Macrowine 2021
IVES 9 IVES Conference Series 9 Optimised extraction and preliminary characterisation of mannoproteins from non-Saccharomyces wine yeasts

Optimised extraction and preliminary characterisation of mannoproteins from non-Saccharomyces wine yeasts

Abstract

The use of non-Saccharomyces yeast species for the improvement of wine technological and oenological properties is a topic that has gained much interest in recent years [1]. Their application as co-starter cultures sequential to the inoculation of Saccharomyces cerevisiae and in aging on the lees has been shown to improve aspects such as protein stability and mouthfeel [2]. These contributions have frequently been associated with higher levels of polysaccharides, specifically the cell wall-derived mannoprotein [3]. Furthermore, mannoprotein structure and composition has been shown to vary between yeast strains, which in turn may influence their behaviour in the wine matrix [4-6]. However, non-Saccharomyces yeasts are typically weak fermentors and are frequently out-competed in the fermentation medium. An alternative strategy to their use as co-starter cultures is the isolation of the compound of interest for exogenous application to wine [7]. Indeed, the addition of exogenous mannoprotein-containing products derived from the cell wall of the wine yeast S. cerevisiae is a fairly common winemaking practice [8]. Nevertheless, the extraction of mannoproteins from non-Saccharomyces yeasts has not yet been well described. AIM: This study aimed to optimise the extraction of mannoproteins from four non-Saccharomyces strains, and to perform a preliminary investigation into the compositional differences of the mannoproteins obtained from the different species.

METHODS: Four non-Saccharomyces wine strains, Saccharomyces cerevisiaeSaccharomyces boulardiiMetschnikowia fructicola and Torulaspora delbrueckii, were exposed to combined methods with varied parameters of ultrasound and enzymatic extraction with β-glucanase to optimise mannoprotein yield. Colorimetric assays were used to quantify protein and carbohydrate concentrations in the extracts.

RESULTS: Yeast cells subjected to 4 min of ultrasound treatment applied at 80% of the maximum amplitude with a 50% duty cycle, followed by an enzymatic treatment of 4000 U lyticase per g dry cells weight, showed the highest yield of mannoproteins from all species. Furthermore, preliminary evaluation of the obtained extracts revealed differences in carbohydrate/protein ratios between species and with increased enzyme incubation time, as demonstrated by the higher ratios obtained for T. Delbrueckii and S. cerevisiae after almost all treatments, in comparison to M. fructicola and S. boulardii.

CONCLUSIONS: The results obtained in this study form an important step towards further characterisation of extraction treatment impact and yeast species effect on the extracted mannoproteins. Their impact on the carbohydrate/protein ratio in particular is an important factor to consider for applications such as wine protein haze reduction and tartrate stabilisation, and requires more in-depth investigation of isolated mannoproteins.

DOI:

Publication date: September 3, 2021

Issue: Macrowine 2021

Type: Article

Authors

Carla Snyman, Benoit DIVOL, Matteo MARANGON, Julie MEKOUE NGUELA, Nathalie SCIECZKOWSKI

South African Grape and Wine Research Institute, Department of Viticulture and Oenology, Private Bag X1, Matieland 7602, South Africa, South African Grape and Wine Research Institute, Department of Viticulture and Oenology, Private Bag X1, Matieland 7602, South Africa, Department of Agronomy, Food, Natural Resources, Animals and Environment (DAFNAE), University of Padova, Viale Dell’Università, 16, 35020, Legnaro, Padova, Italy, Lallemand SAS, 19 rue des briquetiers, BP 59, 31702 Blagnac, France, Lallemand SAS, 19 rue des briquetiers, BP 59, 31702 Blagnac, France

Contact the author

Keywords

mannoprotein; yeast; non-saccharomyces; extraction; wine; ultrasound; β-glucanase

