【作者】 李勤； 黄建安； 李娟； 张玥； 刘仲华；

【Author】 LI QIN~(1,2),HUANG Jian-an~2,LI Juan~(1,2),ZHANG Yue~(1,2),LIU Zhong-hua~(1*) (Functional Components Using National Plant Engineering Technology Research Center;Ministry of Education Key Laboratory of Tea,Changsha 410128,China)

【机构】 国家植物功能成分利用工程技术研究中心； 茶学教育部重点实验室；

【摘要】 构建了一个高效表达大肠杆菌来源的GGT重组质粒pET32α-GGT并转化E·coliBL21(DE3),工程菌株经0.1mmol/L IPTG,20℃诱导表达6h,粗酶液的酶活达到4.4131U/mL(+0.4104),大约是出发菌株E·coliBL21(DE3)的18.7倍。工程菌粗酶液以200mmoL/L谷氨酰胺、1.5moL/L乙胺盐酸盐为底物,pH10于20℃下反应6h,反应结束后用HPLC检测茶氨酸含量,工程菌茶氨酸合成量达到12.6mg/mL,L-谷氨酰胺转化率为41.05%。更多还原

【Abstract】 A recombinant plasmid named pET32a-GGT which could highly produceγ-G lutamyltranspeptidase was constructed.The recombinant plasmid was transformed to E·coliBL21(DE3).γ-Glutamyltranspeptidase was produced at 20℃,which was incubated with 0.2mmol/L IPTG for 6h.The activity of 1mL crude extract of recombinant strain was usually 4.4131U/mL,which was about 18.7 times higher than the activity of E·coliK BL21. When200mmol/L L-Gln and 1.5mol/L ethylamine were used,pH10.0,and incubation at 20℃for 6h.The yield of theanine was 12.6mg/mL,the rate of conversion from L-Gln to theanine was 41.05%.更多还原