CN102978146B - 一株重组大肠杆菌及用其制备溶血性磷脂酶c的方法 - Google Patents
一株重组大肠杆菌及用其制备溶血性磷脂酶c的方法 Download PDFInfo
- Publication number
- CN102978146B CN102978146B CN201210480632.3A CN201210480632A CN102978146B CN 102978146 B CN102978146 B CN 102978146B CN 201210480632 A CN201210480632 A CN 201210480632A CN 102978146 B CN102978146 B CN 102978146B
- Authority
- CN
- China
- Prior art keywords
- liquid
- culture
- phospholipase
- shaking
- enzyme
- 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.)
- Expired - Fee Related
Links
- 241000588724 Escherichia coli Species 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 26
- 102000014384 Type C Phospholipases Human genes 0.000 title abstract description 34
- 108010079194 Type C Phospholipases Proteins 0.000 title abstract description 34
- 230000002949 hemolytic effect Effects 0.000 title abstract description 9
- 239000007788 liquid Substances 0.000 claims abstract description 83
- 102000004190 Enzymes Human genes 0.000 claims abstract description 56
- 108090000790 Enzymes Proteins 0.000 claims abstract description 56
- 238000011218 seed culture Methods 0.000 claims abstract description 31
- 238000000855 fermentation Methods 0.000 claims abstract description 27
- 230000004151 fermentation Effects 0.000 claims abstract description 27
- 238000000605 extraction Methods 0.000 claims abstract description 11
- 239000000203 mixture Substances 0.000 claims description 25
- 230000006798 recombination Effects 0.000 claims description 25
- 238000005215 recombination Methods 0.000 claims description 25
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 22
- 101710101160 Hemolytic phospholipase C Proteins 0.000 claims description 21
- 239000002609 medium Substances 0.000 claims description 21
- 239000001888 Peptone Substances 0.000 claims description 20
- 108010080698 Peptones Proteins 0.000 claims description 20
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 20
- 235000019319 peptone Nutrition 0.000 claims description 20
- 239000000843 powder Substances 0.000 claims description 20
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 18
- 230000001939 inductive effect Effects 0.000 claims description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 14
- 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 claims description 13
- 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 11
- 239000001963 growth medium Substances 0.000 claims description 11
- 239000008101 lactose Substances 0.000 claims description 11
- 239000011780 sodium chloride Substances 0.000 claims description 11
- 239000002054 inoculum Substances 0.000 claims description 10
- 239000006228 supernatant Substances 0.000 claims description 10
- 235000011187 glycerol Nutrition 0.000 claims description 9
- 238000002525 ultrasonication Methods 0.000 claims description 9
- 239000004471 Glycine Substances 0.000 claims description 7
- 238000001556 precipitation Methods 0.000 claims description 5
- 210000001082 somatic cell Anatomy 0.000 claims description 4
- 241000589517 Pseudomonas aeruginosa Species 0.000 abstract description 17
- 230000000694 effects Effects 0.000 abstract description 9
- 230000008569 process Effects 0.000 abstract description 4
- 108090000623 proteins and genes Proteins 0.000 abstract description 4
- 238000007670 refining Methods 0.000 abstract description 4
- 150000003904 phospholipids Chemical class 0.000 abstract description 3
- 238000002360 preparation method Methods 0.000 abstract description 2
- 241001198387 Escherichia coli BL21(DE3) Species 0.000 abstract 1
- 230000003321 amplification Effects 0.000 abstract 1
- 230000004048 modification Effects 0.000 abstract 1
- 238000012986 modification Methods 0.000 abstract 1
- 238000003199 nucleic acid amplification method Methods 0.000 abstract 1
- 230000001580 bacterial effect Effects 0.000 description 12
- 238000004321 preservation Methods 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 8
- 238000011160 research Methods 0.000 description 7
- 239000000047 product Substances 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 5
- 229910021538 borax Inorganic materials 0.000 description 4
- 210000002969 egg yolk Anatomy 0.000 description 4
- 239000013612 plasmid Substances 0.000 description 4
- 239000004328 sodium tetraborate Substances 0.000 description 4