Citation

Related articles…

The interplay between grape ripening and weather anomalies – A modeling exercise

Current climate change is increasing inter- and intra-annual variability in atmospheric conditions leading to grapevine phenological shifts as well altered grape ripening and composition at ripeness. This study aims to (i) detect weather anomalies within a long-term time series, (ii) model grape ripening revealing altered traits in time to target specific ripeness thresholds for four Vitis vinifera cultivars, and (iii) establish empirical relationships between ripening and weather anomalies with forecasting purposes. The Day of the Year (DOY) to reach specific grape ripeness targets was determined from time series of sugar concentrations, total acidity and pH collected from a private company in the period 2009-2021 in North-Eastern Italy. Non-linear models for the DOY to reach the specified ripeness thresholds were assessed for model efficiency (EF) and error of prediction (RMSE) in four grapevine cultivars (Merlot, Cabernet Sauvignon, Glera and Garganega). For each vintage and cultivar, advances or delays in DOY to target specified ripeness thresholds were assessed with respect to the average ripening dynamics. Long-term meteorological series monitored at ground weather station by means of hourly air temperature and rainfall data were analyzed. Climate statistics were obtained and for each time period (month, bimester, quarter and year) weather anomalies were identified. A linear regression analysis was performed to assess a possible correlation that may exist between ripening and weather anomalies. For each cultivar, ripeness advances or delays expressed in number of days to target the specific ripening threshold were assessed in relation to registered weather anomalies and the specific reference time period in the vintage. Precipitation of the warmest month and spring quarter are key to understanding the effect of climate change on sugar ripeness. Minimum temperatures of May-June bimester and maximum temperatures of spring quarter best correlate with altered total acidity evolution and pH increment during the ripening process, respectively.

Phenolic composition of Tempranillo Blanco grapes changes after foliar application of urea

Our research aimed to determine the effect and efficiency of foliar application of urea on the phenolic composition of Tempranillo Blanco grapes. The field experiment was carried out in 2019 and 2020 seasons and the plot was located in D.O.Ca Rioja (North of Spain). The vineyard was Vitis vinifera L. Tempranillo Blanco and grafted on Richter-110 rootstock. The treatments were control (C), whose plants were sprayed with water and three doses of urea: plants were sprayed with urea 3 kg N/ha (U3), 6 kg N/ha (U6) and 9 kg N/ha (U9). The applications were performed in two phenological stages, pre-veraison (Pre) and veraison (Ver). Also, each of the treatments was repeated one week later. Control and treatments were performed in triplicate and arranged in a randomised block design. Grapes were harvested at optimum ripening stage. High-performance liquid chromatography was used to analyse the phenolic composition of the grapes. Finally, the results obtained from the analytical determinations – flavonols, flavanols and non-flavonoid (hydroxybenzoic acids, hydroxycinnamic acids and stilbenes) – were studied statistically by analysis of variance. The results showed that, in 2019, U6-Pre and U9-Pre treatments increased the hydroxybenzoic acid content in grapes, and also all foliar treatments applied at Pre enhanced the stilbene concentration. Moreover, U3-Ver was the only treatment that rose flavonol and stilbene contents in the Tempranillo Blanco grapes. In 2020, all treatments applied at Pre enhanced the flavonol concentration in grapes. Furthermore, U3-Pre and U9-Pre treatments increased stilbene content in grapes. Nevertheless, the hydroxybenzoic acid content was improved by U6-Ver and U9-Ver and besides, hydroxycinnamic acid concentration in grapes was increased by all treatments applied at Ver. In conclusion, the lower and highest dose of urea (U3 and U9), applied at pre-veraison, were the best treatments to improve the Tempranillo Blanco grape phenolic composition.

What are the optimal ranges and thresholds for berry solar radiation for flavonoid biosynthesis?

