IVAS 2022 banner
IVES 9 IVES Conference Series 9 IVAS 9 IVAS 2022 9 Study of the interactions between wine anthocyanins and proline rich proteins

Study of the interactions between wine anthocyanins and proline rich proteins

Abstract

The interaction between tannins and salivary proteins is considered to be the basis of the phenomenon of wine astringency. Recently, some authors have revealed that some anthocyanins can also contribute to this mouthfeel sensation by interacting with proline rich proteins (PRPs). However, more studies are needed in order to elucidate the affinity of anthocyanins with these proteins.

Thus, the general objective of this work was determine the interaction between malvidin-3-O-glucoside, malvidin-3-O-(6-O-acetyl)-glucoside, and malvidin-3-O-(6-O-p-coumaroyl)-glucoside (isolated from grape skin) with a PRP model peptide (IB7-14) and their capacity of precipitate PRPs in a wine model solution. To archived this objectif, several techniques were used: i) mass spectrometry (FIA-ESI-QTOF) and 1H proton NMR to determine the formation of complexes and the stoichiometry of anthocyanins-IB7-14 complexe ii) saturation transfer difference (STD) NMR spectroscopy in order to calculate the dissociation constants (KD) and the affinity of each anthocyanin with the peptide and, iii) HPLC-DAD that was used to evaluate the capacity of anthocyanins to precipitate PRPs (isolated from human saliva).

Our results demonstrate that anthocyanins are able to interact with IB7-14, with different stoichiometries and binding strengths. A stoichiometry of 3:1 for the malvidin-3-O-glucoside-peptide, 1:1 for the acetylated form, and 4:1 for the coumaroylated form were observed. These ratios was also confirmed by 1H proton NMR. According to the obtained dissociation constants, the affinity of malvidin-3-O-glucoside (17.5 mM) was much higher than for malvidin-3-O-(6-O-acetyl)-glucoside (order of hundred mM). Unfortunately, the calculation of KD for malvidin-3-O-(6-O-p-coumaroyl)-glucoside was impossible due to precipitate formation. To finish, when malvidin-3-O-(6-O-p-coumaroyl)-glucoside was mixed with human salivary PRPs we observed that the precipitation of PRPs was much higher (14%) than for malvidin-3-O-glucoside (7%). In the same way, malvidin-3-O-(6-O-acetyl)-glucoside did not lead a significant decrease of their quantities when it was in contact with PRPs, suggesting the absence of interactions. It could be hypothesized that the additional presence of an aromatic group of coumaroylated form of malvidin could provide stronger hydrophobic bonds than malvidin-3-O-glucoside. Likewise, the chemical structural differences between malvidin-3-O-glucoside and malvidin-3-O-(6-O-acetyl)-glucoside can cause a potential loss of hydrogen bonding preventing thus the stabilization between the anthocyanin and the peptide. These findings proved for the first time that wine anthocyanins interact differently with the peptide IB7-14 and that can potentially affect the astringency sensation.

Section for all references

DOI:

Publication date: June 27, 2022

Issue: IVAS 2022

Type: Poster

Authors

Hornedo Ortega Ruth1, Jourdes Michaël, Da Costa Gregory, Pedrot Eric, Richard Tristan and Teissedre Pierre-Louis

1Departamento de Nutrición, Bromatología, Toxicología y Medicina Legal. Facultad de Farmacia. Universidad de Sevilla
2UMR Œnology (OENO), UMR 1366, ISVV, Université de Bordeaux-INRAE-Bordeaux INP, F33882 Villenave d’Ornon France

Contact the author

Keywords

anthocyanin, proline rich protein, astringecy, wine

Tags

IVAS 2022 | IVES Conference Series

Citation

Related articles…

Sustainable fertilisation of the vineyard in Galicia (Spain)

Excessive fertilization of the vineyard leads to low quality grapes, increased costs and a negative impact on the environment. In order to establish an integrated management system aimed at a sustainable fertilization of the vineyards, nutritional reference levels were established. For this purpose, 30 representative vineyards of the Albariño variety were studied, in which soil and petiole analyses were carried out for two years and grape yield and quality at harvest were measured. In both years of study, soil pH, calcium, sodium and cation exchange capacity were positively correlated with calcium content and negatively correlated with manganese in grapes. Irrigated vineyards had higher levels of aluminium in soil and lower levels of calcium in petiole. Climatic conditions were very different in the years of the study. The year 2019 was colder than usual, in 2020 there was a marked water stress with high summer temperatures. This resulted in medium-high acidity in grapes in 2019 and low acidity in 2020, with sugar levels being similar both years. A very marked decrease in must amino nitrogen was observed in 2020, with ammonia nitrogen remaining stable. The correlation of acidity and sugar values in grapes with soil and petiole analysis data made it possible to establish reference levels for the nutritional diagnosis of the Albariño variety in this region. Based on these results, an easy-to-use TIC application is currently being created for grapegrowers, aimed at improving the sustainability of the vineyard through reasoned fertilization. This study has now been extended to other Galician vine varieties.

