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…

Climate change impacts: a multi-stress issue

With the aim of producing premium wines, it is admitted that moderate environmental stresses may contribute to the accumulation of compounds of interest in grapes. However the ongoing climate change, with the appearance of more limiting conditions of production is a major concern for the wine industry economic. Will it be possible to maintain the vineyards in place, to preserve the current grape varieties and how should we anticipate the adaptation measures to ensure the sustainability of vineyards? In this context, the question of the responses and adaptation of grapevine to abiotic stresses becomes a major scientific issue to tackle. An abiotic stress can be defined as the effect of a specific factor of the physico-chemical environment of the plants (temperature, availability of water and minerals, light, etc.) which reduces growth, and for a crop such as the vine, the yield, the composition of the fruits and the sustainability of the plants. Water stress is in many minds, but a systemic vision is essential for at least two reasons. The first reason is that in natural environments, a single factor is rarely limiting, and plants have to deal with a combination of constraints, as for example heat and drought, both in time and at a given time. The second reason is that plants, including grapevine, have central mechanisms of stress responses, as redox regulatory pathways, that play an important role in adaptation and survival. Here we will review the most recent studies dealing with this issue to provide a better understanding of the grapevine responses to a combination of environmental constraints and of the underlying regulatory pathways, which may be very helpful to design more adapted solutions to cope with climate change.

VINIoT: Precision viticulture service for SMEs based on IoT sensors network

The main innovation in the VINIoT service is the joint use of two technologies that are currently used separately: vineyard monitoring using multispectral imaging and deployed terrain sensors. One part of the system is based on the development of artificial intelligence algorithms that are feed on the images of the multispectral camera and IoT sensors, high-level information on water stress, grape ripening status and the presence of diseases. In order to obtain algorithms to determine the state of ripening of the grapes and avoid losing information due to the diversity of the grape berries, it was decided to work along the first year 2020 at berry scale in the laboratory, during the second year at the cluster scale and on the last year at plot scale. Different varieties of white and red grapes were used; in the case of Galicia we worked with the white grape variety Treixadura and the red variety Mencía. During the 2020 and 2021 campaigns, multispectral images were taken in the visible and infrared range of: 1) sets of 100 grapes classifying them by means of densimetric baths, 2) individual bunches. The images taken with the laboratory analysis of the ripening stage were correlated. Technological maturity, pH, probable degree, malic acid content, tartaric acid content and parameters for assessing phenolic maturity, IPT, anthocyanin content were determined. It has been calculated for each single image the mean value of each spectral band (only taking into account the pixels of interest) and a correlation study of these values with laboratory data has been carried out. These studies are still provisional and it will be necessary to continue with them, jointly with the training of the machine learning algorithms. Processed data will allow to determine the sensitivity of the multispectral images and select bands of interest in maturation.

Vineyards and clay minerals: multi-technique analytical approach and correlations with soil properties

Purpose of this research is to quantitatively assess the mineral component of vineyard soils, with particular attention to the mineralogical analysis of clays, which represent an element of high importance in the vineyard culture as well as in general agriculture. An X-ray diffraction (XRD) / thermogravimetric (TG) multi-technique analytical approach was developed, tested on soil samples taken from vineyards around the world. This codified analytical procedure was necessary to obtain precise qualitative and quantitative mineralogical data, globally comparable to distinguish the geopedological identity of the vineyards. Soil samples from vineyards of various locations were analysed, in very different geological conditions. The bulk-rock quantitative phase analysis (QPA) was obtained by the Rietveld method while the detailed composition of the clay-sized fraction was determined by modelling of the oriented X-ray diffraction patterns. The research provided a precise classification of the mineral component of soils, distinguishing the mineral phases of the clays and the so-called mixed-layer clay minerals. We found that the content in mixed layers can be directly correlated with the water retention and the cation exchange capacity ​​of the soil, while the presence of other clayey minerals and phyllosilicates in this research did not affect this CEC parameter, which codes the fertility level of the soils. The study demonstrates that terroir, in particular soils formed in complex or very different geological conditions, can only be effectively interpreted by properly analysing its mineral phases, in particular the mixed-layer clay component. These are characteristic abiotic ecological indicators, which may have specific eco-physiological influences on the plant.

