食品科学2017，Vol.38Issue(14)：1-8,8.DOI:10.7506/spkx1002-6630-201714001
纳豆激酶基因工程菌的构建及酶活力分析
Construction of Genetically Engineered Strain for Nattokinase Production and Enzyme Activity Analysis
摘要
Abstract
Nattokinase (NK),encoded by the aprN gene of Bacillus subtilis natto,has strong fibrinolytic activity both in vitro and in vivo.In this research,the aprN gene from B.subtilis was cloned and the codons which encode the first 30 amino acids were optimized on the basis of the codon preference of B.subtilis.Recombinant vector pHT01-aprN was constructed.Through restriction enzyme digestion analysis,PCR amplification and sequencing,the subcloned gene was confirmed to be aprN.The pHT01-aprN was transformed into B.subtilis 168 by electroporation,and the engineered bacterium (B.s 168/pHT01-aprN) was isolated on LB plates containing chloramphenicol.NK expression was induced by IPTG,and the highest enzyme activity in shaking flask culture was up to (289.00 ± 3.42) U/mL with good stability,which was 3.9 times as high as that of wild-type B.subtilis natto.关键词
纳豆激酶/aprN基因/枯草芽孢杆菌/基因工程菌/酶活力Key words
nattokinase/aprN gene/Bacillus subtilis/enzyme activity/engineered bacteria分类
生物科学引用本文复制引用
崔青,钱炳俊,姚晓敏,季顺利,鲁飞凤,吴静,张建华..纳豆激酶基因工程菌的构建及酶活力分析[J].食品科学,2017,38(14):1-8,8.基金项目
国家自然科学基金面上项目(31171737) （31171737）