In wine grape production, canopy management practices are applied to control the source-sink balance and improve the cluster microclimate to enhance berry composition. The aim of this study was to identify the optimal ranges of berry solar radiation exposure (exposure) for upregulation of flavonoid biosynthesis and thresholds for their degradation, to evaluate how canopy management practices such as leaf removal, shoot thinning, and a combination of both affect the grapevine (Vitis vinifera L. cv. Cabernet Sauvignon) yield components, berry composition, and flavonoid profile under context of climate change. First experiment assessed changes in the grape flavonoid content driven by four degrees of exposure. In the second experiment, individual grape berries subjected to different exposures were collected from two cultivars (Cabernet Sauvignon and Petit Verdot). The third experiment consisted of an experiment with three canopy management treatments (i) LR (removal of 5 to 6 basal leaves), (ii) ST (thinned to 24 shoots per vine), and (iii) LRST (a combination of LR and ST) and an untreated control (UNT). Berry composition, flavonoid content and profiles, and 3-isobutyl 2-methoxypyrazine were monitored during berry ripening. Although increasing canopy porosity through canopy management practices can be helpful for other purposes, this may not be the case of flavonoid compounds when a certain proportion of kaempferol was achieved. Our results revealed different sensitivities to degradation within the flavonoid groups, flavonols being the only monitored group that was upregulated by solar radiation. Within different canopy management practices, the main effects were due to the ST. Under environmental conditions given in this trial, ST and LRST hastened fruit maturity; however, a clear improvement of the flavonoid compounds (i.e., greater anthocyanin) was not observed at harvest. Methoxypyrazine berry content decreased with canopy management practices studied. Although some berry traits were improved (i.e. 2.5° Brix increase in berry total soluble solids) due to canopy management practices (ST), this resulted in a four-fold increase in labor operations cost, two-fold decrease in yield with a 10-fold increase in anthocyanin production cost per hectare that should be assessed together as the climate continues to get hot.

Biodiversity in the vineyard agroecosystem: exploring systemic approaches

Biodiversity conservation and restoration are essential for guarantee the provision of ecosystem services associated to vineyard agroecosystem such as climate regulation trough carbon sequestration and control of pests and diseases. Most of published research dealing with the complexity of the vineyard agroecosystems emphasizes the necessity of innovative approaches, including the integration of information at different temporal and spatial scales and development of systemic analysis based on modelling. A biodiversity survey was conducted in the Franciacorta wine-growing area (Lombardy, Italy), one of the most important Italian wine-growing regions for sparkling wine production, considering a portion of the territory of 112 ha. The area was divided into several Environmental Units (EUs), defined as a whole vineyard or portion of vineyard homogenous in terms of four agronomic characteristics: planting year, planting density, cultivar, and training system. In each EU a set of compartments was identified and characterised by specific variables. The compartments are meteorology, morphology (altitude, slope, aspect, row orientation, and solar irradiance), ecological infrastructures and management. The landscape surrounding EU was also characterised in terms of land-use in a buffer zone of 500 m. For each component a specific methodology was identified and applied. Different statistical approaches were used to evaluate the method to integrate the information related to different compartments within the EU and related to the buffer zone. These approaches were also preliminarily evaluated for their ability to describe the contribution of biodiversity and landscape components to ecosystem services. This methodological exploration provides useful indication for the development of a fully systemic approach to structural and functional biodiversity in vineyard agroecosystems, contributing to promote a multifunctional perspective for the all wine-growing sector.

Projected changes in vine phenology of two varieties with different thermal requirements cultivated in La Mancha DO (Spain) under climate change scenarios

