000011780 sodium chloride Substances 0.000 claims description 11
- 238000003810 ethyl acetate extraction Methods 0.000 claims description 10
- 229940041514 candida albicans extract Drugs 0.000 claims description 8
- 239000012138 yeast extract Substances 0.000 claims description 8
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 7
- 239000007836 KH2PO4 Substances 0.000 claims description 6
- 239000001888 Peptone Substances 0.000 claims description 6
- 108010080698 Peptones Proteins 0.000 claims description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 6
- 235000019319 peptone Nutrition 0.000 claims description 6
- 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 claims description 5
- 239000008101 lactose Substances 0.000 claims description 5
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 5
- 229920001817 Agar Polymers 0.000 claims description 4
- 239000008272 agar Substances 0.000 claims description 4
- 239000007853 buffer solution Substances 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 238000004321 preservation Methods 0.000 claims description 4
- 235000015278 beef Nutrition 0.000 claims description 3
- 239000000284 extract Substances 0.000 claims description 2
- 239000002054 inoculum Substances 0.000 claims description 2
- 239000013028 medium composition Substances 0.000 claims description 2
- 230000001954 sterilising effect Effects 0.000 claims description 2
- 108090000790 Enzymes Proteins 0.000 abstract description 20
- 102000004190 Enzymes Human genes 0.000 abstract description 20
- 241000894006 Bacteria Species 0.000 abstract description 11
- 208000012839 conversion disease Diseases 0.000 abstract description 9
- PZEMWPDUXBZKJN-UHFFFAOYSA-N 4-hydroxy-2-[(2-methylpropan-2-yl)oxycarbonylamino]butanoic acid Chemical compound CC(C)(C)OC(=O)NC(C(O)=O)CCO PZEMWPDUXBZKJN-UHFFFAOYSA-N 0.000 abstract description 7
- 238000003912 environmental pollution Methods 0.000 abstract description 3
- 238000005265 energy consumption Methods 0.000 abstract description 2
- 239000012295 chemical reaction liquid Substances 0.000 abstract 1
- 238000000855 fermentation Methods 0.000 abstract 1
- 230000004151 fermentation Effects 0.000 abstract 1
- 238000009776 industrial production Methods 0.000 abstract 1
- 239000008055 phosphate buffer solution Substances 0.000 abstract 1
- 238000000926 separation method Methods 0.000 abstract 1
- PZEMWPDUXBZKJN-LURJTMIESA-N (2s)-4-hydroxy-2-[(2-methylpropan-2-yl)oxycarbonylamino]butanoic acid Chemical compound CC(C)(C)OC(=O)N[C@H](C(O)=O)CCO PZEMWPDUXBZKJN-LURJTMIESA-N 0.000 description 41
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- 230000001580 bacterial effect Effects 0.000 description 11
- VLXBWPOEOIIREY-UHFFFAOYSA-N dimethyl diselenide Chemical compound C[Se][Se]C VLXBWPOEOIIREY-UHFFFAOYSA-N 0.000 description 11
- 230000000694 effects Effects 0.000 description 11
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 10
- UKAUYVFTDYCKQA-UHFFFAOYSA-N -2-Amino-4-hydroxybutanoic acid Natural products OC(=O)C(N)CCO UKAUYVFTDYCKQA-UHFFFAOYSA-N 0.000 description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- UKAUYVFTDYCKQA-VKHMYHEASA-N L-homoserine Chemical compound OC(=O)[C@@H](N)CCO UKAUYVFTDYCKQA-VKHMYHEASA-N 0.000 description 9
- 238000001514 detection method Methods 0.000 description 9
- 239000000047 product Substances 0.000 description 9
- 125000000659 L-selenomethionine group Chemical class [H]N([H])[C@]([H])(C(=O)[*])C([H])([H])C([H])([H])[Se]C([H])([H])[H] 0.000 description 8
- 239000007789 gas Substances 0.000 description 8
- 150000002596 lactones Chemical class 0.000 description 7
- 238000003786 synthesis reaction Methods 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 239000002253 acid Substances 0.000 description 6
- 229910021529 ammonia Inorganic materials 0.000 description 6
- 230000000813 microbial effect Effects 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 5
- 230000007062 hydrolysis Effects 0.000 description 5
- 238000006460 hydrolysis reaction Methods 0.000 description 5
- 239000000543 intermediate Substances 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 230000004224 protection Effects 0.000 description 5
- GOJUJUVQIVIZAV-UHFFFAOYSA-N 2-amino-4,6-dichloropyrimidine-5-carbaldehyde Chemical group NC1=NC(Cl)=C(C=O)C(Cl)=N1 GOJUJUVQIVIZAV-UHFFFAOYSA-N 0.000 description 4
- BMYNFMYTOJXKLE-UHFFFAOYSA-N 3-azaniumyl-2-hydroxypropanoate Chemical compound NCC(O)C(O)=O BMYNFMYTOJXKLE-UHFFFAOYSA-N 0.000 description 4
- QJPWUUJVYOJNMH-VKHMYHEASA-N L-homoserine lactone Chemical compound N[C@H]1CCOC1=O QJPWUUJVYOJNMH-VKHMYHEASA-N 0.000 description 4
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 4
- 230000001476 alcoholic effect Effects 0.000 description 4
- 238000006555 catalytic reaction Methods 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 4
- 230000014759 maintenance of location Effects 0.000 description 4
- 230000003287 optical effect Effects 0.000 description 4
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 3
- 238000005160 1H NMR spectroscopy Methods 0.000 description 3
- ZNKDXXHRIXGSTF-UHFFFAOYSA-N 2-bromobutanoic acid;hydrobromide Chemical compound Br.CCC(Br)C(O)=O ZNKDXXHRIXGSTF-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- 239000012043 crude product Substances 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000001962 electrophoresis Methods 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 238000004817 gas chromatography Methods 0.000 description 3
- 229910052739 hydrogen Inorganic materials 0.000 description 3
- 239000001257 hydrogen Substances 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 239000012279 sodium borohydride Substances 0.000 description 3
- 229910000033 sodium borohydride Inorganic materials 0.000 description 3
- 239000002689 soil Substances 0.000 description 3
- 238000010189 synthetic method Methods 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- FFRBMBIXVSCUFS-UHFFFAOYSA-N 2,4-dinitro-1-naphthol Chemical compound C1=CC=C2C(O)=C([N+]([O-])=O)C=C([N+]([O-])=O)C2=C1 FFRBMBIXVSCUFS-UHFFFAOYSA-N 0.000 description 2
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- RJFAYQIBOAGBLC-BYPYZUCNSA-N Selenium-L-methionine Chemical compound C[Se]CC[C@H](N)C(O)=O RJFAYQIBOAGBLC-BYPYZUCNSA-N 0.000 description 2
- 241000801924 Sena Species 0.000 description 2
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 2
- 229910052794 bromium Inorganic materials 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 150000002431 hydrogen Chemical class 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 2
- 239000010452 phosphate Substances 0.000 description 2
- 238000007747 plating Methods 0.000 description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 2
- 230000000707 stereoselective effect Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- NXRMEQQSABBWDL-JTQLQIEISA-N (2s)-4-hydroxy-2-[(2-phenylacetyl)amino]butanoic acid Chemical compound OCC[C@@H](C(O)=O)NC(=O)CC1=CC=CC=C1 NXRMEQQSABBWDL-JTQLQIEISA-N 0.000 description 1
- JDLMXICGDYZOJH-UHFFFAOYSA-N (3-bromo-1-carboxypropyl)azanium;bromide Chemical class [Br-].OC(=O)C([NH3+])CCBr JDLMXICGDYZOJH-UHFFFAOYSA-N 0.000 description 1
