IVAS 2022 banner
IVES 9 IVES Conference Series 9 IVAS 9 IVAS 2022 9 Molecular binding mechanisms between grape seed polypeptides and wine anthocyanins by fluorescence spectroscopy and computational techniques

Molecular binding mechanisms between grape seed polypeptides and wine anthocyanins by fluorescence spectroscopy and computational techniques

Abstract

In recent years, proteins endogenous to grape have become of great interest to the wine industry because they represent a new alternative to other biopolymers subjected to more legal restrictions (i.e. animal origin and synthetics) that can be used in technological applications to modulate sensory attributes such as wine color and have a positive impact on wine quality.
Due to their great techno-functional value, some major protein components of grape seed endosperm have been identified and its potential use for color protection purposes have been already tested in wines at experimental scale. Notwithstanding, the capability of proteins to modulate sensory properties is highly dependent on their structural features, their ability to form specific 3D conformations, and the interaction with other molecules present in food matrices such as anthocyanin pigments in wines. Recently, we have confirmed by means of theoretical studies that grape seed  globulins can stablish different types of interactions (hydrogen bonding, alkyl and π-π) with the main grape anthocyanin (malvidin 3-gl).To date, however, such as interactions has been scarcely studied. Although a basic molecular interaction mechanism similar to copigmentation has been suggested, many questions still remain unclarified. In this sense, a deeper molecular understanding of the grape seed protein-anthocyanin interactions is needed to better define their potential uses and technological applications.
Thus, the main objective of this work was to explore new molecular binding mechanisms between grape seed polypeptides and wine anthocyanins in model wine solutions through fluorescence spectroscopy and computational techniques. For this purpose, fluorescence extinction spectroscopy was used to experimentally confirm the possible interaction mechanisms, the affinity constants, the stoichiometry of the complexes and other thermodynamic parameters. In addition, molecular modelling techniques, such as docking studies, were used to model the  most energetically favorable binding sites of wine anthocyanins on the grape seed polypeptide. Results will provide insights and substantial information about protection to colors expressed by anthocyanins in red wine.

DOI:

Publication date: June 23, 2022

Issue: IVAS 2022

Type: Poster

Authors

Chamizo-González Francisco1, Gordillo Arrobas Belén1, Hereida Francisco J.1, Días Ricardo2 and Freitas Víctor2

1Food Colour and Quality Laboratory, Facultad de Farmacia, Universidad de Sevilla, 41012, Sevilla, Spain
2Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade do Porto  

Contact the author

Keywords

wine anthocyanin, grape seed polypeptides, molecular interaction, Quenching of fluorescence, computational techniques

Tags

IVAS 2022 | IVES Conference Series

Citation

Related articles…

Water deficit differentially impacts the performances and the accumulation of grape metabolites of new varieties tolerant to fungi

The use of resistant varieties is a long-term but promising solution to reduce chemical input in viticulture. Several important breeding programs in Europe and abroad are now releasing a range of new hybrids performing well regarding fungi susceptibility and producing good quality wines. Unfortunately, insufficient attention is paid by the breeders to the adaptation of these varieties to climatic changes, notably to the increased climatic demand and water deficit (WD). Thus, prior to the adoption of such varieties by the wine industry in Mediterranean regions, there is a need to consider their suitability to WD. This study aimed to characterize the different drought-strategies adopted by 6 new resistant varieties selected by INRAE in comparison to Syrah. To allow the assessment of long-term impacts of WD, field-grown vines were exposed to contrasted WD from 2018 to 2021 under a semi-arid Mediterranean climate. A gradient of WD was applied in the field and controlled through plant measurements at the single plant level. Grape development was non-destructively monitored to determine the arrest of berry phloem unloading. The impacts of WD on berry composition, including water, primary metabolites (sugars, organic acids), secondary metabolites (anthocyanins, thiols precursors) and main cations contents, were assessed at this specific stage. Results showed different varietal responses during the year and inter-annual acclimation in terms of plant water use efficiency, biomass accumulation, as well as yield components and berry composition. WD differentially reduced the accumulation of primary metabolites at plant and berry levels, but it little changed their concentrations in the fruits at the ripe stage. Moreover, WD differentially impacted the accumulation of secondary metabolites and major cations between the varieties. In the talk, we’ll present the main results regarding the WD impacts on fruit metabolites and enlarge the reflection about the practical assessment of the grapevine acclimation to WD.

Drought effect on aromatic and phenolic potential of seven recovered grapevine varieties in Castilla-La Mancha region (Spain)

The effects of climate change are seriously affecting the quality of wine grapes. High temperatures and drought cause imbalances in the chemical composition of grapes. The result is overripe grapes with low acidity and high sugar content, which produce wines with excessive alcohol content, lacking in freshness and not very aromatic. As a consequence, the search of varieties with capacity of produce quality grapes in adverse climate conditions is a good alternative to preserve the sustainability of vineyards. In this work, quality parameters of seven Vitis vinifera L. cultivars (five whites and two reds) recently recovered from extinction and grown under two different hydric regimes (rainfed and irrigated) were analyzed during the 2020 vintage. At harvest time, weight of 100 berries, must physicochemical parameters (brix degree, total acidity, malic acid, pH), and carbon and oxygen isotope ratios (δ13C, δ18O) were determined. Subsequently, varietal aroma potential index (IPAv) and total polyphenol index (TPI) were analyzed. Quality parameters, IPAv and TPI, showed significant differences between varieties and water regimes. Both red varieties, Moribel and Tinto Fragoso, stood out for their high aromatic and phenolic potential, which was higher under rainfed regime. Regarding to white varieties, Montonera del Casar and Jarrosuelto stood out in terms of varietal aroma potential. Montonera del Casar high acidity in its musts and Jarrosuelto showed the highest berry weights.

