光合细菌中氢化酶的分离纯化及其性质研究

Purification and Characterization of the Hydrogenase Extracted from Photosynthetic Bacteria

以光合细菌为出发菌株,离心得到的菌体经超声破碎后,得到的粗酶液经硫酸铵沉淀、透析、DEAE-52离子交换层析和Sephadex G-200分子筛凝胶层析分离纯化获得电泳纯的氢化酶,且该酶的分子量为62.0 ku和37.0 ku的两个亚基组成的分子量为99.0ku的二聚体,对该酶进行酶学性质的研究得到酶催化最适温度和pH值范围分别为30℃和pH=7.5,且在25～35℃和pH=7.0～8.0范围内催化放氢活力较稳定.

The purification and characteristics of an enzyme extracted from Photosynthetic Bacteria were studied. First, cells were disrupted by ultrasonic treatment. The hydrogenase was purified with a combination of ammonium precipitation, dialysis, DEAE -52 anion exchange chromatography and Sephadex G -200 gel filtration chromatography. The native molecular mass of the enzyme was found out to be the dimer of 99.0ku. The optimum pH and temperature for hydrogen production activity of the hydrogenase is 7.5 and 30 ℃.