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…

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.

Characterization of variety-specific changes in bulk stomatal conductance in response to changes in atmospheric demand and drought stress

In wine growing regions around the world, climate change has the potential to affect vine transpiration and overall vineyard water use due to related changes in atmospheric demand and soil water deficits. Grapevines control their transpiration in response to a changing environment by regulating conductance of water through the soil-plant-atmosphere continuum. Most vineyard water use models currently estimate vine transpiration by applying generic crop coefficients to estimates of reference evapotranspiration, but this does not account for changes in vine conductance associated with water stress, nor differences thought to exist between varieties. The response of bulk stomatal conductance to daily weather variability and seasonal drought stress was studied on Cabernet-Sauvignon, Merlot, Tempranillo, Ugni blanc, and Semillon vines in a non-irrigated vineyard in Bordeaux France. Whole vine sap flow, temperature and humidity in the vine canopy, and net radiation absorbed by the vine canopy were measured on 15-minute intervals from early July through mid-September 2020, together with periodic measurement of leaf area, canopy porosity, and predawn leaf water potential. From this data, bulk stomatal conductance was calculated on 15-minute intervals, and multiple regression analysis was performed to identify key variables and their relative effect on conductance. Attention was focused on addressing multicollinearity and time-dependency in the explanatory variables and developing regression models that were readily interpretable. Variability of vapor pressure deficit over the day, and predawn water potential over the season explained much of the variability in conductance, with relative differences in response coefficients observed across the five varieties. By characterizing this conductance response, the dynamics of vine transpiration can be better parameterized in vineyard water use modeling of current and future climate scenarios.

Grape must quality and mesoclimatic variability in Fruška Gora wine-growing region, Serbia

The Fruška Gora mountain is a traditional wine-growing region in Serbia situated in the Pannonian Basin. Due to such a position, the vicinity of the Danube River and the presence of concave configuration, it is suitable for grape production. This paper provides analyses of spatial variations in meteorological parameters and grape juice quality within Fruška Gora wine region over three consecutive vintages (2018-2020). The examined period can be defined as warm with cool nights during September (AVG 18,9°C; GDD 1918°C; CI 12°CF) and with the presence of mesoclimatic variability. The East part of the study area was somewhat drier and hotter compared to other parts of the region. The analyses of grape must samples (190 in total) of five cultivars (Cabernet-Sauvignon, Merlot, Chardonnay, Sauvignon blanc and Grašac (Welschriesling)) commonly grown across the region (19 sites), were performed using Fourier Transform Infrared Technology (FTIR). Among all cultivars, Sauvignon blanc was harvested first in the East area (DOY=246±5, GDD at harvest=1552±74, 22.2±0.7 °Brix), while the latest harvest was recorded for Cabernet-Sauvignon in the West (DOY=283±5, GDD at harvest=1936±187, 23.4±1.0 °Brix ). Both the red and white cultivars had higher acidity and YAN in the grape must if the vines were grown in the North and East compared to South and West areas. According to PCA analysis, Grašac showed the lowest variation in grape must chemical composition. Thus, the results confirm that Grašac is the most stable cultivar in Fruška Gora. All monitored cultivars reached technological fruit ripeness by the end of the growing season. However, it was difficult to reach full ripeness of red cultivars, mostly beacuse of uncoupling of technolocical and phenolic ripeness. Thus, Cabernet-Sauvignon had higher variations in GDD sums at harvest compared to other cultivars, which probably increased variations in grape must quality.

Understanding graft union formation by using metabolomic and transcriptomic approaches during the first days after grafting in grapevine

Since the arrival of Phyloxera (Daktulosphaira vitifolia) in Europe at the end of the 19th century, grafting has become essential to cultivate Vitis vinifera. Today, grafting provides not only resistance to this aphid, but it used to adapt the cultivars according to the type of soil, environment, or grape production requirements by using a panel of rootstocks. As part of vineyard decline, it is often mentioned the importance of producing quality grafted grapevine to improve vineyard longevity, but, to our knowledge, no study has been able to demonstrate that grafting has a role in this context. However, some scion/rootstock combinations are considered as incompatible due to poor graft union formation and subsequently high plant mortality soon after grafting. In a context of climate change where the creation of new cultivars and rootstocks is at the centre of research, the ability of new cultivars to be grafted is therefore essential. The early identification of graft incompatibility could allow the selection of non-viable plants before planting and would have a beneficial impact on research and development in the nursery sector. For this reason, our studies have focused on the identification of metabolic and transcriptomic markers of poor grafting success during the first days/week after grafting; we have identified some correlations between some specialized metabolites, especially stilbenes, and grafting success, as well as an accumulation of some amino acids in the incompatible combination. The study of the metabolome and the transcriptome allowed us to understand and characterise the processes involved during graft union formation.

Soil quality in Beaujolais vineyard. Importance of pedology and cultural practices

A pedological study was carried out from 2009 to 2017 in Beaujolais vineyard, to improve physical and chemical knowledge of soils. It was completed in 2016 and 2017 by the current study, dealing with microbial aspects, in order to build a reference frame for improved advice in soil management. Microbial biomass was measured on representative plots of the six most common soil types identified in Beaujolais and, for each soil type, on plots with different levels of the main impacting parameters: total organic carbon, pH, cation exchange capacity, extractable copper. A total of 59 soil samples were collected. Confirming the results of various trials carried out in Beaujolais over the past 20 years, the results of the present study showed that the soils were still alive, but exhibited a large variability of biological parameters, which appeared dependant on both pedological and anthropic factors. Therefore, a good interpretation of biological parameters and advice for vine growers must rely on a pedologically-based referential with differentiated main driving factors. For example, the control of pH is of primary importance in granitic soils and in no way organic matter addition can improve soil quality if pH is too low. Conversely, in calcareous soils, biological parameters are more directly affected by direct or indirect (cover crops for example) inputs of organic matter. The use of biological parameters, such as microbial biomass, is of great potential value to improve advice on agro-viticultural practices (soil management, fertilization, liming, etc.), basis of a sustainable wine production on fragile soils.