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Optimization of recombinant cytochrome p450 2C9 protein production in Escherichia coli DH5 alpha by statistically-based experimental design
Hao, Da C.; Zhu, Pan H.; Yang, Sheng L.; Yang, Ling
关键词Cytochrome P4502c9 Heterologous Expression Optimization Taguchi Orthogonal Design Desirability Function Central Composite Design
刊名WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
2006-11-01
DOI10.1007/s11274-006-9158-9
22期:11页:1169-1176
收录类别SCI
文章类型Article
WOS标题词Science & Technology ; Life Sciences & Biomedicine
类目[WOS]Biotechnology & Applied Microbiology
研究领域[WOS]Biotechnology & Applied Microbiology
关键词[WOS]RESPONSE-SURFACE METHODOLOGY ; FERMENTATION CONDITIONS ; DESIRABILITY FUNCTION ; MEDIUM CONSTITUENTS ; ETHANOL ; RESOLUTION ; EXPRESSION ; CYP2C9
英文摘要Cytochrome P450 (CYP) 2C9 is of major importance in drug metabolism. However, the low yield of recombinant CYP2C9 protein in E. coli strains prevents its extensive use in the study of in vitro drug metabolism. In the present study, Taguchi design and desirability function were first used to investigate the effect of medium components (glycerol, delta-ALA, IPTG, ampicillin, chloramphenicol, inoculum density, peptone, thiamine, trace elements, NH4Cl, and MgSO4) on recombinant human CYP2C9 production by E. coli DH5 alpha. An L-12 (2(11)) orthogonal array was used to design the experiments to screen out the most influential factors. The CYP concentration and the specific content of CYP were considered as two product quality variables. A desirability function was applied to combine these two qualities as a single objective function. Optimization via central composite design (CCD) was then undertaken to yield the best performance. The confirmation experiments indicated that the expression performance under the optimized conditions was better than those obtained under other conditions. A compromise between conflicting goals, such as achievement of good yield of recombinant CYP2C9 and facility of the following purification, was found by means of the desirability function D. This is the first report that combined Taguchi design and CCD, and performed experiments in a multiresponse framework to optimize the production of human CYP in a recombinant E. coli strain.
语种英语
WOS记录号WOS:000241905000009
引用统计
被引频次:12[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/140272
专题中国科学院大连化学物理研究所
作者单位1.Chinese Acad Sci, Dalian Inst Chem Phys, Lab Pharmaceut Resource Discovery, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Shanghai Res Ctr Biotechnol, Shanghai, Peoples R China
3.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China
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GB/T 7714
Hao, Da C.,Zhu, Pan H.,Yang, Sheng L.,et al. Optimization of recombinant cytochrome p450 2C9 protein production in Escherichia coli DH5 alpha by statistically-based experimental design[J]. WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY,2006,22(11):1169-1176.
APA Hao, Da C.,Zhu, Pan H.,Yang, Sheng L.,&Yang, Ling.(2006).Optimization of recombinant cytochrome p450 2C9 protein production in Escherichia coli DH5 alpha by statistically-based experimental design.WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY,22(11),1169-1176.
MLA Hao, Da C.,et al."Optimization of recombinant cytochrome p450 2C9 protein production in Escherichia coli DH5 alpha by statistically-based experimental design".WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY 22.11(2006):1169-1176.
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