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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
KeywordCytochrome P4502c9 Heterologous Expression Optimization Taguchi Orthogonal Design Desirability Function Central Composite Design
Source PublicationWORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
2006-11-01
DOI10.1007/s11274-006-9158-9
Volume22Issue:11Pages:1169-1176
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
WOS SubjectBiotechnology & Applied Microbiology
WOS Research AreaBiotechnology & Applied Microbiology
WOS KeywordRESPONSE-SURFACE METHODOLOGY ; FERMENTATION CONDITIONS ; DESIRABILITY FUNCTION ; MEDIUM CONSTITUENTS ; ETHANOL ; RESOLUTION ; EXPRESSION ; CYP2C9
AbstractCytochrome 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.
Language英语
WOS IDWOS:000241905000009
Citation statistics
Cited Times:11[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/140272
Collection中国科学院大连化学物理研究所
Affiliation1.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
Recommended Citation
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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