PURIFICATION OF ALCOHOL-DEHYDROGENASE FROM 4 GENOTYPES OF THE OLIVE FRUIT-FLY BACTROCERA (DACUS) OLEAE

This is the first report describing the purification of alcohol dehydr ogenase (ADH) from four genotypes of the olive fruit fly Bactrocera ol eae, the most important pest of olives in the Mediterranean region. Th e purified enzyme shows a single band after SDS-PAGE analysis, corresp onding to subunit mass of 26 kDa. The native ADH shows a molecular mas s of 48 kDa, after gel filtration HPLC analysis. The purification meth od incorporated a preliminary ammonium sulphate precipitation step, fo llowed by an anion-exchange DEAE chromatography step, a dye affinity c hromatography step on Cibacron blue 3GA, and an anion-exchange DEAE ch romatography step employing the same column of the first step. The pre sent 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 com mercially available chromatography materials.