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…

Exploring resilience and competitiveness of wine estates in Languedoc-Roussillon in the recent past: a multi-level perspective

The Languedoc-Roussillon wineries are facing a decline in wine yields particularly PGI yields due to many factors. Climate change is just ones, but is expected to increase in the future. There is also structurally a large heterogeneity of yield profiles among terroirs, varieties and strategies. This work investigates the link between yield, competitiveness and resilience to explore how resilient winegrowers have been in the recent past. To this end two approaches have been combined; (i) an accountancy database analysis at estate scale and (ii) municipality level competitiveness analysis. A new resilience indicator that characterizes the capacity of an estate to absorb yield variation is also defined. The FADN database between 2000 and 2018 of ex-Languedoc-Roussillon (France) and other data are used to analyse the current situation and the past evolution of competitiveness and resilience by type of estate (type of farm: PGI and/or PDO & type of commercialization: bulk and/or bottles). The net margin, which defines competitiveness, is not correlated to yield for all types but depends on the type of commercialization and the level of specialisation. The resilience indicator shows that the net margin of estates specialized in PGI is particularly sensitive to yield declines. We also show that price evolutions seem to compensate the effect of yield losses for the majority of types. Municipality scale analysis shows the links between local pedoclimate, yield, commercialization strategies and price. Overlapping a PDO with a PGI does not always increase a municipality’s PGI competitiveness. It is difficult to make links between causes and effects due to the complexity of the wine production system. Production diversification may be a solution. Resorting to the two level of analysis helps resolving the data gap that is necessary to explore the links between yield and economic performance of the wine estates in the long term.

A blueprint for managing vine physiological balance at different spatial and temporal scales in Champagne

In Champagne, the vine adaptation to different climatic and technical changes during these last 20 years can be seen through physiological balance disruptions. These disruptions emphasize the general grapevine decline. Since the 2000s, among other nitrogen stress indicators, the must nitrogen has been decreasing. The combination of restricted mineral fertilizers and herbicide use, the growing variability of spring rainfall, the increasing thermal stress as well as the soil type heterogeneity are only a few underlying factors that trigger loss of physiological balance in the vineyards. It is important to weigh and quantify the impact of these factors on the vine. In order to do so, the Comité Champagne uses two key-tools: networking and modelization. The use of quantitative and harmonized ecophysiological indicators is necessary, especially in large spatial scales such as the Champagne appellation. A working group with different professional structures of Champagne has been launched by the Comité Champagne in order to create a common ecophysiology protocol and thus monitor the vine physiology, yearly, around 100 plots, with various cultural practices and types of soil. The use of crop modelling to follow the vine physiological balance within different pedoclimatic conditions enables to understand the present balance but also predict the possible disruptions to come in future climatic scenarios. The physiological references created each year through the working group, benefit the calibration of the STICS model used in Champagne. In return, the model delivers ecophysiology indicators, on a daily scale and can be used on very different types of soils. This study will present the bottom-up method used to give accurate information on the impacts of soil, climate and cultural practices on vine physiology.

Effect of the commercial inoculum of arbuscular mycorrhiza in the establishment of a commercial vineyard of the cultivar “Manto negro

The favorable effect of symbiosis with arbuscular mycorrhizal fungi (AMF) has been known and studied since the 60s. Nowadays, many companies took the chance to start promoting and selling commercial inoculants of AMF, in order to be used as biofertilizers and encourage sustainable biological agriculture. However, the positive effect of these commercial biofertilizers on plant growth is not always demonstrated, especially under field conditions. In this study, we used a commercial inoculum on newly planted grapevines of a local cultivar grafted on a common rootstock R110. We followed the physiological status of vines, growth and productivity and functional biodiversity of soil bacteria during the first and second years of 20 inoculated with commercial inoculum bases on Rhizophagus irregularis and Funeliformis mosseaeAMF at field planting time and 20 non-inoculated control plants. All the parameters measured showed a neutral to negative effect on plant growth and production. The inoculated plants always presented lower values of photosynthesis, growth and grape production, although in some cases the differences did not reach statistical significance. On the contrary, the inoculation supposed an increase of the bacterial functional diversity, although the differences were not statistically significant either. Several studies show that the effect of inoculation with AMF is context-dependent. The non-favorable effects are probably due to inoculation ineffectiveness under complex field conditions and/or that, under certain conditions, AMF presence may be a parasitic association. This puts into question the effectiveness of its application in the field. Therefore, it is recommended to only resort to this type of biofertilizer when the cultivation conditions require it (e.g., very low previous microbial diversity, foreseeable stress due to drought, salinity, or lack of nutrients) and not as a general fertilization practice.

