天津科技大学学报2016，Vol.31Issue(2)：13-19,7.DOI:10.13364/j.issn.1672-6510.20150057
葡萄糖脱氢酶温控表达菌株的构建及其发酵工艺
Construction of a Recombined Bacterium for the Thermo-regulated Expression of Glucose Dehydrogenase and the Optimization of the Fermentation Process
摘要
Abstract
Membrane-bound pyrroloquinoline quinine-dependent glucose dehydrogenase(mPQQGDH,EC1.1.5.2)is a sig-nificant kind of oxidoreductase,and has received considerable attention for its wide application in the fields of clinical diag-nosis and industrial glucose sensors.However,the low expression level limits its mass industrial production and application scope.In this study,temperature sensitive promoter PL-PRwas used to replace the promoter of mPQQGDH,resulting ina recombined bacteriumE.coli B0013-120/pGCd::Km-PL-PR whose glucose dehydrogenase activity was 21.2%, higher than that of the original strain.Subsequently,the fermentation conditions were optimized to determine the suitable carbon source, induction temperature,optical density before induction and the induction time for the expression of mPQQGDH. Finally,by characterizing the enzymatic properties of this enzyme,the temperature and pH optima were determined to be 25,℃ and 6.0,respectively,and this enzyme was stable at 45,℃.These results canlay a solid foundation for the wider range of application and industrial production of mPQQGDH.关键词
葡萄糖脱氢酶/温敏型启动子/高效表达/发酵工艺优化Key words
glucose dehydrogenase/temperature sensitive promoter/overexpression/fermentation process optimization分类
生物科学引用本文复制引用
李杨,张志萌,董自星,王正祥,路福平..葡萄糖脱氢酶温控表达菌株的构建及其发酵工艺[J].天津科技大学学报,2016,31(2):13-19,7.基金项目
国家高技术研究发展计划(863计划)资助项目(2013AA102106-07) （863计划）
