摘要
以茂原链霉菌(Streptomyces mobaraensis)基因组DNA为模板,PCR扩增得到转谷氨酰胺酶酶原基因(pro-transglutaminase,pro-TG),经pMD18-T载体亚克隆到表达载pBEp43(+),然后转化到枯草芽孢杆菌(Bacillus subtilis)WB800中,经过32 h摇瓶发酵,成功表达了转谷氨酰胺酶酶原。采用胰蛋白酶激活转谷氨酰胺酶酶原,成功切除前导肽后得到成熟的转谷氨酰胺酶。牛血清蛋白(BSA)交联表明成熟的转谷氨酰胺酶具有蛋白交联的功能。本研究首次采用枯草芽孢杆菌作为宿主分泌表达了茂原链霉菌的转谷氨酰胺酶酶原,对基因工程异源表达这种特殊的食品用酶提供了新思路。
The pro-transglutaminase（pro-TG） gene was obtained by PCR amplification using Streptomyces mobaraensis genome DNA as template and subcloned into the expression plasmid pBEP43（＋） via pMD18-T vector,then the plasmid was transformed into B.subtilis WB800.After 32 h flask fermentation,pro-TG protein was successfully expressed in the culture broth.Mature-transglutaminase was obtained after cutting the leader peptide by trypsin activation.BSA cross-linking experiment shows that the mature transglutaminase has cross-linking function.In this work,Streptomyces mobaraensis pro-TG was first expressed in Bacillus subtilis.The research provided a new idea for genetic engineering heterologous expression of this particular food enzyme.
出处
《现代食品科技》
EI
CAS
2011年第7期734-737,共4页
Modern Food Science and Technology
基金
广东省科技攻关项目(2006B13001006)
关键词
转谷氨酰胺酶酶原
枯草芽孢杆菌
分泌表达
亲和层析
BSA交联
pro-transglutaminase
Bacillus subtilis
secretory expression
affinity chromatography
BSA cross-linking
