CN103865911A - 青霉素g酰化酶突变体及其在合成头孢类抗生素中的应用 - Google Patents
青霉素g酰化酶突变体及其在合成头孢类抗生素中的应用 Download PDFInfo
- Publication number
- CN103865911A CN103865911A CN201410057240.5A CN201410057240A CN103865911A CN 103865911 A CN103865911 A CN 103865911A CN 201410057240 A CN201410057240 A CN 201410057240A CN 103865911 A CN103865911 A CN 103865911A
- Authority
- CN
- China
- Prior art keywords
- penicillin
- acylase
- cefaclor
- mutant
- synthesis
- 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
- 229930186147 Cephalosporin Natural products 0.000 title claims abstract description 13
- 230000015572 biosynthetic process Effects 0.000 title claims abstract description 13
- 229940124587 cephalosporin Drugs 0.000 title claims abstract description 13
- 150000001780 cephalosporins Chemical class 0.000 title claims abstract description 13
- 238000003786 synthesis reaction Methods 0.000 title claims abstract description 13
- 102000004190 Enzymes Human genes 0.000 title abstract description 46
- 108090000790 Enzymes Proteins 0.000 title abstract description 46
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 title abstract description 17
- 230000010933 acylation Effects 0.000 title abstract 9
- 238000005917 acylation reaction Methods 0.000 title abstract 9
- 229940056360 penicillin g Drugs 0.000 title abstract 6
- 239000003242 anti bacterial agent Substances 0.000 title abstract 3
- 229940088710 antibiotic agent Drugs 0.000 title abstract 3
- 229960005361 cefaclor Drugs 0.000 claims abstract description 29
- QYIYFLOTGYLRGG-GPCCPHFNSA-N cefaclor Chemical compound C1([C@H](C(=O)N[C@@H]2C(N3C(=C(Cl)CS[C@@H]32)C(O)=O)=O)N)=CC=CC=C1 QYIYFLOTGYLRGG-GPCCPHFNSA-N 0.000 claims abstract description 27
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 18
- SZJUWKPNWWCOPG-UHFFFAOYSA-N methyl 2-anilinoacetate Chemical compound COC(=O)CNC1=CC=CC=C1 SZJUWKPNWWCOPG-UHFFFAOYSA-N 0.000 claims abstract description 12
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract description 6
- 108010073038 Penicillin Amidase Proteins 0.000 claims description 48
- 238000006243 chemical reaction Methods 0.000 claims description 27
- 230000003115 biocidal effect Effects 0.000 claims description 10
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 8
- 241000894006 Bacteria Species 0.000 claims description 6
- 239000002994 raw material Substances 0.000 claims description 6
- NVIAYEIXYQCDAN-CLZZGJSISA-N 7beta-aminodeacetoxycephalosporanic acid Chemical compound S1CC(C)=C(C(O)=O)N2C(=O)[C@@H](N)[C@@H]12 NVIAYEIXYQCDAN-CLZZGJSISA-N 0.000 claims description 4
- 239000013598 vector Substances 0.000 claims description 4
- FBIPHLTUVCYGRD-FOUAAFFMSA-N (6R)-4-chloro-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-4-carboxylic acid Chemical compound ClC1(S[C@H]2N(C=C1)C(C2)=O)C(=O)O FBIPHLTUVCYGRD-FOUAAFFMSA-N 0.000 claims description 3
- 208000002991 Ring chromosome 4 syndrome Diseases 0.000 claims description 3
- 230000000968 intestinal effect Effects 0.000 claims description 2
- 150000004702 methyl esters Chemical class 0.000 claims description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims 2
- 235000009508 confectionery Nutrition 0.000 claims 1
- 229960003742 phenol Drugs 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 18
