多二硫键蛋白在超氧化大肠杆菌细胞质中的高效折叠

Highly efficient folding of multi-disulfide proteins in superoxidizing Escherichia coli cytoplasm.

作者信息

Zhang Wenyao, Zheng Wenyun, Mao Miaowei, Yang Yi

机构信息

Synthetic Biology and Biotechnology Laboratory, State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, School of Pharmacy, East China University of Science and Technology, 130 Mei Long Road, Shanghai, 200237, China.

出版信息

Biotechnol Bioeng. 2014 Dec;111(12):2520-7. doi: 10.1002/bit.25309. Epub 2014 Aug 5.

DOI:10.1002/bit.25309

PMID:24917025

Abstract

In this study, we monitored the thiol-disulfide redox potential of different Escherichia coli strains using redox-sensitive variants of green fluorescent protein. The cells with extreme oxidizing cytoplasm were generated by introducing a highly efficient disulfide relay system. The developed cells have exceptionally efficient de novo disulfide bond formation and significantly improve the oxidative folding of the client multi-disulfide proteins. Superoxidizing E. coli strain provides an effective method for the high-level production of recombinant disulfide-containing proteins.

摘要

在本研究中，我们使用绿色荧光蛋白的氧化还原敏感变体监测了不同大肠杆菌菌株的硫醇 - 二硫键氧化还原电位。通过引入高效的二硫键传递系统产生了具有极端氧化性细胞质的细胞。所构建的细胞具有异常高效的从头二硫键形成能力，并显著改善了客户多二硫键蛋白的氧化折叠。超氧化大肠杆菌菌株为高水平生产含重组二硫键的蛋白质提供了一种有效方法。