CN105062997A - 一种酶活提高的l-天冬酰胺酶突变体及其构建方法 - Google Patents
一种酶活提高的l-天冬酰胺酶突变体及其构建方法 Download PDFInfo
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- CN105062997A CN105062997A CN201510526470.6A CN201510526470A CN105062997A CN 105062997 A CN105062997 A CN 105062997A CN 201510526470 A CN201510526470 A CN 201510526470A CN 105062997 A CN105062997 A CN 105062997A
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- asparaginase
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- 108010024976 Asparaginase Proteins 0.000 title claims abstract description 30
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- 102000004190 Enzymes Human genes 0.000 title abstract description 32
- 229940088598 enzyme Drugs 0.000 title abstract description 32
- 230000000694 effects Effects 0.000 title abstract description 19
- 238000010276 construction Methods 0.000 title abstract description 5
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- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 10
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims abstract description 10
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 claims abstract description 6
- 150000001413 amino acids Chemical class 0.000 claims abstract description 6
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- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 claims abstract description 5
- 239000004471 Glycine Substances 0.000 claims abstract description 4
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- 241000894006 Bacteria Species 0.000 claims description 12
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- 238000000034 method Methods 0.000 claims description 5
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- 201000010099 disease Diseases 0.000 claims description 3
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- 229940079593 drug Drugs 0.000 claims description 2
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- 238000000855 fermentation Methods 0.000 abstract description 4
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- 125000000539 amino acid group Chemical group 0.000 abstract description 2
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- 241000276408 Bacillus subtilis subsp. subtilis str. 168 Species 0.000 description 8
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 8
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- 238000006555 catalytic reaction Methods 0.000 description 3
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- 239000011541 reaction mixture Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 2
- 108020004705 Codon Proteins 0.000 description 2
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- 229930027917 kanamycin Natural products 0.000 description 2
- 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 description 2
- 229960000318 kanamycin Drugs 0.000 description 2
- 229930182823 kanamycin A Natural products 0.000 description 2
- 229910000069 nitrogen hydride Inorganic materials 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 2
- 241000700199 Cavia porcellus Species 0.000 description 1
- CKLJMWTZIZZHCS-UHFFFAOYSA-N D-OH-Asp Natural products OC(=O)C(N)CC(O)=O CKLJMWTZIZZHCS-UHFFFAOYSA-N 0.000 description 1
- 102000012410 DNA Ligases Human genes 0.000 description 1
- 108010061982 DNA Ligases Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- CKLJMWTZIZZHCS-UWTATZPHSA-N L-Aspartic acid Natural products OC(=O)[C@H](N)CC(O)=O CKLJMWTZIZZHCS-UWTATZPHSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 229960003272 asparaginase Drugs 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 108010030074 endodeoxyribonuclease MluI Proteins 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000001952 enzyme assay Methods 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 238000002741 site-directed mutagenesis Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 239000013598 vector Substances 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/10—General methods of cooking foods, e.g. by roasting or frying
-
- 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)
- C12N9/82—Asparaginase (3.5.1.1)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/43—Enzymes; Proenzymes; Derivatives thereof
- A61K38/46—Hydrolases (3)
- A61K38/50—Hydrolases (3) acting on carbon-nitrogen bonds, other than peptide bonds (3.5), e.g. asparaginase
-
- 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
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/52—Genes encoding for enzymes or proenzymes
-
- 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)
- C12Y305/01001—Asparaginase (3.5.1.1)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Epidemiology (AREA)
- Public Health (AREA)
- Plant Pathology (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Pharmacology & Pharmacy (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Animal Behavior & Ethology (AREA)