- SRFZPWAZZISKCU-UHFFFAOYSA-N (3R,4S,6S)-6-(hydroxymethyl)piperidine-3,4-diol Natural products OCC1CC(O)C(O)CN1 SRFZPWAZZISKCU-UHFFFAOYSA-N 0.000 description 1
- SRFZPWAZZISKCU-HCWXCVPCSA-N (3r,4s,6s)-6-(hydroxymethyl)piperidine-3,4-diol Chemical compound OC[C@@H]1C[C@H](O)[C@H](O)CN1 SRFZPWAZZISKCU-HCWXCVPCSA-N 0.000 description 1
- YAQLSKVCTLCIIE-UHFFFAOYSA-N 2-bromobutyric acid Chemical compound CCC(Br)C(O)=O YAQLSKVCTLCIIE-UHFFFAOYSA-N 0.000 description 1
- YEJRWHAVMIAJKC-UHFFFAOYSA-N 4-Butyrolactone Chemical compound O=C1CCCO1 YEJRWHAVMIAJKC-UHFFFAOYSA-N 0.000 description 1
- HBAQYPYDRFILMT-UHFFFAOYSA-N 8-[3-(1-cyclopropylpyrazol-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-yl]-3-methyl-3,8-diazabicyclo[3.2.1]octan-2-one Chemical compound C1(CC1)N1N=CC(=C1)C1=NNC2=C1N=C(N=C2)N1C2C(N(CC1CC2)C)=O HBAQYPYDRFILMT-UHFFFAOYSA-N 0.000 description 1
- 241000186063 Arthrobacter Species 0.000 description 1
- 240000006439 Aspergillus oryzae Species 0.000 description 1
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Natural products CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 1
- UKAUYVFTDYCKQA-GSVOUGTGSA-N D-homoserine Chemical compound OC(=O)[C@H](N)CCO UKAUYVFTDYCKQA-GSVOUGTGSA-N 0.000 description 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 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 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- JDLMXICGDYZOJH-DFWYDOINSA-N [(1s)-3-bromo-1-carboxypropyl]azanium;bromide Chemical class Br.OC(=O)[C@@H](N)CCBr JDLMXICGDYZOJH-DFWYDOINSA-N 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 125000003368 amide group Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 238000011953 bioanalysis Methods 0.000 description 1
- 230000031709 bromination Effects 0.000 description 1
- 238000005893 bromination reaction Methods 0.000 description 1
- 238000011088 calibration curve Methods 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000012159 carrier gas Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000005352 clarification Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 238000004807 desolvation Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 229910052564 epsomite Inorganic materials 0.000 description 1
- HVJJYOAPXBPQQV-UHFFFAOYSA-N ethyl 2-azidoacetate Chemical compound CCOC(=O)CN=[N+]=[N-] HVJJYOAPXBPQQV-UHFFFAOYSA-N 0.000 description 1
- XYIBRDXRRQCHLP-UHFFFAOYSA-N ethyl acetoacetate Chemical compound CCOC(=O)CC(C)=O XYIBRDXRRQCHLP-UHFFFAOYSA-N 0.000 description 1
- 238000010812 external standard method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000012847 fine chemical Substances 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 238000006197 hydroboration reaction Methods 0.000 description 1
- GPRLSGONYQIRFK-UHFFFAOYSA-N hydron Chemical compound [H+] GPRLSGONYQIRFK-UHFFFAOYSA-N 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000002633 protecting effect Effects 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 239000013558 reference substance Substances 0.000 description 1
- 229910052711 selenium Inorganic materials 0.000 description 1
- 239000011669 selenium Substances 0.000 description 1
- 229960002718 selenomethionine Drugs 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Inorganic materials [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000009182 swimming Effects 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- IMWMFJMYEKHYKG-ZCFIWIBFSA-N tert-butyl n-[(3r)-2-oxooxolan-3-yl]carbamate Chemical compound CC(C)(C)OC(=O)N[C@@H]1CCOC1=O IMWMFJMYEKHYKG-ZCFIWIBFSA-N 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 229910000368 zinc sulfate Inorganic materials 0.000 description 1
- 239000011686 zinc sulphate Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/06—Arthrobacter
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/04—Alpha- or beta- amino acids
- C12P13/12—Methionine; Cysteine; Cystine
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P41/00—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture
- C12P41/003—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by ester formation, lactone formation or the inverse reactions
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/62—Carboxylic acid esters
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses arthrobacter nicotinovorans WYG001 and application thereof in preparation of N-BOC-L-homoserine lactone. The N-BOC-L-homoserine lactone is prepared by taking a wet bacterium body obtained from the fermentation cultivation of the arthrobacter nicotinovorans WYG001 or a dry bacterium body obtained by freeze-drying the wet bacterium body as a catalyst, N-BOC-DL-homoserine lactone as a substrate and 0.1M of a phosphate buffer solution at pH7.0 as a reaction medium through carrying out a conversion reaction at 20-50 DEG C, and separating and purifying a reaction liquid after the reaction, so as to obtain the N-BOC-L-homoserine lactone. The method disclosed by the invention is strong in enzyme regioselectivity, high in reaction conversion rate, simple in downstream separation, low in energy consumption, slight in environmental pollution and suitable for industrial production.
Description
(1) technical field
The present invention relates to a kind of synthetic method of N-BOC-L- homoserine lactones, more particularly to using thermophilic nicotine arthrobacterium
WYG001 (CCTCC M 2014054) is catalyzed racemic BOC-DL- homoserine lactones, and stereo selective hydrolysis are synthesized light
Pure N-BOC-L- homoserine lactones are learned, then the method that N-BOC-L- homoserine lactones are synthesized L- selenomethionines.
(2) background technology
L- selenomethionines, English is entitled:L- (+)-Selenomethionine, is a kind of important animal foodstuff addition
Agent amino acid, is also to prepare other a kind of important intermediates containing selenium medicine.Its major function has:The essence of animal can be strengthened
Sub- vigor;Increase antioxidant ability of organism, enhance immunity;Effect with cancer-resisting;Body can also be helped to discharge internal
Toxin (such as heavy metal).
The homoserine lactone of optical voidness form is important chiral intermediate, is widely used in medicine and food additives etc.
Field.The present invention is split using microbial enzyme method catalysis and a kind of chiral intermediate N-BOC-L- homoserine lactones is obtained, and with
It is that substrate successfully synthesizes L- selenomethionines.
Because the amino of homoserine lactone is more active, its existence form in different pH reaction solutions is different, to product
The extraction of thing is made troubles, it is therefore desirable to select amido protecting group to protect it.The DL- protected by catalytic amino is high
Serine lactone selective hydrolysis, a configuration is able to catalyzing hydrolysis open loop into acid in realizing raceme, and another configuration is constant,
And then according to the difference of physical property, two kinds of configurations of homoserine lactone are separated, finally obtain optically pure L- Kosé ammonia
Acid lactone.
