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

• Published in: 中国化学工程学报：英文版 no. 2; pp. 185 - 191
• Main Author: LUO Man GUAN Yixin YAO Shanjing
• Format: Journal Article
• Language: English
• Published: 2013
• Subjects: DsbA蛋白; 优化; 发酵液; 离子交换色谱法; 纯化; 肽质量指纹图谱; 设计方法论; 重组大肠杆菌
• ISSN: 1004-9541

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.