Schink B, Schlegel H G
Biochimie. 1978;60(3):297-305. doi: 10.1016/s0300-9084(78)80826-8.
A survey on organisms able to use molecular hydrogen as electron donor in the energy-yielding process is presented. In the group of the aerobic hydrogen-oxidizing bacteria so far two types of hydrogenases have been encountered, a NAD-reducing, soluble enzyme (H2 : NAD oxidoreductase) and a membrane-bound enzyme unable to reduce pyridine nucleotides. With respect to the distribution of both types of hydrogenases three groups of hydrogen-oxidizing bacteria can be diffentiated containing (i) both types (Alcaligenes eutrophus), (ii) a soluble enzyme only (Nocardia opaca lb), and (iii) a membrane-bound hydrogenase only (majority of genera and species). The results of studies on the NAD-specific hydrogenase of A. eutrophus are summarized. Results on the solubilization and purification of the membrane-bound hydrogenase of A. eutrophus are presented in detail. The enzyme was solubilized from purified membranes by Triton X-100 and sodium desoxycholate or phospholipase D. The crude membrane extract was fractionated by ammonium sulfate precipitation and chromatography on carboxymethylcellulose at pH 5.5. The enzyme was stable in potassium phosphate buffer; it resembles the soluble enzyme with respect to stability under oxidizing conditions. Further biochemical and immunological data indicate, however, that both enzymes are different with respect to their native structure.
本文介绍了一项关于能够在产能过程中利用分子氢作为电子供体的生物体的调查。在需氧氢氧化细菌群体中，目前已发现两种类型的氢化酶，一种是还原NAD的可溶性酶（H2：NAD氧化还原酶），另一种是无法还原吡啶核苷酸的膜结合酶。根据这两种类型氢化酶的分布情况，可以区分出三组氢氧化细菌，分别含有（i）两种类型的氢化酶（嗜糖产碱菌），（ii）仅有一种可溶性酶（不透明诺卡氏菌lb），以及（iii）仅有一种膜结合氢化酶（大多数属和种）。总结了对嗜糖产碱菌NAD特异性氢化酶的研究结果。详细介绍了嗜糖产碱菌膜结合氢化酶的溶解和纯化结果。该酶通过Triton X-100、脱氧胆酸钠或磷脂酶D从纯化的膜中溶解出来。粗膜提取物通过硫酸铵沉淀和在pH 5.5的羧甲基纤维素上进行色谱分离。该酶在磷酸钾缓冲液中稳定；在氧化条件下的稳定性方面，它与可溶性酶相似。然而，进一步的生化和免疫学数据表明，这两种酶在其天然结构方面有所不同。