At present, the synthesis N-BOC-L- homoserine lactones of document report mainly pass through L- homoserine and its lactone hydrogen bromine
Hydrochlorate or hydrochloride are substrate, and BOC amido protectings are obtained.The synthetic method of wherein L- homoserine and its lactone salt is main by phase
Answer L-type aspartic acid (such as Han Chong, Jiang Lijian, Zhai Lihai amino acid and the living resources .2008,30 (2) of configuration:33-35)
With L-type methionine (Troels Koch, Ole Buchardt, 1993,1065-1067;Min-Can Wang,et
al.Organic Chemistry.2008,73(1):168-176;Rise Chinese soldier's fine-chemical intermediates .2008,38 (3):20-
21) it is Material synthesis.Other chemical synthesis also have asymmetric with α-ethyl azidoacetate with the furans imines of N-Boc protections
The pure homoserine lactone of synthesizing optical.(Kumaraswamy G,et al.Tetrahedron Letters.2010,51
(50):6500-6502).Chemical synthesis process approach has that low yield, high cost, step are various, equipment requirement is high, product
The problems such as optical purity is relatively low, environmental pollution is serious.
, (M Venkataiah, G Reddipalli, the L Swarnalatha such as Mallam Venkataiah in 2011
Jasti et al.Chemo-enzymatic synthesis of the azasugars1,4-
dideoxyallonojirimycin and 1,4-dideoxymannojirimycin[J].Tetrahedron
Asymmetry.2011,22:Alpha-amido-the gamma-butyrolacton of racemic phenylacetyl first 1855-1860) is used into 10% liquefied ammonia
PH is into alkalescence for alkalization regulation, so that lactone open loop;Then hydrolyzed with Immobilized Pen Gacylase and obtain open loop
(R) derivative of the phenylacetyl of the homoserine of type homoserine and (S)-type;It is last to be acidified by HCl in methanol solution
(S) configuration, the e.e. > 99% of N- phenylacetyl-L- homoserine lactones are obtained, yield is 47% (without conversion ratio).
, (the Kazuya M, Kentaro such as Kazuya Mochizuki, Kentaro Miyazaki in 2007
M.Microbial resolution of DL-homoserine for the production of D-homoserine
using a novel isolate,Arthrobacter nicotinovorans strain 2-3[J].Enzyme and
Microbial Technology.2007,41:318-321) first by racemic homoserine lactone lactone hydrobromate open loop
Homoserine is obtained, then with racemic homoserine as sole carbon source, being filtered out from soil being capable of selective hydrolysis
Fall L-configuration, and obtain the bacterial strain of the homoserine of D configurations.The microbial strains Arthrobacter for screening
Nicotinovorans Strain obtain optics in the racemic homoserine of catalysis of phosphate solution neutral body selectivity
The e.e. of pure enantiomerS>99.9%, D configuration yield are 25%, and the homoserine yield of L- configurations is about 4%, and conversion
Time is 48h, and the reaction time is long, is not easy to industrialized production.
By contrast, the bioanalysis reported of the present invention have that stereoselectivity is high, reaction condition is gentle, high income and product
The advantage of the easily separated purifying of thing.
(3) content of the invention
It is an object of the present invention to provide a kind of synthetic method of N-BOC-L- homoserine lactones, the method utilizes thermophilic nicotine section
Bacillus WYG001 (CCTCC NO:M 2014054) catalysis racemic N-BOC-DL- homoserine lactones, stereo selective hydrolysis
Optical voidness N-BOC-L- homoserine lactones are synthesized, then N-BOC-L- homoserine lactones are synthesized into L- selenomethionines,
The inventive method has the advantages that stereoselectivity is high, reaction condition gentle, high income and the easily separated purifying of product.
The technical solution adopted by the present invention is:
The present invention provides one plant of new strains -- thermophilic nicotine arthrobacterium (Arthrobacter nicotinovorans)
WYG001, is preserved in China typical culture collection center, deposit number CCTCC NO:M2014054, preservation date 2014 2
The moon 28, address is China, Wuhan, Wuhan University, postcode 430072.
The present invention also provides a kind of applications of thermophilic nicotine arthrobacterium WYG001 in N-BOC-L- homoserine lactones are prepared,
Described application is:It is dry after the wet thallus obtained with the fermented cultures of thermophilic nicotine arthrobacterium WYG001 or wet thallus freeze-drying
Thalline is catalyst, with N-BOC-DL- homoserine lactones as substrate, with 0.1M, pH7.0 phosphate buffer as reaction medium,
Conversion reaction (preferably reacting 1-30h) is carried out under the conditions of 20~50 DEG C, 200rpm, after reaction terminates, reaction solution is separated pure
Change, obtain N-BOC-L- homoserine lactones.
The preparation method of catalyst of the present invention is:(1) inclined-plane culture:Thermophilic nicotine arthrobacterium WYG001 is seeded to tiltedly
Face culture medium, 30 DEG C of culture 24h, obtains thalline inclined-plane;Every liter of slant medium (beef-protein medium) composition:Beef
Cream 5g, peptone 10g, NaCl 5g, agar 20g, pH 7.0-7.2, add water to 1000mL;121 DEG C of sterilizing 20min.
(2) seed culture:Seed culture medium is seeded to from the oese thalline of inclined-plane thalline picking one, in 30 DEG C, 200r/
Min cultivates 24h, obtains seed liquor;Seed culture medium is constituted:Peptone 10g/L, yeast extract 5g/L, NaCl 10g/L, solvent is
Water, pH 7.0;
(3) fermented and cultured:Aseptically, seed liquor is inoculated in into producing enzyme with the inoculum concentration of volumetric concentration 6-8% to train
Support in base, 30 DEG C, 200r/min culture 24h obtain zymotic fluid, and zymotic fluid 12000rpm is centrifuged 10min, abandoned by culture after terminating
Supernatant, obtains thermophilic nicotine arthrobacterium WYG001 wet thallus;Culture medium is constituted:Lactose 5-15g/L, yeast extract 10-
14g/L, NaCl 10g/L, MgSO40.22g/L、KH2PO40.136g/L, solvent is water, pH 7.0;
(4) in taking step (3) wet thallus with the phosphate buffer of the amount addition pH 7.0,0.1mol/L of 1.2g/ml, stir
Mix uniform, freeze-drying under the conditions of -80 DEG C obtains dry mycelium.
Further, the consumption of the wet thallus is calculated as 3~150g/L, preferably 10~100g/L with buffer solution volume, optimal
At the beginning of selecting 10~50g/L, the consumption of the dry mycelium to be calculated as 1~50g/L, preferably 5~30g/L, the substrate with buffer solution volume
Beginning concentration is 1~100g/L, preferably 5-50g/L.
The method that reaction solution of the present invention is isolated and purified is:After reaction terminates, reaction solution is centrifuged, takes supernatant, plus
Enter isometric ethyl acetate extraction, take ethyl acetate layer vacuum distillation to dry desolvation, obtain N-BOC-L- homoserine
Lactone.
Further, 1~30h is reacted in the reaction at 20~50 DEG C, and more preferably reaction temperature is 30~35 DEG C.
Culture medium composition of the present invention is preferably:Lactose 10g/L, yeast extract 12g/L, NaCl 10g/L,
MgSO40.22g/L、KH2PO40.136g/L, solvent is water, pH 7.0.