Legacy of land-cover changes on soil erosion and microbiology in Burgundian vineyards

Soils in vineyards are recognized as complex agrosystems whose characteristics reflect complex interactions between natural factors (lithology, climate, slope, biodiversity) and human activities. To date, most of the unknown lies in an incomplete understanding of soil ecosystems, and specifically in the microbial biodiversity even though soil microbiota is involved in many key functions, such as nutrient cycling and carbon sequestration. Soil biological properties are indicative of soil quality. Therefore, understanding how soil communities are related to soil ecosystem functioning is becoming an essential issue for soil strategy conservation. Here, we propose to assess the importance of land-cover history on the present-day microbiological and physico-chemical properties. The studied area was selected in the Burgundian vineyards (Pernand-Vergelesses, Burgundy, France) where land occupation has been reconstructed over the last 40 years. Soil samples were collected in five areas reflecting various land cover history (forest, vineyards, shifting from forest to vineyards). For each area, physico-chemical parameters (pH, C, N, P, grain size) were measured and DNA was extracted to characterize the abundance and diversity of microbial communities. The obtained results show significant differences in the five areas suggesting that present-day microbial molecular biomass and bacterial taxonomic is partly inherited from past land occupation. Over longer period of time, such study of land-uses legacies may help to better assess ecosystem recovery and the impact of management practices for a better soil quality and vineyards sustainability.

Elevational range shifts of mountain vineyards: Recent dynamics in response to a warming climate

Increasing temperatures worldwide are expected to cause a change in spatial distribution of plant species along elevational gradients and there are already observable shifts to higher elevations as a consequence of climate change for many species. Not only naturally growing plants, but also agricultural cultivations are subject to the effects of climate change, as the type of cultivation and the economic viability depends largely on the prevailing climatic conditions. A shift to higher elevations therefore represents a viable adaptation strategy to climate change, as higher elevations are characterized by lower temperatures. This is especially important in the case of viticulture because a certain wine-style can only be achieved under very specific climatic conditions. Although there are several studies investigating climatic suitability within winegrowing regions or longitudinal shifts of winegrowing areas, little is known about how fast vineyards move to higher elevations, which may represent a viable strategy for winegrowers to maintain growing conditions and thus wine-style, despite the effects of climate change. We therefore investigated the change in the spatial distribution of vineyards along an elevational gradient over the past 20 years in the mountainous wine-growing region of Alto Adige (Italy). A dataset containing information about location and planting year of more than 26000 vineyard parcels and 30 varieties was used to perform this analysis. Preliminary results suggest that there has been a shift to higher elevations for vineyards in general (from formerly 700m to currently 850 m a.s.l., with extreme sites reaching 1200 m a.s.l.), but also that this development has not been uniform across different varieties and products (i.e. vitis vinifera vs hybrid varieties and still vssparkling wines). This is important for climate change adaptation as well as for rural development. Mountain areas, especially at mid to high elevations, are often characterized by severe land abandonment which can be avoided to some degree if economically viable and sustainable land management strategies are available.

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…

Updating the Winkler index: An analysis of Cabernet sauvignon in Napa Valley’s varied and changing climate

This study aims to create an updated, agile viticultural climate index (similar to the Winkler Index) by performing in-depth analyses of current and historical data from industry partners in several major winegrowing regions. The Winkler Index was developed in the early twentieth century based on analysis of various grape-growing regions in California. The index uses heat accumulation (i.e. Growing Degree Days) throughout the growing season to determine which grape varieties are best suited to each region. As viticultural regions are increasingly subject to the complexity and uncertainty of a changing climate, a more rigorous, agile model is needed to aid grape growers in determining which cultivars to plant where. For the first phase of this study, 21 industry partners throughout Napa Valley shared historical phenology, harvest, viticultural practice, and weather data related to their Cabernet sauvignon vineyard blocks. To complement this data, berry samples were collected throughout the 2021 growing season from 50 vineyard blocks located throughout 16 American Viticultural Areas that were then analyzed for basic berry chemistry and phenolics. These blocks have been mapped using a Geographic Information System (GIS), enabling analysis of altitude, vineyard row orientation, slope, and remotely sensed climate data. Sampling sites were also chosen based on their proximity to a weather station. By analyzing historical data from industry partners and data specifically collected for this study, it is possible to identify key parameters for further analysis. Initial results indicate extreme variability at a high spatial resolution not currently accounted for in modern viticultural climate indices and suggest that viticultural practices play a major role. Using the structure of data collection and analyses developed for the first phase, this project will soon be expanded to other wine regions globally, while continuing data collection in Napa Valley.

