Macrowine 2021
IVES 9 IVES Conference Series 9 Assessment of wine non-Saccharomyces yeast strains as promising producers of glutathione

Assessment of wine non-Saccharomyces yeast strains as promising producers of glutathione

Abstract

AIM: Glutathione (GSH) is a non-protein thiol naturally present in grape berries and produced by yeasts during fermentation. It has a strong antioxidant activity, thus can be added during winemaking to limit the oxidative phenomena of wine, preserving sensory characteristics and stability, ultimately promoting a healthier product by reducing the need for SO2 addition. A promising alternative approach could be the use of yeast starter cultures high-producers of this compound in situ, during the fermentation process, in substitution of external GSH addition. Since this activity in non-Saccharomyces yeasts is currently poorly investigated, the aim of this research was to evaluate the ability of three strains of non-Saccharomyces (NS) yeasts to produce GSH, both in synthetic media and in fresh grape must.

METHODS: Lachancea thermotolerans SOL13, Metschnikowia sp. FIANO12, and Starmerella bacillaris MALV45 were tested in single or sequential inoculations,with Saccharomyces cerevisiae EC1118, in synthetic grape juice (SGJ) or in Pinot Grigio grape must, under static conditions, or in a medium optimized for GSH production (MGSH) in agitation (200 rpm). GSH concentration was determined using the Glutathione Assay Kit (Sigma-Aldrich). Population dynamics was evaluated by plate count and biomass dry weight, and fermentation kinetics through weight loss measurement.

RESULTS: A variability in GSH production was found among individual strains and growth conditions. Metschnikowia sp. FIANO12 showed the highest intracellular accumulation of GSH when cultivated alone in both synthetic media, and, as expected, higher levels in the optimized MGSH than in SGJ (4.59 vs. 0.19 nmol GSH/mg cells, respectively). In wine, fermentations with S. bacillaris MALV45 had the highest concentration of intracellular GSH, but the lowest content of wine-dissolved GSH. The maximum level of extracellular GSH (21 mg/L) was obtained in the sequential fermentation with L. thermotolerans SOL13, a significant increase compared to the control singly inoculated with S. cerevisiae.

CONCLUSIONS:

This study highlights a new potential interesting feature of NS yeasts to positively modify wine composition. The tested native strains, with previously demonstrated interesting oenological traits, showed a good capacity to accumulate GSH and to increase the concentration of this antioxidant compound in wine. Thus, the strategy of multi-starter fermentation can be a valuable tool to achieve a lower input winemaking. Future investigations are needed to assess the long-term stability of wine made from multi-starter fermentations with NS yeasts producers of GSH.ACKNOWLEDGEMENTS: We thank Nicolò Bersani for laboratory assistance.

DOI:

Publication date: September 3, 2021

Issue: Macrowine 2021

Type: Article

Authors

Renato L. Binati, Wilson J.F. LEMOS JUNIOR, Sandra TORRIANI

Department of Biotechnology, University of Verona, Italy, Department of Biotechnology, University of Verona, Italy, Department of Biotechnology, University of Verona, Italy

Contact the author

Keywords

Glutathione production, multi-starter fermentation, non-saccharomyces yeasts, saccharomycescerevisiae, winemaking, wine quality

Citation

Related articles…

Downscaling of remote sensing time series: thermal zone classification approach in Gironde region

In viticulture, the challenges of local climate modelling are multiple: taking into account the local environment, fine temporal and spatial scales, reliable time series of climate data, ease of implementation and reproducibility of the method. At the local scale, recent studies have demonstrated the contribution of spatialization methods for ground-based climate observation data considering topographic factors such as altitude, slope, aspect, and geographic coordinates (Le Roux et al, 2017; De Rességuier et al, 2020). However, these studies have shown questions in terms of the reproducibility and sustainability of this type of climate study. In this context, we evaluated the potential of MODIS thermal satellite images validated with ground-based climate data (Morin et al, 2020). Previous studies have been encouraging, but questions remain to be explored at the regional scale, particularly in the dynamics of the massive use of bioclimatic indices to classify the climate of wine regions. The results at the local scale were encouraging, but this approach was tested in the current study at the regional scale. Several objectives were set: 1) to evaluate the downscaling method for land surface temperature time series, 2) to identify regional thermal structure variations. We used weekly minimum and maximum surface temperature time series acquired by MODIS satellites at a spatial resolution of 1000 m and downscaled at 500 m using topographical variables. Two types of analyses were performed:

The impact of leaf canopy management on eco-physiology, wood chemical properties and microbial communities in root, trunk and cordon of Riesling grapevines (Vitis vinifera L.)

