Effect of different pH values on the interaction between yeast mannoproteins and grape seed flavanols

Abstract

The consequences of the global climate change in the vitiviniculture are revealed as a gap between phenolic and technological grape maturities, higher grape sugar concentration that leads to high wine alcohols levels, lower acidities and high pH values, among others. The unbalanced phenolic maturity caused in this scenario leads to harsh astringency and to instable colour of wines. Previous studies have reported that the addition of yeast mannoproteins (MPs) to wines may have positive effects on these two organoleptic properties due to their capability to interact with wine polyphenols [1]; however, studies about the effect of the pH on these interactions have not been carried out so far.

 MPs are located in the outer layer of yeast cell wall (Saccharomyces cerevisiae) and they are naturally released into the wine during alcoholic fermentation when yeast is actively growing or during aging when cell wall breaks down in the process known as autolysis. Also, commercial MPs can be added during winemaking and/or ageing. The aim of this work was to study the effect of different pH values (pH 3.0 and 4.0) on the interactions between a flavanol extract from Vitis vinifera L. Tempranillo seeds and the MPs obtained from Saccharomyces cerevisiae. Here, the isolation of MPs from the cell walls of S. cerevisiae was performed using Zymolyase 20T enzyme. MPs were purified by using ethanol, temperature and dialysis. The obtained MPs were characterized by SDS-PAGE and their molecular weights (MWs) were determined by HRSEC-RID [2]. The protein percentage was determined by the Lowry method. The monosaccharide composition was determined by HPLC-MS after derivatisation with 1-phenyl-3-methyl-5-pyrazolone (PMP) [3]. Four main MP fractions were identified: F1 (~2%), with a MW 528.8 kDa, F2 (~12%) (174.1 kDa), F3 (~61 %) (61.0 kDa) and F4 (~25 %) (<10 kDa). The MP–flavanol interactions were performed at pH=3 and pH=4 and studied by means of HPLC-DAD-MS, HRSEC-RID and Isothermal Titration Calorimetry (ITC). The results showed noticeably differences in the interactions between the MPs fractions and the flavanol extract depending on the pH values. 

References

[1] C. Alcalde-Eon, et al. (2019). Food Res. Int., 126; 108650.
[2] E. Manjón, et al. (2020). J. Agric. Food Chem. 25; 13459
[3] Y. Ruiz-García et al. (2014). Carbohydr Polym. 114; 102.