WAC 2022 banner
IVES 9 IVES Conference Series 9 WAC 9 WAC 2022 9 3 - WAC - Oral 9 Impact of aspects of the polysaccharide structure of mannoproteins on their interactions with Enological Tannins

Impact of aspects of the polysaccharide structure of mannoproteins on their interactions with Enological Tannins

Abstract

Mannoproteins (MPs) with different structure of their polysaccharide part (branching, substitutions, …) were used to better understand the impact of characteristics of the usual structure of MPs when interacting with Grape Seed Tannins (ST). 

From four Saccharomyces cerevisiae strains we obtain four MP pools: an enological strain LMD47 (presenting high levels of N-glycosylation and O-Mannosylation), a wild-type BY4742 strain (used as reference), and its mutants ΔMnn4 (with no mannosyl-phosphorylation) and ΔMnn2 (with a linear N-glycosylation backbone). The extraction method applied, with the exclusive enzymatic activity of Endo-β-1,3-Glucanase of Trichoderma sp. (E-LAMSE, Megazym), preserved the indigenous structure of mannoproteins to their utmost extent. Characterizations of the pools confirmed differences among the polysaccharide moieties of the four MPs regarding charge, mannose/glucose ratio, and branching degrees but no differences between their protein moieties.

The formation and evolution of colloidal aggregates due to interactions between MPs and ST at different concentrations were evaluated through Dynamic Light Scattering (DLS), while the number of colloidal aggregates formed and the particle size distribution were assessed by Nanoparticle Tracking Analysis (NTA). The possible differences in the mechanisms of interaction among the four kinds of mannoproteins were analyzed through Isothermal Titration Calorimetry (ITC).

DLS and NTA experiments indicated an immediate formation of colloidal aggregates, in which the final particle size and concentration were dependent on the ST/MP ratio. Whenever the latter was extremely high, a very progressive flocculation related to a reversible aggregation occurred. The kinetics of this instability phenomenon was dependent on the polysaccharide structure of MPs. ITC analysis showed two different kinds of interactions: an intense exothermic one susceptible to temperature, and a much weaker interaction (as for enthalpy release) less thermo-dependent, possibly related to H-bonding and hydrophobic interactions, respectively. 

Neither the absence of mannosyl phosphate groups, the absence of ramifications on the outer chains of the N-glycosylation, nor the protein glycosylation overexpression seem to play a decisive role in those interactions. However, these structural differences affected the stability of MP-ST colloids formed at specific concentrations and slightly changed the enthalpy exchange profiles.

DOI:

Publication date: June 13, 2022

Issue: WAC 2022

Type: Article

Authors

Assunção Bicca, Céline, Poncet-Legrand, Julie, Mekoue Nguela, Thierry, Doco, Aude, Vernhet

Presenting author

Assunção Bicca – Université de Montpellier

Unité Mixte de Recherche Sciences Pour l’OEnologie, Institut Agro, INRAE, Université de Montpellier, Montpellier, France | Lallemand SAS | Unité Mixte de Recherche Sciences Pour l’OEnologie

Contact the author

Keywords

Mannoproteins – Colloidal Stability – Polysaccharide/Polyphenol Interactions – Wine macromolecules

Tags

IVES Conference Series | WAC 2022

Citation

Related articles…

Mobile device to induce heat-stress on grapevine berries

Studying heat stress response of grapevine berries in the field often relies on weather conditions during the growing season. We constructed a mobile heating device, able to induce controlled heat stress on grapes in vineyards. The heater consisted of six 150 W infrared lamps mounted in a profile frame. Heating power of the lamps could be controlled individually by a control unit consisting of a single board computer and six temperature sensors to reach a pre-set temperature. The heat energy applied to individual berries within a cluster decreases by the squared distance to the heat source, enabling the establishment of temperature profiles within individual clusters. These profiles can be measured by infrared thermography once a steady state has been reached. Radiant flux density received by a berry depending on the distance was calculated based on a view factor and measured lamp surface temperature and resulted to 665 Wm-2 at 7cm. Infrared thermography of the fruit surface was in good agreement with measurements conducted with a thermocouple inserted at epidermis level. In combination with infrared thermography, the presented device offers possibilities for a wide range of applications like phenotyping for heat tolerance in the field to proceed in the understanding of the complex response of plants to heat stress. Sunburn necrosis symptoms were artificially induced with the aid of the device for cv. Bacchus and cv. Sylvaner in the 2020 and 2021 growing season. Threshold temperatures for sunburn induction (LT5030min) were derived from temperature data of single berries and visual sunburn assessment, applying logistic regression. A comparison of threshold temperatures for the occurrence of sunburn necrosis confirmed the higher susceptibility of cv. Bacchus. The lower susceptibility of cv. Sylvaner did not seem to be related to its phenolic composition, rendering a thermoprotective role of berry phenolic compounds unlikely.

