Directed Evolution of Oenococcus oeni: optimising yeast-bacteria interactions for improved malolactic fermentation

Several winemaking operations, such as filtration, pumping, and racking, are known to potentially facilitate the incorporation of atmospheric O2 into the wine. Control of grape must oxidation is one key aspect in the management of white wine aroma expression, color stability and shelf-life extension. On the one hand, controlled must oxidation may help to remove highly reactive phenolic compounds, which otherwise could contribute to premature oxidation. And on the other hand, in certain cases of extreme protection of the must from O2 (e.g. pressing under inert atmosphere), it can help to preserve varietal aromas and natural must antioxidants.

Recent studies have demonstrated the role of certain lactones, particularly 2-nonen-4-olide, and volatile thiols (3-sulfanylhexan-1-ol) in the over ripped aromas of noble rot sweet wines (Stamatopoulos et al. 2014ab). These compounds are partly formed during the maturation and under the activity of B. cinerea on grapes. This research was carried out in the vineyard of Sauternes with aim to better understand their genesis depending on the grape over-ripening on two different soil types during 3 vintages. Thus, the study was conducted, with the Sémillon grape, during vintages 2012, 2014 & 2015, at 4 stages of over-maturation of the grapes (healthy, pourri plein, pourri roti, pourri roti + 15 days) considering two vineyard plots with different soil characteristics (calcosol & peyrosol) planted with the 315 Sémillon clone and grafted on 101-14 rootstock respectively in 1981 and 1980 and cultivated with the same vineyard management. Volatile lactones were assayed by liquid-liquid extraction followed by GC/MS analysis and the precursors of 3-sulfanylhexanol by an adaptation of the method by Capone et al. 2010 (SPE-
UPLC/FTMS).

Colour is an important quality parameter in wines and is the result of a complex mixture of pigments
(including anthocyanins and their derivatives, quinones, xanthyllium compounds, etc.). Red wine colour changes over time as pigments react between themselves and with other wine macromolecules
(particularly polyphenols). During wine tasting, colour is normally assessed on the outer rim of the wine profile in a tilted glass, since most wines are too opaque to be analysed in the middle of the glass. Therefore, depending on the depth of observation considered, the perception of wine colour can be different.

Glutathione is a tripeptide (γ-Glu-Cys-Gly), which can occur in grapes, in must and in wine prevalently in the reduced form as well as in the oxidized form as glutathione disulfide. The importance of the reduced form of glutathione lies in its antioxidant activity. In must, it limits browning by reducing o-quinones produced by polyphenol oxidase activity on hydroxycinnamic acids; in wine, it exerts a protective effect on various aromatic compounds. Glutathione concentration in wine is lower than in grape juice and variable as it depends on several factors, ranging from the native content of grapes to winemaking technique.

The effectiveness of enzyme-mediated maceration processes in red winemaking relies on a clear picture of the target (berry cell wall structure) to achieve the optimum combination of specific enzymes to be used. However, we lack the information on both essential factors of the reaction (i.e. specific activities in commercial enzyme preparation and the cell wall structure of berry tissue). In this study, the different combinations of pure recombinant enzymes and the recently validated high throughput cell wall profiling tools were applied to extend our knowledge on the grape berry cell wall polymeric deconstruction during the winemaking following a combinatorial enzyme treatment design.