生物加工过程Issue(3)：20-25,6.DOI:10.3969/j.issn.1672-3678.2015.03.004
蜡样芽胞杆菌蛋白酶的克隆及在枯草芽胞杆菌系统中的高效表达
Cloning and over-expression of protease gene from Bacillus cereus WQ9-2 in Bacillus subtilis
摘要
Abstract
Based on the liquid chromatography⁃mass⁃mass spectrometry ( LC⁃MS⁃MS) analysis of trypsin⁃digested protein fragments of the solvent⁃stable protease WQ from a solvent⁃stable strain Bacillus cereus WQ9⁃2,solvent⁃stable protease gene was successfully cloned. The gene contains an open reading frame of 1 701 bp,encoding a pre⁃pro⁃protein enzyme of 566 amino acids(28⁃aa pre⁃signal peptide,220⁃aa pro⁃peptide and 318⁃aa mature protein with the molecular mass of 3�7×104). The expression plasmid pMA5/aprWQ was constructed by inserting the aprWQ gene into the vector pMA5 without native signal peptide sequence. The maximum concentration of the recombinant protease was 17 400 U/mL,5 fold higher than the natural production level. Molecular weight, tolerance in organic solvent of recombinant solvent⁃stable protease was identical to the native protease. The findings provides the basis for further expansion of the catalytic applications of organic solvent⁃stable protease.关键词
耐有机溶剂蛋白酶/枯草芽胞杆菌/高表达/发酵优化Key words
solvent-stable protease/Bacillus subtilis/over-expression/fermentation optimization分类
生物科学引用本文复制引用
承龙飞,朱富成,刘可可,何冰芳..蜡样芽胞杆菌蛋白酶的克隆及在枯草芽胞杆菌系统中的高效表达[J].生物加工过程,2015,(3):20-25,6.基金项目
国家高技术研究发展计划（863计划）重大项目（2012AA022205）；国家重点基础研究发展计划（973计划） （）
