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…

Is wine terroir a valid concept under a changing climate?

The OIV[i] defines terroir as a concept referring to an area in which collective knowledge of the interactions between the physical and biological environment (soil, topography, climate, landscape characteristics and biodiversity features) and vitivinicultural practices develops, providing distinctive wine characteristics. Those are perceptible in the taste of wine, which drives consumer preference and, therefore, wine’s value in the marketplace. Geographical indications (GI) are recognized regulatory constructs formalizing and protecting the nexus between wine taste and the terroir generating it. Despite considering updates, GIs do not consider the nexus as a dynamic one and do not anticipate change, namely of climate. Being climate a fundamental feature of terroir, it strongly impacts wine characteristics, such as taste. According to IPCC[ii], many widespread, rapid and unprecedented changes of climate occurred, some being irreversible over hundreds to thousands of years. Climatic shifts and atmospheric-driven extreme events have been widely reported worldwide. Recent climatic trends are projected to strengthen in upcoming decades, whereas extremes are expected to increase in frequency and intensity, forcing wines away from GI definitions. Geographical shifts of viticultural suitability are projected, often moving into regions and countries different from current ones. Some authors propose adaptation in viticulture, winemaking and product innovation. We show evidence of climate changing wine characteristics in the Douro valley, home of 270-year-old Port GI. We discuss herein resist or adapt stances for when climate changes the nexus between terroir and wine characteristics. Using the MED-GOLD[iii] dashboard, a tool allowing for easy visual navigation of past and future climates, we demonstrate how policymakers can identify future moments, throughout the 21st century under different emission scenarios, when GI specifications will likely need updates (e.g., boundaries, varieties) to reduce climate-change impacts.

The effects of alternative herbicide free cover cropping systems on soil health, vine performance, berry quality and vineyard biodiversity in a climate change scenario in Switzerland

There is an urgent need in viticulture to adopt alternative herbicide-free soil management strategies to mitigate climate change, increase biodiversity, reduce plant protection products and improve soil quality while minimizing detrimental effects on grapevine’s stress tolerance and fruit quality. To propose sustainable solutions, adapted to different pedoclimatic conditions in Switzerland, we developed a multidisciplinary 4-year project, started in 2020. Objectives of the project are to a) evaluate the impact of green covers (spontaneous flora, winter cover crop and permanent ground cover) on environmental and agronomic parameters and b) develop subsequently innovative strategies for different viticultural contexts of Switzerland. The project is divided into 3 phases: 1) diagnosis, 2) on-farm and 3) on-station experiments. Phase 1) consisted in an assessment of 30 commercial vineyards all over Switzerland, where growers already use different herbicide-free soil management strategies. The most promising practices identified in this exploratory phase will be replicated in commercial vineyards across Switzerland (“on-farm”) as well as in a classical randomized block design in an experimental plot (“on-station”). For phase 1), measurements consisted in evaluation of soil status (compaction, structure, roots development), soil microbial diversity (metagenomics), plant diversity and biomass, vine physiology (water stress, vigor, leaf nitrogen) and berry quality (acidity, sugar, available nitrogen). Interestingly, the permanent ground cover resulted in a higher Shannon index thus a higher biodiversity as compared to the other itineraries. The winter cover crop increased vine nitrogen and vigor while deteriorating soil quality, leaving the soil more exposed and compacted likely due to more frequent tillage. The spontaneous flora led to higher berry sugar accumulation, less nitrogen and higher malic acid concentration putatively due to a higher water retention of the flora in a particularly wet vintage. Phases 2) and 3) are required to confirm those tendencies, over the 3 next vintages and different climatic conditions.

