Yagi T, Kimura K, Daidoji H, Sakai F, Tamura S
J Biochem. 1976 Mar;79(3):661-71. doi: 10.1093/oxfordjournals.jbchem.a131111.
The properties of purified hydrogenase [EC 1.12.2.1] solubilized from particulate fraction of sonicated Desulfovibrio vulgaris cells are described. The enzyme was a brownish iron-sulfur protein of molecular weight 89,000, composed of two different subunits (mol. wt.: 28,000 and 59,000), and it contained 7-9 iron atoms and 7-8 labile sulfide ions. Molybdenum was not detected in the preparation. The absorption spectrum of the enzyme was characteristic of iron-sulfur proteins. The millimolar absorbance coefficients of the enzyme were about 164 at 280nm, and 47 at 400nm. The absorption spectrum of the enzyme in the visible region changed upon incubating the enzyme under H2 in the presence of cytochrome c3, but not in its absence. This spectral change was due to the reduction of the enzyme. The absorbance ratio at 400nm of the reduced and the oxidized forms of the enzyme was 0.66. The activity of the enzyme was hardly affected by metal-complexing agents such as cyanide, azide, 1,10-phenanthroline, etc., except for CO, which was a strong inhibitor of the enzyme. The activity was inhibited by SH-reagents such as p-chloromercuribenzenesulfonate. The enzyme was significantly resistant to urea, but susceptible to sodium dodecyl sulfate. These properties were very similar to those of clostridial hydrogenase [EC 1.12.7.1], in spite of differences in the acceptor specificity and subunit structure.
本文描述了从超声破碎的普通脱硫弧菌细胞颗粒部分中溶解得到的纯化氢化酶[EC 1.12.2.1]的性质。该酶是一种分子量为89,000的褐色铁硫蛋白，由两种不同的亚基组成（分子量分别为28,000和59,000），含有7 - 9个铁原子和7 - 8个不稳定硫化物离子。在该制剂中未检测到钼。该酶的吸收光谱具有铁硫蛋白的特征。该酶在280nm处的毫摩尔吸光系数约为164，在400nm处为47。在细胞色素c3存在下，将该酶在H2中孵育时，其可见区域的吸收光谱会发生变化，而在没有细胞色素c3时则不会。这种光谱变化是由于酶的还原所致。该酶还原态和氧化态在400nm处的吸光度比值为0.66。除了作为该酶强抑制剂的CO外，该酶的活性几乎不受氰化物、叠氮化物、1,10 - 菲咯啉等金属络合剂的影响。该酶的活性受到对氯汞苯磺酸盐等巯基试剂的抑制。该酶对尿素具有显著抗性，但对十二烷基硫酸钠敏感。尽管在受体特异性和亚基结构上存在差异，但这些性质与梭菌氢化酶[EC 1.12.7.1]非常相似。