How does aromatic composition of red wines, resulting from varieties adapted to climate change, modulate fruity aroma?

One of the major issues for the wine sector is the impact of climate change linked to the increasing temperatures which affects physicochemical parameters of the grape varieties planted in Bordeaux vineyard and consequently, the quality of wine. In some varietals, the attenuation of their fresh fruity character is accompanied by the accentuation of dried-fruit notes [1]. As a new adaptive strategy on climate change, some winegrowers have initiated changes in the Bordeaux blend of vine varieties [2]. This study intends to explore the fruitiness in wines produced from grape varieties adapted to the future climate of Bordeaux. 10 commercial single–varietal wines from 2018 vintage made from the main grape varieties in the Bordeaux region (Cabernet franc, Cabernet-Sauvignon and Merlot) as well as from indigenous grape varieties from the Mediterranean basin, such as Cyprus (Yiannoudin), France (Syrah), Greece (Agiorgitiko and Xinomavro), Portugal (Touriga Nacional) and Spain (Garnacha and Tempranillo), were selected among 19 samples using sensory descriptive analyses. Both sensory and instrumental analyses were coupled, to investigate their fruity aroma expression. For sensory analysis, samples were prepared from wine, using a semi preparative HPLC method which preserves wine aroma and isolates fruity characteristics in 25 specific fractions [3,4]. Fractions of interest with intense fruity aromas were sensorially selected for each wine by a trained panel and mixed with ethanol and microfiltered water to obtain fruity aromatic reconstitutions (FAR) [5]. A free sorting task was applied to categorize FAR according to their similarities or dissimilarities, and different clusters were highlighted. Instrumental analysis of the different FAR and wines demonstrated variations in their molecular composition. Results obtained from sensory and gas chromatography analysis enrich the knowledge of the fruity expression of red wines from “new” grape varieties opening up new perspectives in wine technology, including blending, thus providing new tools for producers.

Measurement of redox potential as a new analytical winegrowing tool

Excell laboratory has initiated the development of an analytical method based on electrochemistry to evaluate the ability of wines to undergo or resist to oxidative phenomena. Electrochemistry is a powerful tool to probe reactions involving electron transfers and offers possibility of real-time measurements. In that context, the laboratory has implemented electrochemical analysis to assess oxidation state of different wine matrices but also in order to evaluate oxidative or reduced character of leaf and soil. Initially, our laboratory focused on dosage of compounds involved in responses of plant stresses and we were also interested in microbiological activity of soils. These analyses were compared with the measurement of redox potential (Eh) and pH which are two fundamental variables involved in the modulation of plant metabolism. Indeed, the variation of redox states of the plant reflects its biological activity but also its capacity to absorb nutriments. The Eh-pH conditions mainly determine metabolic processes involved in soil and leaf and our goal is to determine if this combined analytical approach will be sufficiently precise to detect biological evolutions (plant health, parasitic attack…).

Influence of agronomic practices in soil water content in mid-mountain vineyards

In the context of LIFE project MIDMACC (LIFE18 CCA/ES/001099), several pilots have been installed in vineyards in mid mountain areas of Catalonia (NE Spain) to test well stablished agronomic practices to increase the adaptation of Mediterranean mid mountain to climate change. Soil water content (SWC) at three different depths (15, 30 and 45cm) was measured in continuum from August 2020. One pilot (WC) included a well-established green cover (GC), a new GC (NC) and a conventional soil management (CM, tilling+herbicides). NC presented an intermediate state between WC and CM, responding similarly to CM in autumn but quickly reaching similar SWC to WC, then following the same evolution till next spring, with CM presenting lower values along autumn and winter. Then vegetation activation decreased SWC in all plots, (much slower in CM, lacking GC). Sensibility to spring rains is again intermediate for NC, which joins SWC evolution of CM by the end of spring till next autumn. It is expected that NC will resemble WC more and more as its GC develops. In the pilot combining vine training (VSP vs Gobelet) and hillside management (slope vs terrace), no clear pattern could be related with these conditions. However, both terraces seem to be more sensitive to spring rains. A third pilot included new vineyards (7 and 1 year old). In the new vineyard (N), higher canopy development, a spontaneous green cover and row straw resulted in a slower SWC dynamic, not so sensitive to rains but conserving more soil water in spring and most of summer, even with presumably a higher water extraction by vines. In the newest vineyard (VN) the deepest sensor is still sensitive to rain events all over the year and SWC is always highest at this depth, revealing small water capture by vines.

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.