从四种基因型的橄榄果蝇（油橄榄实蝇）中纯化乙醇脱氢酶。
Purification of alcohol dehydrogenase from four genotypes of the olive fruit fly Bactrocera (Dacus) oleae.
作者信息
Mazi V E, Cosmidis N, Clonis Y D, Loukas M
机构信息
Laboratories of Enzyme Technology and Genetics, Department of Agricultural Biotechnology, The Agricultural University of Athens, Iera Odos 75, 118 55 Athens, Greece.
出版信息
Biotechnol Prog. 1998 Mar-Apr;14(2):294-9. doi: 10.1021/bp970114n.
This is the first report describing the purification of alcohol dehydrogenase (ADH) from four genotypes of the olive fruit fly Bactrocera oleae, the most important pest of olives in the Mediterranean region. The purified enzyme shows a single band after SDS-PAGE analysis, corresponding to subunit mass of 26 kDa. The native ADH shows a molecular mass of 48 kDa, after gel filtration HPLC analysis. The purification method incorporated a preliminary ammonium sulphate precipitation step, followed by an anion-exchange DEAE chromatography step, a dye affinity chromatography step on Cibacron blue 3GA, and an anion-exchange DEAE chromatography step employing the same column of the first step. The present method offers good overall recovery (40%) and high enzyme purity, and it is applicable to different genotypes. Furthermore, the method is rapid and economical, as it employs two cheap, widely used, and commercially available chromatography materials.
这是首篇描述从四种基因型的地中海实蝇（橄榄在地中海地区最重要的害虫）中纯化乙醇脱氢酶（ADH）的报告。经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析，纯化后的酶呈现单一条带，对应亚基质量为26 kDa。经凝胶过滤高效液相色谱（HPLC）分析，天然ADH的分子量为48 kDa。纯化方法包括初步硫酸铵沉淀步骤，随后是阴离子交换DEAE色谱步骤、基于Cibacron blue 3GA的染料亲和色谱步骤，以及使用第一步相同色谱柱的阴离子交换DEAE色谱步骤。本方法具有良好的总体回收率（40%）和高酶纯度，且适用于不同基因型。此外，该方法快速且经济，因为它使用了两种廉价、广泛使用且可商购的色谱材料。
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