Late season canopy management practices to reduce sugar loading and improve color profile of Cabernet-Sauvignon grapes and wines in the high irradiance and hot conditions of California Central Valley

Global warming is accelerating grape ripening, leading to unbalanced wines from fruit with high sugar content but poor aroma and colour development. Reducing the size of the photosynthetic apparatus after veraison has been shown to delay technological ripeness in cool climates, but methods have not been tested in areas with high irradiance and temperature where fruit exposure could have disastrous effects on berry composition. In this Cabernet-Sauvignon trial, we compared the application of an antitranspirant (pinolene), to severe canopy topping and above bunch zone leaf removal, all performed at mid-ripening, with an untouched control. We monitored the vines weekly by measuring stem water potential, gas exchange, fruit zone light exposure. We sampled berries to measure berry weight, total soluble solids, pH, titratable acidity, and the anthocyanin profile. At harvest, we assessed yield components, measured carbon isotope discrimination, rated sunburn on clusters, and produced experimental wines. We submitted harvest samples to metabolomic profiling through PFP-Q Exactive MS/MS and wines to sensory analysis. Application of the antitranspirant significantly reduced stomatal conductance and assimilation rate but did not affect the stem water potential. Inversely, leaf removal and topping increased water potential but did not affect leaf gas exchange. The late topping was the only treatment able to decrease sugar content (up to 2Bx), increase titratable acidity and pH, and improve anthocyanin content because of lower degradation of di-hydroxylated forms. Late leaf removal above the bunch zone increased lightning conditions in the canopy and produced the most significant damage on fruits. Yield components were not affected. This work suggests that late-season canopy management can effectively control ripening speeds and improve grapes and wines. Still, the effect on grape exposure in a critical time must be well balanced to avoid problems with the appropriate technique.

Terroir traceability in grapes, musts and wine: results of research on Gewürztraminer and Sauvignon Blanc grape varieties in northern Italy

In the study of terroir, a separate analysis of its many component factors can be of great help in accurately identifying a vineyard’s natural elements that impact wine quality and typicity. This research used a dedicated pluri-disciplinary approach to investigate the ecological characteristics, including geology and geographical features, of 14 vineyards that produce Gewürztraminer and Sauvignon Blanc cultivars in the alpine Alto Adige DOC wine region. Both the geopedological method using Vineyards Geological Identity (VGI) and the new Solar Radiaton Identity (SRI) topoclimatic classification method were used to provide analytical measurements and qualitative/quantitative characterisations. In addition, wide-ranging targeted and untargeted oenological and chemical analyses were carried out on grapes, musts and wines to correlate the soils’ geomineral and physical conditions with the biochemical properties of their fruits and wines. The research identified strong correlations between vineyard geo-identity and wine biofingerprint, confirming a mineral traceability of strontium rubidium ratio and some minerals distinctive to the local geology, such as K, Ca, Ag, Ba and Mn.  The study also discovered that particular geomineral and physical soil conditions of the studied vineyards are related to the different amount of amino acids, primary varietal aromas and polyphenols found in grapes, musts and wines. The research confirmed that winemaking technologies support oenological quality, although in some cases, human practices can overpower certain characteristic elements in wine, erasing the typical imprint left by the vineyards’ natural terroir, which becomes less traceable. Terroir abiotic ecological factors and vineyard identity can be classified in detail using the new VGI and SRI analysis methods to discover interrelationships between geo-pedological and topoclimatic conditions that impact wine quality. These methods are also helpful in identifying which ecological elements are exclusive to a particular vineyard or wine sub-region.