The plantation frame as a measure of adaptation to climate change

The mechanization of vineyard work originally led to a reduction in planting densities due to the lack of machinery adapted to the vineyard. The current availability of specific machinery makes it possible to establish higher planting densities. In this work, three planting densities (1.40×0.80 m, 1.80×1 m and 2.20×1.20 m, corresponding to 8928, 5555 and 3787 plants/ha respectively) were studied with four varieties autochthonous of Galicia (northwestern Spain): Albariño and Treixadura (white), Sousón and Mencía (red). The vines were trained in a vertical shoot positioning system using a single Royat cordon, and pruned to spurs with two buds each. Agronomic data (yield, pruning wood weight, Ravaz index) and oenological data in must were collected. The higher planting density (1.40×0.80 m) had no significant effect on grape yield per vine in white varieties, although production per hectare was much higher due to the greater number of plants. In red varieties, this planting density resulted in a significantly lower production per vine, compensated by the greater number of plants. In addition, it significantly reduced the Brix degree in the must of the Albariño, Treixadura and Sousón varieties, and increased the total acidity in the latter two and Mencía. It also caused an increase in extractable and total anthocyanins and IPT in red grapes. The effects of high planting density on grapes are of great interest for the adaptation of varieties in the context of climate change. In the future, it could be advisable to modify the limits imposed by the appellations of origin on the planting density of these varieties in order to obtain more balanced wines.

Sustaining wine identity through intra-varietal diversification

With contemporary climate change, cultivated Vitis vinifera L. is at risk as climate is a critical component in defining ecologically fitted plant materiel. While winegrowers can draw on the rich diversity among grapevine varieties to limit expected impacts (Morales-Castilla et al., 2020), replacing a signature variety that has created a sense of local distinctiveness may lead to several challenges. In order to sustain wine identity in uncertain climate outcomes, the study of intra-varietal diversity is important to reflect the adaptive and evolutionary potential of current cultivated varieties. The aim of this ongoing study is to understand to what extent can intra-varietal diversity be a climate change adaptation solution. With a focus on early (Sauvignon blanc, Riesling, Grolleau, Pinot noir) to moderate late (Chenin, Petit Verdot, Cabernet franc) ripening varieties, data was collected for flowering and veraison for the various studied accessions (from conservatory plots) and clones. For these phenological growing stages, heat requirements were established using nearby weather stations (adapted from the GFV model, Parker et al., 2013) and model performances were verified. Climate change projections were then integrated to predict the future behaviour of the intra-varietal diversity. Study findings highlight the strong phenotypic diversity of studied varieties and the importance of diversification to enhance climate change resilience. While model performances may require improvements, this study is the first step towards quantifying heat requirements of different clones and how they can provide adaptation solutions for winegrowers to sustain local wine identity in a global changing climate. As genetic diversity is an ongoing process through point mutations and epigenetic adaptations, perspective work is to explore clonal data from a wide variety of geographic locations.

The potential of multispectral/hyperspectral technologies for early detection of “flavescence dorée” in a Portuguese vineyard

“Flavescence dorée” (FD) is a grapevine quarantine disease associated with phytoplasmas and transmitted to healthy plants by insect vectors, mainly Scaphoideus titanus. Infected plants usually develop symptoms of stunted growth, unripe cane wood, leaf rolling, leaf yellowing or reddening, and shrivelled berries. Since plants can remain symptomless up to four years, they may act as reservoirs of FD contributing to the spread of the disease. So far, conventional management strategies rely mainly on the insecticide treatments, uprooting of infected plants and use of phytoplasma-free propagation material. However, these strategies are costly and could have undesirable environmental impacts. Thus, the development of sustainable and noninvasive approaches for early detection of FD and its management are of great importance to reduce disease spread and select the best cultural practices and treatments. The present study aimed to evaluate if multispectral/hyperspectral technologies can be used to detect FD before the appearance of the first symptoms and if infected grapevines display a spectral imaging fingerprint. To that end, physiological parameters (leaf area, chlorophyll content and photosynthetic rate) were collected in concomitance to the measurements of plant reflectance (using both a portable apparatus and a remote sensing drone). Measurements were performed in two leaves of 8 healthy and 8 FD-infected grapevines, at four timepoints: before the development of disease symptoms (21st June); and after symptoms appearance (ii) at veraison (2nd August); at post-veraison (11th September); and at harvest (25th September). At all timepoints, FD infected plants revealed a significant decrease in the studied physiological parameters, with a positive correlation with drone imaging data and portable apparatus analyses. Moreover, spectra of either drone imaging and portable apparatus showed clear differences between healthy and FD-infected grapevines, validating multispectral/ hyperspectral technology as a potential tool for the early detection of FD or other grapevine-associated diseases.

Influence of climatic conditions on grape composition of Tempranillo in La Mancha DO (Spain)

The aim of this work was to analyze the variability in grape composition of the Tempranillo cultivar related to climatic conditions, in La Mancha Designation of Origin. Grape composition (sugar content, total acidity, pH, malic acid, and total and extractable anthocyanins) recorded during ripening, were analysed for the period 2000-2019. The weather conditions at daily time scale, recorded during the same period, were also evaluated. The relationships between grape parameters with climatic variables related to temperature and to water deficits, referring different periods between phenological events along the growing cycle, were evaluated using regression analysis. High variability in grape composition was observed in the period analysed. Total acidity varied between 3.7 and 7.3 gL-1 while malic acid varied between 1.2 and 4 gL-1. The extractable anthocyanins ranged between 526 and 972 mgL-1, and total anthocyanins ranged between 922 and 1388 mgL-1, being the lowest values recorded in the hottest year (2017). Total acidity decreased 0.77 gL-1 for an increase of 100 GDD, while malic acid decrease in 0.42 gL-1 for the same GDD increase, being the period between veraison and harvest the one that seemed to have higher influence on acidity. In addition, it was confirmed that increasing water deficits decreased acidity. Total and extractable anthocyanins increased in about 210 and 105 mgL-1, respectively, with an increase of 100 GDD from veraison to harvest, and the increase in water deficits favour the increase of anthocyanins, both total and extractable anthocyanins. Total and extractable anthocyanins concentration increased in 35 and 22 mgL-1 per an increase of 10 mm in the water deficit. These results can be of interest to understand the potential changes that grapes composition may suffer under future warmer climates.