Macrowine 2021
IVES 9 IVES Conference Series 9 Impact of mannoproteins structural features on the colloid stability when facing different kinds of wine polyphenols

Impact of mannoproteins structural features on the colloid stability when facing different kinds of wine polyphenols

Abstract

The aim was to study the impact of structural features in the polysaccharide moiety of mannoproteins on their interaction with polyphenols and the formation of colloidal aggregates. To this end, mannoproteins fractions were extracted from four different yeast strains: a commercial enological strain (MP-com), the wild-type BY4742 strain (MP-WT) and its mutants ΔMnn4 (MP-Mnn4) and ΔMnn2 (MP-Mnn2). The Mnn4p and Mnn2p are responsible for mannosyl-phosphorylation and branching of the N-glycosylation backbone [1]. Enzymatic extraction was performed using a commercial Endo-beta-1,3-Glucanase of Trichoderma sp. (E-LAMSE, Megazym)[2]. Mannoprotein fractions were thoroughly characterized by composition of their polysaccharide and protein moieties, branching degree, net charge, molecular weight distribution, static and dynamic molecular parameters [3]. Their interactions with seed tannins and a pool of red wine polyphenols and the formation of colloidal aggregates were studied in model solutions at different polyphenol/mannoprotein ratios through Dynamic Light Scattering (DLS). Model solutions were followed during one month. The number and size distribution of colloidal aggregates was determined by Nanoparticle Tracking Analysis (NTA).The four Mannoprotein fractions had broad and high molecular weight distributions, as well as similar protein, polysaccharide mass % and amino acid composition. However, they showed different proportions of mannose and glucose and the structural characterization of the polysaccharide moiety confirmed the expected differences between MP-WT, MP-Mnn2, and MP-Mnn4. DLS and NTA experiments indicated a two-step interaction process between seed tannins and mannoproteins: an immediate formation of colloidal aggregates (150-300 nm), followed by a very progressive evolution related to a reversible aggregate flocculation. The number, dispersity and extent of flocculation were dependent on the tannin/MP ratio. So far, no notable differences were evidenced between the four MP fractions. With the polyphenol pool of red wine, neither DLS nor NTA experiments were able to evidence the formation of colloidal aggregates. This does not mean that interactions do not exist[4,5]. Although the mannoproteins used had different polysaccharide compositions, structures, and properties, no difference in terms of colloidal behavior when in solution with tannins or wine polyphenols was evidenced by the methods applied. Thus, neither the absence of mannosyl phosphate groups (MP-Mnn4) nor the absence of branching of the outer chains of the N-glycosylated carbohydrate structures (MP-Mnn2) seems to play a determining role in the colloidal behavior of mannoproteins in the presence of seed tannins or red wine polyphenols.

DOI:

Publication date: September 14, 2021

Issue: Macrowine 2021

Type: Article

Authors

Saul Assunção Bicca

UMR-SPO, University of Montpellier, INRAE, Montpellier Supagro, Montpellier, France,Thierry, DOCO, UMR-SPO, University of Montpellier, INRAE, Montpellier Supagro, Montpellier, France Céline, PONCET-LEGRAND, UMR-SPO, University of Montpellier, INRAE, Montpellier Supagro, Montpellier, France Pascale, WILLIAMS, UMR-SPO, University of Montpellier, INRAE, Montpellier Supagro, Montpellier, France Julie MEKOUE N’GUELA, UMR-SPO, University of Montpellier, INRAE, Montpellier Supagro, Montpellier, France & Lallemand SAS, Blagnac, France Aude VERNHET, UMR-SPO, University of Montpellier, INRAE, Montpellier Supagro, Montpellier, France

Contact the author

Keywords

mannoproteins, colloidal stability, wine interactions

Citation

Related articles…

Comparison of imputation methods in long and varied phenological series. Application to the Conegliano dataset, including observations from 1964 over 400 grape varieties

