作　　者： ; ; (仪朝印）;

机构地区： 黑龙江八一农垦大学

出　　处： 《中国食品学报》 2014年第2期42-46,共5页

摘　　要： 目的:构建高产转谷氨酰胺酶且具有活性的基因工程菌。方法:以茂原链霉菌总DNA为模板,利用PCR技术扩增目的基因,构建重组质粒pET-22b-MTG,使其在大肠杆菌中得到高效表达并优化表达时间和温度。结果:成功构建产转谷氨酰胺酶的基因工程菌,酶活为15.9 U/mL,最优表达时间5 h,温度25℃。结论:成功构建TG基因工程菌,TG基因在大肠杆菌中得到高效表达,为后续试验提供原材料,为微生物发酵生产TG打下基础。 Objective： To construct an active gene engineering bacteria which produces transglutaminase（transglutaminase, TG）. Method： In this experiment, the TG target gene was amplified by PCR with the total genomic DNA of Streptomyces mobaraensis（Streptomyces mobaraensis） as a template, recover the target gene fragment and expression vector pET-22b with gel, structure the expression plasmid of pET-MTG to make it effectively expressed in Escherichia coli, optimize the express time and temperature.Result： The gene engineering bacteria which produces transglutaminase was successfully constructed, enzyme activity was 13.1 U/ml, the appropriate expression of time and temperature were determined by 25 ℃ and 5 h. Conclusion： The gene engineering bacteria constructed get effectively expressed, laying the foundation for providing raw materials as well as microbial fermentation production of TG for the following experiments.

关 键 词： 转谷氨酰胺酶 大肠杆菌 表达 改性 大豆分离蛋白

领　　域： [轻工技术与工程] [轻工技术与工程]