CN113699086B - Recombinant bacterium for producing sulforecombinant hirudin and preparation method and application thereof - Google Patents
Recombinant bacterium for producing sulforecombinant hirudin and preparation method and application thereof Download PDFInfo
- Publication number
- CN113699086B CN113699086B CN202010444223.2A CN202010444223A CN113699086B CN 113699086 B CN113699086 B CN 113699086B CN 202010444223 A CN202010444223 A CN 202010444223A CN 113699086 B CN113699086 B CN 113699086B
- Authority
- CN
- China
- Prior art keywords
- hirudin
- gene
- nucleotide sequence
- recombinant
- vector
- 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
- 108010007267 Hirudins Proteins 0.000 title claims abstract description 86
- WQPDUTSPKFMPDP-OUMQNGNKSA-N hirudin Chemical compound C([C@@H](C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C=CC(OS(O)(=O)=O)=CC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CCCCN)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H]1NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCC(O)=O)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H]2CSSC[C@@H](C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(=O)N[C@H](C(NCC(=O)N[C@@H](CCC(N)=O)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N2)=O)CSSC1)C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]1NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=2C=CC(O)=CC=2)NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)C(C)C)[C@@H](C)O)CSSC1)C(C)C)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 WQPDUTSPKFMPDP-OUMQNGNKSA-N 0.000 title claims abstract description 75
- 102000007625 Hirudins Human genes 0.000 title claims abstract description 59
- 229940006607 hirudin Drugs 0.000 title claims abstract description 56
- 241000894006 Bacteria Species 0.000 title claims abstract description 30
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 239000002773 nucleotide Substances 0.000 claims abstract description 32
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 32
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 21
- 238000000034 method Methods 0.000 claims abstract description 20
- 238000004519 manufacturing process Methods 0.000 claims abstract description 17
- 241000588724 Escherichia coli Species 0.000 claims abstract description 9
- 238000000746 purification Methods 0.000 claims abstract description 8
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 25
- 230000001580 bacterial effect Effects 0.000 claims description 23
- 230000028327 secretion Effects 0.000 claims description 19
- 239000013598 vector Substances 0.000 claims description 19
- 239000007788 liquid Substances 0.000 claims description 17
- 239000013604 expression vector Substances 0.000 claims description 12
- 239000000243 solution Substances 0.000 claims description 12
- 229960005091 chloramphenicol Drugs 0.000 claims description 11
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 claims description 11
- 229930027917 kanamycin Natural products 0.000 claims description 11
- 229960000318 kanamycin Drugs 0.000 claims description 11
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 claims description 11
- 229930182823 kanamycin A Natural products 0.000 claims description 11
- 239000013600 plasmid vector Substances 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 108090001033 Sulfotransferases Proteins 0.000 claims description 9
- 239000002244 precipitate Substances 0.000 claims description 9
- 239000012153 distilled water Substances 0.000 claims description 8
- 108010090127 Periplasmic Proteins Proteins 0.000 claims description 7
- 238000012258 culturing Methods 0.000 claims description 7
- 238000000502 dialysis Methods 0.000 claims description 7
- 239000001963 growth medium Substances 0.000 claims description 7
- 230000003321 amplification Effects 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 6
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 6
- 239000012460 protein solution Substances 0.000 claims description 6
- 238000012360 testing method Methods 0.000 claims description 6
- JBGHTSSFSSUKLR-UHFFFAOYSA-N 4-nitrophenyl hydrogen sulfate Chemical compound OS(=O)(=O)OC1=CC=C([N+]([O-])=O)C=C1 JBGHTSSFSSUKLR-UHFFFAOYSA-N 0.000 claims description 5
- 230000014509 gene expression Effects 0.000 claims description 5
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 claims description 5
- 230000008569 process Effects 0.000 claims description 5
- 230000003248 secreting effect Effects 0.000 claims description 5
- 229930006000 Sucrose Natural products 0.000 claims description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 4
- 239000007983 Tris buffer Substances 0.000 claims description 4
- 238000010276 construction Methods 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 239000005720 sucrose Substances 0.000 claims description 4
- 239000010414 supernatant solution Substances 0.000 claims description 4
- 230000009466 transformation Effects 0.000 claims description 4
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims description 4
- 238000004108 freeze drying Methods 0.000 claims description 3
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 2
- 102000004896 Sulfotransferases Human genes 0.000 claims description 2
- 238000001556 precipitation Methods 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 16
- 238000010353 genetic engineering Methods 0.000 abstract description 2
- 108090000790 Enzymes Proteins 0.000 description 11
- 102000004190 Enzymes Human genes 0.000 description 11
- 229940088598 enzyme Drugs 0.000 description 11
- 239000000047 product Substances 0.000 description 11
- 108020004414 DNA Proteins 0.000 description 9
- 230000000052 comparative effect Effects 0.000 description 9
