一种高效表达碱性蛋白酶的新型启动子的筛选及研究  

Screening and Study of a New Promoter with Effective Expression of Alkaline Protease

作　　者：陈坤[1] 袁飞燕 柴昊男 刘焕 王兴吉 张会图[1] 路福平[1] 

机构地区：[1]工业发酵微生物教育部重点实验室天津科技大学生物工程学院,天津300457 [2]山东隆科特酶制剂有限公司,沂水276400

出　　处：《生物技术通报》2018年第1期208-214,共7页Biotechnology Bulletin

基　　金：国家重点研发计划(2017YFB0308401)

摘　　要：碱性蛋白酶广泛的应用价值及市场供不应求的现状,旨在构建碱性蛋白酶高效表达系统。通过蛋白电泳和质谱检测的方法,由地衣芽孢杆菌中筛选获得一种新型启动子p Chi,以两种已知强组成型启动子p Shuttle-09和pyxi E为对照,研究其对克劳氏芽孢杆菌碱性蛋白酶与地衣芽孢杆菌碱性蛋白酶基因的表达活性。成功构建了6种重组表达载体及重组菌,结果表明,p Chi的启动强度是p Shuttle-09的1.25倍,pyxi E的2倍,从而为介导枯草芽孢杆菌表达系统中异源基因的表达奠定基础。The alkaline protease has extensive applications but in short supply, for which the efficient expression system of alkaline protease is constructed, A new type of promoter pChi was screened fi＇om the Bacillus lieheniformis by SDS-PAGE and mass speetrqmetrie detection. With the other two strong constitutive promoters pShuttle-09 and pyxiE as the control, pChi＇s expression activities in the alkaline proteases from B, elausii and B. licheniformis was investigated. Further, six types of reemnhinant expression vectors and bacteria were eonstmcted. The results showed that the functional expression of pChi was 1.25 times of pShuttle-09 and 2 times of pyxiE, which offers the foundation for the gene heterologous expression in mediated B. subtilis expression system.

关 键 词：碱性蛋白酶 高效表达系统 启动子 异源表达 

分 类 号：Q78[生物学—分子生物学]