Aging of wine in wooden barrels improves the final product both visually and olfactorily. However, common contamination yeasts such as Brettanomyces bruxellensis (Brett) burrowed in the wooden barrels can render the wine undrinkable. Brett uses its vinylphenol reductase enzyme to reduce natural, desirable vinylphenols to ethylphenols, causing the wine to acquire an unpleasing odor. Our research marks the first protein expression and purification protocols for recombinant Brett vinylphenol reductase. Ligation independent cloning was used to place the vinylphenol reductase gene into an inducible protein expression vector. Following cloning confirmation through PCR and DNA sequencing, the enzyme was purified using affinity and gel filtration chromatography. Currently, full structural and functional characterization is being pursued through protein X-ray crystallography and reductase assay development.  By understanding this protein at the molecular level, Brett's detailed mechanism for wine spoilage will be revealed, which may lead to solutions for eradicating Brett wine contamination.

Understanding wine contamination by studying a new <i>Brettanomyces </i>recombinant enzyme

243rd National Meeting (2012)

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Understanding wine contamination by studying a new Brettanomyces recombinant enzyme