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.福建省自然科学基金(C0410002)资
