Evelyne Aguera

Impact of oxygen addition during enological fermentation on sterol contents in yeast lees and their reactivity towards oxygen.

During enological fermentations, superfluous oxygen consumption by yeast cells is observed. The superfluous oxygen consumed by the yeast cells is mainly related to the operation of non-respiratory oxygen consumption pathways resulting in an overall decrease in the total sterol fraction in yeast. On the other hand, yeast lees remaining at the end of alcoholic fermentations exhibit specific oxygen utilization rates ranging from 1 to 4 micromol O2 h- 10(-10) cells from the second to the thirteenth month of wine aging. This oxygen consumption capacity of yeast lees was independent of residual cell viability. In this study, we investigated the potential relationship between the oxygen added to commercial yeast strains during enological fermentation and the capacity of the corresponding yeast lees to interact with oxygen. Additions of low (7 mg l(-)) and excess (37 mg l(-1)) amounts of oxygen at the end of the cell growth phase were compared in terms of repercussions on the oxygen consumption activity of the corresponding yeast lees. As expected, the superfluous oxygen consumption by yeast cells during fermentation had a positive influence on the fermentation kinetics and increased cell biomass formation. Oxygen consumption rates and the total capacity of oxygen consumption by the corresponding yeast lees clearly decreased when oxygen was added during fermentation. This marked decrease in yeast lees reactivity towards oxygen was concomitantly related to an increase in ergosterol synthesis and to oxygen-dependent sterol degradation. Such degradation occurred when oxygen was added in excess. Therefore, oxygenation control during fermentation appears to be a potential way to optimize both the fermentation kinetics and control yeast lees reactivity towards oxygen. For practical applications, oxygenation control during alcoholic fermentation may be considered as a general tool for decreasing the highly reductive effect of yeast lees during wine aging.

A comparison of laboratory and pilot-scale fermentations in winemaking conditions

We investigated the influence of the fermenter size on alcoholic fermentation. Experiments were carried out at pilot scale, in 100-L fermenters, and at laboratory scale, in stirred and static 1-L fermenters. Two musts, Grenache blanc and Sauvignon, were fermented with and without the addition of solid particles from grape musts. Highly clarified must fermentation kinetics was strongly affected by the scale of the experiment, with slower fermentation occurring in the 100-L fermenter. Alcohol, ester, and thiol synthesis in clarified sauvignon must fermentation was also strongly correlated with the fermentation scale. Addition of solid particles from grape tended to reduce the effects on kinetics associated with increasing the scale of the fermentation, by increasing the maximum rate of CO2 production, and by shortening the duration of fermentation. The addition of such particles also decreased the effects of scaling up the fermentation on the concentration of some volatile compounds, i.e., isoamyl acetate, ethyl octanoate, but did not decrease this effect for other compounds, such as isobutyl acetate, isobutanol, and 3-mercaptohexanol.

Pilot-scale evaluation the enological traits of a novel, aromatic wine yeast strain obtained by adaptive evolution

In the competitive context of the wine market, there is a growing interest for novel wine yeast strains that have an overall good fermentation capacity and that contribute favorably to the organoleptic quality of wine. Using an adaptive evolution strategy based on growth on gluconate as sole carbon source, we recently obtained wine yeasts with improved characteristics in laboratory-scale fermentations. The characteristics included enhanced fermentation rate, decreased formation of acetate and greater production of fermentative aroma. We report an evaluation of the potential value of the evolved strain ECA5™ for winemaking, by comparing its fermentation performance and metabolite production to those of the parental strain in pilot-scale fermentation trials, with various grape cultivars and winemaking conditions. We show that the evolved strain has outstanding attributes relative to the parental wine yeast strain, and in particular the production of less volatile acidity and greater production of desirable volatile esters, important for the fruity/flowery character of wines. This study highlights the potential of evolutionary engineering for the generation of strains with a broad range of novel properties, appropriate for rapid application in the wine industry.

Dynamics and quantitative analysis of the synthesis of fermentative aromas by an evolved wine strain of Saccharomyces cerevisiae

We performed a dynamic and quantitative analysis of the synthesis of fermentative aromas by an aromatic wine yeast, ECA5, obtained by adaptive evolution. During fermentation at pilot scale on synthetic and natural musts, ECA5 produced volatile compounds (higher alcohols and their acetates, ethyl esters) at higher rates than the ancestral strain, with the exception of propanol. Marked differences in the chronology of synthesis of several compounds were observed between the two strains. Overproduction of phenyl ethanol occurred mainly during the growth phase for ECA5, consistent with its higher flux through the pentose phosphate pathway, which plays a key role in biosynthetic processes. The kinetics of production of isobutanol and isoamyl alcohol were differently affected by different media (synthetic or natural must) and, in particular, according to the nature of the sterols in the media (ergosterol or phytosterols). We also observed differences in the chronology of synthesis of ethyl acetate and isoamyl acetate or ethyl esters, suggesting that the regulation of the synthesis of these compounds in the evolved strain differs from that in the ancestral strain. This study shows that a dynamic analysis of volatile compounds, using high acquisition frequency online gas chromatography, can provide novel insights into the synthesis and regulation of aromas and is thus a potentially powerful tool for strain characterization. Copyright

Comprehensive Study of the Evolution of the Gas–Liquid Partitioning of Acetaldehyde during Wine Alcoholic Fermentation

Determining the gas-liquid partitioning ( Ki) of acetaldehyde during alcoholic fermentation is an important step in the optimization of fermentation control with the aim of minimizing the accumulation of this compound, which is responsible for the undesired attributes of green apples and fresh-cut grass in wines. In this work, the effects of the main fermentation parameters on the Ki of acetaldehyde were assessed. Ki values were found to be dependent on the temperature and composition of the medium. A nonlinear correlation between the evolution of the Ki and fermentation progress was observed, attributable to the strong retention effect of ethanol at low concentrations, and it was demonstrated that the partitioning of this specific molecule was not influenced by the CO2 production rate. A model was developed that quantifies the Ki of acetaldehyde with a very accurate prediction, as the difference between the observed and predicted values did not exceed 9%.

Interest in Online Higher Alcohol and Ester Determinations during Winemaking Fermentations

To access the kinetics of synthesis of higher alcohols and esters during the alcoholic fermentation process, we developed an original online gas chromatography device. The high frequency measurement made it possible, for the first time, to calculate rates and specific rates of production. In parallel, it was possible to perform dynamic gas–liquid balances. Losses highly depended on the studied compound. They were negligible for the higher alcohols but high for the esters, even at moderate temperatures.