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本文证明在提纯白养性大豆根瘤菌膜结合态氢酶过程中,为了保持酶活性和防止酶蛋白的沉淀,必须排除O_2、加入还原剂和去垢剂Genapol。纯化的氢酶在H_2气氛下储存于—80℃,半年后仍保留78％的活性,最纯的氢酶其比活性为142μmol亚甲蓝还原·min~(-1)·mg蛋白~(-1)。纯化的氢酶经SDS凝胶电泳呈现二条主要蛋白带,其分子量分别接近于60000和30000。不同提纯阶段的氢酶其比活性与细胞色素b的含量不成正相关。最高比活性的氢酶不含细胞色素b。文中还讨论了自养性大豆根瘤菌膜结合态氢酶的其它特性。
This paper proves that during the process of purifying the white nodulated Rhizobium japonicum membrane-bound hydrogenase, in order to maintain the enzyme activity and prevent the precipitation of the enzyme protein, O2 must be excluded and the reducer and the detergent Genapol should be added. The purified hydrogenase was stored at -80 ℃ in H 2 atmosphere and still retained 78% activity after six months. The specific activity of the pure hydrogenase was 142μmol Methylene blue reduced · min -1 · mg protein ~ (- 1). Purified hydrogenase SDS gel electrophoresis showed two major protein bands, the molecular weights were close to 60,000 and 30,000. In different purification stage, the specific activity of hydrogenase is not positively correlated with the content of cytochrome b. The highest specific activity of hydrogenase does not contain cytochrome b. The paper also discussed the autotrophic soybean Rhizobium other characteristics of membrane-bound hydrogenase.