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

Powdery mildew, caused by Erysiphe necator, is a widespread disease that causes high economical losses in viticulture. The main strategy to control the disease is the recurrent application of sulphur based phytochemical compounds. However, in order to reduce their accumulation in the environment and promote the sustainability of the sector, the European Commission has applied restrictions to the number of pesticide treatments and the maximum quantity of fungicides to be applied in viticulture. Seaweeds, in particular macroalgae, are marine resources rich in sulphated polysaccharides with bio-protective potential for the plant, representing an environmentally-friendly alternative approach for sustainable wine production.

Hydroxytyrosol (HT) is a phenolic compound present in olives, virgin olive oil and wine. HT has attracted great scientific interest due to its biological activities which have been related with the ortho-dihydroxy conformation in the aromatic ring. In white and red wines, HT has been detected at concentrations ranging from 0.28 to 9.6 mg/L and its occurrence has been closely related with yeast metabolism of aromatic amino acids by Ehrlich pathway during alcoholic fermentation. One of the most promising properties of this compound is the neuroprotective activity against pathological mechanisms related with neurode-generative disorders including Alzheimer’s and Parkinson’s disease.

Aging wine on lees is a consolidated practice during which some yeast components (e.g., polysaccharides,
proteins, peptides) are released and solubilized in wine thus, affecting its stability and quality.
Apart from the widely studied mannoproteins, the role of other yeast components in modulating wine
characteristics is still scarce. Wine peptides have been studied for their contribution to taste, antioxidant,
and antihypertensive potentials. However, the peptides detected in wine can be influenced by the
interaction between yeasts and grape components.

Most of the effects of ultrasound (US) result from the collapse of bubbles due to cavitation. The shockwave produced is associated with shear forces, along with high localised temperatures and pressures. However, the high-speed stream, radical species formation, and heat generated during sonication may also affect the stability of some enzymes and proteins, depending on their chemical structure. Recently, Ce-lotti et al. (2021) reported the effects of US on protein stability in wines. To investigate this further, the effect of temperature (40°C and 70°C; 60s), sonication (20 kHz and 100 % amplitude, for 20s and 60s, leading to the same temperatures as above, respectively), in combination with Aspergillopepsins I (AP-I) supplementation (100 μg/L), was studied on unstable protein concentration (TLPs and chitinases) using HPLC with an UV–Vis detector in a TLPs-supplemented model system and in an unstable white wine.

In a recent study several genes controlling the acidification properties of the wine yeast Saccharomyces cerevisiae have been identified by a QTL approach [1]. Many of these genes showed allelic variations that affect the metabolism of malic acid and the pH homeostasis during the alcoholic fermentation. Such alleles have been used for driving genetic selection of new S. cerevisiae starters that may conversely acidify or deacidify the wine by producing or consuming large amount of malic acid [2]. This particular feature drastically modulates the final pH of wine with difference of 0.5 units between the two groups.