- 230000007062 hydrolysis Effects 0.000 abstract description 12
- 238000006460 hydrolysis reaction Methods 0.000 abstract description 12
- 229930182555 Penicillin Natural products 0.000 abstract description 5
- 229940049954 penicillin Drugs 0.000 abstract description 5
- 239000002773 nucleotide Substances 0.000 abstract description 4
- 125000003729 nucleotide group Chemical group 0.000 abstract description 4
- WDLWHQDACQUCJR-ZAMMOSSLSA-N (6r,7r)-7-[[(2r)-2-azaniumyl-2-(4-hydroxyphenyl)acetyl]amino]-8-oxo-3-[(e)-prop-1-enyl]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@@H]3N(C2=O)C(=C(CS3)/C=C/C)C(O)=O)=CC=C(O)C=C1 WDLWHQDACQUCJR-ZAMMOSSLSA-N 0.000 abstract 1
- 229960004841 cefadroxil Drugs 0.000 abstract 1
- NBFNMSULHIODTC-CYJZLJNKSA-N cefadroxil monohydrate Chemical compound O.C1([C@@H](N)C(=O)N[C@H]2[C@@H]3N(C2=O)C(=C(CS3)C)C(O)=O)=CC=C(O)C=C1 NBFNMSULHIODTC-CYJZLJNKSA-N 0.000 abstract 1
- 229960002580 cefprozil Drugs 0.000 abstract 1
- 230000008859 change Effects 0.000 description 20
- 239000000758 substrate Substances 0.000 description 16
- 239000007788 liquid Substances 0.000 description 13
- 238000006555 catalytic reaction Methods 0.000 description 12
- 238000000034 method Methods 0.000 description 11
- 108700023418 Amidases Proteins 0.000 description 10
- 102000005922 amidase Human genes 0.000 description 10
- 239000000243 solution Substances 0.000 description 10
- 239000000047 product Substances 0.000 description 9
- 230000035772 mutation Effects 0.000 description 7
- 238000005516 engineering process Methods 0.000 description 6
- 235000018102 proteins Nutrition 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 5
- 238000002703 mutagenesis Methods 0.000 description 5
- 231100000350 mutagenesis Toxicity 0.000 description 5
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 4
- ZGUNAGUHMKGQNY-ZETCQYMHSA-N L-alpha-phenylglycine zwitterion Chemical compound OC(=O)[C@@H](N)C1=CC=CC=C1 ZGUNAGUHMKGQNY-ZETCQYMHSA-N 0.000 description 4
- 235000001014 amino acid Nutrition 0.000 description 4
- 150000001413 amino acids Chemical class 0.000 description 4
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 4
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 4
- 235000011130 ammonium sulphate Nutrition 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- -1 heterocyclic amino group amides acid amides Chemical class 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 238000001228 spectrum Methods 0.000 description 4
- ZYLDQHILNOZKIF-DHLUJLSBSA-N (6r,7r)-7-azaniumyl-8-oxo-3-[(e)-prop-1-enyl]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate Chemical compound S1CC(/C=C/C)=C(C(O)=O)N2C(=O)[C@@H](N)[C@@H]12 ZYLDQHILNOZKIF-DHLUJLSBSA-N 0.000 description 3
- 229920000936 Agarose Polymers 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 3
- 102100035460 Polynucleotide 5'-hydroxyl-kinase Human genes 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 150000001408 amides Chemical class 0.000 description 3
- 238000004140 cleaning Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000001257 hydrogen Substances 0.000 description 3
- 229910052739 hydrogen Inorganic materials 0.000 description 3
- 150000002460 imidazoles Chemical class 0.000 description 3
