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…

An analytical framework to site-specifically study climate influence on grapevine involving the functional and Bayesian exploration of farm data time series synchronized using an eGDD thermal index

Climate influence on grapevine physiology is prevalent and this influence is only expected to increase with climate change. Although governed by a general determinism, climate influence on grapevine physiology may present variations according to the terroir. In addition, these site-specific differences are likely to be enhanced when climate influence is studied using farm data. Indeed, farm data integrate additional sources of variation such as a varying representativity of the conditions actually experienced in the field. Nevertheless, there is a real challenge in valuing farm data to enable grape growers to understand their own terroir and consequently adapt their practices to the local conditions. In such a context, this article proposes a framework to site-specifically study climate influence on grapevine physiology using farm data. It focuses on improving the analysis of time series of weather data. The analytical framework includes the synchronization of time series using site-specific thermal indices computed with an original method called Extended Growing Degree Days (eGDD). Synchronized time series are then analyzed using a Bayesian functional Linear regression with Sparse Steps functions (BLiSS) in order to detect site-specific periods of strong climate influence on yield development. The article focuses on temperature and rain influence on grape yield development as a case study. It uses data from three commercial vineyards respectively situated in the Bordeaux region (France), California (USA) and Israel. For all vineyards, common periods of climate influence on yield development were found. They corresponded to already known periods, for example around veraison of the year before harvest. However, the periods differed in their precise timing (e.g. before, around or after veraison), duration and correlation direction with yield. Other periods were found for only one or two vineyards and/or were not referred to in literature, for example during the winter before harvest.

Genotypic variability in root architectural traits and putative implications for water uptake in grafted grapevine

Root system architecture (RSA) is important for soil exploration and edaphic resources acquisition by the plant, and thus contributes largely to its productivity and adaptation to environmental stresses, particularly soil water deficit. In grafted grapevine, while the degree of drought tolerance induced by the rootstock has been well documented in the vineyard, information about the underlying physiological processes, particularly at the root level, is scarce, due to the inherent difficulties in observing large root systems in situ. The objectives of this study were to determine genetic differences in the root architectural traits and their relationships to water uptake in two Vitis rootstocks genotypes (RGM, 140Ru) differing in their adaptation to drought. Young rootstocks grafted upon the Riesling variety were transplanted into cylindrical tubes and in 2D rhizotrons under two conditions, well watered and moderate water stress. Root traits were analyzed by digital imaging and the amount of transpired water was measured gravimetrically twice a week. Root phenotyping after 30 days reveal substantial variation in RSA traits between genotypes despite similar total root mass; the drought-tolerant 140Ru showed higher root length density in the deep layer, while the drought-sensitive RGM was characterised by shallow-angled root system development with more basal roots and a larger proportion of fine roots in the upper half of the tube. Water deficit affected canopy size and shoot mass to a greater extent than root development and architectural-related traits for both 140Ru and RGM, suggesting vertical distribution of roots was controlled by genotype rather than plasticity to soil water regime. The deeper root system of 140Ru as compared to RGM correlated with greater daily water uptake and sustained stomata opening under water-limited conditions but had little effect on above-ground growth. Our results highlight that grapevine rootstocks have constitutively distinct RSA phenotypes and that, in the context of climate change, those that develop an extensive root network at depth may provide a desirable advantage to the plant in coping with reduced water resources.

δ13C : A still underused indicator in precision viticulture  

The first demonstration of the interest of carbon isotope composition of sugars in grapevine, as an integrated indicator of vineyard water status, dates back to 2000 (Gaudillère et al., 1999; Van Leeuwen et al., 2001). Thanks to the isotopic discrimination of Carbon that takes place during plant photosynthesis, under hydric stress conditions, it is possible to accurately estimate the photosynthetic activity. Ever since, δ13C has been widely applied with success to zonation, terroir studies and vine physiology research, but is still not widely used by viticulturists. This is quite astonishing by considering the impact of global warming on viticulture and the need to improve water management, that would justify a widespread use of δ13C.
The lack of private laboratories proposing the analysis, the cost of the technology, as well as the long analytical delays, have been detrimental to its development. Some laboratories tried to overcome the analytical difficulties of isotopic analysis by using fourier transformed infrared spectroscopy, as a fast and cheap alternative to the official OIV method (IRMS). These claimed FTIR models have never been published or peer reviewed and cannot be considered robust. In this work, thanks to the recent acquisition of IRMS technology, new modern and robust applications of δ13C for viticulture are proposed. This includes the use of the analysis to make parcel separations at harvesting, the possibility to increase the precision of hydric stress cartography and the potential cost reduction when compared with Scholander pressure bomb analysis.

How distinctive are single vineyard Gewürztraminer musts and wines from Alto Adige (Italy) based on untargeted analysis, sensory profiling, and chemometric elaboration?

