Purification of Alcohol Dehydrogenase from Four Genotypes of the Olive Fruit Fly Bactrocera (Dacus) oleae

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• 作者：Vasiliki E. Mazi ; Nikos Cosmidis ; Yannis D. Clonis ; and Michalis Loukas
• 刊名：Biotechnology Progress
• 出版年：1998
• 出版时间：April 1998
• 年：1998
• 卷：14
• 期：2
• 页码：294 - 299
• 全文大小：105K
• 年卷期：v.14,no.2(April 1998)
• ISSN：1520-6033

文摘

This is the first report describing the purification of alcoholdehydrogenase (ADH)from four genotypes of the olive fruit fly Bactrocera oleae,the most important pest ofolives in the Mediterranean region. The purified enzyme shows asingle band afterSDS-PAGE analysis, corresponding to subunit mass of 26 kDa. Thenative ADH showsa molecular mass of 48 kDa, after gel filtration HPLC analysis.The purificationmethod incorporated a preliminary ammonium sulphate precipitation step,followedby an anion-exchange DEAE chromatography step, a dye affinitychromatography stepon Cibacron blue 3GA, and an anion-exchange DEAE chromatography stepemployingthe same column of the first step. The present method offers goodoverall recovery(40%) and high enzyme purity, and it is applicable to differentgenotypes. Furthermore,the method is rapid and economical, as it employs two cheap, widelyused, andcommercially available chromatography materials.