The present invention prepares the method for L- selenomethionines using N-BOC-L- homoserine lactones:
(1) N-Boc-L- homoserine lactones are mixed with the acetic acid solution of the hydrobromic acid of mass concentration 33%, 100 DEG C of heating
Backflow 10h, continues stirring reaction 10h, is slowly dropped to room temperature, stands overnight, and filters, and has solid to separate out, and is filtrated to get product L-
2- amino -4- bromobutanoic acid hydrobromides, and with ethyl acetate 30mL washed products, dry and obtain white solid, yield is 85%;
The volumetric usage of the acetic acid solution of the hydrobromic acid is calculated as 2.5ml/g with N-Boc-L- homoserine lactone quality;
(2) dimethyl diselenide ether is mixed with absolute methanol, sodium borohydride (hydroboration is dividedly in some parts under nitrogen protection
Sodium:Dimethyl diselenide ether=mass ratio 2:1), it is stirred at room temperature 0.5h-2.5h, solution is become colorless transparent by golden yellow, is obtained
CH3SeNa alcoholic solutions;To CH3The bromobutanoic acid hydrobromide of 2- amino -4 is added in SeNa alcoholic solutions, oil bath heating to 45 DEG C -65 DEG C,
Continue to stir 2-10h, filtering can obtain L- selenomethionine crude products after reaction terminates;The bromo-butyric acid hydrogen bromine of the 2- amino -4
Hydrochlorate is 1.4 with dimethyl diselenide ether mass ratio:1, the volumetric usage of the absolute methanol is calculated as with dimethyl diselenide ether quality
8ml/g。
N-BOC-L- homoserine lactones (I), obtain in the S-2- amino bromo- butyric acid of -4- (III) through hydrobromic acid open loop bromination,
With the L- selenomethionines (V) of dimethyl diselenide ether reaction system in the presence of sodium borohydride.
The beneficial effect major embodiment of the preparation method of medicine intermediate N-BOC-L- homoserine lactones of the present invention
:The present invention provides one plant of new strains -- thermophilic nicotine arthrobacterium (Arthrobacter nicotinovorans) WYG001, utilizes
Thermophilic nicotine arthrobacterium WYG001 prepares N-BOC-L- homoserine lactones, and the regioselectivity of enzyme is strong, and reaction conversion ratio is high, downstream
Separate simply, energy consumption is low, and environmental pollution is small, be adapted to industrialized production.
(4) illustrate
Fig. 1 racemic modification N-BOC-L- homoserine lactone reference substance gas chromatograms, peak a is N-Boc-D- homoserine
Lactone, retention time 16.831 minutes, peak area 489.4, peak height 51.2, peak width 0.1465, symmetrical factor 1.16, peak b is N-
Boc-L- homoserine lactones, retention time 17.797 minutes, peak area 485.9, peak height 50.1, peak width 0.1501, it is symmetrical because
Son 0.905;
The gas chromatogram of Fig. 2 N-BOC-L- homoserine lactone standard items, peak b is N-Boc-L- homoserine lactones,
Retention time 17.791 minutes, peak area 677.1, peak height 73.1, peak width 0.1111, symmetrical factor 0.788;
The gas chromatogram of Fig. 3 microorganism catalysis product N-BOC-L- homoserine lactones, peak b is N-Boc-L- Kosé ammonia
Acid lactone, retention time 17.792 minutes, peak area 628.2, peak height 67.4, peak width 0.1327, symmetrical factor 0.815.
Fig. 4 is reacting flow chart of the present invention.
Fig. 5 is the colonial morphology figure of bacterial strain WYG001.
Fig. 6 is light micrographs of the bacterial strain WYG001 under 100 times.
Fig. 7 is the 16S rDNA electrophoretograms of bacterial strain WYG001, and A is the electrophoretogram of bacterial strain WYG001, and left side swimming lane is bacterial strain
WYG001, right lanes are standard molecular weight, and B is the enlarged drawing of A.
(5) specific embodiment
With reference to specific embodiment, the present invention is described further, but protection scope of the present invention is not limited in
This:
Embodiment 1
1st, the screening of microorganism
The present invention relates to microorganism screen isolated by following procedure:Weigh 1~2g of wet soil sample (from the whole nation not
Obtained with soil under environmental condition) it is suspended in the SPSSs of 10mL 0.5%, vibration is mixed;Connect 1~2mL suspensions
In 30mL liquid enriched mediums, after cultivating 2~3d under 30 DEG C, 200r/min, sterile working switching 1mL muddy bacterium solution
To fresh liquid enriched medium, continuous enrichment 3 times.It is unique with N-BOC-DL- homoserine lactones in enriched medium
Carbon source, is formulated as follows:NaNO34.0g/L, KH2PO42.0g/L, MgSO4·7H2O 0.5g/L, KCl 1.0g/L,
ZnSO40.05g/L, solvent is water, pH 7.0.The concentration 1g/L of the N-BOC-DL- homoserine lactones of first round enrichment, second
The concentration of wheel is 3g/L, and the concentration of third round is 5g/L, pH 7.0.Bacterium solution is through gradient dilution, coating point after third round is enriched with
From plating medium, separating for several times obtains single bacterium colony.Separating plating medium composition is:Enriched medium adds agar dense eventually
Degree 20g/L, substrate N-BOC-DL- homoserine lactones final concentration of 5g/L, pH 7.0.
The isolated single bacterium colony of picking is inoculated in slant medium, and 30 DEG C are cultivated 1-2 days, obtain inclined-plane thalline.From oblique
The oese thalline of face thalline picking one is seeded to the seed culture medium of 50mL, in 30 DEG C, 200r/min culture 24h, obtains seed
Liquid.Aseptically, take 3mL seed liquors to be inoculated in the culture medium of 50mL, 30 DEG C, 200r/min culture 24h are obtained
Zymotic fluid.Every liter of slant medium constitutes beef extract 5g, peptone 10g, NaCl 5g, agar 20g, pH 7.0-7.2, adds water to
1000mL;Seed culture based formulas are:Peptone 10g/L, yeast extract 5g/L, NaCl 10g/L, solvent is water, pH 7.0;Produce
Enzyme culture medium is constituted:Lactose 10g/L, yeast extract 12g/L, NaCl 10g/L, MgSO40.22g/L、KH2PO40.136g/L, it is molten
Agent is water, pH 7.0.15mL zymotic fluids 12000rpm centrifugations 10min in 50mL centrifuge tubes is taken, supernatant is abandoned, 5mL is added
The phosphate buffer of 0.1mol/L pH 7.0, vibration is mixed, and adds final concentration 8g/L substrate N-Boc-DL- homoserine
Lactone, under 30 DEG C, 200r/min, conversion reaction 24h.Centrifuge tube is taken out, the reaction solution of 1mL is taken, 2mL ethyl acetate is added,
Fully vibration, 6000rpm centrifugations 5min.Upper organic phase 1mL is taken, is removed water with a small amount of anhydrous sodium sulfate drying, with efficient liquid phase
Chromatogram detects the enantiomeric excess value of N-Boc-L- homoserine lactones
(e.e.), reaction conversion ratio C, screening enantiomeric excess value (e.e.) is more than 90%, and reaction conversion ratio C is (45%-
60%) microbial strains, are designated as bacterial strain WYG001.
2nd, the identification of microbial strains
Physiological and biochemical property:Bacterial strain WYG001 is Gram-negative bacteria, obligate aerobic, and growth changes more sensitive to pH, most
Suitable pH is 7.0, shown in colonial morphology as Fig. 5 and Fig. 6.Poor using inorganic nitrogen-sourced ability, growth and producing enzyme are subject to Cu2+、Fe2 +、Al3+、Zn2+Deng suppression, by Mg2+、K+、Na+Deng promotion.