In the last decades, climate change required already adaptation of vineyard management. Increase in temperature and unexpected weather events cause changes in all phenological stages requiring new management tools. For example, defoliation can be a useful tool to reduce the sugar content in the berries creating differences in the wine profiles. In a ten-year field experiment using Riesling (Vitis vinifera L, planted 1986, Geisenheim, Germany), various mechanical defoliation strategies and different intensities were trialed until 2016 before the vineyard was uprooted. Wood was sampled from the plant compartments root, trunk, cordon and shoot for analyses of physicochemical properties (e.g. lignin and element content, pH, diameter), nonstructural carbohydrates and the microbial communities. The aim of the study was to investigate the influence of reduced canopy leaf area on the sink-source allocation into different compartments and potential changes of the fungal and prokaryotic wood-inhabiting community using a metabarcoding approach. Severe summer pruning (SSP) of the canopy and mechanical defoliation (MDC) above the bunch zone decreased the leaf area by 50% compared to control (C). SSP reduced the photosynthetic capacity, which resulted in an altered source-sink allocation and carbohydrate storage. With lower leaf area, less carbohydrates are allocated. This for example resulted in a decreased trunk diameter. Further, it affected the composition of the grapevine wood microbiota. SSP and MDC management changed significantly the prokaryotic community composition in wood of the root samples, but had no effect in other compartments. In general, this study found strong compartment and less management effects of the microbial community composition and associated physicochemical properties. The highest microbial diversities were identified in the wood of the trunk, and several species were recorded the first time in grapevine.

Metabolomic discrimination of grapevine water status for Chardonnay and Pinot noir

Water status impact in viticulture has been widely explored, as it strongly affects grapevine physiology and grape chemical composition. It is considered as a key component of vitivinicultural terroir. Most of the studies concerning grapevine water status have focused on either physiological traits, or berry compounds, or traits involved in wine quality. Here, the response of grapevine to water availability during the ripening period is assessed through non-targeted metabolomics analysis of grape berries by ultra-high resolution mass spectrometry. The grapevine water status has been assessed during 2 consecutive years (2019 & 2020), through carbon isotope discrimination on juices from berries collected at maturity (21.5 brix approx.) for 2 Vitis vinifera cv. Pinot noir (PN) and Chardonnay (CH). A total of 220 grape juices were collected from 5 countries worldwide (Italy; Argentina; France; Germany; Portugal). Measured δ13C (‰) varied from -28.73 to -22.6 for PN, and from -28.79 to -21.67 for CH. These results also clearly revealed higher water stress for the 2020 vintage. The same grape juices have been analysed by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR-MS) and Liquid Chromatography coupled to Mass Spectrometry (LC-qTOF-MS), leading to the detection of up to 4500 CHONS containing elemental compositions, and thus likely tens of thousands of individual compounds, which include fatty acids, organic acids, peptides, phenolics, also with high levels of glycosylation. Multivariate statistical analysis revealed that up to 160 elemental compositions, covering the whole range of detected masses (100 –1000 m/z), were significantly correlated to the observed gradients of water status. Examples of chemical markers, which are representative of these complex fingerprints, include various derivatives of the known abscisic acid (ABA), such as phaesic acid or abscisic acid glucose ester, which are significantly correlated with higher water stress, regardless of the variety. Cultivar-specific behaviours could also be identified from these fingerprints. Our results provide an unprecedented representation of the metabolic diversity, which is involved in the water status regulation at the grape level, and which could contribute to a better knowledge of the grapevine mitigation strategy in a climate change context.

Towards adaptation to climate change in Rioja: Quality evaluation of wines obtained from Grenache x Tempranillo selections

The wine sector is of great relevance and tradition in Mediterranean countries, however, it may be most susceptible to climate change. In recent years, wine production is facing changes worldwide, both at environmental as well as commercial levels, due to global warming and the shift in consumers’ preferences. Wine growers and wine makers are in search of solutions that allow to face these new challenges. One of the most promising initiatives in the long term is the introduction of new plant materials, specifically intraspecific hybridizations between premium varieties that may improve traditional germplasm in its adaptation to climate change. These inter-varietal crosses have the potential to generate quality wines, whilst maintaining the regional typicity, and constitute an attractive alternative for the consumer due to their sensory attributes. In this study, we have evaluated wines from 29 intraspecific Garnacha x Tempranillo hybrids in two different locations, with the aim to assess their oenological potential and sensory attributes. Thirteen of the selections were white and 16 were red. Microvinifications were conducted with two or three replications depending on grape availability. Conventional oenological parameters were determined for all wines. The sensory evaluation and hedonic scores were given by five experts. Red selections obtained higher quality scores than white ones. Among the white selections with higher quality scores, GT-41 Varea and GT-159 Varea outstand, due to their high total acidity and high malic acid content. Regarding red selections, GT-57 Varea and GT-57 UR were perceived as higher in quality, highlighted for their moderate alcoholic and high anthocyanin content. Our results indicate that intraspecific hybridization may be a powerful tool for adapting traditional cultivars to climate change in Rioja.

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.