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

Sforzato di Valtellina DOCG is a special dry red wine produced from partially dehydrated Nebbiolo wine-grapes growing in the Rhaetian Alps valley of Valtellina (Lombardy, Italy). Valtellina terraced vineyards are located at an altitude of 350–800 m according to ‘heroic’ viticulture on steep slopes. The harvested grape bunches are naturally dehydrated indoors, where a slow and continuous withering occurs (about 20% w/w of weight loss), until at least 1st December when the grapes reach the desired sugar content and can be processed following a normal winemaking with maceration.

The presence of grape-derived heat unstable proteins can lead to haze formation in white wines [1], an instability prevented by removing these proteins by adding bentonite, a hydrated aluminum silicate that interacts electrostatically with wine proteins leading to their flocculation. Despite effective, using bentonite has several drawbacks as the costs associated with its use, the potential negative effects on wine quality, and its environmental impact, so that alternative solutions are needed.

Hanseniaspora vineae (Hv) is a fermenting non-Saccharomyces yeast that compared to Saccharomyces cerevisiae (Sc) present some peculiar features on its metabolism that make it attractive for its use in wine production. Among them, it has been reported a faster yeast lysis and release of polysaccharides, as well as increased ß-glucosidase activity. Hv also produces distinctive aroma compounds, including elevated levels of fermentative compounds such as ß-phenylethyl acetate and norisoprenoids like safranal. However, it is known for its high nutritional requirements, resulting in prolonged and sluggish fermentations, even when complemented with Sc strain and nutrients.

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.

Haze formation in wine during transportation and storage is an important issue for winemakers, since turbid wines are unacceptable for sale. Such haze often results from aggregation of unstable grape proteinaceous colloids. To date, foreseeably unstable wines need to be treated with bentonite to remove these, while excessive quantities, which are often required, affect the wine volume and quality (Cosme et al. 2020). One solution to avoid these drawbacks might be the use of peptidases. Marangon et al. (2012) reported that Aspergillopepsins I and II were able to hydrolyse the respective haze-relevant proteins in combination with a flash pasteurisation. In 2021, the OIV approved this enzymatic treatment for wine stabilisation (OIV-OENO 541A and 541B).