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Am. J. Enol. Vitic. 41:4:295-300 (1990)
Copyright © 1990 by the American Society for Enology and Viticulture.
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Wine Benzyl Alcohol Oxidase: Isolation and Characterization

M. Goetghebeur 1, M. Nicolas 2, A. Blaise 1, S. Brun 1, and P. Galzy 3

1 Centre de Formation et de Recherche en Oenologie, Faculté de Pharmacie, 15 Av. Charles Flahault, 34060 Montpellier, France
2 Laboratoire de Biologie Cellulaire et Moléculaire, INSERM U 65 USTL, Place E. Bataillon, 34095 Montpellier, France
3 Laboratoire de Microbiologie Industrielle et de Génétique des Microorganismes, Place P. Viala, 34060 Montpellier, France

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.

Key words: bitter almond flavor, benzyl alcohol oxidase, enzyme extraction, wine flavor

Submitted on April 16, 1990






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Copyright © 1990 by the American Society for Enology and Viticulture.