酵母醇脱氢酶ADHI的纯化及动力学研究  

Purification And Kinetic Studies of Yeast Alcohol Dehydrogenase

作　　者：廉德君[1] 李林[1] 许根俊 

机构地区：[1]中国科学院上海生物化学研究所,中国科学院生物物理研究所生物大分子国家重点实验室

出　　处：《生物化学与生物物理学报》1996年第4期396-403,共8页

基　　金：国家自然科学基金;国家攀登计划项目资助

摘　　要：本文报道了啤酒酵母醇脱氢酶组成型同工酶ＡＤＨＩ的快速高效的纯化方法。通过活性蓝色染料柱（Ｂｌｕｅ－Ｓｅｐｈａｒｏｓｅ）亲和层析的方法将该酶纯化至电泳均一，收率达４７％。对该酶的产物抑制及端点抑制动力学研究结果支持Ｗｒａｔｔｅｎ和Ｃｌｅｌａｎｄ提出的序列有序机制。高浓度的乙醇对酶具有激活作用。二氧六环可作为乙醇的竟争性抑制剂。综合以上实验结果，我们提出酵母醇脱氢酶催化反应机制的复杂性是由于其底物与活性中心结合复杂性所致．An efficient procedure for the purification of yeast alcohol dehydrogenase I(ADH I)was developed.By using Blue-Sepllarose 4B affinity chromatography,ADH I from yeast(S.cerevisiae)was purified 200-fold with an overall yield of47% to homogeneity as judged by sodium dodecy1 sulfate polyacrylamide gel electrophorsis and starch gel electrophoresis.Results of studies on the product and dead-end inhibition kinetics were in agreement with an orderde Bi Bi mechanism as proposed by Wratten and Cleland.Dioxane was found to be a competitive idhibitor of this enzyme.The complexity of the reaction mechanism of this enzyme is discussed.

关 键 词：酵母 醇脱氢酶 同功酶 提纯 动力学 

分 类 号：Q949.326.1[生物学—植物学] Q554.9