Vitis vinifera L. ‘Gewürztraminer’ is a historical grape variety of Alto Adige (Südtirol), Italy, which is widely grown in the area of Tramin an der Weinstraße, but is also grown globally. It produces highly aromatic wines that are strongly influenced by the terroir of the vineyard sites where they are grown. This study looked at musts and young wines from ‘Gewürztraminer’ grapes harvested in seven distinct vineyards near Tramin and then processed at Cantina di Termeno, minimizing winemaking protocol variability. Samples were profiled using bidimensional gas chromatography–time-of-flight mass spectrometry, liquid chromatography coupled to electrochemical detection, and near-IR spectrometry. The data were subjected to Principle Component Analysis and Hierarchical Clustering Analysis. Sensory discriminant testing was undertaken using the sorting method with a semi-trained panel, and the data were processed using Multidimensional Scaling. Seven must/wine pairs could be distinguished based on their untargeted volatilome profiles and on sensory evaluation. As expected, there were greater differences in the volatile compounds between the wines than between the musts. The wines from vineyards 4 and 5 were nonetheless quite homogenous in terms of chemical and sensory analyses, as were the wines from vineyards 1 and 3. For the phenolic profile, differences were noted between the musts and wines of vineyards 2, 3, and 4, but the musts from vineyards 5 and 7 were similar. Sensory analysis showed the wines from vineyards 6 and 7 to be distinct from the rest. These results reinforce that the composition of ‘Gewürztraminer’ musts and wines is strongly determined by vineyard site, even in a small geographic area with high variability of the terroir (soil and microclimate), and that these differences are apparent in the flavours and aromas of the finished wines. Further confirmation would require a larger sample of wines, preferably from several vintages.

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.

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…

Green berries on Gewürztraminer (Vitis vinifera L.) in South Tyrol (Italy)

The grape variety Gewürztraminer is known to be affected by two physiological disorders namely berry shrivel and bunch stem necrosis. During the season 2014 we noticed a new symptomatology type of ripening disorder on the variety. The new symptom showed not all berries fallowing the normal maturation stages, but single berries remaining at a soft but green stage till harvest. The broad distribution of these so called “green berries” symptoms in different production sites of our region, caused huge damage due to the difficulty of eliminating single berries per bunch before harvesting. Therefore, the Research Centre Laimburg began to investigate the reasons and origins of this new symptom. This work shows the results of first attempts to find causes for the symptom as well as the resulting approach to mitigate symptoms. Applications of magnesium leaf fertilizer showed first promising results against this putative disorder. To study the causal effect of the green berries 30 symptomatic vineyards in 2014 have been selected for a monitoring during the season 2016. To evaluate the foliar nutrient treatment two vineyards have been selected for application of magnesium sulfate and magnesium chloride. Leaf and berry nutrient analysis, as well as the main quality parameters during ripening have been performed. As soon as “green berries” symptoms appeared, incidence and severity have been evaluated. Most of the symptomatic vineyards of the 2016 monitoring showed light to clear magnesium deficit symptoms on their foliage. Only during the seasons 2020 and 2021 “green berries” symptoms could be found in the leaf fertilizer treatment vineyards. Both seasons showed a significant effect of the magnesium treatments to reduce the incidence and severity of the symptom. It seems that the appearance of the “green berries” symptom on Gewürztraminer is correlated to a disturbed uptake of magnesium of the vines.

Grapevine xylem embolism resistance spectrum reveals which varieties have a lower mortality risk in a future dry climate

Wine growing regions have recently faced intense and frequent droughts that have led to substantial economical losses, and the maintenance of grapevine productivity under warmer and drier climate will rely notably on planting drought-resistant cultivars. Given that plant growth and yield depend on water transport efficiency and maintenance of photosynthesis, thus on the preservation of the vascular system integrity during drought, a better understanding of drought-related hydraulic traits that have a significant impact on physiological processes is urgently needed. We have worked towards this end by assessing vulnerability to xylem embolism in 30 grapevine commercial varieties encompassing red and white Vitis vinifera varieties, hybrid varieties characterized by a polygenic resistance for powdery and downy mildew, and commonly used rootstocks. These analyses further allowed a global assessment of wine regions with respect to their varietal diversity and resulting vulnerability to stem embolism. Hybrid cultivars displayed the highest vulnerability to embolism, while rootstocks showed the greatest resistance. Significant variability also arose among Vitis vinifera varieties, with Ψ12 and Ψ50 values ranging from -0.4 to -2.7 MPa and from -1.8 to -3.4 MPa, respectively. Cabernet franc, Chardonnay and Ugni blanc featured among the most vulnerable varieties while Pinot noir, Merlot and Cabernet Sauvignon ranked among the most resistant. In consequence, wine regions bearing a significant proportion of vulnerable varieties, such as Poitou-Charentes, France and Marlborough, New Zealand, turned out to be at greater risk under drought. These results highlight that grapevine varieties may not respond equally to warmer and drier conditions, outlining the importance to consider hydraulic traits associated with plant drought tolerance into breeding programmes and modeling simulations of grapevine yield maintenance under severe drought. They finally represent a step forward to advise the wine industry about which varieties and regions would have the lowest risk of drought-induced mortality under climate change.

