启动子及信号肽筛选提高普鲁兰酶在地衣芽孢杆菌中的表达  

Improving the Production of Pullulanase in Bacillus licheniformis by Screening Promoters and Signal Peptides

作　　者：王倩[1,2] 初晓宇 朱宝成[1] 伍宁丰[2] WANG Qian;CHU Xiaoyu;ZHU Baocheng;WU Ningfeng(College of Life Sciences, Hebei Agricultural University, Hebei Baoding 071000;Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China)

机构地区：[1]河北农业大学生命科学学院,河北保定071000 [2]中国农业科学院生物技术研究所,北京100081

出　　处：《中国农业科技导报》2019年第6期61-69,共9页Journal of Agricultural Science and Technology

基　　金：国家863计划项目(2013AA102804)资助

摘　　要：普鲁兰酶(pullulanase)是一种淀粉脱支酶,在淀粉糖化加工等工业生产中具有重要的应用价值。选用地衣芽孢杆菌(Bacillus licheniformis) BL10作为表达宿主,为了提高其中的普鲁兰酶产量,对来自枯草芽孢杆菌(Bacillus subtilis) 168和地衣芽孢杆菌BL10中的15种不同信号肽以及4种不同启动子进行筛选,构建了不同的重组工程菌株,最终发现在以P43为启动子、apr为信号肽的条件下,上清液中普鲁兰酶的表达量最高,酶活可达12 U/m L,且测得其最适反应温度为60℃,最适p H为4.5。结果表明通过对启动子及信号肽进行筛选是提高重组工程菌株中目的蛋白表达量的一种有效手段。Pullulanase is a well-known starch debranching enzyme with high application value in industrial production such as starch saccharification. In this study, in order to improve the production of pullulanase in Bacillus licheniformis BL10,fifteen signal peptides and four promoters from B. licheniformis BL10 and Bacillus subtilis 168 were screened,and the recombinant strains were constructed. The results showed that the recombinant strain with P43 as the promoter and apr as the signal peptide displayed the highest expression level of pullulanase in supernatant and its enzyme activity reached 12 U/m L. The optimal reaction temperature and p H of pullulanase were 60℃ and 4.5,respectively. The results indicated that it was an effective method to enhance the expression of a target protein by screening promoters and signal peptides in recombinant strain.

关 键 词：地衣芽孢杆菌 普鲁兰酶 启动子 信号肽 

分 类 号：Q78[生物学—分子生物学]