- Physics & Mathematics (AREA)
- Veterinary Medicine (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
Description
Claims (8)
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201510526470.6A CN105062997A (zh) | 2015-08-25 | 2015-08-25 | 一种酶活提高的l-天冬酰胺酶突变体及其构建方法 |
PCT/CN2015/094554 WO2017031839A1 (zh) | 2015-08-25 | 2015-11-13 | 一种酶活提高的l-天冬酰胺酶突变体及其构建方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
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CN201510526470.6A CN105062997A (zh) | 2015-08-25 | 2015-08-25 | 一种酶活提高的l-天冬酰胺酶突变体及其构建方法 |
Publications (1)
Publication Number | Publication Date |
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CN105062997A true CN105062997A (zh) | 2015-11-18 |
Family
ID=54492515
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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CN201510526470.6A Withdrawn CN105062997A (zh) | 2015-08-25 | 2015-08-25 | 一种酶活提高的l-天冬酰胺酶突变体及其构建方法 |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN105062997A (zh) |
WO (1) | WO2017031839A1 (zh) |
Cited By (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106434612A (zh) * | 2016-10-21 | 2017-02-22 | 江南大学 | 一种天冬酰胺酶突变体及其应用 |
CN107828768A (zh) * | 2017-12-13 | 2018-03-23 | 江南大学 | 一种l‑天冬酰胺酶突变体及其构建方法 |
CN107988194A (zh) * | 2017-12-15 | 2018-05-04 | 江南大学 | 一种酶活提高的L-天冬氨酸α-脱羧酶变体及其构建方法 |
CN108070581A (zh) * | 2017-12-15 | 2018-05-25 | 江南大学 | 一种酶活提高的L-天冬氨酸β-脱羧酶突变体及其应用 |
CN108094976A (zh) * | 2017-12-15 | 2018-06-01 | 江南大学 | 一株嗜热l-天冬酰胺酶在高温油炸食品中的应用 |
CN108559734A (zh) * | 2018-01-15 | 2018-09-21 | 江南大学 | 一种催化效率提高的l-乳酸脱氢酶突变体及其应用 |
CN109266635A (zh) * | 2018-11-20 | 2019-01-25 | 江南大学 | 一种酶活提高的l-天冬酰胺酶突变体及其构建方法 |
WO2019113965A1 (zh) * | 2017-12-15 | 2019-06-20 | 江南大学 | 一种嗜热l-天冬酰胺酶突变体及其筛选和发酵方法 |
CN112941059A (zh) * | 2021-02-23 | 2021-06-11 | 江南大学 | 一种l-天冬酰胺酶突变体及其在枯草芽孢杆菌中的表达 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103060299A (zh) * | 2011-10-19 | 2013-04-24 | 北京安百胜生物科技有限公司 | 活性提高的l-天冬酰胺酶变体 |
WO2014170811A2 (en) * | 2013-04-16 | 2014-10-23 | Sonawane Avinash | Novel mutant l-asparaginases |
CN104371993A (zh) * | 2014-10-22 | 2015-02-25 | 江南大学 | 一种酶活提高的天冬酰胺酶突变体 |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102864163A (zh) * | 2012-09-20 | 2013-01-09 | 江南大学 | 一种高效表达枯草芽孢杆菌l-天冬酰胺酶的方法 |
CN103243063A (zh) * | 2013-05-24 | 2013-08-14 | 江南大学 | 枯草芽孢杆菌L-天冬酰胺酶(ansZ)在大肠杆菌中的高效表达 |
CN104611317B (zh) * | 2015-03-09 | 2018-07-06 | 江南大学 | 一种提高l-天冬酰胺酶分泌表达的方法 |
-
2015
- 2015-08-25 CN CN201510526470.6A patent/CN105062997A/zh not_active Withdrawn
- 2015-11-13 WO PCT/CN2015/094554 patent/WO2017031839A1/zh active Application Filing
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103060299A (zh) * | 2011-10-19 | 2013-04-24 | 北京安百胜生物科技有限公司 | 活性提高的l-天冬酰胺酶变体 |
WO2014170811A2 (en) * | 2013-04-16 | 2014-10-23 | Sonawane Avinash | Novel mutant l-asparaginases |
CN104371993A (zh) * | 2014-10-22 | 2015-02-25 | 江南大学 | 一种酶活提高的天冬酰胺酶突变体 |
Non-Patent Citations (2)
Title |
---|
SHUIQING LONG ET AL.: "Amino acid residues adjacent to the catalytic cavity of tetramer l-asparaginase II contribute significantly to its catalytic efficiency and thermostability", 《ENZYME AND MICROBIAL TECHNOLOGY》 * |
陈建华等: "E.coli L-天冬酰胺酶B细胞抗原表位氨基酸残基Lys196对其抗原性的影响", 《中国药科大学学报》 * |
Cited By (16)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106434612A (zh) * | 2016-10-21 | 2017-02-22 | 江南大学 | 一种天冬酰胺酶突变体及其应用 |
CN107828768A (zh) * | 2017-12-13 | 2018-03-23 | 江南大学 | 一种l‑天冬酰胺酶突变体及其构建方法 |
CN107828768B (zh) * | 2017-12-13 | 2020-10-09 | 江南大学 | 一种l-天冬酰胺酶突变体及其构建方法 |
CN108094976A (zh) * | 2017-12-15 | 2018-06-01 | 江南大学 | 一株嗜热l-天冬酰胺酶在高温油炸食品中的应用 |
CN108070581A (zh) * | 2017-12-15 | 2018-05-25 | 江南大学 | 一种酶活提高的L-天冬氨酸β-脱羧酶突变体及其应用 |
WO2019113965A1 (zh) * | 2017-12-15 | 2019-06-20 | 江南大学 | 一种嗜热l-天冬酰胺酶突变体及其筛选和发酵方法 |
CN108094976B (zh) * | 2017-12-15 | 2020-03-06 | 江南大学 | 一株嗜热l-天冬酰胺酶在高温油炸食品中的应用 |
CN107988194B (zh) * | 2017-12-15 | 2020-08-04 | 江南大学 | 一种酶活提高的L-天冬氨酸α-脱羧酶变体及其构建方法 |
CN108070581B (zh) * | 2017-12-15 | 2020-09-04 | 江南大学 | 一种酶活提高的L-天冬氨酸β-脱羧酶突变体及其应用 |
CN107988194A (zh) * | 2017-12-15 | 2018-05-04 | 江南大学 | 一种酶活提高的L-天冬氨酸α-脱羧酶变体及其构建方法 |
US11001825B2 (en) | 2017-12-15 | 2021-05-11 | Jiangnan University | Thermophilic L-asparaginase mutant and screening and fermentation methods thereof |
CN108559734A (zh) * | 2018-01-15 | 2018-09-21 | 江南大学 | 一种催化效率提高的l-乳酸脱氢酶突变体及其应用 |
CN108559734B (zh) * | 2018-01-15 | 2020-09-04 | 江南大学 | 一种催化效率提高的l-乳酸脱氢酶突变体及其应用 |
CN109266635A (zh) * | 2018-11-20 | 2019-01-25 | 江南大学 | 一种酶活提高的l-天冬酰胺酶突变体及其构建方法 |
CN109266635B (zh) * | 2018-11-20 | 2020-12-01 | 江南大学 | 一种酶活提高的l-天冬酰胺酶突变体及其构建方法 |
CN112941059A (zh) * | 2021-02-23 | 2021-06-11 | 江南大学 | 一种l-天冬酰胺酶突变体及其在枯草芽孢杆菌中的表达 |
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WO2017031839A1 (zh) | 2017-03-02 |
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