重组大肠杆菌表达豹蛙酶发酵培养基和发酵条件的优化  

Optimization of Fermentation Medium and Conditions for Recombinant E. coli Expressing Onconase

作　　者：夏祯 赵兴聪 王学东[1] XIA Zhen;ZHAO Xingcong;WANG Xuedong(State Key Laboratory of Bioreactor Engineering,East China University of Science and Technology,Shanghai 200237,China)

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

出　　处：《华东理工大学学报（自然科学版）》2021年第5期592-597,共6页Journal of East China University of Science and Technology

基　　金：国家科技重大专项“生物技术药物中试放大及分离纯化技术平台”(2012ZX09304009)。

摘　　要：采用Plackett-Burman(PB)实验设计,考察了培养基各组分对重组大肠杆菌表达豹蛙酶的影响,优化了重组大肠杆菌表达豹蛙酶的发酵培养基,并进一步优化了诱导表达条件。在此基础上,采用指数补料和pH-Stat方式在7 L发酵罐中进一步验证优化结果。结果表明:摇瓶水平豹蛙酶表达量由优化前的9%提高到优化后的36%,OD_(600)值由优化前的4.80提升到优化后的5.47。优化后7 L发酵罐中重组菌豹蛙酶表达水平可达到55%,OD_(600)可达到35。Ranpirnase,a multi-functional protein drug,was first isolated from the oocytes and early embryos of the Northern Leopard Frog,and belonged to the ribonuclease A(RNase A)superfamily.Ranpirnase not only inhibits protein biosynthetic pathway,but also independently induces tumor cell apoptosis.It also has antiviral activity.In this study,in order to improve the expression level of ranpirnase in E.coli,a Plackett-Burman(PB)experimental design was applied to examine the effect of various factors on ranpirnase expression in recombinant E.coli.The recombinant E.coli fermentation medium and inducing conditions were also optimized.On the basis of these results,the ranpirnase expression in recombinant E.coli was carried out in a 7 L fermentor with the combined exponential and pH-Stat feeding strategy.With the optimized fermentation medium,the protein expression at the flask level increased from 9%to 36%,and the OD_(600) value increased from 4.80 to 5.47.The expression level of ranpirnase at the 7 L fermentor level reached 55%,and the OD_(600) value was increased to 35 after the optimization.The use of the optimized medium for high-density fermentation of ranpirnase leads to a lowered production cost,facilitating its commercialization.

关 键 词：豹蛙酶 大肠杆菌 蛋白表达 发酵优化 PLACKETT-BURMAN 

分 类 号：Q815[生物学—生物工程]