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

Grape (Vitis vinifera) is one of the world’s oldest agricultural fruit crops, grown for wine, table grape, raisin, and other products. One of the factors that can cause a reduction in the grape growing area is temperature rise due to climate change. Elevated temperature causes changes in grapevine phenology and fruit chemical composition. Previous studies showed that grape varieties respond differently to a temperature shift of 1.5°C; few varieties had difficulties in the fruit development or could not reach the desired Brix level.

Wine production is a complex biochemical process that involves a heterogeneous microbiota consisting of different microorganisms such as yeasts, bacteria, and filamentous fungi. Among these microorganisms, yeasts play a predominant role in the chemistry of wine, as they actively participate in alcoholic fermentation, a biochemical process that transforms the sugars in grapes into ethanol and carbon dioxide while producing additional by-products. The quality of the final product is greatly influenced by the microbiota present in the grape berry, and the demand for indigenous yeast starters adapted to specific grape must and reflecting the biodiversity of a particular region is increasing. This supports the concept that indigenous yeast strains can be associated with a “terroir”.

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).

Infection of grapes (Vitis vinifera) by Botrytis cinerea (grey mould) is a frequent occurrence in vineyards and during prolonged wet and humid conditions can lead to significant detrimental impact on yield and overall quality. Growth of B. cinerea causes oxidisation of phenolic compounds resulting in a loss of colour and formation of a suite of off-flavours and odours in wine made from excessively infected fruit. Apart from wine grapes, developing post-harvest B. cinerea infection in high-value horticultural products during storage, shipment and marketing may cause significant loss in fresh fruits, vegetables and other crops. A rapid and sensitive assessment method to detect, screen and quantify fungal infection would greatly assist viticultural growers and winemakers in determining fruit quality.

Nowadays, the rapid growth of vineyards with organic practices and the use of copper as the only fun-gicide against downy mildew raises again the question of the effect of copper on varietal thiols in wine, especially 3-sulfanylhexan-1-ol (3SH) and its acetate (3SHA). A few decades ago, several works indicated that the use of copper in the vineyard had a negative effect on the content of varietal thiols in Sauvignon blanc wines [1, 2]. However, these studies only considered the concentration of the reduced form (RSH) of varietal thiols, without quantifying the oxidised ones. For this purpose, we proposed to monitor both reduced and oxidised forms of varietal thiols in wine under copper stress during alcoholic fermentation to have a more complete picture of the biological and chemical mechanisms.