Macrowine 2021
IVES 9 IVES Conference Series 9 Assay of distinct modes of polysaccharidases dosage in vinification with cv. Malbec. Effects on microbiological evolution, color and skin depletion

Assay of distinct modes of polysaccharidases dosage in vinification with cv. Malbec. Effects on microbiological evolution, color and skin depletion

Abstract

In the maceration stage of winemaking, enzymes can be used to degrade the polysaccharides present in the cell walls and middle sheets, and thus facilitate the extraction of juice and the release of polyphenols and aroma precursors retained in the grape skins. This work aims to analyze the influence of two enzyme complexes produced by autochthonous yeasts on the red winemaking process, in order to evaluate their effect on the chemical composition of the wines obtained, as well as on the extraction of color and polyphenols, and the depletion of the skin. Two strains previously selected for the effect of their enzymatic complex on the color extraction and improvement in the technological properties of the grape must were used (Longhi et al., 2019). A multi-enzymatic extract from Aureobasidium pullulans m11-2 was obtained by inoculating the microorganism in a broth according to Moyo et al. (2003) with modifications (pH 3.8) and incubated with stirring at 28°C for 72 h. Pectinase, xylanase, cellulase and amylase activities were quantified by determining reducing sugars by DNS, modified by Qian Li et al. (2015). Likewise, Torulaspora delbrueckii m7-2 was used for the production of the enzyme complex during vinification. Malbec red grapes (Vitis vinifera L.) from San Rafael (Mendoza) wine region, vintage 2021, were used to conduct the vinifications. The must obtained by crushing 60 Kg of grapes was corrected in acidity, sulfited (50 ppm) and distributed in 5 L containers. Four winemaking assays were performed, in duplicate: (1) inoculation with a native strain of Sacchromyces cerevisiae (SR1), at 108 cell/mL as inoculum, conducted at 20°C (control, C); (2) sequential inoculation of T. delbrueckii m7-2, with an initial cellular concentration of 107 cells/mL, followed by SR1 inoculation at 4th day (Td); (3) cold pre-fermentation maceration (CPM, 8°C-4 days) with m11-2 enzyme extract and SR1 inoculation (Ap); and (4) CPM without enzymatic treatment and SR1 inoculation (E). Growth kinetics of total yeasts were determined on YPD and DRBC agar, and of non-Saccharomyces yeasts in lysine medium. All enzymatic activities were monitored at pH 3.80 and 20°C. The pectinolytic activity was the main one, showing a level of 1.80 U/mL in the m11-2 extract and an initial level of 1.47 U/mL for the in situ producer strain (m7-2). Microscopic observations of the extracted skins in Ap and E vinifications were carried out to evaluate the effect of the enzymatic complex m11-2 on the cell wall, and were also compared with the fresh grape skins. Differences were observed between the skins enzymatically treated (Ap) and the control (E); the former showed cell emptying, greater rupture of the epidermis layers and less firmness, unlike the control that exhibited almost intact epidermal layers. These images allowed us to know the cell morphology of the varietal cv. Malbec and the enzymatic hydrolysis of its cell walls.

DOI:

Publication date: September 10, 2021

Issue: Macrowine 2021

Type: Article

Authors

Sara Jaquelina Longhi 

 Biotechnology Laboratory, Department of Biology and Food, Faculty of Sciences Applied to Industry, National University of Cuyo. Bernardo de Irigoyen 375, San Rafael, Mendoza, Argentina. National Council for Scientific and Technical Research (CONICET), Godoy Cruz 2290, Autonomous City of Buenos Aires, Argentina. ,María Carolina MARTÍN (1,2)  María Belén AVENDAÑO(1) María Gabriela MERÍN (1,2)   Luciana Paola PRENDES (1,2) Juliana GARAU (1,2) Vilma Inés MORATA DE AMBROSINI (1,2) (1) Biotechnology Laboratory, Department of Biology and Food, Faculty of Sciences Applied to Industry, National University of Cuyo. Bernardo de Irigoyen 375, San Rafael, Mendoza, Argentina. (2) National Council for Scientific and Technical Research (CONICET), Godoy Cruz 2290, Autonomous City of Buenos Aires, Argentina

Contact the author

Keywords

polysaccharidases, winemaking, polyphenols, malbec, grape cell wall

Citation

Related articles…

‘Cabernet Sauvignon’ (Vitis vinifera L.) berry skin flavonol and anthocyanin composition is affected by trellis systems and applied water amounts

Trellis systems are selected in wine grape vineyards to mainly maximize vineyard yield and maintain berry quality. This study was conducted in 2020 and 2021 to evaluate six commonly utilized trellis systems including a vertical shoot positioning (VSP), two relaxed VSPs (VSP60 and VSP80), a single high wire (SH), a high quadrilateral (HQ), and a guyot (GY), combined with three levels of irrigation regimes based on different crop evapotranspiration (ETc) replacements, including a 25% ETc, 50% ETc, and 100% ETc. The results indicated SH yielded the most fruits and accumulated the most total soluble solids (TSS) at harvest in 2020, however, it showed the lowest TSS in the second season. In 2020, SH and HQ showed higher concentrations in most of the anthocyanin derivatives compared to the VSPs. Similar comparisons were noticed in 2021 as well. SH and HQ also accumulated more flavonols in both years compared to other trellis systems. Overall, this study provides information on the efficacy of trellis systems on grapevine yield and berry flavonoid accumulation in a currently warming climate.

