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

Malolactic fermentation (MLF)¹, the decarboxylation of L-malic acid into L-lactic acid, is performed by lactic acid bacteria (LAB). MLF has a deacidifying effect that may compromise freshness or microbiological stability in wines² and can be inhibited by fumaric acid [E297] (FA). In wine, can be added at a maximum allowable dose of 0.6 g/L³. Its inhibition with FA is being studied as an alternative strategy to minimize added doses of SO₂⁴. In addition, wine yeasts are capable of metabolizing and storing small amounts of FA and during alcoholic fermentation (AF).

Flow cytometry (FCM) is a powerful technique allowing the detection, characterization and quantification of microbial populations in different fields of application (medical environment, food industry, enology, etc.). Depending on the fluorescent markers and specific probes used, FCM provides information on the physiological state of the cell and allows the quantification of a microorganism of interest within a mixed population. For 15 years, the enological sector has shown growing interest in this technique, which is now used to determine the populations present (of interest or spoilage) and the physiological state of microorganisms at the different stages of winemaking.

The aging potential of Burgundy chardonnay wines is considered as quality indicator. However, some of them exhibit higher oxidative sensitivity and premature oxidative aging symptoms, which are potentially induced by no-enzymatic oxidation such as Fenton-type reaction (Danilewicz, 2003). This chemical mechanism involves the action of transition metal, native phenolic compounds and oxygen which promote the generation of highly reactive oxygen species (ROS) such as hydroxyl radicals (OH) or 1-hydroxyethyl radicals (1-HER) from oxidation of ethanol. Such mechanism is involved in the radical oxidation occurring during bottle aging. According to Elias et al.,(2009a), the 1-HER is the most abundant radical in forced oxidation treated wines. Consequently, understanding its evolution kinetic in dry white wines is of great importance.

During alcoholic fermentation, nitrogen is one of essential nutrient for yeast as it plays a key role in sugar transport and biosynthesis of and wine aromatic compounds (thiols, esters, higher alcohols). The main issue of a lack in yeast assimilable nitrogen (YAN) in winemaking is sluggish or stuck fermentations promoting the growth of alteration species and leads to economic losses. Currently, grape musts are often characterized by low YAN concentration and an increase of sugars concentration due to global warming, making alcoholic fermentations even more difficult. YAN depletion can be corrected by addition of inorganic (ammonia) or organic (yeast derivatives products) nitrogen during alcoholic fermentation.

The light exposure of wine can be detrimental as a relevant loss of aromas takes place [1] and light-induced reactions can occur. The latter involves riboflavin (RF), a photosensitive compound, that is fully reduced by acquiring two electrons. When the electron-donor is methionine, the light-struck taste (LST) can appear leading to cooked cabbage, onion and garlic odours-like [2]. The use of oenological tannins can limit the appearance of LST in both model wine [3] and white wine [4]. This research aimed to evaluate the impact of certain oenological tannins, selected in a previous study as the most effective against LST [5], in both white and rosé wines.