一种用于proCPB可溶性表达的新型表达系统  

A Novel Expression System for Soluble Expression of Recombinant Procarboxypeptidase B

作　　者：文良柱 方宏清 韦炎龙 窦俊 WEN Liangzhu;FANG Hongqing;WEI Yanlong;DOU Jun(Wanbang Pharmatech Co.,Ltd.,Xuzhou 221004,China)

机构地区：[1]江苏万邦医药科技有限公司,江苏徐州221004

出　　处：《生物化工》2022年第2期46-51,共6页Biological Chemical Engineering

摘　　要：目的:构建一种新型可溶性表达系统,实现包括重组猪羧肽酶原B(proCPB)在内的目标蛋白的可溶性表达。方法:通过筛选不同来源的蛋白质二硫键异构酶并与巯基氧化酶进行共表达,并添加N末端标签对这两种蛋白的表达进行优化。结果:在大肠杆菌中,ERV1-TrxsPDI系统与目的蛋白共表达可实现目的蛋白的可溶性表达,且不受到启动子和诱导剂的限制,在重组蛋白生产和新酶工程挖掘等领域具有应用前景。Objective: A novel soluble expression system was constructed to achieve the soluble expression of target proteins including recombinant porcine procarboxypeptidase B(proCPB). Method: The protein disulfide isomerase from different sources were screened and co-expressed with sulfhydryl oxidase, and the expression of the two proteins was optimized by adding nitrogen terminal tags. Results: In Escherichia coli, the system(ERV1-TrxsPDI) was co-expressed with the recombinant porcine procarboxypeptidase B, and the soluble expression of the target protein was achieved without the restriction of promoters and inducers, and the formation of inclusion bodies was avoided. It has a promising application in the fields of recombinant protein production and new enzyme engineering mining.

关 键 词：可溶性表达系统 巯基氧化酶 蛋白质二硫键异构酶 猪羧肽酶原B 大肠杆菌 

分 类 号：Q819[生物学—生物工程]