Green berries on Gewürztraminer (Vitis vinifera L.) in South Tyrol (Italy)

The grape variety Gewürztraminer is known to be affected by two physiological disorders namely berry shrivel and bunch stem necrosis. During the season 2014 we noticed a new symptomatology type of ripening disorder on the variety. The new symptom showed not all berries fallowing the normal maturation stages, but single berries remaining at a soft but green stage till harvest. The broad distribution of these so called “green berries” symptoms in different production sites of our region, caused huge damage due to the difficulty of eliminating single berries per bunch before harvesting. Therefore, the Research Centre Laimburg began to investigate the reasons and origins of this new symptom. This work shows the results of first attempts to find causes for the symptom as well as the resulting approach to mitigate symptoms. Applications of magnesium leaf fertilizer showed first promising results against this putative disorder. To study the causal effect of the green berries 30 symptomatic vineyards in 2014 have been selected for a monitoring during the season 2016. To evaluate the foliar nutrient treatment two vineyards have been selected for application of magnesium sulfate and magnesium chloride. Leaf and berry nutrient analysis, as well as the main quality parameters during ripening have been performed. As soon as “green berries” symptoms appeared, incidence and severity have been evaluated. Most of the symptomatic vineyards of the 2016 monitoring showed light to clear magnesium deficit symptoms on their foliage. Only during the seasons 2020 and 2021 “green berries” symptoms could be found in the leaf fertilizer treatment vineyards. Both seasons showed a significant effect of the magnesium treatments to reduce the incidence and severity of the symptom. It seems that the appearance of the “green berries” symptom on Gewürztraminer is correlated to a disturbed uptake of magnesium of the vines.

Local ancient grapevine cultivars to face future viticulture

Among the different strategies to cope with the negative impacts of climate change on viticulture, the exploitation of genetic diversity is one of the most promising to adapt to new conditions and maintain wine production and quality. One of the biggest concerns in the context of climate change is to improve water use efficiency (WUE). In this way, the use of genotypes that present a better response to drought and high WUE is a key issue. In this work, physiological performance analysis was conducted to compare the water deficit stress (WDS) responses of local and widespread grapevines cultivars. Leaf gas exchange, water use efficiency (WUE) at different levels (leaf and long-term WUE (∆13C)), leaf osmotic adjustment and other water relations parameters were determined in plants under well-watered and WDS conditions alongside assessment of the levels of foliar hormones concentrations. Results denote that local cultivars displayed better physiological performance under WDS as compared to the widely-distributed ones. he results corroborate the hypothesis that better stomatal control allows increasing leaf WUE under drought as occurred in the local Callet cv.; but the minority local cultivar Escursac cv. showed high WUE under both treatments. In this case, high WUE can be related to maintaining higher photosynthetic activity under drought. The different mechanisms underlying the better performance under WDS and high WUE of minority local cultivars are discussed.

Effect of the commercial inoculum of arbuscular mycorrhiza in the establishment of a commercial vineyard of the cultivar “Manto negro

The favorable effect of symbiosis with arbuscular mycorrhizal fungi (AMF) has been known and studied since the 60s. Nowadays, many companies took the chance to start promoting and selling commercial inoculants of AMF, in order to be used as biofertilizers and encourage sustainable biological agriculture. However, the positive effect of these commercial biofertilizers on plant growth is not always demonstrated, especially under field conditions. In this study, we used a commercial inoculum on newly planted grapevines of a local cultivar grafted on a common rootstock R110. We followed the physiological status of vines, growth and productivity and functional biodiversity of soil bacteria during the first and second years of 20 inoculated with commercial inoculum bases on Rhizophagus irregularis and Funeliformis mosseaeAMF at field planting time and 20 non-inoculated control plants. All the parameters measured showed a neutral to negative effect on plant growth and production. The inoculated plants always presented lower values of photosynthesis, growth and grape production, although in some cases the differences did not reach statistical significance. On the contrary, the inoculation supposed an increase of the bacterial functional diversity, although the differences were not statistically significant either. Several studies show that the effect of inoculation with AMF is context-dependent. The non-favorable effects are probably due to inoculation ineffectiveness under complex field conditions and/or that, under certain conditions, AMF presence may be a parasitic association. This puts into question the effectiveness of its application in the field. Therefore, it is recommended to only resort to this type of biofertilizer when the cultivation conditions require it (e.g., very low previous microbial diversity, foreseeable stress due to drought, salinity, or lack of nutrients) and not as a general fertilization practice.