- 235000010339 sodium tetraborate Nutrition 0.000 description 4
- 239000013598 vector Substances 0.000 description 4
- 241000193755 Bacillus cereus Species 0.000 description 3
- 102000012410 DNA Ligases Human genes 0.000 description 3
- 108010061982 DNA Ligases Proteins 0.000 description 3
- 239000002585 base Substances 0.000 description 3
- 239000006161 blood agar Substances 0.000 description 3
- 230000002255 enzymatic effect Effects 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 108091008146 restriction endonucleases Proteins 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- 102000002322 Egg Proteins Human genes 0.000 description 2
- 108010000912 Egg Proteins Proteins 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 102000003960 Ligases Human genes 0.000 description 2
- 108090000364 Ligases Proteins 0.000 description 2
- 102000006486 Phosphoinositide Phospholipase C Human genes 0.000 description 2
- 108010044302 Phosphoinositide phospholipase C Proteins 0.000 description 2
- 102000003923 Protein Kinase C Human genes 0.000 description 2
- 108090000315 Protein Kinase C Proteins 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 239000013599 cloning vector Substances 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000000354 decomposition reaction Methods 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 235000013345 egg yolk Nutrition 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 231100000219 mutagenic Toxicity 0.000 description 2
- 230000003505 mutagenic effect Effects 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 2
- MEIRRNXMZYDVDW-MQQKCMAXSA-N (2E,4E)-2,4-hexadien-1-ol Chemical compound C\C=C\C=C\CO MEIRRNXMZYDVDW-MQQKCMAXSA-N 0.000 description 1
- 102000005701 Calcium-Binding Proteins Human genes 0.000 description 1
- 108010045403 Calcium-Binding Proteins Proteins 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 108010042407 Endonucleases Proteins 0.000 description 1
- 102000004533 Endonucleases Human genes 0.000 description 1
- 241000672609 Escherichia coli BL21 Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 206010018910 Haemolysis Diseases 0.000 description 1
- 108010006464 Hemolysin Proteins Proteins 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 208000034530 PLAA-associated neurodevelopmental disease Diseases 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 101000730481 Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1) Hemolytic phospholipase C Proteins 0.000 description 1
- 241001240958 Pseudomonas aeruginosa PAO1 Species 0.000 description 1
- 241000220324 Pyrus Species 0.000 description 1
- 206010047249 Venous thrombosis Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- JUGOREOARAHOCO-UHFFFAOYSA-M acetylcholine chloride Chemical compound [Cl-].CC(=O)OCC[N+](C)(C)C JUGOREOARAHOCO-UHFFFAOYSA-M 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 230000000702 anti-platelet effect Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127218 antiplatelet drug Drugs 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 235000008429 bread Nutrition 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000003518 caustics Substances 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000005754 cellular signaling Effects 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 239000006781 columbia blood agar Substances 0.000 description 1
- 230000008602 contraction Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 230000008588 hemolysis Effects 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 230000007918 pathogenicity Effects 0.000 description 1
- 235000021017 pears Nutrition 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 125000001095 phosphatidyl group Chemical group 0.000 description 1
- 101150066982 plcH gene Proteins 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
Description
Claims (1)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201210480632.3A CN102978146B (zh) | 2012-11-23 | 2012-11-23 | 一株重组大肠杆菌及用其制备溶血性磷脂酶c的方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201210480632.3A CN102978146B (zh) | 2012-11-23 | 2012-11-23 | 一株重组大肠杆菌及用其制备溶血性磷脂酶c的方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN102978146A CN102978146A (zh) | 2013-03-20 |
