文献类型：期刊文献

中文题名：氧化葡萄糖酸杆菌胞内脱氢酶Gox0525的酶学性质研究

英文题名：Enzymatic Characterization of Recombinant Gox0525 from Gluconobacter oxydans

作者：王嘉乐[1];魏东芝[1];林金萍[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

年份：2011

卷号：42

期号：10

起止页码：742

中文期刊名：中国医药工业杂志

外文期刊名：Chinese Journal of Pharmaceuticals

收录：CSTPCD;;北大核心:【北大核心2008】；CSCD:【CSCD2011_2012】；

语种：中文

中文关键词：氧化葡萄糖酸杆菌;Gox0525;蛋白表达;酶学性质

外文关键词：Gluconobacter oxydans; Gox0525; protein expression; enzymatic characterization

摘要：将氧化葡萄糖酸杆菌胞内脱氢酶推测基因Gox0525克隆至表达载体pET-28a(+)中,并在大肠杆菌中得到可溶性表达的蛋白,纯化后经HPLC以及SDS-PAGE检测,表明其活性形式为四聚体。重组蛋白Gox0525属于短链脱氢酶家族,选择该家族的催化底物谱测试,结果表明Gox0525具有还原二酮的能力,且专一性利用辅酶NADPH,存pH 9.0、30℃条件下活性较高,适合保存在pH 7.0、4℃条件下。酶反应的发生对金属离子无依赖性,添加Ca^(2+)对酶活力有促进作用,而Fe^(3+)有抑制作用。
A putative dehydrogenase gene Gox0525 from Gluconobacter oxydans was cloned into the expression vector pET-28a （＋） and then expressed in Escherichia coli in soluble form. The purified protein Gox0525 was analyzed via HPLC and SDS-PAGE, showing homotetramer as its active form. Gox0525 belonging to short-chain dehydrogenase superfamily and could reduce diketone with NADPH as the cofactor. The maximum activity was detected at pH 9.0 at 30 ℃ and it should be stored at pH 7.0 and 4 ℃.The activity of Gox0525 was independent of metal ions, whereas was activated by Ca2＋ and inhibited by Fe3＋.