- 239000003242 anti bacterial agent Substances 0.000 description 8
- 229940088710 antibiotic agent Drugs 0.000 description 8
- 239000006228 supernatant Substances 0.000 description 8
- 230000010100 anticoagulation Effects 0.000 description 7
- 238000001976 enzyme digestion Methods 0.000 description 7
- 230000006698 induction Effects 0.000 description 5
- 239000000758 substrate Substances 0.000 description 5
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 4
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 4
- 108090000190 Thrombin Proteins 0.000 description 4
- 239000003146 anticoagulant agent Substances 0.000 description 4
- 229940127219 anticoagulant drug Drugs 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 210000001322 periplasm Anatomy 0.000 description 4
- 108091008146 restriction endonucleases Proteins 0.000 description 4
- 229960004072 thrombin Drugs 0.000 description 4
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 3
- 230000029087 digestion Effects 0.000 description 3
- 229960002897 heparin Drugs 0.000 description 3
- 229920000669 heparin Polymers 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 101000942941 Arabidopsis thaliana DNA ligase 6 Proteins 0.000 description 2
- 241000186394 Eubacterium Species 0.000 description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 239000012362 glacial acetic acid Substances 0.000 description 2
- 239000012474 protein marker Substances 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000006277 sulfonation reaction Methods 0.000 description 2
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 2
- 102000004411 Antithrombin III Human genes 0.000 description 1
- 108090000935 Antithrombin III Proteins 0.000 description 1
- 206010008190 Cerebrovascular accident Diseases 0.000 description 1
- 241001198387 Escherichia coli BL21(DE3) Species 0.000 description 1
- 208000032843 Hemorrhage Diseases 0.000 description 1
- 102000004032 Heparin Cofactor II Human genes 0.000 description 1
- 108090000481 Heparin Cofactor II Proteins 0.000 description 1
- 241000545744 Hirudinea Species 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 208000006011 Stroke Diseases 0.000 description 1
- 101000712605 Theromyzon tessulatum Theromin Proteins 0.000 description 1
- 229940122388 Thrombin inhibitor Drugs 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 102000004357 Transferases Human genes 0.000 description 1
- 108090000992 Transferases Proteins 0.000 description 1
- 238000003916 acid precipitation Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 229960005348 antithrombin iii Drugs 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 150000001491 aromatic compounds Chemical class 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 210000002249 digestive system Anatomy 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 208000009190 disseminated intravascular coagulation Diseases 0.000 description 1
- 230000002888 effect on disease Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000012215 gene cloning Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- OTQCKZUSUGYWBD-BRHMIFOHSA-N lepirudin Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCCN)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O)C(C)C)[C@@H](C)O)[C@@H](C)O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC(C)C)[C@@H](C)O)C1=CC=C(O)C=C1 OTQCKZUSUGYWBD-BRHMIFOHSA-N 0.000 description 1
- 229960004408 lepirudin Drugs 0.000 description 1
- 238000006011 modification reaction Methods 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 230000014508 negative regulation of coagulation Effects 0.000 description 1
- 230000004481 post-translational protein modification Effects 0.000 description 1
- 238000000751 protein extraction Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 210000003079 salivary gland Anatomy 0.000 description 1
- 239000003868 thrombin inhibitor Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/81—Protease inhibitors
- C07K14/815—Protease inhibitors from leeches, e.g. hirudin, eglin
-
- 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/70—Vectors or expression systems specially adapted for E. coli
-
- 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
- C12N9/10—Transferases (2.)
- C12N9/13—Transferases (2.) transferring sulfur containing groups (2.8)
-
- 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
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y208/00—Transferases transferring sulfur-containing groups (2.8)
- C12Y208/02—Sulfotransferases (2.8.2)
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/036—Fusion polypeptide containing a localisation/targetting motif targeting to the medium outside of the cell, e.g. type III secretion
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Gastroenterology & Hepatology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
Claims (10)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010444223.2A CN113699086B (en) | 2020-05-22 | 2020-05-22 | Recombinant bacterium for producing sulforecombinant hirudin and preparation method and application thereof |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010444223.2A CN113699086B (en) | 2020-05-22 | 2020-05-22 | Recombinant bacterium for producing sulforecombinant hirudin and preparation method and application thereof |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113699086A CN113699086A (en) | 2021-11-26 |
CN113699086B true CN113699086B (en) | 2023-03-28 |
Family
ID=78646436
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202010444223.2A Active CN113699086B (en) | 2020-05-22 | 2020-05-22 | Recombinant bacterium for producing sulforecombinant hirudin and preparation method and application thereof |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113699086B (en) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113789335B (en) * | 2021-11-16 | 2022-03-15 | 江苏省中国科学院植物研究所 | Leech tyrosine sulfotransferase gene and application thereof |