Through sequencing identification, the 16S rDNA sequences of bacterial strain WYG001 are following (SEQ ID NO.1), and electrophoretogram is as shown in Figure 7:
TGCTTACACATGCAAGTCGAACGATGATCCCAGCTTGCTGGGGGATTAGTGGCGAACGGGTGAGTAACA
CGTGAGTAACCTGCCCTTGACTCTGGGATAAGCCTGGGAAACTGGGTCTAATACCGGATATGACTCCTCATCGCATG
GTGGGGGGTGGAAAGCTTTTGTGGTTTTGGATGGACTCGCGGCCTATCAGCTTGTTGGTGGGGTAATGGCCTACCAA
GGCGACGACGGGTAGCCGGCCTGAGAGGGTGACCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGG
CAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCGACGCCGCGTGAGGGATGACGGCCTTCGGGTTG
TAAACCTCTTTCAGTAGGGAAGAAGCGTAAGTGACGGTACCTGCAGAAGAAGCGCCGGCTAACTACGTGCCAGCAGC
CGCGGTAATACGTAGGGCGCAAGCGTTATCCGGAATTATTGGGCGTAAAGAGCTCGTAGGCGGTTTGTCGCGTCTGC
TGTGAAAGACCGGGGCTCAACTCCGGTTCTGCAGTGGGTACGGGCAGACTAGAGTGCAGTAGGGGAGACTGGAATTC
CTGGTGTAGCGGTGAAATGCGCAGATATCAGGAGGAACACCGATGGCGAAGGCAGGTCTCTGGGCTGTAACTGACGC
TGAGGAGCGAAAGCATGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGTTGGGCACTAGGTGT
GGGGGACATTCCACGTTTTCCGCGCCGTAGCTAACGCATTAAGTGCCCCGCCTGGGGAGTACGGCCGCAAGGCTAAA
ACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGCGGATTAATTCGATGCAACGCGAAGAACCTTACC
AAGGCTTGACATGAACCGGAAAGACCTGGAAACAGGTGCCCCGCTTGCGGTCGGTTTACAGGTGGTGCATGGTTGTC
GTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGTTCTATGTTGCCAGCGGTTCGGC
CGGGGACTCATAGGAGACTGCCGGGGTCAACTCGGAGGAAGGTGGGGACGACGTCAAATCATCATGCCCCTTATGTC
TTGGGCTTCACGCATGCTACAATGGCCGGTACAAAGGGTTGCGATACTGTGAGGTGGAGCTAATCCCAAAAAGCCGG
TCTCAGTTCGGATTGGGGTCTGCAACTCGACCCCATGAAGTCGGAGTCGCTAGTAATCGCAGATCAGCAACGCTGCG
GTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCAAGTCACGAAAGTTGGTAACACCCGAAGCCGGTGGCCTAA
CCCTTGTGGGGGGAGCCGTCGAAGGTGGGACCGGCGATGGGACTAAGTC
According to physio-biochemical characteristics and molecular biology identification, the bacterial strain is accredited as thermophilic nicotine arthrobacterium bacterium
(Arthrobacter nicotinovorans), is named as thermophilic nicotine arthrobacterium bacterium (Arthrobacter
Nicotinovorans) WYG001, be preserved in China typical culture collection center (address China, Wuhan, Wuhan University,
430072), deposit number CCTCC NO:M 2014054, preservation date 2014 year 2 month 28 days.
Embodiment 2:
Inclined-plane culture:Thermophilic nicotine arthrobacterium WYG001 is seeded to slant medium, 30 DEG C of culture 24h obtain thalline oblique
Face;Slant medium is constituted with embodiment 1.
Seed culture:100mL seed culture mediums are seeded to from the oese thalline of inclined-plane thalline picking one, in 30 DEG C, 200r/
Min cultivates 24h, obtains seed liquor;Seed culture medium is constituted with embodiment 1.
Fermented and cultured:Aseptically, 10mL seed liquors are taken to be inoculated in the culture medium of 150mL, 30 DEG C,
200r/min cultivates 24h, obtains zymotic fluid, and zymotic fluid 12000rpm is centrifuged 10min, abandons supernatant, obtained by culture after terminating
Thermophilic nicotine arthrobacterium WYG001 wet thallus.Take the phosphate-buffered that wet thallus add pH 7.0,0.1mol/L with the amount of 1.2g/ml
In liquid, stir, freeze-drying obtains dry mycelium under the conditions of -80 DEG C.
Embodiment 3:
The wet thallus 0.34g that the method for Example 2 is obtained, addition N-BOC-DL- homoserine lactones 50mg and 0.1M,
PH7.0 phosphate buffer 5mL, under the conditions of 30 DEG C, 200rpm, stirring reaction 7.5h.Reacting liquid filtering is removed into enzyme, filtrate is taken
The content of the N-Boc-DL- homoserine lactones after conversion is determined with gas chromatographic analysis, enzyme activity, the zymotic fluid specific enzyme activity is calculated
Power is 36.85U/L.
Enzyme activity is defined as:U is 1 enzyme activity unit, and 1 described enzyme activity unit is every point under conditions of regulation
Clock the N-Boc- homoserine lactones of 1 μm of ol are hydrolyzed into N-Boc- homoserine needed for enzyme amount.
Analytical conditions for gas chromatography:Using the type gas chromatographs of gas chromatograph Agilent 6890;Chiral gas chromatography
Post BGB-175 (0.25mm × 30m);Carrier gas N2(14.54kPa), split ratio 20:1,250 DEG C of injector temperature, detector temperature
220℃;Column temperature rise program:180 DEG C of initial temperature, keeps 20min;The μ L of sample size 1.0, hydrogen flowing quantity:30mL/min air
Flow:300mL/min;Make-up gas flow:N225mL/min;Hydrogen ion flame detector.N-BOC-D- homoserine lactones and
N-BOC-L- homoserine lactones are respectively in 16.831min and 17.797min appearances.
For the N-BOC-DL- homoserine lactone calibration curve equations of enzyme activity determination:
The racemic N-Boc-DL- that compound concentration is respectively 0.1%, 0.4%, 0.5%, 0.6%, 0.8%, 1.0% is high
Serine lactone (w/v), is detected using gas-chromatography, draws the linear relationship between the concentration of lactone and peak area, with
The mass concentration of N-Boc-DL- homoserine lactones is ordinate, and gas chromatographic detection gained peak area is marked for abscissa
Directrix curve equation:Y=1352x -26.79, in 0.1%-2% (w/v) concentration range, coefficient R2=0.999, reach
Requirement needed for external standard method, the analysis that the curve can be used for N-Boc-DL- homoserine lactones is determined.
Enzyme activity computing formula:
A=[(a1-a)/a1]×[C×V/(M×t)]×106Formula (1)
A --- the vigor (U) of enzyme in sample;
A --- conversion reaction terminates the peak area of N-BOC-DL- homoserine lactones in rear conversion fluid;
a1--- calculated according to above-mentioned standard curvilinear equation with initial substrate concentration identical N-BOC-DL- Kosé ammonia
The peak area of acid lactone standard specimen;
C --- in conversion fluid, the mass concentration of substrate, g/L;
V --- conversion fluid volume, L;
M --- the molal weight of substrate N-BOC-DL- homoserine lactones, g/mol;
T --- transformation time, Min.
