作　　者： ; ; ; ;

机构地区： 西北农林科技大学生命科学学院

出　　处： 《生物技术通讯》 2005年第2期144-146,共3页

摘　　要： 利用大肠杆菌-生黑醋杆菌穿梭载体pUG18构建了山梨糖脱氢酶基因sdh和氨甲酰化酶基因hyuC的融合表达质粒pUG18-SDH-HyuC。在JM109/pUG18-SDH-HyuC中检测到山梨糖脱氢酶(SDH)和氨甲酰化酶(HyuC)的酶活性,大量的SDH和HyuC不以融合蛋白形式存在。将质粒pUG18-SDH-HyuC电转化至生黑醋杆菌H24,检测到HyuC活性,但尚未检测到SDH酶活性。从生黑醋杆菌转化子中提取质粒,重新转化至JM109,又可同时检测到SDH和HyuC活性,传代实验和对质粒的鉴定结果也说明该质粒在H24中稳定存在。 Fusion expression shuttle vector pUG18-SDH-HyuC for sdh gene and hyuC gene was constructed and the enzyme activities of sorbose dehydrogenase(SDH) and N-carbamoylase(HyuC) in Escherichia coli JM109/pUG18-SDH-HyuC could be detected. Western blot result showed that most of SDH and HyuC did not exist in fusion protein style. After electroporating pUG18-SDH-HyuC into Acetobacter melanogenum H24, it was of HyuC activity, but not of SDH activity. Plasmid was extracted from H24 and retransformed into JM109 where enzyme activities of SDH and HyuC could be detected again. The plasmid restriction map indicated that it was stable in H24.

关 键 词： 山梨糖脱氢酶 氨甲酰化酶 融合表达 维生素 两步发酵法 生黑醋杆菌

领　　域： [轻工技术与工程] [化学工程] [轻工技术与工程]