IMPACT OF MANNOPROTEIN N-GLYCOSYL PHOSPHORYLATION AND BRANCHING ON WINE POLYPHENOL INTERACTIONS WITH YEAST CELL WALLS

Botrytis cinerea Pers., the causal agent of grey mould disease, is responsible for substantial economic losses, as it causes reduction of grape and wine quality and quantity. Exploitation of antagonistic yeasts is a promising strategy for controlling grey mould incidence and limiting the usage of synthetic fungicides. In our previous studies, 119 different indigenous yeasts were screened for putative multidimensional modes of action against filamentous fungus B. cinerea [1]. The most promissing biocontrol yeast was Pichia guilliermondii ZIM624, which exhibited several anatagonistic traits (production of cell wall degrading enzymes, chitinase and β-1,3-glucanase; demonstration of in vitro inhibitory effect on B. cinerea mycelia radial growth; production of antifungal volatiles, assimilation of a broad diversity of carbon sources, contributing to its competitivnes in inhabiting grapes in nature).

Non-Saccharomyces yeasts not only increase microbial diversity during wine fermentation, but also have a positive effect on improving wine aroma. Among these non-Saccharomyces yeast species, Metschnikowia pulcherrima is often studied and used in winemaking in recent years, but its application in icewine has been rarely reported. In this study, indigenous M. pulcherrima strains and Saccharomyces cerevisiae strains (commercial and indigenous strains) were sequentially inoculated for icewine fermentations; meanwhile, pure S. cerevisiae fermentations were used as the control; indigenous strains used above were screened from spontaneous fermentations of Vidal blanc icewine.

The composition of grape berry cell walls was studied on two grape varieties, two years and two maturation levels at the same time as the extraction of anthocyanins and tannins. The chemical composition of skins, seeds, and pulps, focused on polyphenols and polysaccharides, was compared to the chemical composition in polyphenols after extraction from the skins in model solutions or after wine making of the berries. Polyphenols were mainly characterized by UPLC-MS and HPLC-SEC. Polysaccharides were characterized by analysis of the neutral sugar compositions, and also by the CoMPP (comprehensive micropolymer profiling) analysis, a new method which targets the functional groups of cell wall polysaccharides.

The positive contribution of toasted vine-shoots (SEGs, Shoot from vines – Enological – Granule) used in winemaking to the chemical and sensory profile of wines has been widely proven. However, the combination of this new enological tool with other winemaking technologies, such as micro-oxygenation (MOX), has not been studied so far. It is known that micro-oxygenation is used in wineries to stabilizes color, improves structure or combining with oak alternatives products to achieve a more effective aroma integration of wines. For that, its implementation in combination with SEGs could result in differentiated wines.

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.