Impact of mannoprotein N-glycosyl phosphorylation and branching on the sorption of wine polyphenols by yeasts and yeast cell walls.

Yeast cell walls (CWs) may adsorb red wine polyphenols with a significant impact on wine quality. This adsorption is mainly attributed to physicochemical interactions with mannoproteins. However, the mannoprotein structural features that promote polyphenol adsorption are not clearly established yet. The impact of mannosylphosphorylation and mannan backbone branching on polyphenol adsorption by yeast was studied through adsorption isotherms with Saccharomyces cerevisiae BY4742 wild-type and mnn4 and mnn2 mutants (EUROSCARF). Polymeric and oligomeric tannins and derived pigments were preferentially adsorbed by the yeasts and their corresponding CWs. Mannoprotein N-glycosyl phosphorylation appeared as the key groupments involved in polyphenol adsorption by CWs. When dealing with whole yeasts, behaviors were different, due to the coupled phenomena of polyphenol adsorption at cell wall level and their diffusion inside the yeast cytoplasm. The key role of phosphorylation observed in this work is of interest in the modulation of wine color and astringency by yeasts.

Effect of grape must polyphenols on yeast metabolism during alcoholic fermentation.

In red winemaking, polyphenols from grape berry pericarp and seed are extracted during fermentation and their interactions with yeast have been widely demonstrated. However, information concerning the impact of extracted polyphenols on yeast metabolism during fermentation is missing. The aim of this study was to further explore interactions between yeasts and polyphenols and to identify their effects on yeast metabolism and fermentation kinetics. This impact was studied in synthetic musts for four commercial Saccharomyces cerevisiae wine strains, using polyphenols purified from a thermovinification must, in both stressed (phytosterol deficient medium) and non-stressed conditions. Interactions between grape polyphenols and yeast cells were substantiated from the early stage of fermentation by means of epifluorescence and confocal microscopy. If these interactions were limited to yeast cell walls in non-stressed conditions, the passage of polyphenols through yeast envelope and their accumulation in the intracellular space of living cells was shown in phytosterol-deficient medium. Whatever the conditions used (stressed and non-stressed conditions) and for all strains, the presence of polyphenols led to a significant decrease of cell growth (50%), CO2 production rate (60 to 80%) and nitrogen consumption (3 to 4 times less), resulting in increased fermentation lengths. The perturbation of yeast growth and metabolism due to polyphenol compounds was likely mostly linked to their interactions with the yeast plasma membrane. From the mid-stationary phase to the end of the fermentation, an adaptive response was exhibited by yeast, resulting in lower mortality. This work evidenced a strong impact of polyphenols on yeast fermentative capacity and highlighted the importance of a better knowledge of the mechanisms involved to improve the management of fermentations in the context of red winemaking.

Interactions of grape tannins and wine polyphenols with a yeast protein extract, mannoproteins and ß-glucan.

At present, there is a great interest in enology for yeast derived products to replace aging on lees in winemaking or as an alternative for wine fining. These are yeast protein extracts (YPE), cell walls and mannoproteins. Our aim was to further understand the mechanisms that drive interactions between these components and red wine polyphenols. To this end, interactions between grape skin tannins or wine polyphenols or tannins and a YPE, a mannoprotein fraction and a ß-glucan were monitored by binding experiments, ITC and DLS. Depending on the tannin structure, a different affinity between the polyphenols and the YPE was observed, as well as differences in the stability of the aggregates. This was attributed to the mean degree of polymerization of tannins in the polyphenol fractions and to chemical changes that occur during winemaking. Much lower affinities were found between polyphenols and polysaccharides, with different behaviors between mannoproteins and ß-glucans.

Sorption of grape proanthocyanidins and wine polyphenols by yeasts, inactivated yeasts, and yeast cell walls.

Inactivated yeast fractions (IYFs) can be used in enology to improve the stability and mouthfeel of red wines. However, information concerning the mechanisms involved and the impact of the IYF characteristics is scarce. Adsorption isotherms were used to investigate interactions between grape proanthocyanidin fractions (PAs) or wine polyphenols (WP) and a commercial yeast strain (Y), the inactivated yeast (IY), the yeast submitted to autolyzis and inactivation (A-IY), and the cell walls obtained by mechanical disruption (CW). High affinity isotherms and high adsorption capacities were observed for grape PAs and whole cells (Y, IY, and A-IY). Affinity and adsorbed amount were lower with wine PAs, due to chemical changes occurring during winemaking. By contrast to whole cells, grape PAs and WP adsorption on CW remained very low. This raises the issue of the part played by cell walls in the interactions between yeast and proanthocyanidins and suggests the passage of the latter through the wall pores and their interaction with the plasma membrane.