CN113789293B (en) * | 2021-11-16 | 2022-03-15 | 江苏省中国科学院植物研究所 | Escherichia coli engineering strain for high yield of natural hirudin and application thereof |
CN114736289B (en) * | 2022-03-17 | 2023-07-18 | 华南理工大学 | Chemical synthesis method of hirudin with tyrosine sulfation modification |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6514730B1 (en) * | 1991-03-21 | 2003-02-04 | Consortium für elektrochemische Industrie GmbH | Secretion of hirudin derivatives |
CN103059130A (en) * | 2012-12-22 | 2013-04-24 | 浙江工业大学 | Hirudin mutant and genetically engineered bacterium thereof |
-
2020
- 2020-05-22 CN CN202010444223.2A patent/CN113699086B/en active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6514730B1 (en) * | 1991-03-21 | 2003-02-04 | Consortium für elektrochemische Industrie GmbH | Secretion of hirudin derivatives |
CN103059130A (en) * | 2012-12-22 | 2013-04-24 | 浙江工业大学 | Hirudin mutant and genetically engineered bacterium thereof |
Non-Patent Citations (2)
Title |
---|
The effects of overexpression of cytoplasmic chaperones on secretory production of hirudin-PA in E.coli;Fin等;《Protein Exoression and Purification》;20191231;全文 * |
重组水蛭素的原核表达及分离纯化;黄孔威等;《中国畜牧兽医》;20180120(第01期);全文 * |
Also Published As
Publication number | Publication date |
---|---|
CN113699086A (en) | 2021-11-26 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN113699086B (en) | Recombinant bacterium for producing sulforecombinant hirudin and preparation method and application thereof | |
US11377674B2 (en) | Recombinant strain expressing phospholipase D and application thereof | |
WO2008100833A2 (en) | Production of recombinant collagenases colg and colh in escherichia coli | |
JP6302415B2 (en) | Method for producing human epidermal growth factor | |
US11807883B2 (en) | Polypeptide tag, highly soluble recombinant nitrilase and application thereof in synthesis of pharmaceutical chemicals | |
CN112608933B (en) | High-purity preparation method of recombinant blue copper peptide precursor-oligopeptide | |
CN110283797B (en) | Tyrosinase, gene, engineering bacterium and preparation method thereof | |
KR20090039239A (en) | Method for over-expressing human epidermal growth factor as bioactive form in escherichia. coli | |
CN115141763B (en) | Yeast engineering strain of efficient exocrine protein, construction method and application thereof | |
RU2447151C1 (en) | ALKALINE PHOSPHATASE CmAP SYNTHESIS-DETERMINING 40Ph PLASMID, E. coli rosetta(DE3)/40Ph STRAIN - PRODUCER OF CHIMERIC PROTEIN, CONTAINING AMINO ACID SEQUENCE OF RECOMBINANT ALKALINE PHOSPHATASE CmAP, AND PRODUCTION METHOD THEREOF | |
CN113265345B (en) | Natto kinase eukaryotic high-efficiency expression dual-promoter system recombinant genetic engineering bacterium, construction method and application thereof | |
CN113025599B (en) | Recombinant clostridium histolyticum type I collagenase as well as preparation method and application thereof | |
WO2023019832A1 (en) | Expression and purification method for main protease of coronavirus | |
CN114058606A (en) | Application of bacillus licheniformis with deleted xpt gene in production of heterologous protein | |
RU2453604C1 (en) | Hybrid protein (versions), bacterial strain escherichia coli - hybrid protein producer (versions) and method for producing methionine-free human interferon alpha-2 | |
RU2441072C1 (en) | FUSION PROTEIN, ESCHERICHIA COLI STRAIN BEING FUSION PROTEIN PRODUCER AND METHOD FOR PRODUCING METHIONINE-FREE HUMAN INTERFERON ALPHA-2b OF SUCH FUSION PROTEIN | |
KR101261852B1 (en) | Agarase expression vector, transgenic organism transformed with the same vector and method for the production of agarase using the same transgenic organism | |
JPH06311884A (en) | Plasmid and escherichia coli transformed with the same | |
US20070141698A1 (en) | Microbial protein expression system utilizing plant viral coat protein | |
CN114702571B (en) | Fibronectin capable of promoting stem cell colonization and preparation method thereof | |
CN116064628B (en) | Construction method of escherichia coli surface display system | |
KR930003913B1 (en) | Method for preparation of polypeptide having thrombin inhibiting activity | |
CN117229988B (en) | Engineering strain free of antibiotics and capable of efficiently and stably expressing and construction method thereof | |
CN116715746A (en) | Method for secretory expression of human epidermal growth factor in pichia pastoris | |
JPH08103278A (en) | Production of active human alt |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant | ||
TR01 | Transfer of patent right | ||
TR01 | Transfer of patent right |
Effective date of registration: 20240221 Address after: 266101 Shandong Province, Qingdao city Laoshan District Songling Road No. 189 Patentee after: Shandong Energy Research Institute Country or region after: China Address before: 266101 Shandong Province, Qingdao city Laoshan District Songling Road No. 189 Patentee before: QINGDAO INSTITUTE OF BIOENERGY AND BIOPROCESS TECHNOLOGY, CHINESE ACADEMY OF SCIENCES Country or region before: China |
|
TR01 | Transfer of patent right |
Effective date of registration: 20240523 Address after: Room 312, Building 6, No. 61 Guangsheng Road, High tech Zone, Qingdao City, Shandong Province, 266000 Patentee after: Qingdao High Energy Hecheng Biotechnology Co.,Ltd. Country or region after: China Address before: 266101 Shandong Province, Qingdao city Laoshan District Songling Road No. 189 Patentee before: Shandong Energy Research Institute Country or region before: China |