Embodiment 4:
The dry mycelium 0.2g that the method for Example 2 is obtained, addition N-BOC-DL- homoserine lactones 0.1g, 0.1M,
PH7.0 phosphate buffer 10mL, under the conditions of 30 DEG C, 200rpm, stirring reaction 7.5h.Enzyme activity is determined using the method for embodiment 3,
The dry mycelium specific enzyme activity power is 5.53U/g.
Embodiment 5:
The method of Example 2 gained wet thallus 3.4g, addition N-BOC-DL- homoserine lactones 0.25g and 0.1M,
PH7.0 phosphate buffer 50mL, under the conditions of 30 DEG C, 200rpm, stirring reaction 4h after reaction terminates, reaction solution is centrifuged, and is taken
Supernatant, adds isometric ethyl acetate extraction, after taking ethyl acetate layer vacuum distillation to dry desolventizing, obtains N-BOC-L-
Homoserine lactone 0.12g.N-BOC-L- homoserine lactones are calculated using GC (testing conditions are with embodiment 3) detections
Enantiomeric excess value e.e.s99.9%, conversion ratio C are 51.8%, and yield is 96.4%.
Enantiomeric excess value (enantiomeric excesses) is the important indicator for weighing enzyme selectivity, N-BOC-L-
The enantiomeric excess value abbreviation e.e. of homoserine lactones, its computing formula is:
Formula (2)
Wherein S represents substrate, SRAnd SSRespectively (D)-and (L)-configuration substrate peak area.
The computing formula of reaction conversion ratio is:
Formula (3)
The computing formula of the yield (Yield) of N-BOC-L- homoserine lactones is
Wherein, SR0And SS0Respectively (D)-and (L)-configuration substrate initial peak area.
Embodiment 6:
Thermophilic nicotine arthrobacterium WYG001 dry mycelium 1g obtained in the method for Example 2, add in N-BOC-DL- homoserine
Ester 0.5g and 0.1M, pH7.0 phosphate buffer 100mL, under the conditions of 30 DEG C, 200rpm, stirring reaction 7.5h, reaction terminates
Afterwards, reaction solution is centrifuged, takes supernatant, add isometric ethyl acetate extraction, taken ethyl acetate layer vacuum distillation and taken off to dry
After solvent, N-BOC-L- homoserine lactones 0.24g is obtained.N- is calculated using GC (testing conditions are with embodiment 3) detections
The enantiomeric excess value e.e. of BOC-L- homoserine lactonessIt is 99.9%, conversion ratio C is 52.0%, and yield is 96.0%.
Embodiment 7
Thermophilic nicotine arthrobacterium WYG001 dry mycelium 4g obtained in the method for Example 2, add in N-BOC-DL- homoserine
Ester 2g and 0.1M, pH7.0 phosphate buffer 100mL, under the conditions of 30 DEG C, 200rpm, stirring reaction 8h, after reaction terminates, will
Reaction solution is centrifuged, and takes supernatant, adds isometric ethyl acetate extraction, takes ethyl acetate layer vacuum distillation to dry desolventizing
Afterwards, N-BOC-L- homoserine lactones 0.97g is obtained.The mapping of N-BOC-L- homoserine lactones is detected using the method for embodiment 3
Body excessive value e.e.99.9%, conversion ratio C are 51.0%, and yield is 97.0%.
Embodiment 8
Thermophilic nicotine arthrobacterium WYG001 dry mycelium 1.0g obtained in the method for Example 2, add N-BOC-DL- homoserine
Lactone 0.5g and 0.1M, pH7.0 phosphate buffer 20mL, under the conditions of 35 DEG C, 200rpm, stirring reaction 14h, reaction terminates
Afterwards, reaction solution is centrifuged, takes supernatant, add isometric ethyl acetate extraction, taken ethyl acetate layer vacuum distillation and taken off to dry
After solvent, N-BOC-L- homoserine lactones 0.233g is obtained.N-BOC-L- Kosé ammonia is calculated using the detection of the method for embodiment 3
The enantiomeric excess value e.e. of acid lactonesIt is 99.9%, conversion ratio C is 52.5%, and yield is 93.2%.
Embodiment 9
Thermophilic nicotine arthrobacterium WYG001 dry mycelium 1.0g obtained in the method for Example 2, add N-BOC-DL- homoserine
Lactone 1g and 0.1M, pH7.0 phosphate buffer 20mL, under the conditions of 35 DEG C, 200rpm, after stirring reaction 30h, reaction terminates
Afterwards, reaction solution is centrifuged, takes supernatant, add isometric ethyl acetate extraction, taken ethyl acetate layer vacuum distillation and taken off to dry
After solvent, N-BOC-L- homoserine lactones 0.453g is obtained.N-BOC-L- homoserine lactones are calculated using GC detections
Enantiomeric excess value e.e.sIt is 95.6%, conversion ratio C is 52.0%, and yield is 90.6%.
Embodiment 10:
Thermophilic nicotine arthrobacterium WYG001 dry mycelium 0.2g obtained in the method for Example 2, add N-BOC-DL- homoserine
Lactone 0.15g and 0.1M, pH7.0 phosphate buffer 10mL, under the conditions of 30 DEG C, 200rpm, stirring reaction 13h, reaction terminates
Afterwards, reaction solution is centrifuged, takes supernatant, add isometric ethyl acetate extraction, taken ethyl acetate layer vacuum distillation and taken off to dry
After solvent, N-BOC-L- homoserine lactones 0.144g is obtained.Using the method for embodiment 3 detection N-BOC-L- homoserine lactones
Enantiomeric excess value e.e.99.9%, conversion ratio C are 50.8%, and yield is 96.0%.
Embodiment 11:
Thermophilic nicotine arthrobacterium WYG001 dry mycelium 1.5g obtained in the method for Example 1, add N-BOC-DL- homoserine
Lactone 1.5g and 0.1M, pH7.0 phosphate buffer 100mL, under the conditions of 30 DEG C, 200rpm, stirring reaction 13h, reaction terminates
Afterwards, reaction solution is centrifuged, takes supernatant, add isometric ethyl acetate extraction, taken ethyl acetate layer vacuum distillation and taken off to dry
After solvent, N-BOC-L- homoserine lactones 0.725g is obtained.N-BOC-L- homoserine lactones are calculated using GC detections
Enantiomeric excess value e.e.sIt is 99.9%, conversion ratio C is 50.7%, and yield is 96.6%.
Embodiment 12:
Thermophilic nicotine arthrobacterium WYG001 dry mycelium 5g obtained in the method for Example 1, add in N-BOC-DL- homoserine
Ester 10g and 0.1M, pH7.0 phosphate buffer 100mL, under the conditions of 40 DEG C, 200rpm, stirring reaction 15h, after reaction terminates,
Reaction solution is centrifuged, supernatant is taken, isometric ethyl acetate extraction is added, ethyl acetate layer vacuum distillation to dry desolventizing is taken
Afterwards, N-BOC-L- homoserine lactones 4.70g is obtained.The enantiomer of N-BOC-L- homoserine lactones is calculated using GC detections
Excessive value e.e.sIt is 99.0%, conversion ratio C is 51.2%, and yield is 94.0%.