- DKLZRLRMFNWRFB-UHFFFAOYSA-N methyl 2-(4-hydroxyanilino)acetate Chemical compound COC(=O)CNC1=CC=C(O)C=C1 DKLZRLRMFNWRFB-UHFFFAOYSA-N 0.000 description 3
- 230000002194 synthesizing effect Effects 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 150000003952 β-lactams Chemical class 0.000 description 3
- NGHVIOIJCVXTGV-ALEPSDHESA-N 6-aminopenicillanic acid Chemical compound [O-]C(=O)[C@H]1C(C)(C)S[C@@H]2[C@H]([NH3+])C(=O)N21 NGHVIOIJCVXTGV-ALEPSDHESA-N 0.000 description 2
- NGHVIOIJCVXTGV-UHFFFAOYSA-N 6beta-amino-penicillanic acid Natural products OC(=O)C1C(C)(C)SC2C(N)C(=O)N21 NGHVIOIJCVXTGV-UHFFFAOYSA-N 0.000 description 2
- 101000925662 Enterobacteria phage PRD1 Endolysin Proteins 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- NPKSPKHJBVJUKB-UHFFFAOYSA-N N-phenylglycine Chemical compound OC(=O)CNC1=CC=CC=C1 NPKSPKHJBVJUKB-UHFFFAOYSA-N 0.000 description 2
- 108010024026 Nitrile hydratase Proteins 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 125000000539 amino acid group Chemical group 0.000 description 2
- FCPVYOBCFFNJFS-LQDWTQKMSA-M benzylpenicillin sodium Chemical compound [Na+].N([C@H]1[C@H]2SC([C@@H](N2C1=O)C([O-])=O)(C)C)C(=O)CC1=CC=CC=C1 FCPVYOBCFFNJFS-LQDWTQKMSA-M 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- YGBFLZPYDUKSPT-MRVPVSSYSA-N cephalosporanic acid Chemical compound S1CC(COC(=O)C)=C(C(O)=O)N2C(=O)C[C@H]21 YGBFLZPYDUKSPT-MRVPVSSYSA-N 0.000 description 2
- 239000000460 chlorine Substances 0.000 description 2
- 229910052801 chlorine Inorganic materials 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 230000002349 favourable effect Effects 0.000 description 2
- 239000003205 fragrance Substances 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 2
- 239000007791 liquid phase Substances 0.000 description 2
- 230000008707 rearrangement Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 210000001082 somatic cell Anatomy 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- OQSAFIZCBAZPMY-AWFVSMACSA-N (6r,7r)-7-azaniumyl-3-chloro-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate Chemical compound S1CC(Cl)=C(C([O-])=O)N2C(=O)[C@@H]([NH3+])[C@H]21 OQSAFIZCBAZPMY-AWFVSMACSA-N 0.000 description 1
- 125000000954 2-hydroxyethyl group Chemical group [H]C([*])([H])C([H])([H])O[H] 0.000 description 1
- HSHGZXNAXBPPDL-HZGVNTEJSA-N 7beta-aminocephalosporanic acid Chemical compound S1CC(COC(=O)C)=C(C([O-])=O)N2C(=O)[C@@H]([NH3+])[C@@H]12 HSHGZXNAXBPPDL-HZGVNTEJSA-N 0.000 description 1
- 241000588624 Acinetobacter calcoaceticus Species 0.000 description 1
- 241000588625 Acinetobacter sp. Species 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 101000911390 Homo sapiens Coagulation factor VIII Proteins 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 101710141454 Nucleoprotein Proteins 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 229920002684 Sepharose Polymers 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 239000003905 agrochemical Substances 0.000 description 1
- SRBFZHDQGSBBOR-QMKXCQHVSA-N alpha-L-arabinopyranose Chemical compound O[C@H]1CO[C@@H](O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-QMKXCQHVSA-N 0.000 description 1
