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光合细菌 Chromatium vinosum可溶性氢酶经 5次柱层析 ( DE-2 3,TSK-DEAE( ) ,Ultragel Ac A-4 4 ,TSK-DEAE( ) ,Superdex TM75 )分离后被纯化 365倍 ,得率为 32 % ,其催化放氢的活性为 8.4μmol H2 /( min· mg prot) .氧化态可溶性氢酶在 4 5 K下 ,产生了 Ni( )的典型 EPR信号 ( gx,y,z=2 .37,2 .1 6,2 .0 1 6和 gx,y,z=2 .30 ,2 .2 3,2 .0 1 6) ;在 1 0 K下 ,没有出现膜结合态氢酶中存在的 [3Fe-4 S]簇特征 EPR信号 .可溶性氢酶被 H2 还原后 ,Ni( )的特征 EPR信号消失 ,同时出现一个 [4 Fe-4 S]簇的特征 EPR信号 ( gx,y,z=1 .88,1 .90 ,2 .0 4 5 ) .研究结果表明 ,C.vinosum可溶性氢酶在结构和功能上不同于膜结合态氢酶 ,是一种新的催化放氢的 Ni Fe-氢酶
The photosynthetic bacteria Chromatium vinosum soluble hydrogenase was purified 365 times after separation by five column chromatography (DE-2 3, TSK-DEAE (), Ultragel Ac A-4 4, TSK-DEAE (), Superdex ™ 75) Was 32%, and its catalytic hydrogen evolution activity was 8.4 μmol H2 / (min · mg prot) .A typical EPR signal of Ni () was generated at 45 K by oxidation state soluble hydrogenase (gx, y, z = 2 .37, 2 .1 6,2 .0 1 6 and gx, y, z = 2 .30, 2 .2 3,2 .0 1 6); at 10 K, no membrane bound hydrogenase The characteristic EPR signal of [3Fe-4S] cluster exists in the presence of [3Fe-4S] clusters, and the characteristic EPR signal of Ni () disappears after the soluble hydrogenase is reduced by H2. z = 1 .88,1 .90, 2.04 5). The results show that, C.vinosum soluble hydrogenase in the structure and function is different from membrane-bound hydrogenase, is a new catalytic hydrogenation of Ni Fe-hydrogenase