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…

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.

A better understanding of the climate effect on anthocyanin accumulation in grapes using a machine learning approach

The current climate changes are directly threatening the balance of the vineyard at harvest time. The maturation period of the grapes is shifted to the middle of the summer, at a time when radiation and air temperature are at their maximum. In this context, the implementation of corrective practices becomes problematic. Unfortunately, our knowledge of the climate effect on the quality of different grape varieties remains very incomplete to guide these choices. During the Innovine project, original experiments were carried out on Syrah to study the combined effects of normal or high air temperature and varying degrees of exposure of the berries to the sun. Berries subjected to these different conditions were sampled and analyzed throughout the maturation period. Several quality characteristics were determined, including anthocyanin content. The objective of the experiments was to investigate which climatic determinants were most important for anthocyanin accumulation in the berries. Temperature and irradiance data, observed over time with a very thin discretization step, are called functional data in statistics. We developed the procedure SpiceFP (Sparse and Structured Procedure to Identify Combined Effects of Functional Predictors) to explain the variations of a scalar response variable (a grape berry quality variable for example) by two or three functional predictors (as temperature and irradiance) in a context of joint influence of these predictors. Particular attention was paid to the interpretability of the results. Analysis of the data using SpiceFP identified a negative impact of morning combinations of low irradiance (lower than about 100 μmol m−2 s−1 or 45 μmol m−2 s−1 depending on the advanced-delayed state of the berries) and high temperature (higher than 25oC). A slight difference associated with overnight temperature occurred between these effects identified in the morning.

Impact of long term agroecological and conventional practices on subsurface soil microbiota in Macabeu and Xarel·lo vineyards

There is a growing trend on the transition from conventional to agroecological management of vineyards. However, the impact of practices, such as reduced-tillage, organic fertilization and cover crops, is not well-understood regarding the soil microbial diversity, and its relationship with the soil physicochemical properties in the subsurface depth near the rooting zone. Soil bacterial diversity is an important contributor towards plant health, productivity and response to environmental stresses. A field experiment was conducted by sampling subsurface soil bacterial community (NGS and qPCR) near to the root zone of Macabeu and Xarel·lo vineyards, located at the Penedes. 3 organic (ECO) and 3 conventional (CON) vineyards, with more than 10 years of respective management were sampled (n=5 each plot). ECO practices did not affect bacterial and fungal abundance but increased significantly the ammonium oxidizing bacteria and alpha-diversity (Inv.Simpson). Interestingly beta-diversity was significantly affected by the management strategy. ANOSIM-tests revealed a significative effect of the management (ecological vs conventional) and plot, on the soil microbial structure (ASV abundance). Main phyla depicted were Proteobacteria, Actinobacteria and Acidobacteria, whose relative abundances were not affected by the management. EdgeR assay revealed a significant increase of Cyanobacteria and decrease of Gemmatimonadetes and Firmicutes phyla in ECO. Interestingly, the grapevine variety was not correlated with the soil microbial community structure. Mantel-test revealed an important correlation (Spearman) of some physicochemical parameters with the soil microbiota structure, in order of importance: texture, EC, pH Ca/Mg, Mg/P, K+, Mg2+, Ca2+, SO42-, and OM. N-NH4 and NTK, which were higher in the ECO managed soils, did not correlated significantly with the soil microbiome population. The results revealed the importance of combining a deep physicochemical characterization of each replicate with the microbial diversity assessment to gain better insights on the relationship between soil microbiome and vineyard management.

Influence of weather and climatic conditions on the viticultural production in Croatia

The research includes an analysis of the impact of weather conditions on phenological development of the vine and grape quality, through monitoring of four experimental cultivars (Chardonnay, Graševina, Merlot and Plavac mali) over two production years. In each experimental vineyard, which were evenly distributed throughout the regions of Slavonia and The Croatian Danube, Croatian Uplands,

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.