- 125000003368 amide group Chemical group 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- KLOHDWPABZXLGI-YWUHCJSESA-M ampicillin sodium Chemical compound [Na+].C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C([O-])=O)(C)C)=CC=CC=C1 KLOHDWPABZXLGI-YWUHCJSESA-M 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000003782 beta lactam antibiotic agent Substances 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 150000001735 carboxylic acids Chemical class 0.000 description 1
- 238000005266 casting Methods 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 238000006482 condensation reaction Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000010612 desalination reaction Methods 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 102000057593 human F8 Human genes 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- SZBDOFWNZVHVGR-MRVPVSSYSA-N methyl (2r)-2-amino-2-(4-hydroxyphenyl)acetate Chemical compound COC(=O)[C@H](N)C1=CC=C(O)C=C1 SZBDOFWNZVHVGR-MRVPVSSYSA-N 0.000 description 1
- BHFLUDRTVIDDOR-MRVPVSSYSA-N methyl (2r)-2-amino-2-phenylacetate Chemical compound COC(=O)[C@H](N)C1=CC=CC=C1 BHFLUDRTVIDDOR-MRVPVSSYSA-N 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000003032 molecular docking Methods 0.000 description 1
- 238000000329 molecular dynamics simulation Methods 0.000 description 1
- 125000000896 monocarboxylic acid group Chemical group 0.000 description 1
- 230000000869 mutational effect Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 235000008729 phenylalanine Nutrition 0.000 description 1
- 150000002994 phenylalanines Chemical class 0.000 description 1
- 150000003016 phosphoric acids Chemical class 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 230000007096 poisonous effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000001742 protein purification Methods 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 229940047431 recombinate Drugs 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000002132 β-lactam antibiotic Substances 0.000 description 1
- 229940124586 β-lactam antibiotics Drugs 0.000 description 1
Images
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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
- C12N9/80—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5) acting on amide bonds in linear amides (3.5.1)
- C12N9/84—Penicillin amidase (3.5.1.11)
-
- 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
- C12P35/00—Preparation of compounds having a 5-thia-1-azabicyclo [4.2.0] octane ring system, e.g. cephalosporin
- C12P35/04—Preparation of compounds having a 5-thia-1-azabicyclo [4.2.0] octane ring system, e.g. cephalosporin by acylation of the substituent in the 7 position
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y305/00—Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5)
- C12Y305/01—Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5) in linear amides (3.5.1)
- C12Y305/01011—Penicillin amidase (3.5.1.11), i.e. penicillin-amidohydrolase
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
Description
10×reaction buffering | 5μl |
dNTPs(各2mM) | 5μl |
Mg2+(25mM) | 3μl |
模板 | 1μl |
PHE TO ALA-F(10mM) | 1.5μl |
PHE TO ALA-ANTI(10mM) | 1.5μl |
KOD enzyme(1U/μl) | 1μl |
ddH2O | 32μl |
Total | 50μl |
