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	花生根瘤菌类菌体氢酶的分离提纯及特性; PuriFication and Some Properties of the Membrane-Bound Hydrogenase From the Bacteroids of Peanut Root Nodules
	黄南仲 ; 许良树 ; 张凤章 ; 龙敏南
	1995
关键词	花生 类菌体 氨酶 提纯 特性 penut bacteroid hydrogenase puriFication properties
英文摘要	花生根瘤菌类菌体经超声波破碎，TrITOnX-100溶解，正已烷-硫酸铵处理后，再经dEAE-纤维素和SEPHACryl凝胶柱层析等纯化步骤，获得凝胶电泳纯的膜结合态氢酶，比活为71.4μMOlH2Mg-1PrOTMIn-1，为类菌体吸H2活性的211倍。纯化的氢酶分子量为110kd。经SdS-PAgE后，呈现两个蛋白带，分子量分利为65kd和35kd。纯酶的nI含量为0.62MOlnI/MOl氢酶。在磷酸缓冲液中其活性的最适PH为6.5。dCIP、亚甲蓝、铁氰化钾、细胞色素C均可作为氢酶的电子受体，其中以dCIP为最适。; The H2-uptake hydrogenase Fromthe bacteroids of peanut nodules hasbeen puriFied and characterized.Bacteroids were prepared, then broken bysonication.The membrane-bound hydrogenase was solubilized by treatmentwith Triton X-100 and Followed byammonium sulFate-he-cane extractionto remove lipids and detergent..Thehydrogenase was Further puriFied byDEAE-cellulars column chromatography, eluted with Tris-HCl buFFer containing NaCl 0.3 mol/L and GenapolX-080 0.2%.The enzyme was puriFied to homogeneity by runningSephacryl H200 column chromatogtaphy twice.The speciFic activity was71.4 pmol H2 oxidiZed Per mg proteinper mid and was incrsased 211 Foldsrelative to that in hacteroids.Themolecular weisht of native enzyme wasabout 1 10 kD as determined by PAGEin undenatured Form.SDS-PAGEShowed two bands with molecularweight of 65 kD and 35 kD, indicatingthat the membrane-bound hydsogenasewas an a, 6 dimer.The nickel contentof puriFied hydrogenase was Found tobe 0.62 mole Ni/mol hydrogenase.The optimum PH For the activity of thepuriFied enzyme was Found to be near6.5 in potassium phosphate buFFer.Suitable electron acceptors are DCIP,methylene blue, Ferricyanide and CytC.The best one is DCIP.Benzyl viologen, methyl viologen and NAD werealmost not reduced.; 国家自然科学基金
语种	zh_CN
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/118986]
专题	生命科学－已发表论文
推荐引用方式 GB/T 7714	黄南仲,许良树,张凤章,等. 花生根瘤菌类菌体氢酶的分离提纯及特性, PuriFication and Some Properties of the Membrane-Bound Hydrogenase From the Bacteroids of Peanut Root Nodules[J],1995.
APA	黄南仲,许良树,张凤章,&龙敏南.(1995).花生根瘤菌类菌体氢酶的分离提纯及特性..
MLA	黄南仲,et al."花生根瘤菌类菌体氢酶的分离提纯及特性".(1995).

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