RESUMO
AIMS: Dekkera bruxellensis and Pichia guilliermondii are contaminating yeasts in wine due to the production of phenolic aromas. Although the degradation pathway of cinnamic acids, precursors of these phenolic compounds has been described in D. bruxellensis, no such pathway has been described in P. guilliermondii. METHODS AND RESULTS: A molecular and physiological characterization of 14 D. bruxellensis and 15 P. guilliermondii phenol-producing strains was carried out. Both p-coumarate decarboxylase (CD) and vinyl reductase (VR) activities, responsible for the production of volatile phenols, were quantified and the production of 4-vinylphenol and 4-ethylphenol were measured. All D. bruxellensis and some P. guilliermondii strains showed the two enzymatic activities, whilst 11 of the 15 strains of this latter species showed only CD activity and did not produce 4-EP in the assay conditions. Furthermore, PCR products obtained with degenerated primers showed a low homology with the sequence of the gene for a phenyl acrylic acid decarboxylase activity described in Saccharomyces cerevisiae. CONCLUSIONS: D. bruxellensis and P. guilliermondii may share a similar metabolic pathway for the degradation of cinnamic acids. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first work that analyses the CD and VR activities in P. guilliermondii, and the results suggest that within this species, there are differences in the metabolization of cinnamic acids.
Assuntos
Cinamatos/metabolismo , Dekkera/metabolismo , Pichia/metabolismo , Vinho/microbiologia , Carboxiliases/metabolismo , Dekkera/genética , Contaminação de Alimentos , Oxirredutases/metabolismo , Fenóis/análise , Pichia/genética , Reação em Cadeia da Polimerase , Técnica de Amplificação ao Acaso de DNA PolimórficoRESUMO
AIMS: To investigate whether the presence of Pichia guilliermondii impacts on the production of volatile phenols from mixed wine fermentations with Dekkera bruxellensis and Saccharomyces cerevisiae. METHODS AND RESULTS: Four inoculation strategies were performed in small-scale fermentations involving P. guilliermondii, D. bruxellensis and S. cerevisiae using Syrah grape juice supplemented with 100 mg l(-1) of p-coumaric acid. High pressure liquid chromatography was used for the quantification or volatile phenols. Significant high levels of 4-ethylphenol and 4-ethylguaicol (720 and 545 microg l(-1), respectively), as well as the highest levels of 4-vinylphenol (>4500 microg l(-1)), were observed when P. guilliermondii species was inoculated from the beginning of the fermentation. CONCLUSIONS: The metabolic interaction occurring between the high vinylphenol producer species P. guilliermondii and D. bruxellensis exhibiting a high vinylphenol reductase activity resulted in an increased production of volatile phenols in wine. SIGNIFICANCE AND IMPACT OF THE STUDY: Pichia guilliermondii must be considered a very important spoilage yeast in the wine industry capable of producing large amounts of volatile phenols.