A large varietal collection including over 1700 varieties was maintained in Conegliano, ITA, since the 1950s. Phenological data on a subset of 400 grape varieties including wine grapes, table grapes, and raisins were acquired at bud break, flowering, veraison, and ripening since 1964. Despite the efforts in maintaining and acquiring data over such an extensive collection, the data set has varying degrees of missing cases depending on the variety and the year. This is ubiquitous in phenology datasets with significant size and length. In this work, we evaluated four state-of-the-art methods to estimate missing values in this phenological series: k-Nearest Neighbour (kNN), Multivariate Imputation by Chained Equations (mice), MissForest, and Bidirectional Recurrent Imputation for Time Series (BRITS). For each phenological stage, we evaluated the performance of the methods in two ways. 1) On the full dataset, we randomly hold-out 10% of the true values for use as a test set and repeated the process 1000 times (Monte Carlo cross-validation). 2) On a reduced and almost complete subset of varieties, we varied the percentage of missing values from 10% to 70% by random deletion. In all cases, we evaluated the performance on the original values using normalized root mean squared error. For the full dataset we also obtained performance statistics by variety and by year. MissForest provided average errors of 17% (3 days) at budbreak, 14% (4 days) at flowering, 14.5% (7 days) at veraison, and 17% (3 days) at maturity. We completed the imputations of the Conegliano dataset, one of the world’s most extensive and varied phenological time series and a steppingstone for future climate change studies in grapes. The dataset is now ready for further analysis, and a rigorous evaluation of imputation errors is included.

The plantation frame as a measure of adaptation to climate change

The mechanization of vineyard work originally led to a reduction in planting densities due to the lack of machinery adapted to the vineyard. The current availability of specific machinery makes it possible to establish higher planting densities. In this work, three planting densities (1.40×0.80 m, 1.80×1 m and 2.20×1.20 m, corresponding to 8928, 5555 and 3787 plants/ha respectively) were studied with four varieties autochthonous of Galicia (northwestern Spain): Albariño and Treixadura (white), Sousón and Mencía (red). The vines were trained in a vertical shoot positioning system using a single Royat cordon, and pruned to spurs with two buds each. Agronomic data (yield, pruning wood weight, Ravaz index) and oenological data in must were collected. The higher planting density (1.40×0.80 m) had no significant effect on grape yield per vine in white varieties, although production per hectare was much higher due to the greater number of plants. In red varieties, this planting density resulted in a significantly lower production per vine, compensated by the greater number of plants. In addition, it significantly reduced the Brix degree in the must of the Albariño, Treixadura and Sousón varieties, and increased the total acidity in the latter two and Mencía. It also caused an increase in extractable and total anthocyanins and IPT in red grapes. The effects of high planting density on grapes are of great interest for the adaptation of varieties in the context of climate change. In the future, it could be advisable to modify the limits imposed by the appellations of origin on the planting density of these varieties in order to obtain more balanced wines.

VINIoT – Precision viticulture service

The project VINIoT pursues the creation of a new technological vineyard monitoring service, which will allow companies in the wine sector in the SUDOE space to monitor plantations in real time and remotely at various levels of precision. The system is based on spectral images and an IoT architecture that allows assessing parameters of interest viticulture and the collection of data at a precise scale (level of grape, plant, plot or vineyard) will be designed. In France, three subjects were specifically developed: evaluation of maturity, of water stress, and detection of flavescence dorée. For the evaluation of maturity, it has been decided first to work at the berry scale in the laboratory, then at the bunch scale and finally in the vineyard. The acquisition of the spectral hyperstal image as well as the reference analyzes to measure the maturity, were carried out in the laboratory after harvesting the berries in a maturity monitoring context. This work focuses on a case study to predict sugar content of three different grape varieties: Syrah, Fer Servadou and Mauzac. A robust method called Roboost-PLSR, developed in the framework of this work (Courand et al., 2022), to improve prediction model performance was applied on spectra after the acquirement of hyperspectral images. Regarding the evaluation of water stress, to work with a significant variability in terms of water status, it has been worked first with potted plants under 2 different water regimes. The facilities have allowed the supervision of irrigation and micro-climatic conditions. The regression models on agronomic variables (stomatal conductance, water potential, …) are studied. To detect flavescence dorée, the experimental plan has consisted of work at leaf scale in the laboratory first, and then in the field. To detect the disease from hyper-spectral imaging, a combination of multivariate curve resolution-alternating least squares (MCR-ALS) and factorial discriminant analysis (FDA) was proposed. This strategy proved the potential towards the discrimination of healthy and infected leaves by flavescence dorée based on the use of hyperspectral images (Mas Garcia et al., 2021).