Estimating bulk stomatal conductance of grapevine canopies

In response to changes in their environment, grapevines regulate transpiration using various physiological mechanisms that alter conductance of water through the soil-plant-atmosphere continuum. Expressed as bulk stomatal conductance at the canopy scale, it varies diurnally in response to changes in vapor pressure deficit and net radiation, and over the season to changes in soil water deficits and hydraulic conductivity of both soil and plant. It is necessary to characterize the response of conductance to these variables to better model how vine transpiration also responds to these variables. Furthermore, to be relevant for vineyard-scale modeling, conductance is best characterized using data collected in a vineyard setting. Applying a crop canopy energy flux model developed by Shuttleworth and Wallace, bulk stomatal conductance was estimated using measurements of individual vine sap flow, temperature and humidity within the vine canopy, and estimates of net radiation absorbed by the vine canopy. These measurements were taken on several vines in a non-irrigated vineyard in Bordeaux France, using equipment that did not interfere with ongoing vineyard operations. An inverted Penman-Monteith equation was then used to calculate bulk stomatal conductance on 15-minute intervals from July to mid-September 2020. Time-series plots show significant diurnal variation and seasonal decreases in conductance, with overall values similar to those in the literature. Global sensitivity analysis using non-parametric regression found transpiration flux and vapor pressure deficit to be the most important input variables to the calculation of bulk stomatal conductance, with absorbed net radiation and bulk boundary layer conductance being much less important. Conversely, bulk stomatal conductance was one of the most important inputs when calculating vine transpiration, further emphasizing the need for characterizing its response to environmental changes for use in vineyard water use modeling.

Terroir analysis and its complexity

Terroir is not only a geographical site, but it is a more complex concept able to express the “collective knowledge of the interactions” between the environment and the vines mediated through human action and “providing distinctive characteristics” to the final product (OIV 2010). It is often treated and accepted as a “black box”, in which the relationships between wine and its origin have not been clearly explained. Nevertheless, it is well known that terroir expression is strongly dependent on the physical environment, and in particular on the interaction between soil-plant and atmosphere system, which influences the grapevine responses, grapes composition and wine quality. The Terroir studying and mapping are based on viticultural zoning procedures, obtained with different levels of know-how, at different spatial and temporal scales, empiricism and complexity in the description of involved bio-physical processes, and integrating or not the multidisciplinary nature of the terroir. The scientific understanding of the mechanisms ruling both the vineyard variability and the quality of grapes is one of the most important scientific focuses of terroir research. In fact, this know-how is crucial for supporting the analysis of climate change impacts on terroir resilience, identifying new promised lands for viticulture, and driving vineyard management toward a target oenological goal. In this contribution, an overview of the last findings in terroir studies and approaches will be shown with special attention to the terroir resilience analysis to climate change, facing the use and abuse of terroir concept and new technology able to support it and identifying the terroir zones.

1H-NMR-based Metabolomics to assess the impact of soil type on the chemical composition of Mediterranean red wines

The aim of this study was to evaluate the effects of different soil types on the chemical composition of Mediterranean red wines, through untargeted and targeted 1H-NMR metabolomics. One milliliter of raw wine was analyzed by means of a Bruker Avance II 400 spectrometer operating at 400.15 MHz. The spectra were recorded by applying the NOESYGPPS1D pulse sequency, to achieve water and ethanol signals suppression. No modification of the pH was performed to avoid any chemical alteration of the matrix. The generation of input variables for untargeted analysis was done via bucketing the spectra. The resulting dataset was preprocessed prior to perform unsupervised PCA, by means of MetaboAnalyst web-based tool suite. The identification of compounds for the targeted analysis was performed by comparison to pure compounds spectra by means of SMA plug-in of MNova 14.2.3 software. The dataset containing the concentrations (%) of identified compounds was subjected to one-way analysis of variance (ANOVA) to highlight significant differences among the wines. The untargeted analysis, carried out through the PCA, revealed a clear differentiation among the wines. The fragments of the spectra contributing mostly to the separation were attributed to flavonoids, aroma compounds and amino acids. The targeted analysis leaded to the identification of 68 compounds, whose concentrations were significant different among the wines. The results were related to soils physical-chemical analysis and showed that: 1) high concentrations of flavan-3-ols and flavonols are correlated with high clay content in soils; 2) high concentrations of anthocyanins, amino acids, and aroma compounds are correlated with neutral and moderately alkaline soil pH; 3) low concentrations of flavonoids and aroma compounds are correlated with high soil organic matter content and acidic pH. The 1H-NMR metabolomic analysis proved to be an excellent tool to discriminate between wines originating from grapes grown on different soil types and revealed that soils in the Mediterranean area exert a strong impact on the chemical composition of the wines.