摘要
目的利用基因工程菌合成谷胱甘肽。方法克隆γ-谷氨酰转肽酶,构建重组质粒pET30a-GGT,乳糖诱导融合蛋白的表达,利用pET30a-GGT/E.coliBL21和pET32a-GSH-II/E.coliBL21分两步法合成γ-谷氨酰半胱氨酸与谷胱甘肽。结果工程菌的最佳乳糖诱导条件为:浓度3mmoL.L-1、时间6h、温度28℃。工程菌经最佳条件诱导后,γ-谷氨酰转肽酶的酶活大约是出发菌株E.coliDH5α的21倍,酶活力得到明显提高,谷胱甘肽的产量达到0.524g.L-1。结论成功地为今后利用基因工程菌催化合成谷胱甘肽寻找到一种新的方法 。
Objective To produce glutathione by engineering bacteria. Methods The gene of γ-glutamy/transpeptidase was cloned into vector pET30a. The recombinant protein was induced by lactose. E. coli BL21（pET30a-GGT） together with E. coli BL21（pET32a- GSH-II） used to produce GSH by two-step faction. Results The optimial lactose induction conditions of ）γ-glutamyl transpeptidase were as follows：concentration 3 mmol.L^-1 , time 6 h, and temperature 28℃. After lactose induction, the activity of γ-glutamyl transpeptidase of the engineered strain was 21 times higher than that of the starting bacterium. The yield of glutathione was 0. 524 g.L^-1. Conelusion These results found a new rneathod for glutathione synthesis by engineering bacteria.
出处
《西北药学杂志》
CAS
2010年第4期288-291,共4页
Northwest Pharmaceutical Journal
关键词
Γ-谷氨酰转肽酶
克隆
表达
谷胱甘肽合成
γ-glutamyl transpeptidase
Clone
Expression
glutathione synthesis
