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)HP1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.
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.
出处
《厦门大学学报（自然科学版）》
CAS
CSCD
北大核心
2006年第5期709-712,共4页
Journal of Xiamen University：Natural Science
基金
国家自然科学基金(30470395)
厦门大学新世纪优秀人才支持计划(X07114)资助
