文献类型：期刊文献

中文题名：氧化葡萄糖酸杆菌DSM 2003膜结合乙醇脱氢酶的纯化鉴定和性质研究

英文题名：Purification, Identification and Characterization of Membrane-bound Alcohol Dehydrogenase from Gluconobacter oxydans DSM 2003

作者：韦柳静[1];林金萍[1];杨雪鹏[2];魏东芝[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室,鲁华生物技术研究所,上海200237;[2]郑州轻工业大学食品与生物工程学院,河南郑州450002

年份：2010

卷号：31

期号：13

起止页码：164

中文期刊名：食品科学

外文期刊名：Food Science

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

基金：国家“973”计划项目(2009CB724703);国家自然科学基金项目(20976053/B060804);生物反应器国家重点实验室专用基金项目(2060204)

语种：中文

中文关键词：氧化葡萄糖酸杆菌;膜结合乙醇脱氢酶;纯化;鉴定;酶学特性

外文关键词：Gluconobacter oxydans; alcohol dehydrogenase; purification; identification; characterization

摘要：通过超速离心收集膜组分、表面活性剂溶解膜蛋白、CM-纤维素柱层析纯化等步骤,从氧化葡萄糖酸杆菌DSM2003中获得电泳纯的具有1,2-丙二醇脱氢酶活性的脱氢酶,此酶由两个亚基组成,其表观相对分子质量分别为80000和50000。经MALDI-TOFMS-MS质谱分析与肽质量指纹图谱检索鉴定,证明纯化得到的酶是乙醇脱氢酶。酶学性质分析表明,该酶的最适反应温度为30℃,最适pH值为5.5～6.0;该酶能催化多种一元醇、二元醇,但对包含3个以上羟基的多元醇基本无氧化活性;其催化活性随着底物碳链长度的增加而减小;大多数金属离子及抑制剂(Cu2+、Fe3+、Ca2+、EDTA等)对此酶活性均有抑制作用。
The membrane fraction of Gluconobacter oxydans DSM 2003 was collected by ultra-centrifugation, and an enzyme with dehydrogenase activity to 1,2-propanediol was isolated from the membrane fraction and purified by CM-cellulose column chromatography. The purified enzyme was composed of two subunits with molecular weights of approximately 80 and 50 kD, respectively. Meanwhile, this enzyme was confirmed to be alcohol dehydrogenase （ADH） by mass spectrometry. The optimal reaction pH and temperature for this enzyme were 5.5 - 6.0 and 30 ℃, respectively. Although this enzyme exhibited a broad substrate spectrum including primary and secondary alcohols, alcohols containing 3 hydroxyl groups and more failed to be oxidized by it. The activity of this enzyme was inhibited by most of the selected metal ions, especially by Cu2＋ and Fe3＋.