Embodiment 13:
Thermophilic nicotine arthrobacterium WYG001 dry myceliums in embodiment 11 are substituted into (dry bacterium with the dry mycelium 0.2g in table 1 respectively
The preparation of body is with embodiment 2), N-BOC-DL- homoserine lactones 0.1g and 0.1M, pH7.0 phosphate buffer 10mL is added,
30 DEG C, under the conditions of 200rpm, stirring reaction 24h after reaction terminates, reaction solution is centrifuged, and takes supernatant, adds isometric second
Acetoacetic ester is extracted, and after taking ethyl acetate layer vacuum distillation to dry desolventizing, obtains N-BOC-L- homoserine lactones.Detected using GC
It is calculated the enantiomeric excess value e.e. of N-BOC-L- homoserine lactonessWith conversion ratio C, 1 is shown in Table.
Table 1
The result of table 1 shows that the bacterial strain such as aspergillus oryzae wzz007 has certain catalytic effect and stereoselectivity to substrate, but
It is to be not so good as thermophilic nicotine arthrobacterium WYG001 of the invention in effect.
Embodiment 14:
(1) thermophilic nicotine arthrobacterium WYG001 dry mycelium 10g obtained in the method for Example 2, add N-BOC-DL- Kosé ammonia
Acid lactone 5g and 0.1M, pH7.0 phosphate buffer 200mL, under the conditions of 30 DEG C, 200rpm, stirring reaction 11h, terminating reaction,
Reaction solution is centrifuged off thalline, and supernatant is extracted with ethyl acetate (50mL × 5), takes ethyl acetate layer vacuum distillation to dry precipitation
After agent, optically pure N-Boc-L- homoserine lactones 2.40g (detection method is with embodiment 3) is obtained, yield is 96.0%;N-
The enantiomeric excess value e.e. of BOC-L- homoserine lactonessIt is 99.9%, its data structure characterizes as follows:
1H NMR(600MHz,CDCl3) δ 7.34 (d, J=8.4Hz, 1H), 4.37-4.28 (m, 2H), 4.19-4.15 (m,
1H),2.39-2.34(m,1H),2.17-2.12(m,1H),1.39(s,9H).13C NMR(150MHz,CDCl3))δ174.5,
155.6,78.5,64.7,49.1,29.2,27.3.Mp 123-125℃.[α]D=+8.6 ° of (c=1, CHCl3)
(2) synthesis of 2- amino -4- bromobutanoic acid hydrobromides
In the round mouth flask of 50mL, N-Boc-L- homoserine lactone 2.40g are added, be subsequently adding 6mL mass concentrations
The acetic acid solution of 33% hydrobromic acid, oil bath heating continues stirring reaction 10h to 100 DEG C, and reaction solution fades to clarification by muddiness,
Room temperature is slowly dropped to, is stood overnight, there is solid to separate out, filtered, obtain the crude product of the bromobutanoic acid hydrobromide of 2- amino -4, be used in combination
Ethyl acetate washed product, is finally dried to obtain white solid 2.65g, and yield is 85%.
1H NMR(500MHz,D2O) δ 2.22-2.41 (m, 2H), 3.59-3.71 (m, 2H), 4.00 (d, J=5.6Hz,
1H).13C NMR(126MHz,D2O)δ174.6,51.7,32.9,28.3.
(3) synthesis of L- selenomethionines hydrobromate
In two mouthfuls of flasks of 50mL, 1.88g dimethyl diselenide ethers are added, be subsequently adding the absolute methanol of 15mL, make it
Mixing, one end is connected with nitrogen protection device.Sodium borohydride 3.76g is dividedly in some parts under nitrogen protection, to prevent reaction excessively acute
It is strong.(25 DEG C) of room temperature is stirred 10 minutes, and solution is become colorless transparent by golden yellow, obtains CH3SeNa alcoholic solutions.By what is prepared
CH3The bromobutanoic acid hydrobromide of 2.65g 2- amino -4 is added in SeNa alcoholic solutions, oil bath heating continues to stir 5h to 60 DEG C.Instead
There is solid to separate out after should terminating, filtering can obtain the crude product of L- selenomethionines and then use ethanol washed product again, dry
Obtain white solid.Yield is 61.2%.
1H NMR(500MHz,D2O)δ3.73-3.71(m,1H),2.52-2.49(m 2H),2.15-2.04(m,2H),
1.92-1.90(m,3H).13C NMR(125MHz,D2O)δ174.2,54.9,31.0,19.4,3.4.MS(ESI):m/z[M+H]
+:198.Mp 267.2-269.3℃.[α]D=+18.8 ° (c=1,1N HCl).
Claims (10)
1. thermophilic nicotine arthrobacterium (Arthrobacter nicotinovorans) WYG001, it is preserved in Chinese Typical Representative culture guarantor
Tibetan center, deposit number CCTCC NO:M 2014054, preservation date 2014 year 2 month 28 days, preservation address is Chinese, Wuhan,
Wuhan University, postcode 430072.
2. prepared by thermophilic nicotine arthrobacterium WYG001 described in a kind of claim 1NApplication in-BOC-L- homoserine lactones.
3. application as claimed in claim 2, it is characterised in that described application is:It is fermented with thermophilic nicotine arthrobacterium WYG001
The dry mycelium cultivated after the wet thallus or wet thallus freeze-drying for obtaining is catalyst, is with N-BOC-DL- homoserine lactones
Substrate, with 0.1M, pH7.0 phosphate buffer as reaction medium, carries out converting instead under the conditions of 30 ~ 35 DEG C or 40 DEG C, 200rpm
Should, after reaction terminates, reaction solution is isolated and purified, obtainN- BOC-L- homoserine lactones.
4. application as claimed in claim 3, it is characterised in that the preparation method of the catalyst is:
(1)Inclined-plane culture:Thermophilic nicotine arthrobacterium WYG001 is seeded to slant medium, 30 DEG C of 24 h of culture obtain thalline oblique
Face;Every liter of slant medium composition:The g of beef extract 5, peptone 10 g, NaCl 5 g, agar 20 g, pH 7.0-7.2, add water
To 1000 mL;121 C sterilizings, 20 min;
(2)Seed culture:Seed culture medium is seeded to from the oese thalline of inclined-plane thalline picking one, in 30 DEG C, 200 r/min
24 h are cultivated, seed liquor is obtained;Seed culture medium is constituted:The g/L of peptone 10, yeast extract 5 g/L, NaCl 10 g/L, solvent
It is water, pH 7.0;
(3)Fermented and cultured:Aseptically, seed liquor is inoculated in by culture medium with the inoculum concentration of volumetric concentration 6-8%
In, 30 DEG C, 200 r/min culture 24h obtain zymotic fluid, and the rpm of zymotic fluid 12000 is centrifuged 10 min, abandoned by culture after terminating
Supernatant, obtains thermophilic nicotine arthrobacterium WYG001 wet thallus;Culture medium is constituted:Lactose 5-15 g/L, yeast extract 10-14
G/L, NaCl 10 g/L, MgSO4 0.22g/L、KH2PO40.136g/L, solvent is water, pH 7.0;
(4)Take step(3)Wet thallus are added in pH 7.0, the phosphate buffer of 0.1 mol/L with the amount of 1.2g/ml, stirring
Uniformly, freeze-drying under the conditions of -80 DEG C, obtains dry mycelium.
