硫酸盐还原菌氢化酶的分离纯化及酶学性质研究  

Purification and Characterization of the Hydrogenase from Sulfate Reducing Bacteria

作　　者：常磊峰[1] 邱广亮[1] 陈香君[1] 

机构地区：[1]内蒙古师范大学生命科学与技术学院,内蒙古呼和浩特010022

出　　处：《内蒙古师范大学学报（自然科学汉文版）》2008年第3期412-416,共5页Journal of Inner Mongolia Normal University(Natural Science Edition)

基　　金：国家自然科学基金资助项目(50066002)

摘　　要：硫酸盐还原菌氢化酶上清液经硫酸铵沉淀、透析、DEAE-52阴离子交换层析和Sephadex G-150凝胶过滤层析纯化出氢化酶,纯化倍数为34.7,酶回收率为34.1%.对纯化酶性质进行研究,结果表明：该酶是由分子量为46 KD和34 KD的两个亚基构成的总分子量为80 KD的αβ异二聚体;酶催化最适温度和pH值范围分别为45℃和6.5～8.5,在34～55℃和pH=6～9.2范围内具有较稳定的催化放氢活力;光谱扫描分析和激活剂及抑制剂测定表明该酶属含铁硫中心的唯铁氢化酶.A hydrogenase from the Sulfate Reducing Bacteria is purified to homogeneity using ammonium sulfate preeipitation,dialysis,DEAE-Sepharose Fast Flow anion exchange chromatography and Sephadex G150 gel filtration chromatography. This purification protocol resulted in a 34.7-fold purification of lipase with 34. 1% final yield,and the relative molecular weight of the enzyme is determined to be approximately a 1 ： 1 αβ-heterodimer of molecular mass =80 KD,and two subunits （α=46 KD,β=34 KD）. The optimum pH and temperature for hydrogen production activity of the hydrogenase is 6.5～8.5 and 46℃, respectively. The absorption spectrum of the enzyme showed that the enzyme has an iron-sulfur cluster. The activator and inhibitor of the enzyme showed that the enzyme might belong to Fe-only hydrogenase.

关 键 词：硫酸盐还原菌 唯铁氢化酶 分离纯化 酶学性质 

分 类 号：O629.8[理学—有机化学]