CN102978146B true CN102978146B (zh) | 2014-10-08 |
Family
ID=47852488
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201210480632.3A Expired - Fee Related CN102978146B (zh) | 2012-11-23 | 2012-11-23 | 一株重组大肠杆菌及用其制备溶血性磷脂酶c的方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN102978146B (zh) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103525744A (zh) * | 2013-07-07 | 2014-01-22 | 江南大学 | 一种重组大肠杆菌及制备磷脂酶c的方法和应用 |
CN103881955A (zh) * | 2013-07-07 | 2014-06-25 | 江南大学 | 一种重组大肠杆菌及制备磷脂酶c的方法和脱胶工业中的应用 |
CN103525745A (zh) * | 2013-07-07 | 2014-01-22 | 江南大学 | 一种重组大肠杆菌及制备磷脂酶c的方法和大豆毛油脱胶方法 |
CN104726356B (zh) * | 2013-12-20 | 2019-08-13 | 丰益(上海)生物技术研发中心有限公司 | 一株产生磷脂酶的枯草芽孢杆菌诱变菌株 |
-
2012
- 2012-11-23 CN CN201210480632.3A patent/CN102978146B/zh not_active Expired - Fee Related
Non-Patent Citations (9)
Title |
---|
Characterization of the phospholipase C gene of Pseudomonas aeruginosa cloned in Escherichia coli;Stephen Lory et al;《Gene》;19830430;第22卷(第1期);95-101 * |
Cloning, and expression in Escherichia coli K-12, of the chromosomal hemolysin (Phospholipase C) determinant of Pseudomonas aeruginosa;K Coleman et al;《Journal of bacteriology》;19830228;第153卷(第2期);909-915 * |
K Coleman et al.Cloning, and expression in Escherichia coli K-12, of the chromosomal hemolysin (Phospholipase C) determinant of Pseudomonas aeruginosa.《Journal of bacteriology》.1983,第153卷(第2期),909-915. |
Stephen Lory et al.Characterization of the phospholipase C gene of Pseudomonas aeruginosa cloned in Escherichia coli.《Gene》.1983,第22卷(第1期),95-101. |
Stiver C K et al.GenBank:AE004091.2.《GenBank:AE004091.2》.2010,全文. * |
嗜热子囊菌角质酶—纤维素结合域融合蛋白的构建、 及其在纺织工业中的应用;张瑶;《中国博士学位论文全文数据库工程科技I辑》;20120615;B024-15 * |
复合与合成培养基对大肠杆菌胞外生产α-环糊精葡萄糖基转移酶的影响;程婧 等;《中国生物工程杂志》;20101231;第30卷(第9期);36-42 * |
张瑶.嗜热子囊菌角质酶—纤维素结合域融合蛋白的构建、 及其在纺织工业中的应用.《中国博士学位论文全文数据库工程科技I辑》.2012,B024-15. |
程婧 等.复合与合成培养基对大肠杆菌胞外生产α-环糊精葡萄糖基转移酶的影响.《中国生物工程杂志》.2010,第30卷(第9期),36-42. |
Also Published As
Publication number | Publication date |
---|---|
CN102978146A (zh) | 2013-03-20 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN102978146B (zh) | 一株重组大肠杆菌及用其制备溶血性磷脂酶c的方法 | |
CN111206025B (zh) | 一种比活提高的溶菌酶突变体 | |
CN103695341B (zh) | 一种由海洋细菌分泌的褐藻胶裂解酶及其制备方法 | |
CN104130951A (zh) | 一种重组毕赤酵母工程菌和代谢的重组木聚糖酶及其制备 | |
CN105385609A (zh) | 一株高产葡萄糖氧化酶的黑曲霉及其应用 | |
CN102584977B (zh) | 一种猪防御素pBD2多肽及其在抑制猪病原菌中应用 | |
CN102994406B (zh) | 一株产葡萄糖氧化酶的基因工程菌及其构建与应用 | |
CN110117601A (zh) | 灰树花葡聚糖合成酶、其编码基因及应用 | |
CN108795837A (zh) | 一种高效表达磷脂酶d的枯草芽孢杆菌工程菌 | |
CN103509748B (zh) | 一种产磷脂酶c的重组大肠杆菌及制备磷脂酶c的方法和应用 | |
CN104480083A (zh) | 一种脂肪酶、工程菌及其制备方法 | |
CN103436575B (zh) | 一种毕赤酵母重组菌高密度发酵培养基及高密度发酵方法 | |
CN103525782A (zh) | 一种突变的有机磷农药降解酶基因及其应用 | |
CN102220276B (zh) | 一种产胆盐水解酶的基因工程菌及其构建方法和应用 | |
CN103525745A (zh) | 一种重组大肠杆菌及制备磷脂酶c的方法和大豆毛油脱胶方法 | |
CN104328095A (zh) | 一种最适pH在酸性范围的磷脂酶A2及其应用 | |
CN107022591A (zh) | 一种提高胰岛素及其类似物前体表达的毕赤酵母发酵方法 | |
CN115806886B (zh) | 粗糙脉孢菌木酮糖激酶基因敲除突变株及其应用 | |
CN103525744A (zh) | 一种重组大肠杆菌及制备磷脂酶c的方法和应用 | |
CN104560910B (zh) | 提高毕赤酵母发酵产磷脂酶c产量的培养基及方法 | |
CN100432213C (zh) | 一种编码高温植酸酶基因的重组菌和该基因及其合成、克隆和表达 | |
CN103146726A (zh) | 一种黑曲霉α-葡萄糖苷酶基因及其高效表达方法 | |
CN103074290B (zh) | 一株重组大肠杆菌及用该菌株制备磷脂酶a1的方法 | |
CN102807958A (zh) | 一种可分泌纤维素酶的菌株及其纤维素酶提取方法与应用 | |
CN102242092A (zh) | 一种家蚕肠道益生菌的分离鉴定及其酶基因的克隆与表达 |
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 | ||
C41 | Transfer of patent application or patent right or utility model | ||
CB03 | Change of inventor or designer information |
Inventor after: Zhang Liang Inventor after: Shi Guiyang Inventor after: Zhao Jinxing Inventor after: Ding Zhongyang Inventor after: Gu Zhenghua Inventor after: Chen Guoan Inventor before: Zhang Liang Inventor before: Shi Guiyang Inventor before: Zhao Jinxing Inventor before: Ding Zhongyang Inventor before: Gu Zhenghua |
|
COR | Change of bibliographic data | ||
TR01 | Transfer of patent right |
Effective date of registration: 20151223 Address after: No. 1800 road 214122 Jiangsu Lihu Binhu District City of Wuxi Province Patentee after: Jiangnan University Patentee after: Southern Yangze University Biotech Address before: No. 1800 road 214122 Jiangsu Lihu Binhu District City of Wuxi Province Patentee before: Jiangnan University |
|
CF01 | Termination of patent right due to non-payment of annual fee | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20141008 |