5. application as claimed in claim 3, it is characterised in that the consumption of the wet thallus is calculated as 3 ~ 150 g with buffer solution volume
/L。
6. application as claimed in claim 3, it is characterised in that the consumption of the dry mycelium is calculated as 1 ~ 50 g with buffer solution volume
/L。
7. application as claimed in claim 3, it is characterised in that the initial substrate concentration is 1 ~ 100g/L.
8. application as claimed in claim 3, it is characterised in that the method that the reaction solution is isolated and purified is:After reaction terminates,
Reaction solution is centrifuged, supernatant is taken, isometric ethyl acetate extraction is added, ethyl acetate layer vacuum distillation is taken molten to dry removing
Agent, obtainsN- BOC-L- homoserine lactones.
9. application as claimed in claim 3, it is characterised in that the reaction reacts 1 ~ 30h at 30 ~ 35 DEG C or 40 DEG C.
10. application as claimed in claim 4, it is characterised in that culture medium constitutes and is:The g/L of lactose 10, yeast extract 12
G/L, NaCl 10 g/L, MgSO4 0.22g/L、KH2PO40.136g/L, solvent is water, pH 7.0.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201410800564.3A CN104531577B (en) | 2014-12-19 | 2014-12-19 | Arthrobacter nicotinovorans WYG001 and application thereof in preparation of N-BOC-L-homoserine lactone |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201410800564.3A CN104531577B (en) | 2014-12-19 | 2014-12-19 | Arthrobacter nicotinovorans WYG001 and application thereof in preparation of N-BOC-L-homoserine lactone |
Publications (2)
Publication Number | Publication Date |
---|---|
CN104531577A CN104531577A (en) | 2015-04-22 |
CN104531577B true CN104531577B (en) | 2017-05-17 |
Family
ID=52847212
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201410800564.3A Active CN104531577B (en) | 2014-12-19 | 2014-12-19 | Arthrobacter nicotinovorans WYG001 and application thereof in preparation of N-BOC-L-homoserine lactone |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN104531577B (en) |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106399191B (en) * | 2016-10-31 | 2019-07-16 | 杭州朗境环保科技有限公司 | The application of one plant of arthrobacterium B2 and its nitric wastewater of degrading |
CN109868248A (en) * | 2019-04-09 | 2019-06-11 | 中国科学院沈阳应用生态研究所 | A kind of arthrobacter bacteria high density fermentation culture medium |
CN110642768A (en) * | 2019-10-22 | 2020-01-03 | 河北凡博医药科技有限公司 | Preparation method of L-selenomethionine |
CN113698331B (en) * | 2020-05-23 | 2023-12-15 | 普济生物科技(台州)有限公司 | Synthesis method of L-selenium-methyl selenocysteine |
CN115466695B (en) * | 2022-09-19 | 2024-02-20 | 苏州科技大学 | Arthrobacter nicotinovorans and application thereof |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1085095A1 (en) * | 1997-09-24 | 2001-03-21 | NSC Technologies Company, L.L.C. | Synthesis of optically active phenylalanine analogs through microbial transformations |
CN101374806A (en) * | 2006-01-28 | 2009-02-25 | 赢创德固赛有限责任公司 | Method for producing methionine from homoserine |
CN104242738A (en) * | 2013-06-21 | 2014-12-24 | 中国石油集团渤海石油装备制造有限公司 | Frequency conversion and energy saving control device and method for cluster well pumping units |
-
2014
- 2014-12-19 CN CN201410800564.3A patent/CN104531577B/en active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1085095A1 (en) * | 1997-09-24 | 2001-03-21 | NSC Technologies Company, L.L.C. | Synthesis of optically active phenylalanine analogs through microbial transformations |
CN101374806A (en) * | 2006-01-28 | 2009-02-25 | 赢创德固赛有限责任公司 | Method for producing methionine from homoserine |
CN104242738A (en) * | 2013-06-21 | 2014-12-24 | 中国石油集团渤海石油装备制造有限公司 | Frequency conversion and energy saving control device and method for cluster well pumping units |
Non-Patent Citations (1)
Title |
---|
Microbial resolution of dl-homoserine for the production of d-homoserine using a novel isolate, Arthrobacter nicotinovorans strain 2-3;Kazuya Mochizuki等;《Enzyme and Microbial Technology》;20070802;第41卷(第3期);318-321 * |
Also Published As
Publication number | Publication date |
---|---|
CN104531577A (en) | 2015-04-22 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN104531577B (en) | Arthrobacter nicotinovorans WYG001 and application thereof in preparation of N-BOC-L-homoserine lactone | |
CN110527646B (en) | Tropical bacillus WZZ018 and application thereof | |
CN104372017B (en) | A kind of method and application for improving genetic engineering bacterium isoprene and its derivative yield | |
US10196660B2 (en) | Xylene monooxygenase-producing strain Arthrobacter woluwensis and its application | |
CN103509816B (en) | Produce the construction process of Coenzyme Q10 99.0 engineering bacteria, engineering bacteria and application thereof | |
CN109504634B (en) | Bacillus WZZ006 and application thereof | |
CN109385379A (en) | A kind of engineering bacteria of high yield rhodioside and its application | |
CN103898037A (en) | Genetically engineered bacterium co-generating geraniol and nerol and construction method and application thereof | |
CN103509728B (en) | Produce the construction process of Coenzyme Q10 99.0 engineering bacteria, engineering bacteria and application method | |
CN104263713A (en) | Tistrella mobilis, halohydrin dehalogenase, gene, vector, recombinant strain and application of halohydrin dehalogenase | |
CN105925519B (en) | A kind of method, Co-Q10 superior strain and its application reducing or eliminating by-product D in Co-Q10 production bacterial strain SZ | |
CN103045504B (en) | Microorganism catalysis prepared (2S,3R)-2-benzoyl aminomethyl-3-hydroxybutyric acid ester and bacterial strain | |
CN112063531B (en) | Geotrichum candidum ZJPH1907 and application thereof in preparing (S) -2-chloro-1- (3, 4-difluorophenyl) ethanol | |
CN106083811B (en) | (R)-alpha-lipoic acid preparation method | |
CN111411134B (en) | Preparation method for producing purine by fermenting Bacillus sp.JIN118 | |
US9322040B2 (en) | Pseudonocardia sp. and method for preparing deoxynyboquinone by utilizing same | |
CN103589665B (en) | Celebrating sheng, a reed pipe wind instrument rhodococcus and the application in preparation (S)-4-chloro-3-hydroxyl ethyl butyrate thereof | |
CN102851238B (en) | Sphingobacterium and method for preparing levetiracetam acid by utilizing same | |
CN107881202A (en) | A kind of biological preparation method of Levonorgestrel key chiral intermediate | |
CN105802872B (en) | Pseudomonas fluorescens, method for producing phenazine amide and application thereof | |
CN102120977A (en) | Microbacterium chocolatum and method for preparing (4S,5R)-half ester by using same | |
CN108441433B (en) | Rhodotorula mucilaginosa NQ1 and application thereof in preparation of chiral alcohol | |
CN103074239B (en) | Candida sorboxylosa and application thereof in preparation of (S)-4-chloro-3-hydroxybutanoate | |
CN107142227B (en) | Acid-producing klebsiella and application thereof | |
CN106591155B (en) | Fusarium oxysporum strain and its 3 β, 7 α are being prepared, the application in 15 α-trihydroxyandrost -5- alkene -17- ketone |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |