Optimization of DsbA Purification from Recombinant Escherichia coli Broth Using Box-Behnken Design Methodolog

作     者：LUO Man GUAN Yixin YAO Shanjing 

作者机构：Department of Chemical and Biological Engineering Zhejiang University Hangzhou 310027 China 

出 版 物：《Chinese Journal of Chemical Engineering》 (中国化学工程学报（英文版）)

年 卷 期：2013年第21卷第2期

页      面：185-191页

核心收录：

学科分类：0710[理学-生物学] 1007[医学-药学(可授医学、理学学位)] 0830[工学-环境科学与工程（可授工学、理学、农学学位）] 071010[理学-生物化学与分子生物学] 100705[医学-微生物与生化药学] 081704[工学-应用化学] 07[理学] 0817[工学-化学工程与技术] 08[工学] 0703[理学-化学] 10[医学] 

基　　金：Supported by the National Natural Science Foundation of China (21036005) 

主　　题：disulfide bond formation protein A protein purification Box-Behnken experiment design response surface methodology multi-object programming 

摘      要：Disulfide bond formation protein A （DsbA） is one of the important helper proteins for folding in protein synthesis in vivo. In this study, purification of recombinant DsbA was investigated by examining four important factors with Box-Behnken design method, a statistic-based design of experiments. The optimal operation conditions were obtained by adopting the effectiveness coefficient method on the multi-objective problem, which takes the protein recovery, purification efficiency and throughput of ion-exchange chromatography into account. After the optimization, protein recovery of 96.8% and purity higher than 95% DsbA was achieved, and the productivity was （377.9±1.7） mg soluble DsbA per liter broth. The purified protein was identified by peptide mass fingerprinting matching the record of gil2624856, a mutant of DsbA. The DsbA was preliminarily applied to the refolding of denatured lysozyme in vitro.