Volatile sulfur compounds can be formed at various stages during wine production and storage, and some may impart unpleasant "reduced" aromas to wine when present at sensorially significant concentrations. Quantitative data are necessary to understand factors that influence the formation of volatile sulfur compounds, but their analysis is not a trivial undertaking. A rapid and selective method for determining 10 volatile sulfur-containing aroma compounds in wine that have been linked to "off-odors" has been developed. The method utilizes static headspace injection and cool-on-column gas chromatography coupled with sulfur chemiluminescence detection (GC-SCD). Validation demonstrated that the method is accurate, precise, robust, and sensitive, with limits of quantitation around 1 microg/L or better, which is below the aroma detection thresholds for the analytes. Importantly, the method does not form artifacts, such as disulfides, during sample preparation or analysis. To study the contribution of volatile sulfur compounds, the GC-SCD method was applied to 68 commercial wines that had reductive sensory evaluations. The analytes implicated as contributors to reductive characters were hydrogen sulfide, methanethiol, and dimethyl sulfide, whereas carbon disulfide played an uncertain role.

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ABSTRACTSaccharomyces cerevisiaehas evolved a highly efficient strategy for energy generation which maximizes ATP energy production from sugar. This adaptation enables efficient energy generation under anaerobic conditions and limits competition from other microorganisms by producing toxic metabolites, such as ethanol and CO2. Yeast fermentative and flavor capacity forms the biotechnological basis of a wide range of alcohol-containing beverages. Largely as a result of consumer demand for improved flavor, the alcohol content of some beverages like wine has increased. However, a global trend has recently emerged toward lowering the ethanol content of alcoholic beverages. One option for decreasing ethanol concentration is to use yeast strains able to divert some carbon away from ethanol production. In the case of wine, we have generated and evaluated a large number of gene modifications that were predicted, or known, to impact ethanol formation. Using the same yeast genetic background, 41 modifications were assessed. Enhancing glycerol production by increasing expression of the glyceraldehyde-3-phosphate dehydrogenase gene,GPD1, was the most efficient strategy to lower ethanol concentration. However, additional modifications were needed to avoid negatively affecting wine quality. Two strains carrying several stable, chromosomally integrated modifications showed significantly lower ethanol production in fermenting grape juice. Strain AWRI2531 was able to decrease ethanol concentrations from 15.6% (vol/vol) to 13.2% (vol/vol), whereas AWRI2532 lowered ethanol content from 15.6% (vol/vol) to 12% (vol/vol) in both Chardonnay and Cabernet Sauvignon juices. Both strains, however, produced high concentrations of acetaldehyde and acetoin, which negatively affect wine flavor. Further modifications of these strains allowed reduction of these metabolites.

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During alcoholic fermentation of grape sugars, wine yeasts produce a range of secondary metabolites that play an important role in the aroma profile of wines. In this study, we have explored the ability of a large number of wine yeast strains to modulate wine aroma composition, focusing on the release of the “fruity” thiols 3-mercaptohexan-1-ol (3-MH) and 4-mercapto-4-methylpentan-2-one (4-MMP) from their respective cysteinylated nonvolatile precursors. The role of the yeast geneIRC7in thiol release has been well established, and it has been shown that a 38-bp deletion found in many wine strains cause them to express a truncated version of Irc7p that does not possess cysteine-S-conjugate β-lyase activity. In our data, we find thatIRC7allele length alone does not fully explain the capacity of a strain to release thiols. Screening of a large number of strains coupled with analysis of genomic sequence data allowed us to identify several previously undescribed single-nucleotide polymorphisms (SNPs) inIRC7that, when coupled with allele length, more robustly explain the ability of a particular yeast strain to release thiols from their cysteinylated precursors. We also demonstrate that allelic variation ofIRC7not only affects the release of thiols but modulates the formation of negative volatile sulfur compounds from the amino acid cysteine. The results of this study provide winemakers with an improved understanding of the genetic determinants that affect wine aroma and flavor, which can be used to guide the choice of yeast strains that are fit for purpose.IMPORTANCEVolatile sulfur compounds contribute to wine aromas that may be considered pleasant, such as “tropical,” “passionfruit,” and “guava,” as well as aromas that are considered undesirable, such as “rotten eggs,” “onions,” and “sewer.” During fermentation, wine yeasts release some of these compounds from odorless precursor molecules, a process that is most efficient when performed by yeasts that express active forms of the protein Irc7p. We show that most wine yeasts carry mutations that reduce activity of this protein, affecting the formation of volatile sulfur compounds that impart both pleasant and unpleasant aromas. The results provide winemakers with guidance on the choice of yeasts that can emphasize or deemphasize this particular contribution to wine quality.