Claims (10)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201410057240.5A CN103865911B (zh) | 2014-02-20 | 2014-02-20 | 青霉素g酰化酶突变体及其在合成头孢类抗生素中的应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201410057240.5A CN103865911B (zh) | 2014-02-20 | 2014-02-20 | 青霉素g酰化酶突变体及其在合成头孢类抗生素中的应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN103865911A true CN103865911A (zh) | 2014-06-18 |
CN103865911B CN103865911B (zh) | 2015-10-21 |
Family
ID=50904921
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201410057240.5A Active CN103865911B (zh) | 2014-02-20 | 2014-02-20 | 青霉素g酰化酶突变体及其在合成头孢类抗生素中的应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN103865911B (zh) |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105063158A (zh) * | 2015-08-18 | 2015-11-18 | 浙江东盈药业有限公司 | 一种头孢丙烯的合成方法 |
CN105385621A (zh) * | 2015-11-20 | 2016-03-09 | 浙江昌明药业有限公司 | 琼氏不动杆菌zjutfet-1及其应用 |
CN105483105A (zh) * | 2016-02-23 | 2016-04-13 | 上海星维生物技术有限公司 | 一种青霉素g酰化酶突变体 |
CN106222230A (zh) * | 2016-08-03 | 2016-12-14 | 广州白云山医药集团股份有限公司白云山化学制药厂 | 一种绿色酶法合成头孢克洛的方法 |
CN107099523A (zh) * | 2017-06-15 | 2017-08-29 | 清华大学 | 头孢拉定合成酶突变体及其编码基因 |
CN109266713A (zh) * | 2018-11-12 | 2019-01-25 | 齐鲁安替制药有限公司 | 一种适用于工业生产的头孢克洛的制备方法 |
CN113567607A (zh) * | 2021-07-23 | 2021-10-29 | 河北冀安检测服务有限公司 | 一种头孢克洛中青霉素g酰基转移酶含量的检测方法 |
CN115433701A (zh) * | 2022-11-08 | 2022-12-06 | 南京农业大学 | 一株变形杆菌及其菌剂和在降解头孢类抗生素中的应用 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1028541C (zh) * | 1989-10-26 | 1995-05-24 | 中国科学院微生物研究所 | 微生物发酵法生产青霉素酰化酶 |
CN1224471A (zh) * | 1997-04-22 | 1999-07-28 | 吉斯特-布罗卡迪斯有限公司 | 一种制备β-内酰胺抗生素的方法 |
CN1608130A (zh) * | 2001-12-27 | 2005-04-20 | Dsmip资产有限公司 | β-内酰胺抗生素的制备方法 |
WO2007138148A1 (es) * | 2006-05-30 | 2007-12-06 | Consejo Superior De Investigaciones Científicas | Enzima penicilina g acilasa mutada inmovilizada y estabilizada, procedimiento de obtención y sus aplicaciones |
-
2014
- 2014-02-20 CN CN201410057240.5A patent/CN103865911B/zh active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1028541C (zh) * | 1989-10-26 | 1995-05-24 | 中国科学院微生物研究所 | 微生物发酵法生产青霉素酰化酶 |
CN1224471A (zh) * | 1997-04-22 | 1999-07-28 | 吉斯特-布罗卡迪斯有限公司 | 一种制备β-内酰胺抗生素的方法 |
CN1608130A (zh) * | 2001-12-27 | 2005-04-20 | Dsmip资产有限公司 | β-内酰胺抗生素的制备方法 |
WO2007138148A1 (es) * | 2006-05-30 | 2007-12-06 | Consejo Superior De Investigaciones Científicas | Enzima penicilina g acilasa mutada inmovilizada y estabilizada, procedimiento de obtención y sus aplicaciones |
Non-Patent Citations (3)
Title |
---|
CERQUEIRA, G.等: "penicillin amidase [Acinetobacter gyllenbergii CIP 110306]", 《GENBANK DATABASE》 * |
FRANTISEK SKROB等: "Novel penicillin G acylase from Achromobacter sp. CCM 4824", 《ENZYME AND MICROBIAL TECHNOLOGY》 * |
周政等: "利用DNA家族重排提高青霉素G酰化酶合成活力(英文)", 《生物化学与生物物理学报》 * |
Cited By (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105063158A (zh) * | 2015-08-18 | 2015-11-18 | 浙江东盈药业有限公司 | 一种头孢丙烯的合成方法 |
CN105385621A (zh) * | 2015-11-20 | 2016-03-09 | 浙江昌明药业有限公司 | 琼氏不动杆菌zjutfet-1及其应用 |
CN105483105A (zh) * | 2016-02-23 | 2016-04-13 | 上海星维生物技术有限公司 | 一种青霉素g酰化酶突变体 |
CN105483105B (zh) * | 2016-02-23 | 2018-08-31 | 上海星维生物技术有限公司 | 一种青霉素g酰化酶突变体 |
CN106222230A (zh) * | 2016-08-03 | 2016-12-14 | 广州白云山医药集团股份有限公司白云山化学制药厂 | 一种绿色酶法合成头孢克洛的方法 |
CN107099523A (zh) * | 2017-06-15 | 2017-08-29 | 清华大学 | 头孢拉定合成酶突变体及其编码基因 |
CN107099523B (zh) * | 2017-06-15 | 2019-09-27 | 清华大学 | 头孢拉定合成酶突变体及其编码基因 |
CN109266713A (zh) * | 2018-11-12 | 2019-01-25 | 齐鲁安替制药有限公司 | 一种适用于工业生产的头孢克洛的制备方法 |
