Occurrence of Brettanomyces/Dekkera in Brazilian red wines and its correlation with ethylphenols

The yeast Brettanomyces/Dekkeracan cause important spoilage in wines, with the production of ethylphenols and other off-flavor compounds. This study aimed at determining the presence of this yeast and the ethylphenols produced by them in Brazilian red wines, establishing their relationship with other chemical characteristics. Isolates of Brettanomyces/Dekkerawere quantified by plating 126 samples of dry red wine in selective culture medium, while ethylphenols were analyzed by solid phase extraction and GC/FID. Free and total SO2, alcohol, total dry extract, residual sugar, total and volatile acidity, and pH were also determined. Brettanomyces/Dekkerawas present in 27% of samples. Ethylphenols were detected in most samples, with amounts higher than the threshold limit of 426 mg/L found in 46.03% of samples. The majority of wine samples showed inadequate levels of SO2and residual sugars, facts that might facilitate microbial spoilage. The passage in barrels and the grape varieties (Cabernet Sauvignon and Merlot), did not show any influence on the levels of contamination or ethylphenols contents. The prevalence of Brettanomyces/Dekkeraand the concentrations of ethylphenols were high considering the sensory impact they can cause. The growth of Brettanomyces/Dekkerawas dependent on the levels of SO2and alcohol of wines. Knowledge of the contamination, the presence of ethylphenols, and their relationship with the chemical characteristics of wines can entice effective measures to prevent Brettanomyces/Dekkeraand contribute to improve the general quality of Brazilian red wines.

Rapid yeast DNA extraction by boiling and freeze-thawing without using chemical reagents and DNA purification

The purpose of this work was to study a rapid yeast DNA extraction by boiling and freeze-thawing processes without using chemical reagents or any purification procedures, to obtain a high grade PCR-product. A specific DNA fragment of the 18S region of Dekkera bruxellensis and Saccharomyces cerevisiae was chosen. The described boiling and freeze-thawing protocols generated the PCR-grade product preparations and could be used to process many samples. The amplification of the fragments could be observed after 30 and 35 cycles. These processes of extraction without using any kind of chemical reagents, especial water, and purification procedures proved to be efficient, reproducible, simple, fast, and inexpensive.

Cinnamic acid, ethanol and temperature interaction on coumarate decarboxylase activity and the relative expression of the putative cd gene in D. bruxellensis

Dekkera bruxellensis is one of the main contaminating yeasts in wine due to its ability to metabolize cinnamic acids into volatile phenols. This yeast metabolizes p-coumaric acid into 4-vinylphenol through a coumarate decarboxylase (CD) and then transforms it into to 4-ethylphenol (EF) through a vinylphenol reductase. In this work we investigated the influence of the interaction between the concentration of p-coumaric acid, ferulic acid and ethanol as well as growth temperature on the production of CD activity and the expression of a putative gene that codes for this enzymatic activity. For this, a Box Behnken experimental design was used. The concentration of p-coumaric acid (5-26 ppm) and ferulic acid (3-9 ppm) alone did not show any significant effect on any of the studied response variables. However, the interaction between (ethanol concentration * cinnamic acid concentration) and (ethanol concentration * temperature) had a significant statistical effect on the production of CD activity. Additionally, a higher growth temperature negatively affected the expression of the putative cd gene and the production of CD activity. This is the first work that studies the effect of cinnamic acids on the production of CD activity and the relative expression of its putative gene, using natural concentrations of cinnamic acid found in wine.

Influence of Brettanomyces custersianus upon the activity of Saccharomyces cerevisiae strains during the tumultuous phase of vinification

The aim of this work was to evaluate the influence of Brettanomyces custersianus on the metabolic activity of Saccharomyces cerevisiae during the tumultuous stage of wine production. The Cabernet Sauvignon grape must with the skin was inoculated with individual cultures of Sacch. cerevisiae and with mixed cultures of Sacch. cerevisiae and Br. custersianus. During the 6-day tumultuous phase of fermentation, the highest ethanol production and the highest sugar consumption were obtained with the strains without B. custersianus. Fermentations carried out with the addition of Brettanomyces metabolites, acetic acid and 4-ethylphenol, showed that only the former inhibited the growth of both Sacch. cerevisiae strains used. In some cases, Br. custersianus could affect the rate higher alcohols production and their final concentrations during the tumultuous phase of vinification.