1H-NMR-based Metabolomics to assess the impact of soil type on the chemical composition of Mediterranean red wines

The aim of this study was to evaluate the effects of different soil types on the chemical composition of Mediterranean red wines, through untargeted and targeted 1H-NMR metabolomics. One milliliter of raw wine was analyzed by means of a Bruker Avance II 400 spectrometer operating at 400.15 MHz. The spectra were recorded by applying the NOESYGPPS1D pulse sequency, to achieve water and ethanol signals suppression. No modification of the pH was performed to avoid any chemical alteration of the matrix. The generation of input variables for untargeted analysis was done via bucketing the spectra. The resulting dataset was preprocessed prior to perform unsupervised PCA, by means of MetaboAnalyst web-based tool suite. The identification of compounds for the targeted analysis was performed by comparison to pure compounds spectra by means of SMA plug-in of MNova 14.2.3 software. The dataset containing the concentrations (%) of identified compounds was subjected to one-way analysis of variance (ANOVA) to highlight significant differences among the wines. The untargeted analysis, carried out through the PCA, revealed a clear differentiation among the wines. The fragments of the spectra contributing mostly to the separation were attributed to flavonoids, aroma compounds and amino acids. The targeted analysis leaded to the identification of 68 compounds, whose concentrations were significant different among the wines. The results were related to soils physical-chemical analysis and showed that: 1) high concentrations of flavan-3-ols and flavonols are correlated with high clay content in soils; 2) high concentrations of anthocyanins, amino acids, and aroma compounds are correlated with neutral and moderately alkaline soil pH; 3) low concentrations of flavonoids and aroma compounds are correlated with high soil organic matter content and acidic pH. The 1H-NMR metabolomic analysis proved to be an excellent tool to discriminate between wines originating from grapes grown on different soil types and revealed that soils in the Mediterranean area exert a strong impact on the chemical composition of the wines.

Frost risk projections in a changing climate are highly sensitive in time and space to frost modelling approaches

Late spring frost is a major challenge for various winegrowing regions across the world, its occurrence often leading to important yield losses and/or plant failure. Despite a significant increase in minimum temperatures worldwide, the spatial and temporal evolution of spring frost risk under a warmer climate remains largely uncertain. Recent projections of spring frost risk for viticulture in Europe throughout the 21st century show that its evolution strongly depends on the model approach used to simulate budburst. Furthermore, the frost damage modelling methods used in these projections are usually not assessed through comparison to field observations and/or frost damage reports.
The present study aims at comparing frost risk projections simulated using six spring frost models based on two approaches: a) models considering a fixed damage threshold after the predicted budburst date (e.g BRIN, Smoothed-Utah, Growing Degree Days, Fenovitis) and b) models considering a dynamic frost sensitivity threshold based on the predicted grapevine winter/spring dehardening process (e.g. Ferguson model). The capability of each model to simulate an actual frost event for the Vitis vinifera cv. Chadonnay B was previously assessed by comparing simulated cold thermal stress to reports of events with frost damage in Chablis, the northernmost winegrowing region of Burgundy. Models exhibited scores of κ > 0.65 when reproducing the frost/non-frost damage years and an accuracy ranging from 0.82 to 0.90.
Spring frost risk projections throughout the 21st century were performed for all winegrowing subregions of Bourgogne-Franche-Comté under two CMIP5 concentration pathways (4.5 and 8.5) using statistically downscaled 8×8 km daily air temperature and humidity of 13 climate models. Contrasting results with region-specific spring frost risk trends were observed. Three out of five models show a decrease in the frequency of frost years across the whole study area while the other two show an increase that is more or less pronounced depending on winegrowing subregion. Our findings indicate that the lack of accuracy in grapevine budburst and dehardening models makes climate projections of spring frost risk highly uncertain for grapevine cultivation regions.

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.

Inhibition of Oenococcus oeni during alcoholic fermentation by a selected Lactiplantibacillus plantarum strain

The use of selected cultures of the species Lactiplantibacillus plantarum in Oenology has grown in prominence in recent years. While initial applications of this species centred very much around malolactic fermentation (MLF), there is strong evidence to show that certain strains can be harnessed for their bio-protective effects. Unwanted spontaneous MLF during alcoholic fermentation (AF), driven by rogue Oenococcus oeni, is a winemaking deviation that is very difficult to manage when it occurs. This work set out to determine the efficacy of one particular strain of Lactiplantibacillus plantarum(Viniflora® NoVA™ Protect), against this problem in Cabernet Sauvignon must. The work was carried out at commercial scale and in a winery environment and compared the bio-protective culture with the more traditional approach of reducing must pH by the addition of tartaric acid. The combination of both was also investigated. The concentration of both Oenococcus oeni and Lactiplantibacillus plantarum was determined using qPCR. The adventitious Oenococcus oeni showed the most growth during AF in the control wine, whereas in the wines treated with Lactiplantibacillus plantarum a bacteriostatic effect against this species was observed. This effect was comparable to the wines treated with tartaric acid. This has particular commercial relevance for controlling the flora in musts with high pH, or when the addition of tartaric acid is either not permitted or is prohibitive for other reasons.