聚球藻PCC 7942氢酶的分离纯化及特性研究  

Purification and characterization of hydrogenase from Synechococcus sp. PCC 7942

作　　者：徐惠娟[1] 邬小兵[1] 李筱泉[1] 龙敏南[1] 梁世中[2] 

机构地区：[1]厦门大学生命科学学院,厦门361005 [2]华南理工大学生物科学与工程学院,广州510006

出　　处：《生态科学》2008年第4期227-231,共5页Ecological Science

基　　金：福建省自然科学基金(C0410002)资助

摘　　要：报道了室温、空气环境下聚球藻Synechococcus sp.PCC 7942氢酶的分离纯化。经过超声破碎、超速离心、离子交换层析、疏水层析及凝胶层析等步骤,氢酶被纯化了218倍,得率为6.5%,比活为1.46 U·mg-1蛋白。纯化氢酶的SDS-PAGE图显示五条蛋白带,分子量约为83kDa,60kDa,47kDa,30kDa和27kDa。该氢酶为可溶性的双向氢酶,其催化放氢的最佳电子供体为还原态的甲基紫精,最适温度50℃,最适pH 8.0。Hydrogenase from Synechococcus sp. PCC 7942 was purified to close homogeneity aerobically at room temperature. The hydrogenase-containing crude extract was collected after ultrasonic disruption and removal of cell debris by ultracentrifugation. Subsequently, three steps of column chromatographies （anion exchange, hydrophobic interaction and gel filtration） were performed. Hydrogenase was purified about 218-fold with a yield of 6.5% finally. The purified enzyme has a specific activity for hydrogen evolution of 1.46 U·mg^-1 protein. SDS-PAGE gel of the purified enzyme revealed five predominant protein bands with estimated molecular weights of 83, 60, 47, 30 and 27 kDa, respectively. The enzyme is a soluble bidirectional hydrogenase and shows maximum activity while using reduced methyl viologen as an electron donor. The optimum temperature and pH value for hydrogen evolution catalyzed by the purified hydrogenase are 50 ℃ and pH 8.0.

关 键 词：氢酶 放氢 聚球藻 纯化 特性 

分 类 号：Q936[生物学—微生物学]