The aim of this work was to analyze the phenology variability of Tempranillo and Chardonnay cultivars, related to the climatic characteristics in La Mancha Designation of Origin, and their potential changes under climate change scenarios. Phenological dates referred to budbreak, flowering, veraison and harvest were analyzed for the period 2000-2019. The weather conditions at daily time scale, recorded during the same period, were also evaluated. The thermal requirements to reach each of these phenological stages were calculated and expressed as the GDD accumulated from DOY=60. Changes in phenology were projected by 2050 and 2070 taking into account those values and the projected temperatures and precipitation, simulated under two Representative Concentration Pathway (RCP) scenarios –RCP4.5 and RCP8.5– using an ensemble of models. The average phenological dates during the period under study were, April 16th ± 6.6 days and April 5th ± 6.0 days for budbreak, May 31st ± 6.0 days and May 27th ± 5.3 days for flowering, July 26th ± 5.6 days and July 25th ± 5.8 days for veraison, and Ago 23rd ± 10.8 days and Ago 17th ± 9.0 days for harvest, respectively, for Tempranillo and Chardonnay. The projected changes in temperature imply an average change in the maximum growing season (April-August) temperatures of 1.2 and 1.9°C by 2050, and 1.6 and 2.6°C by 2070, under the RCP4.5 and RCP8.5 scenarios, respectively. A reduction in precipitation is predicted, which vary between 15% for 2050 under RCP4.5 scenario and up to 30% by 2070 under RCP8.5. The advance of the phenological dates for 2050, could be of 6, 7, 7, and 8 days for Tempranillo and 4, 6, 6 and 9 days for Chardonnay, respectively for budbreak, flowering, veraison and harvest under the RCP4.5 scenario. Under the RCP8.5 emission scenario, the advance could be up to 30% higher.

Macrowine 2021
IVES 9 IVES Conference Series 9 Optimised extraction and preliminary characterisation of mannoproteins from non-Saccharomyces wine yeasts

Optimised extraction and preliminary characterisation of mannoproteins from non-Saccharomyces wine yeasts

Abstract

Content of the article

References

Section for all references

DOI:

Publication date: September 3, 2021

Issue: (ex: Issue: Terclim 2023)

Type: typeofpublication

Authors

author1, author2, author3

Presenting author

Description

List of affiliations ¹ ² ³

Contact the author

Email address (with mailto: link)

Keywords

List of different keywords (keyword1, keyword2, keyword3)

Tags

Citation

Related articles…

A spatial explicit inventory of EU wine protected designation of origin to support decision making in a changing climate

Winemaking areas recognized as protected designations of origin (PDOs) shape important economic, environmental and cultural values that are tied to closely defined geographic locations. To preserve wine products and wine-growing practices adopted in different PDOs these areas are strictly regulated by legal specifications. However, quality viticulture is increasingly under pressure from climate change, which is altering the local conditions of many winegrowing areas. Therefore, maintaining traditional wine products will require the adoption of tailored adaptation strategies, including possible changes in the legal regulation of protected wines. To this end, it is necessary to have a comprehensive knowledge on PDOs including their extension, products and allowed practices. While there have been efforts to build databases that summarize the characteristics for individual wine PDO areas and to quantify the related effects of climate change, much information is still included only in the official documentation of the EU geographical indication register and has never been collected in a comprehensive manner. With this study we aim at filling this gap by building a spatial inventory of European wine PDOs that supports decision making in viticulture in the context of climate change. To map and characterize European wine PDOs, we analysed their legal documents and extracted relevant information useful for climate change adaptation. The output consists of a comprehensive geographical dataset that identifies the boundaries of all 1200 European wine PDOs at unprecedented spatial resolution and includes a set of legally binding regulations, such as authorized vine varieties, maximum yields and planting density. The inventory will allow researchers to analyse the impacts of climate change on European wine PDOs and support decision makers in developing tailored adaptation strategies. This includes, among others, the evaluation of new vineyard site selection, the expansion of cultivated varieties or the authorization of irrigation in vineyards.

