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…

Influence of a spontaneous cover crop on the vineyard and soil erosion under Mediterranean climate

Sixty five % of the agricultural area of the Basque Country located in the DO Ca Rioja corresponds to vineyards. More than 40% of it has an average slope greater than 10%, which makes it sensitive to erosive processes. Furthermore, it is foreseeable that extreme weather events (storms, hail, extreme heat and cold, etc.) will be favored due to climate change. Cover cropping can mitigate this risk, and therefore the objective of this work is to evaluate the impact that a vegetable cover has on the agronomic behavior of the vineyard, the quality of the grape and soil erosion. For this, a trial has been carried out with a Graciano variety vineyard with a slope between 10% -20% during the years 2020 and 2021. Conventional tillage management in the area has been compared (4-6 passes per year of tillage machinery) versus spontaneous vegetation cover management in the vineyard. This implies not tilling and allowing the grass of the land to colonize the range between the lines of vines, controlling their height through 1-3 mowing passes per year, always trying to affect the surface of the land as little as possible. The vegetative growth, yield and quality of the grape and wine was measured. Furthermore, erosion has been measured using Gerlasch boxes. The yield was lower in the second year of the trial in the cover crop treatment, but erosion was significantly reduced.

Amino nitrogen content in grapes: the impact of crop limitation

As an essential element for grapevine development and yield, nitrogen is also involved in the winemaking process and largely affects wine composition. Grape must amino nitrogen deficiency affects the alcoholic fermentation kinetics and alters the development of wine aroma precursors. It is therefore essential to control and optimize nitrogen use efficiency by the plant to guarantee suitable grape nitrogen composition at harvest. Understanding the impact of environmental conditions and cultural practices on the plant nitrogen metabolism would allow us to better orientate our technical choices with the objective of quality and sustainability (less inputs, higher efficiency). This trial focuses on the impact of crop limitation – that is a common practice in European viticulture – on nitrogen distribution in the plant and particularly on grape nitrogen composition. A wide gradient of crop load was set up in a homogeneous plot of Chasselas (Vitis vinifera) in the experimental vineyard of Agroscope, Switzerland. Dry weight and nitrogen dynamics were monitored in the roots, trunk, canopy and grapes, during two consecutive years, using a 15N-labeling method. Grape amino nitrogen content was assessed in both years, at veraison and at harvest. The close relationship between fruits and roots in the maintenance of plant nitrogen balance was highlighted. Interestingly, grape nitrogen concentration remained unchanged regardless of crop load to the detriment of the growth and nitrogen content of the roots. Meanwhile, the size and the nitrogen concentration of the canopy were not affected. Leaf gas exchange rates were reduced in response to lower yield conditions, reducing carbon and nitrogen assimilation and increasing intrinsic water use efficiency. The must amino nitrogen profiles could be discriminated as a function of crop load. These findings demonstrate the impact of plant balance on grape nitrogen composition and contribute to the improvement of predictive models and sustainable cultural practices in perennial crops.

Elucidating vineyard site contributions to key sensory molecules: Identification of correlations between elemental composition and volatile aroma profile of site-specific Pinot noir wines

The reproducibility of elemental profile in wines produced across multiple vintages has been previously reported using grapes from a single scion clone of Vitis vinifera L. cv. Pinot noir. The grapevines were grown on fourteen different vineyard sites, from Oregon to southern California in the U.S.A., which span distances from approximately hundreds of meters to 1450 km, while elevations range from near sea level to nearly 500 m. In addition, sensorial (i.e. aroma, taste, and mouthfeel) and chemical (i.e. polyphenolic and volatile) differences across the different vineyard sites have also been observed among these wines at two aging time points. While strong evidence exists to support that grapes grown in different regions can produce wines with unique chemical and sensorial profiles, even when a single clone is used, the understanding of growing site characteristics that result in this reproducible differentiation continues to emerge. One hypothesis is that the elemental profile that a vineyard site imparts to the grape berries and the resulting wine is an important contributor to this differentiation in chemistry and sensory of wines. For example, various classes of enzymes that catalyze the formation of key aroma compounds or their precursors require specific metals. In this work, we begin to report correlations between elemental and volatile aroma profiles of site-specific Pinot noir wines, made under standardized winemaking conditions, that have been previously shown to be distinguished separately by these chemical analyses.

The concept of terroir: what place for microbiota?

Microbes play key roles on crop nutrient availability via biogeochemical cycles, rhizosphere interactions with roots as well as on plant growth and health. Recent advances in technologies, such as High Throughput Sequencing Techniques, allowed to gain deeper insight on the structure of bacterial and fungal communities associated with soil, rhizosphere and plant phyllosphere. Over the past 10 years, numerous scientific studies have been carried out on the microbial component of the vineyard. Whether the soil or grape compartments have been taken into account, many studies agree on the evidence of regional delineations of microbial communities, that may contribute to regional wine characteristics and typicity. Some authors proposed the term “microbial terroir” including “yeast terroir” for grapes to describe the connection between microbial biogeography and regional wine characteristics. Many factors are involved in terroir including climate, soil, cultivar and human practices as well as their interactions. Studies considering “microbial terroir” greatly contributed to improve our knowledge on factors that shape the vineyard microbial structure and diversity. However, the potential impact of “microbial terroir” on wine composition has yet not received strong scientific evidence and many questions remain to be addressed, related to the functional characterization of the microbial community and its impact on plant physiology and grape composition, the origins and interannual stability of vineyard microbiota, as well as their impact on wine sensorial attributes. The presentation will give an overview on the role of microbiota as a terroir component and will highlight future perspectives and challenges on this key subject for the wine industry.

Phenological characterization of a wide range of Vitis Vinifera varieties

In order to study the impact of climate change on Bordeaux grape varieties and to assess the adaptation capacities of candidates to the grape varieties of this wine region to the new climatic conditions, an experimental block design composed of 52 grape varieties was set up in 2009 at the INRAE Bordeaux Aquitaine center. Among the many parameters studied, the three main phenological stages of the vine (budburst, flowering and veraison) have been closely monitored since 2012. Observations for each year, stage and variety were carried out on four independent replicates. Precocity indices have been calculated from the data obtained over the 2012-2021 period (Barbeau et al. 1998). This work allowed to group the phenological behaviour of the grapevine varieties, not only based on the timing of the subsequent developmental stages, but also on the overall precocity of the cycle and the total length of the cycle between budburst and veraison. Results regarding the variability observed among the different grape varieties for these phenological stages are presented as heat maps.