OPTIMIZATION, VALIDATION AND APPLICATION OF THE EPR SPIN-TRAPPING TECHNIQUE TO THE DETECTION OF FREE RADICALS IN CHARDONNAY WINES

Brettanomyces bruxellensis is considered as the main spoilage yeast in oenology. Its presence in red wine leads to off-flavour due to the production of volatile phenols such as 4-vinylphenol, 4-vinylguaiacol, 4-ethylphenol and 4-ethylguaiacol, whose aromatic notes are unpleasant (e.g. animal, leather, horse or pharmaceutical). Beside wine, B. bruxellensis is commonly isolated from beer, kombucha and bioethanol production, where its role can be described as negative or positive. Recent genomic studies unveiled the existence of various populations.

Cognac is a famous spirit produced in southwest France in the region of the eponymous town from wines mainly from Vitis vinifera cv. Ugni blanc. This variety gives very acidic and poorly aromatic base wines for distillation which are produced according to a very specific procedure. Grapes are picked at low sugar concentrations ranging 13-21 °Brix and musts with high turbidity (>500 NTU) are fermented without sulphite addition [1]. Fermentative aromas, as esters and higher alcohols, are currently the main quality markers considered in Cognac spirits.

Oenococcus oeni is the main Lactic Acid Bacteria responsible for malolactic fermentation, converting malic acid into lactic acid and carbon dioxide in wines. Following the alcoholic fermentation, this second fermentation ensures a deacidification and remains essential for the release of aromatic notes and the improvement of microbial stability in many wines. Nevertheless, wine is a harsh environment for microbial growth, especially because of its low pH (between 2.9 and 3.6 depending on the type of wine) and nutrient deficiency. In order to maintain homeostasis and ensure viability, O. oeni possesses different cellular mechanisms including organic acid metabolisms which represent also the major pathway to synthetize energy in wine.

Botrytis cinerea Pers., the causal agent of grey mould disease, is responsible for substantial economic losses, as it causes reduction of grape and wine quality and quantity. Exploitation of antagonistic yeasts is a promising strategy for controlling grey mould incidence and limiting the usage of synthetic fungicides. In our previous studies, 119 different indigenous yeasts were screened for putative multidimensional modes of action against filamentous fungus B. cinerea [1]. The most promissing biocontrol yeast was Pichia guilliermondii ZIM624, which exhibited several anatagonistic traits (production of cell wall degrading enzymes, chitinase and β-1,3-glucanase; demonstration of in vitro inhibitory effect on B. cinerea mycelia radial growth; production of antifungal volatiles, assimilation of a broad diversity of carbon sources, contributing to its competitivnes in inhabiting grapes in nature).

Yeast cell walls (CWs) may adsorb wine components with a significant impact on wine quality. When dealing with red wines, this adsorption is mainly related to physicochemical interactions between wine polyphenols and cell wall mannoproteins. However, mannoproteins are a heterogeneous family of complex peptidoglycans including long and highly branched N-linked oligosaccharides and short linear O-linked oligosaccharides, resulting in a huge structural diversity.