多二硫键蛋白在大肠杆菌中的表达研究  

Research on the Expression of Proteins with Multiple Disulfide Bonds in E.coli

作　　者：王彦婷[1] 黄智刚[1] 郭西英 汪盛[1] 

机构地区：[1]华中科技大学生命科学与技术学院分子生物物理教育部重点实验室,中国湖北武汉430074

出　　处：《生命科学研究》2015年第3期258-264,共7页Life Science Research

基　　金：国家重点基础研究发展计划项目(2010CB529804);中央高校基本科研业务费专项资金资助项目(2014TS087)

摘　　要：真核生物的许多蛋白富含二硫键。由于大肠杆菌细胞质中含有二硫键还原酶,利用大肠杆菌生产具有生物活性的重组二硫键蛋白充满挑战。近年来,SHuffle菌株、超氧化性细胞的相继问世,利用分子伴侣或引入二硫键从头形成体系,使多二硫键蛋白在大肠杆菌中的表达成为可能。简要概述了野生型大肠杆菌细胞周质和经遗传改造的工程菌细胞质中二硫键的形成机制,并着重介绍了近年来重组二硫键蛋白表达策略的最新研究进展,对利用大肠杆菌反应器生产富含二硫键蛋白起指导意义。A large number of proteins in eukaryotes are rich in disulfide bonds. Since the cells have enzymes dedicated to reducing disulfide bonds in their cytoplasm, production of soluble and active disulfidebonded proteins in E. coli is a challenge. Recently, SHuffle strain and superoxidizing cells have been successfully developed; molecular chaperones have been applied in prokaryotic expression system; and the de novo disulfide bond formation system has been introduced in the cytoplasm of E. coli. These developments made it possible to express proteins with multiple disulfide bonds in E. coli. The mechanisms responsible for disulfide bond formation in E. coli, both in its native periplasm of wild-type strains and in the genetically modified cytoplasm of engineered strains are summarized briefly. Advances of expressing recombinant disulfide bonded proteins in E. coli are highlighted. It may be important for producing recombinant proteins with multiple disulfide bonds in E. coli as a microbial cell factory.

关 键 词：二硫键蛋白 大肠杆菌 Dsb蛋白 分子伴侣 巯基氧化酶 超氧化性细胞 

分 类 号：Q786[生物学—分子生物学]