中国科学院大连化学物理研究所机构知识库
Advanced  
DICP OpenIR  > 中国科学院大连化学物理研究所  > 会议论文
学科主题: 物理化学
题名: Optimization of Recombinant Cytochrome P450 2C9 Protein Production in Escherichia Coli DH5α by Statistically-based Experimental Design
作者: Hao DC(郝大程) ;  Zhu PH(朱盼鸿) ;  Yang SL(杨胜利) ;  Yang L(杨凌)
会议名称: The 1st asia Pacific ISSX Meeting
会议日期: 2006-5-24
出版日期: 2006-05-24
会议地点: 韩国
通讯作者: 杨凌
部门归属: 十八室
摘要: Objective: To optimize Cytochrome P450 (CYP) 2C9 recombinant protein production in E. coli DH5α. Methods: Taguchi design and desirability function were first used to investigate the effect of medium components (glycerol, δ-ALA, IPTG, ampicillin, chloramphenicol, inoculum density, peptone, thiamine, trace element, NH4Cl, and MgSO4) on recombinant human CYP2C9 production by E. coli DH5α. An L12 (211) 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. Amp, IPTG, Cm, and peptone were found to be four significant factors. Optimization via the central composite design (CCD) was then undertaken to yield the best performance. Results: The location of optimum was determined to be Amp 169.5 μg/ml, Cm 1.92 μg/ml, IPTG 1.22 mM, and peptone 1.84 g/l obtained by the differentiation of the quadratic model. 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. Conclusion: This is the first report that combined Taguchi design and CCD, and performed experiments in a multiresponse framework to optimize the human CYP production in a recombinant E. coli strain. (Supported by the 973 program 2003CB716006 of the Ministry of Science & Technology of China)
语种: 中文
内容类型: 会议论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/112182
Appears in Collections:中国科学院大连化学物理研究所_会议论文

Files in This Item:

There are no files associated with this item.


Recommended Citation:
Hao DC,Zhu PH,Yang SL,et al. Optimization of Recombinant Cytochrome P450 2C9 Protein Production in Escherichia Coli DH5α by Statistically-based Experimental Design[C]. 见:The 1st asia Pacific ISSX Meeting. 韩国. 2006-5-24.
Service
 Recommend this item
 Sava as my favorate item
 Show this item's statistics
 Export Endnote File
Google Scholar
 Similar articles in Google Scholar
 [郝大程]'s Articles
 [朱盼鸿]'s Articles
 [杨胜利]'s Articles
CSDL cross search
 Similar articles in CSDL Cross Search
 [郝大程]‘s Articles
 [朱盼鸿]‘s Articles
 [杨胜利]‘s Articles
Related Copyright Policies
Null
Social Bookmarking
  Add to CiteULike  Add to Connotea  Add to Del.icio.us  Add to Digg  Add to Reddit 
所有评论 (0)
暂无评论
 
评注功能仅针对注册用户开放,请您登录
您对该条目有什么异议,请填写以下表单,管理员会尽快联系您。
内 容:
Email:  *
单位:
验证码:   刷新
您在IR的使用过程中有什么好的想法或者建议可以反馈给我们。
标 题:
 *
内 容:
Email:  *
验证码:   刷新

Items in IR are protected by copyright, with all rights reserved, unless otherwise indicated.

 

 

Valid XHTML 1.0!
Powered by CSpace