Cao C, Shi C, Li P, Ma Q
Institute of Biotechnology, Academy of Military Medical Sciences, Beijing, China.
Chin J Biotechnol. 1995;11(2):79-86.
A secretion expression plasmid vector pMC05S was constructed taking advantage of the promoter, signal peptide, and transcriptional terminator of cholera toxin B subunit gene and beta-galactosidase was overexpressed in E. coli and most of the expressed enzyme was secreted into periplasma when the lacZ gene was inserted downstream of the signal peptide sequence of pMC05S. The yield of beta-galactosidase by engineered E. coli reached 30 mg/L and most of the beta-galactosidase retained the activity of the enzyme. The appropriate host strain and medium were also investigated. This system provided a new approach for the expression of proteins that easily form inclusion bodies.
利用霍乱毒素B亚基基因的启动子、信号肽和转录终止子构建了分泌表达质粒载体pMC05S,当将lacZ基因插入pMC05S信号肽序列下游时,β-半乳糖苷酶在大肠杆菌中过表达,且大部分表达的酶分泌到周质中。工程化大肠杆菌产生的β-半乳糖苷酶产量达到30 mg/L,且大部分β-半乳糖苷酶保留了酶活性。还研究了合适的宿主菌株和培养基。该系统为表达易形成包涵体的蛋白质提供了一种新方法。