Grapevine yield-gap: identification of environmental limitations by soil and climate zoning in Languedoc-Roussillon region (south of France)

Grapevine yield has been historically overlooked, assuming a strong trade-off between grape yield and wine quality. At present, menaced by climate change, many vineyards in Southern France are far from the quality label threshold, becoming grapevine yield-gaps a major subject of concern. Although yield-gaps are well studied in arable crops, we know very little about grapevine yield-gaps. In the present study, we analysed the environmental component of grapevine yield-gaps linked to climate and soil resources in the Languedoc Roussillon. We used SAFRAN data and IGP Pays d’Oc wine yields from 2010 to 2018. We selected climate and soil indicators proving to have a significant effect on average wine yield-gaps at the municipality scale. The most significant factors of grapevine yield were the Soil Available Water Capacity; followed by the Huglin Index and the Climatic Dryness Index. The Days of Frost; the Soil pH; and the Very Hot Days were also significant. Then, we clustered geographical zones presenting similar indicators, facilitating the identification of resources yield-gaps. We discussed the number of zones with the experts of IGP Pays d’Oc label, obtaining 7 zones with similar limitations for grapevine yield. Finally, we analysed the main resources causing yield-gaps and the grapevine varieties planted on each zone. Mapping grapevine resource yield-gaps are the first stage for understanding grapevine yield-gaps at the regional scale.

Estimating bulk stomatal conductance of grapevine canopies

In response to changes in their environment, grapevines regulate transpiration using various physiological mechanisms that alter conductance of water through the soil-plant-atmosphere continuum. Expressed as bulk stomatal conductance at the canopy scale, it varies diurnally in response to changes in vapor pressure deficit and net radiation, and over the season to changes in soil water deficits and hydraulic conductivity of both soil and plant. It is necessary to characterize the response of conductance to these variables to better model how vine transpiration also responds to these variables. Furthermore, to be relevant for vineyard-scale modeling, conductance is best characterized using data collected in a vineyard setting. Applying a crop canopy energy flux model developed by Shuttleworth and Wallace, bulk stomatal conductance was estimated using measurements of individual vine sap flow, temperature and humidity within the vine canopy, and estimates of net radiation absorbed by the vine canopy. These measurements were taken on several vines in a non-irrigated vineyard in Bordeaux France, using equipment that did not interfere with ongoing vineyard operations. An inverted Penman-Monteith equation was then used to calculate bulk stomatal conductance on 15-minute intervals from July to mid-September 2020. Time-series plots show significant diurnal variation and seasonal decreases in conductance, with overall values similar to those in the literature. Global sensitivity analysis using non-parametric regression found transpiration flux and vapor pressure deficit to be the most important input variables to the calculation of bulk stomatal conductance, with absorbed net radiation and bulk boundary layer conductance being much less important. Conversely, bulk stomatal conductance was one of the most important inputs when calculating vine transpiration, further emphasizing the need for characterizing its response to environmental changes for use in vineyard water use modeling.

Heatwaves and grapevine yield in the Douro region, crop model simulations

Heatwaves or extreme heat events can be particularly harmful to agriculture. Grapevines grown in the Douro winemaking region are particularly exposed to this threat, due to the specificities of the already warm and dry climatic conditions. Furthermore, climate change simulations point to an increase in the frequency of occurrence of these extreme heat events, therefore posing a major challenge to winegrowers in the Mediterranean type climates. The current study focuses on the application of the STICS crop model to assess the potential impacts of heatwaves in grapevine yields over the Douro valley winemaking region. For this purpose, STICS was applied to grapevines using high-resolution weather, soil and terrain datasets over the Douro. To assess the impact of heatwaves, the weather dataset (1989-2005) was artificially modified, generating periods with anomalously high temperatures (+5 ºC), at certain onset dates and with specific durations (from 5 to 9 days). The model was run with this modified weather dataset and results were compared to the original unmodified runs. The results show that heatwaves can have a very strong impact on grapevine yields, strongly depending on the onset dates and duration of the heatwaves. The highest negative impacts may result in a decrease in the yield by up to -35% in some regions. Despite some uncertainties inherent to the current modelling assessment, the present study highlights the negative impacts of heatwaves on viticultural yields in the Douro region, which is critical information for stakeholders within the winemaking sector for planning suitable adaptation measures.

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.