Effect of vigour and number of clusters on eonological parameters and metabolic profile of Cabernet Sauvignon red wines

Vegetative growth and yield are reported to affect grape and wine quality. They can be controlled through different techniques linked to vine management. The objective of this research was to determine the effect of vine vigour and number of clusters per vine on physicochemical composition and phenolic profile of red wines. The experiment was carried out during two vegetative cycles, with cv. Cabernet Sauvignon grafted onto Paulsen 1103. Three vine vigour were defined, according to shoot weight at previous harvests, being low, medium and high. Five treatments of number of clusters were used for each vigour, with 15, 22, 29, 36, and 45 clusters per vine. Grapes from all treatments were harvested in the same day from Brix and total acidity criteria. Thirty days after bottling, classical analyzes and phenolic compounds were performed. As results, different responses were obtained from each vintage. In 2020, a dry season from veraison to harvest, grapes and wines obtained from low vigour treatment and 45 clusters per vine was the highest in sugar and alcohol content respectively, while grapes and wines from high vigour and 15 clusters presented the lowest sugar and alcohol content. Total anthocyanins were higher in treatment with low vigour and 15 clusters, while the lowest amounts were found in low vigour with 45 clusters, as well as medium and high vigour with 36 clusters per vine. Total tannins were higher in high vigour with 22 clusters and medium vigour with 29 clusters, while were lower in low vigour with 36 clusters. In 2021, a wet season at harvest, responses were different, and great variations were observed between treatments. As conclusions, yield and vine vigour had strong influence on grape and wine quality, promoting different enological potentials on which can be indicated/used for aging strategies of red and even rosé wines.

Adaptability of grapevines to climate change: characterization of phenology and sugar accumulation of 50 varieties, under hot climate conditions

Climate is the major factor influencing the dynamics of the vegetative cycle and can determine the timing of phenological periods. Knowledge of the phenology of varieties, their chronological duration, and thermal requirements, allows not only for the better management of interventions in the vineyard, but also to predict the varieties’ behaviour in a scenario of climate change, giving the wine producer the possibility of selecting the grape varieties that are best adapted to the climatic conditions of a certain terroir. In 2014, Symington Family Estates, Vinhos, established two grape variety libraries in two different places with distinctive climate conditions (Douro Superior, and Cima Corgo), with the commitment of contributing to a deeper agronomic and oenological understanding of some grape varieties, in hot climate conditions. In these research vineyards are represented local varieties that are important in the regional and national viticulture, but also others that have over time been forgotten — as well as five international reference cultivars. From 2017 to 2021, phenological observations have been made three times a week, following a defined protocol, to determine the average dates of budbreak, flowering and veraison. With the climate data of each location, the thermal requirements of each variety and the chronological duration of each phase have been calculated. During maturation, berry samples have been gathered weekly to study the dynamics of sugar accumulation, between other parameters. The data was analysed applying phenological and sugar accumulation models available in literature. The results obtained show significant differences between the varieties over several parameters, from the chronological duration and thermal requirements to complete the various stages of development, to the differences between the two locations, confirming the influence of the climate on phenology and the stages of maturation, in these specific conditions.

Aromatic maturity is a cornerstone of terroir expression in red wine

Harvesting grapes at adequate maturity is key to the production of high-quality red wines. Enologists and wine makers define several types of maturity, including technical maturity, phenolic maturity and aromatic maturity. Technical maturity and phenolic maturity are relatively well documented in the scientific literature, while articles on aromatic maturity are scarcer. This is surprising, because aromatic maturity is, without a doubt, the most important of the three in determining wine quality and typicity (including terroir expression). Optimal terroir expression can be obtained when the different types of maturity are reached at the same time, or within a short time frame. This is more likely to occur when the ripening takes place under mild temperatures, neither too cool, nor too hot. Aromatic expression in wine can be driven, from low to high maturity, by green, herbal, fresh fruit, ripe fruit, jammy fruit, candied fruit or cooked fruit aromas. Green and cooked fruit aromas are not desirable in red wines, while the levels of other aromatic compounds contribute to the typicity of the wine in relation to its origin. Wines produced in cool climates, or on cool soils in temperate climates, are likely to express herbal or fresh fruit aromas; while wines produced under warm climates, or on warm soils in temperate climates, may express ripe fruit, jammy fruit or candied fruit aromas. Growers can optimize terroir expression through their choice of grapevine variety. Early ripening varieties perform better in cool climates and late ripening varieties in warm climates. Additionally, maturity can be advanced or delayed by different canopy management practices or training systems.