H Muratsubaki, K Enomoto, Y Ichijoh, T Tezuka, T Katsume
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引用次数: 13
Abstract
The rapid and effective purification of soluble fumarate reductase from baker's yeast achieved by Blue Sepharose CL-6B chromatography. Cibacron Blue F3GA, the chromophore of Blue Sepharose, inhibited the activity of fumarate reductase. The enzyme bound to the column was selectively eluted by flavin adenine dinucleotide (FAD), flavin mononucleotide (FMN) or riboflavin. The purified enzyme was essentially homogeneous as indicated by polyacrylamide gel electrophoresis under nondenaturing conditions and under denaturing conditions in sodium dodecylsulfate. By this procedure, the enzyme could be rapidly purified with high yield from yeast cells.
采用Blue Sepharose CL-6B色谱法对面包酵母中可溶性富马酸还原酶进行了快速有效的纯化。蓝色Sepharose的发色团Cibacron Blue F3GA抑制富马酸还原酶的活性。结合柱上的酶被黄素腺嘌呤二核苷酸(FAD)、黄素单核苷酸(FMN)或核黄素选择性洗脱。在非变性条件下和在十二烷基硫酸钠中变性条件下，聚丙烯酰胺凝胶电泳表明纯化的酶基本均匀。该方法可快速、高产地从酵母细胞中纯化酶。
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