天津科技大学学报2016，Vol.31Issue(6)：11-16,6.DOI:10.13364/j.issn.1672-6510.20160044
枯草芽胞杆菌谷氨酰胺转氨酶在谷氨酸棒杆菌中的分泌表达及诱导条件的优化
Expression of the Transglutaminase from Bacillussubtilisand Optimization of Fermentation Conditions for Corynebacterium glutamicum
摘要
Abstract
Overlap PCR was used to produceSprodbtg encode with transglutaminase(BTG)fromBacillus subtilis.The recombined geneSprodbtg contains signal peptide ΔS0949 fromCorynebacterium glutamicum,SD sequence and the proD-BTG gene.Thesprodbtg was cloned into pXMJ19 to construct secretion expression vector pXMJ19-Sprodbtg.After that,the recombined plasmid was transformed intoC.glutamicumATCC13032 to express proD-BTG.The optimized fermentation conditions were also studied.The results showed that the recombined strainC.glutamicumATCC13032/pXMJ19-Sprodbtg accumulated proD-BTG up to a level of(41.23±2.01)U/L.The optimized conditions for the induction of expression ofthe recombined protein proD-BTG in strain 13032/pXMJ19-Sprodbtg were as follows:growing in MMTG medium for 12 h, adding IPTG with a final concentration of 0.8 mmol/L and the subsequent incubation for 40 h.The activity of BTG reached (55.62±2.34)U/L,a 34.90%, increase compared with the non-optimized control.关键词
谷氨酰胺转氨酶/谷氨酸棒杆菌/枯草芽胞杆菌/分泌表达/条件优化Key words
transglutaminase/Corynebacterium glutamicum/Bacillus subtilis/secretion expression/condition optimi-zation分类
生物科学引用本文复制引用
黄琳,刘逸寒,李明杰,李瑞,郭伟,桂爽,路福平..枯草芽胞杆菌谷氨酰胺转氨酶在谷氨酸棒杆菌中的分泌表达及诱导条件的优化[J].天津科技大学学报,2016,31(6):11-16,6.基金项目
天津市应用基础与前沿技术研究计划资助项目(14JCYBJC23800)；天津市科技特派员资助项目(15JCTPJC56500) （14JCYBJC23800）