Spatial variability of temperature is linked to grape composition variability in the Saint-Emilion winegrowing area

Elevated temperature during the grape maturation period is a major threat for grape quality and thus wine quality. Therefore, characterizing the grape composition response to temperature at a larger scale would represent a crucial step towards adaptation to climate change. In response to changes in temperature, various physiological mechanisms regulate grape composition. Primary and secondary metabolisms are both involved in this response, with well-known effects, for example on anthocyanins, and lesser known effects, for example on aromas or aroma precursors. At the field scale or at the regional scale, however, numerous environmental or plant-specific factors intervene to make the effects of temperature difficult to distinguish from overall variability. In this study, it was attempted to overcome this difficulty by selecting well-characterized situations with differing temperatures.
A long-term study of air temperature variability across several Merlot vineyards in the Saint-Emilion and Pomerol wine producing area found significant temperature differences and gradients at various time scales linked to environmental factors. From this study area, a few sites were selected with similar age, soil and training system conditions, and with repeated and contrasted temperature differences during the maturation period. The average temperature difference during the maturation period was about 2°C between cooler and warmer sites, a difference similar to that expected under future climate change scenarios. In close vicinity to the temperature sensors at each site, grape berries were sampled at different times until full maturity during 2019 and 2020. Also, berries from bunches on either side of the row were analyzed separately, allowing an investigation of bunch exposure effect associated with the coupling of berry temperature and solar radiation. Four replicates of pooled berries for each time – site – bunch exposure combination were obtained and analyzed for biochemical composition. Analyses of variance of the biochemical composition data collected at different sampling times reveal significant effects associated with temperature, site, and bunch azimuth. For instance, anthocyanins in grape skins are clearly influenced by temperature and solar radiation exposure, with up to 30% reduction in warmer conditions.

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.

Spatiotemporal patterns of chemical attributes in Vitis vinifera L. cv. Cabernet Sauvignon vineyards in Central California

Spatial variability of vine productivity in winegrapes is important to characterise as both yield and quality are relevant for the production of different wine styles and products. The objectives were to understand how patterns of variability of Cabernet Sauvignon fruit composition changed over time and space, how these patterns could be characterised with indirect measurements, and how spatial patterns of the variation in fruit compositional attributes can aid in improving management. Prior to the 2017 vintage, 125 data vines were distributed across each of four vineyards in the Lodi American Viticultural Area (AVA) of California. Each data vine was sampled at commercial harvest in 2017, 2018, and 2019. Yield components and fruit composition were measured at harvest for each data vine, and maps of yield and fruit composition were produced for eight ‘objective measures of fruit quality’: total anthocyanins, polymeric tannins, quercetin glycosides, malic acid, yeast assimilable nitrogen, β-damascenone, C6 alcohols and aldehydes, and 3-isobutyl-2-methoxypyrazine. Patterns of variation in anthocyanins and phenolic compounds were found to be most stable over time. Given this relative stability, management decisions focused on fruit quality could be based on zonal descriptions of anthocyanins or phenolics to increase profitability in some vineyards. In each vineyard, dormant season pruning weights and soil cores were collected at each location, elevation and soil apparent electrical conductivity surveys were completed, and remotely sensed imagery was captured by fixed wing aircraft and two satellite platforms at major phenological stages. The data collected were used to develop relationships among biophysical data, soil, imagery, and fruit composition. The standardised and aggregated samples from four vineyards over three seasons were included in the estimation of ‘common variograms’ to assess how this technique could aid growers in producing geostatistically rigorous maps of fruit composition variability without cumbersome, single season sampling efforts.

Metabolomic discrimination of grapevine water status for Chardonnay and Pinot noir

Water status impact in viticulture has been widely explored, as it strongly affects grapevine physiology and grape chemical composition. It is considered as a key component of vitivinicultural terroir. Most of the studies concerning grapevine water status have focused on either physiological traits, or berry compounds, or traits involved in wine quality. Here, the response of grapevine to water availability during the ripening period is assessed through non-targeted metabolomics analysis of grape berries by ultra-high resolution mass spectrometry. The grapevine water status has been assessed during 2 consecutive years (2019 & 2020), through carbon isotope discrimination on juices from berries collected at maturity (21.5 brix approx.) for 2 Vitis vinifera cv. Pinot noir (PN) and Chardonnay (CH). A total of 220 grape juices were collected from 5 countries worldwide (Italy; Argentina; France; Germany; Portugal). Measured δ13C (‰) varied from -28.73 to -22.6 for PN, and from -28.79 to -21.67 for CH. These results also clearly revealed higher water stress for the 2020 vintage. The same grape juices have been analysed by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR-MS) and Liquid Chromatography coupled to Mass Spectrometry (LC-qTOF-MS), leading to the detection of up to 4500 CHONS containing elemental compositions, and thus likely tens of thousands of individual compounds, which include fatty acids, organic acids, peptides, phenolics, also with high levels of glycosylation. Multivariate statistical analysis revealed that up to 160 elemental compositions, covering the whole range of detected masses (100 –1000 m/z), were significantly correlated to the observed gradients of water status. Examples of chemical markers, which are representative of these complex fingerprints, include various derivatives of the known abscisic acid (ABA), such as phaesic acid or abscisic acid glucose ester, which are significantly correlated with higher water stress, regardless of the variety. Cultivar-specific behaviours could also be identified from these fingerprints. Our results provide an unprecedented representation of the metabolic diversity, which is involved in the water status regulation at the grape level, and which could contribute to a better knowledge of the grapevine mitigation strategy in a climate change context.