Diagnosis of soil quality and evaluation of the impact of viticultural practices on soil biodiversity in a vineyard in southwestern France

Viticulture is facing two major changes – climate change and agroecological transition. In both cases, soil quality is seen as a lever to move towards a more sustainable viticulture. However, soil biological quality is little considered in the implementation of viticultural practices. Gascogn’Innov (2017-2022) is an Operational Group funded by the European Innovation Partnership for Agriculture. As such, it brings together winegrowers from the south-west of France, scientists, advisors and technicians, around a project focused on viticultural soil biological functioning and the design of technical routes more respectful toward soil heritage. To achieve this, the project aims to acquire references on the impact of viticultural practices on soil biology from a dynamic way, and to test a methodology to integrate information provided by the soil bioindicators to manage farming systems. A set of indicators of soil biological quality are evaluated in the project: microorganisms (bacteria and fungi abundance and diversity), fauna (abundance and diversity of nematodes and earthworms), physico-chemical characteristics, soil structure assessment and degradation rate of organic matter. Based on a network of 13 plots that have been subject to an initial diagnosis in 2017, several agronomical practices to restore soil fertility are experimented to redesign the cropping system (for instance plant cover, organic matter inputs, reduction of herbicides, mineral fertilizers). System redesign was made in collaboration by winegrowers and an interdisciplinary group of experts (agronomists, biologists). Several indicators are measured on vine and soil at each vintage to assess vine health and productivity. At the end of the project (2021), a final diagnosis was carried out. Gascogn’Innov allowed to create a regional database on the quality of wine-growing soils, which permitted to evaluate the effect of practices according to soil types. Especially, decreasing the intensity of tillage and increasing the duration and diversity of grass coverage tends to increase the abundance of all the organisms studied. This project confirmed the value of soil biological quality indicators to drive the sustainability of practices, but also highlighted the key-role of expertise, in both agronomy and soil biology, to help winegrowers understand and appropriate their soil quality diagnoses.

Sustainable fertilisation of the vineyard in Galicia (Spain)

Excessive fertilization of the vineyard leads to low quality grapes, increased costs and a negative impact on the environment. In order to establish an integrated management system aimed at a sustainable fertilization of the vineyards, nutritional reference levels were established. For this purpose, 30 representative vineyards of the Albariño variety were studied, in which soil and petiole analyses were carried out for two years and grape yield and quality at harvest were measured. In both years of study, soil pH, calcium, sodium and cation exchange capacity were positively correlated with calcium content and negatively correlated with manganese in grapes. Irrigated vineyards had higher levels of aluminium in soil and lower levels of calcium in petiole. Climatic conditions were very different in the years of the study. The year 2019 was colder than usual, in 2020 there was a marked water stress with high summer temperatures. This resulted in medium-high acidity in grapes in 2019 and low acidity in 2020, with sugar levels being similar both years. A very marked decrease in must amino nitrogen was observed in 2020, with ammonia nitrogen remaining stable. The correlation of acidity and sugar values in grapes with soil and petiole analysis data made it possible to establish reference levels for the nutritional diagnosis of the Albariño variety in this region. Based on these results, an easy-to-use TIC application is currently being created for grapegrowers, aimed at improving the sustainability of the vineyard through reasoned fertilization. This study has now been extended to other Galician vine varieties.