CN113567607A (zh) * | 2021-07-23 | 2021-10-29 | 河北冀安检测服务有限公司 | 一种头孢克洛中青霉素g酰基转移酶含量的检测方法 |
CN115433701A (zh) * | 2022-11-08 | 2022-12-06 | 南京农业大学 | 一株变形杆菌及其菌剂和在降解头孢类抗生素中的应用 |
Also Published As
Publication number | Publication date |
---|---|
CN103865911B (zh) | 2015-10-21 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN103865911B (zh) | 青霉素g酰化酶突变体及其在合成头孢类抗生素中的应用 | |
CN108424900B (zh) | 一种腈水解酶突变体及其构建方法和应用 | |
Yamada et al. | Nitrile hydratase and its application to industrial production of acrylamide | |
US11001823B2 (en) | Nitrilase mutants and application thereof | |
Sio et al. | Improved β-lactam acylases and their use as industrial biocatalysts | |
CN103834631B (zh) | 一种青霉素g酰化酶突变体及其编码基因和应用 | |
JP7117792B2 (ja) | 酵素法によってr-3-アミノ酪酸を製造する方法 | |
CN102533705B (zh) | 一种腈水解酶及其基因和应用 | |
JP2009538118A (ja) | 2−ヒドロキシ−2−メチルカルボン酸の酵素的な製造法 | |
CN105483105B (zh) | 一种青霉素g酰化酶突变体 | |
CN101701222B (zh) | 一种编码产碱杆菌腈水解酶的基因及其用于制备扁桃酸单一对映体的方法 | |
CN105274082B (zh) | 一种合成用青霉素g酰化酶突变体及其在制备阿莫西林中的应用 | |
Ma et al. | High-level expression of glutaryl-7-aminocephalosporanic acid acylase from Pseudomonas diminuta NK703 in Escherichia coli by combined optimization strategies | |
CN104130967A (zh) | 一株共表达l-乳酸脱氢酶和甲酸脱氢酶的大肠杆菌及其构建方法与应用 | |
CN104152500A (zh) | 一种生物合成(r)-3-羟基戊二酸单酯的新方法 | |
CN109852593A (zh) | 一种重组酮还原酶及在制备r-3-羟基丁酸及其盐中的应用 | |
CN110358751B (zh) | 一种重组脂肪酶突变体、编码基因、重组工程菌及应用 | |
JP2012143183A (ja) | L−ホモセリン及びl−ホモセリンラクトンの製造法 | |
CN103409402B (zh) | 醛缩酶突变体 | |
Shen et al. | Stereoselective nitrile hydratase | |
KR100463966B1 (ko) | 시아노카르복실산 에스테르로부터 디카르복실산모노에스테르의 제조 | |
CN110628800A (zh) | 一种手性醇高效生产重组菌的构建方法及其应用 | |
CN116478974B (zh) | 一种谷氨酸脱羧酶及其基因和应用 | |
CN116515802A (zh) | 天冬氨酸酶突变体及其工程菌的合成与应用 | |
CN103998618A (zh) | 光学活性的α-取代-β-氨基酸的制造方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
TR01 | Transfer of patent right |
Effective date of registration: 20200629 Address after: 322118, 519, Zhenjiang South Road, Dongyang, Zhejiang, Jinhua, Hengdian Co-patentee after: APELOA PHARMACEUTICAL Co.,Ltd. Patentee after: ZHEJIANG APELOA TOSPO PHARMACEUTICAL Co.,Ltd. Address before: 322118 No. 519 Jiangnan Road, Hengdian Industrial Zone, Zhejiang, Jinhua, China Patentee before: ZHEJIANG APELOA TOSPO PHARMACEUTICAL Co.,Ltd. |
|
TR01 | Transfer of patent right | ||
PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Penicillin G acylase mutant and its application in the synthesis of cephalosporin antibiotics Effective date of registration: 20220917 Granted publication date: 20151021 Pledgee: Dongyang sub branch of Bank of China Ltd. Pledgor: ZHEJIANG APELOA TOSPO PHARMACEUTICAL Co.,Ltd. Registration number: Y2022330002320 |
|
PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20230925 Granted publication date: 20151021 Pledgee: Dongyang sub branch of Bank of China Ltd. Pledgor: ZHEJIANG APELOA TOSPO PHARMACEUTICAL Co.,Ltd. Registration number: Y2022330002320 |
|
PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Mutants of penicillin G acylase and their application in the synthesis of cephalosporin antibiotics Effective date of registration: 20231007 Granted publication date: 20151021 Pledgee: Dongyang sub branch of Bank of China Ltd. Pledgor: ZHEJIANG APELOA TOSPO PHARMACEUTICAL Co.,Ltd. Registration number: Y2023980059566 |