RED WINE AGING THROUGH ¹H-NMR METABOLOMICS

Enzymatic browning (BE) of must is caused by polyphenol oxidases (PPOs), tyrosinase and laccase. Both PPOs can oxidize diphenols such as hydroxycinnamic acids (HA) to quinones, which can later polymerize to form melanins [1], which are responsible of BE in white wines and of oxidasic haze in red wines. SO₂ is the main tool used to protect must from BE thanks to its capacity to inhibit PPOs [2]. However, the current trend in winemaking is to reduce and even eliminate this unfriendly additive. Among the different possible alternatives for protecting must against BE, the inoculation with a selected Metschnikowia pulcherrima MP1 is without any doubt one of the most promising ones.

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).

The fresh mushroom off-flavour (FMOff) has been appearing in wines since the 2000s. Some C8 compounds such as 1-octen-3-one, 1-octen-3-ol, 1-hydroxyoctan-3-one, 3-octanol and others are involved in this specific off-flavour [1-3]. At the same time, glycosidic precursors of some FMOff compounds have been identified in musts contaminated by Crustomyces subabruptus [4], highlighting the role of aroma precursors in this specific taint. However, the fate of these volatile molecules and glycosidic fractions during fermentation is not well known.

Right after the pouring of wine in a glass, a myriad of volatile organic compounds, including ethanol, overwhelm the glass headspace, thus causing the so-called wine’s bouquet [1]. Otherwise, it is worth noting that during wine tasting, most people automatically swirl their glass to enhance the release of aromas in the glass headspace [1]. About a decade ago, Swiss researchers revealed the complex fluid mechanics underlying wine swirling [2]. However, despite mechanically repeated throughout wine tasting, the consequences of glass swirling on the chemical space found in the headspace of wine glasses are still barely known.

In this work we briefly present a microfluidic device aiming to sort yeast cells according to their morphology. The technology is based upon microfluidic chips made out of Polydimethylsiloxane and glass using soft lithography processes and replica molding. The microfluidic device was used for encapsulating single yeast cells in liquid droplets containing growth medium. Liquid droplet containing yeast cells were sorted using a real time imaging and decision-making process.