TY - JOUR T1 - Wine Benzyl Alcohol Oxidase: Isolation and Characterization JF - American Journal of Enology and Viticulture JO - Am J Enol Vitic. SP - 295 LP - 300 DO - 10.5344/ajev.1990.41.4.295 VL - 41 IS - 4 AU - M. Goetghebeur AU - M. Nicolas AU - A. Blaise AU - S. Brun AU - P. Galzy Y1 - 1990/01/01 UR - http://www.ajevonline.org/content/41/4/295.abstract N2 - A benzyl alcohol oxidase, responsible for bitter almond taste in wines, was extracted from wine with dithiothreitol (DTT: 1 mM) and glycerol (20%, v/v) present in the extraction medium. The enzyme was purified 1440-fold with 43% yield by ultrafiltration, ion exchange (Q Sepharose®fast flow), and gel filtration (Sephacryl® S 300) chromatographies. The enzyme so obtained was homogeneous in SDS PAGE gel electrophoresis. Optimum pH was 5 and optimium temperature was 30°C. The molecular weight values estimated by gel filtration chromatography and SDS PAGE gel electrophoresis are indicative of an oligomeric structure for this enzyme constituted by three identical subunits of 67 kD. Inhibition studies show that SH groups are essential for enzyme activity and that metallic cofactors are not necessary. Sulfur dioxide inhibits the enzyme activity irreversibly, and the inhibition effect of ethanol is reversible. ER -