INVESTIGATION OF MALIC ACID METABOLIC PATHWAYS DURING ALCOHOLIC FERMENTATION USING GC-MS, LC-MS, AND NMR DERIVED 13C-LABELED DATA

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.

The alteration in the rainfall pattern and the increase in the temperatures associated to global climate change are already affecting wine production in many viticultural regions all around the world (1). In fact, grapes are nowadays ripening earlier from a technological point of view than in the past, but they are not necessarily mature from a phenolic point of view. Consequently, the wines made from these grapes can be unbalanced or show high alcohol content. Dramatic shifts in viticultural areas are currently being projected for the future (2).

In recent years, consumer demand for products without chemical additives increased, becoming a priority for the wine sector. SO₂ is widely used for its multiple properties including antiseptics, antioxidants and antioxidasics and the strategy of bioprotection in winemaking represents now an alternative to this chemical additive. In oenology, results have highlighted the interest of bioprotection to limit the development of microorganisms like Hanseniaspora uvarum and thus reduce the doses of sulphite. Indeed, this species is considered because of its acetic acid and methyl butyl acetate production, the latter can cover the varietal character of wines.

Understanding the composition of wine and how it is influenced by climate or wine-making practices is a challenging issue. Two approaches are typically used to explore this issue. The first approach uses chemical
fingerprints, which require advanced tools such as high-resolution mass spectrometry and multidimensional chromatography. The second approach is the targeted method, which relies on the widely available 1-D GC/MS, but involves integrating the areas under a few peaks which ends up using only a small fraction of the chromatogram.

During red wine vinification, maceration allows the must, and consequently the wine, to be enriched with several compounds that contribute to the creation of the typical organoleptic characteristics of red wines. Among these, extraction of polyphenols (PPs) during maceration is a major process of enological interest.
The purpose of this study was the evaluate the suitability of a rapid analytical approach based in linear sweep voltammetry to monitor PPs extraction during vinification.