大肠杆菌副产物合成途径删除提高外源蛋白表达水平  

Elimination of byproduct synthesis in Escherichia coli to improve recombinant protein production

作　　者：夏颖[1] 沈微[1] 周丽[1] 陈献忠[1] 樊游[1] 王正祥[1] 

机构地区：[1]江南大学生物工程学院,江苏无锡214122

出　　处：《工业微生物》2014年第3期35-40,共6页Industrial Microbiology

基　　金：国家自然科学基金项目(编号:21006039

摘　　要：大肠杆菌是应用最广泛的外源基因表达宿主。为探索阻断副产物产生途径对提高大肠杆菌表达外源蛋白的能力,本实验以野生型大肠杆菌菌株为基础,删除其乳酸脱氢酶基因(ldhA),磷酸烯醇式丙酮酸合成酶基因(pps)和丙酮酸甲酸裂解酶基因(pflB)。在此基础上,以甘露聚糖酶基因man为报告基因,考察阻断以上代谢途径对大肠杆菌产酶能力的影响。结果显示,以上述三个基因叠加删除的三重突变株为宿主时,重组茵产酶水平最高,比酶活达到158.3 U/mg,相比野生出发菌株提高82.3%。Escherichia coli was one of the most common hosts for recombinant protein production. In this study, multiplegene deletions with mutation in D-lactate dehydrogenase （ldhA）, phosphoenolpyruvate synthase （pps）, pyruvate formate lyase （pflB）, were tested for their effects on recombinant protein productivity in E. coli, and a mannanase gene （man） was used as the reporter gene. As a result, the CICIM B0013 -003 was obtained from deletions of all three genes in E. coli CICIM B0013 and its activity was as high as 158.3 U/mg, which was increased by 82.3 % if compared with that of the wild strain.

关 键 词：大肠杆菌 基因删除 蛋白表达 

分 类 号：Q78[生物学—分子生物学]