中文摘要：

采用硫酸铵沉淀和柱层析（DEAE-Sepharose F．F．，Sephacryl S-200，TSK-DEAE）初步分离纯化了产酸克雷伯氏菌（Klebsiella oxytoca）HPI可溶性氢酶，研究了温度、pH值、电子载体等对氢酶催化放氢活性的影响．研究表明，可溶性氢酶催化放氢的最适温度为30℃，催化放氢的最适pH值为7.5，甲基紫晶（MV）是氢酶催化放氢的最适人工电子载体，氧对氢酶催化活性有较大的抑制作用．

英文摘要：

The hydrogenase-containing crude extraction of Klebsiella oxytoca HP1 was firstly precipitated by 20% and 60% of ammonium sulfate solution in water, respectively. It was subsequently purified by three steps of column chromatography （DEAE- Sepharose F. F. ,Sephacryl S-200 and TSK-DEAE）. After three steps of purification,the soluble hydrogense of K. oxytoca HP1 was purified about 200 folds with a yield of 15.3% and a specific activity of 17.68μmol H2/min·mg prot. The catalytic property of soluble hydrogenase was studied. The optimal conditions for hydrogen production were obtained as follow： temperature 30℃, pH 7.5. Oxygen was an inhibitor for the hydrogen production of soluble hydrogenase. MV was the optimal electron carrier for the soluble hydrogenasee＇s activity.