Impact of rare yeasts in Saccharomyces cerevisiae wine fermentation performance: Population prevalence and growth phenotype of Cyberlindnera fabianii, Kazachstania unispora, and Naganishia globosa.

Saccharomyces cerevisiae is a highly fermentative species able to complete the wine fermentation. However, the interaction with other non-Saccharomyces yeasts can determine the fermentation performance of S. cerevisiae. We have characterised three rare non-Saccharomyces yeasts (Cyberlindnera fabianii, Kazachstania unispora and Naganishia globosa), studying their impact on S. cerevisiae fitness and wine fermentation performance. Using a wide meta-taxonomic dataset of wine samples, analysed through ITS amplicon sequencing, we show that about a 65.07% of wine samples contains Naganishia spp., a 27.21% contains Kazachstania spp., and only a 4.41% contains Cyberlindnera spp; in all cases with average relative abundances lower than 1% of total fungal populations. Although the studied N. globosa strain showed a limited growth capacity in wine, both K. unispora and C. fabianii showed a similar growth phenotype to that of S. cerevisiae in different fermentation conditions, highlighting the outstanding growth rate values of K. unispora. In mixed fermentations with S. cerevisiae, the three yeast species affected co-culture growth parameters and wine chemical profile (volatile compounds, polysaccharides and proteins). K. unispora DN201 strain presents an outstanding capacity to compete with S. cerevisiae strains during the first stage of wine fermentation, causing stuck fermentations in both synthetic and natural grape musts.

Copigmentation of Malvidin-3-O-Monoglucoside by Oenological Tannins: Incidence on Wine Model Color in Function of Botanical Origin, pH and Ethanol Content.

The effect of the botanical origin, pH level and ethanol content of different oenological tannins on the color of malvidin-3-O-monoglucoside solution, including their effectiveness as copigments, was studied. Briefly, a model wine solution (4 g/L of tartaric acid, pH 3.5 and 12% ethanol) containing 50 mg/L of malvidin-3-O-monoglucoside was prepared and supplemented with 0.1, 0.2 and 0.4 g/L of commercial tannins using (-)-epicatechin as reference control copigment. Six additional model wine solutions (12% ethanol at pH 3.1, 3.5 or 3.9, and 10%, 12% or 14% ethanol at pH 3.5) were prepared as previously described. Samples were stored under airtight conditions. After a week the full absorbance spectrum in the visible range (400-800 nm) was measured and CIELAB color space was determined. These measurements, including an increase in a* (redness), a decrease in b* (yellowness) and a decrease in L* (lightness), indicated that all oenological tannins had a clear positive effect on color copigmentation. Moreover, hydrolysable tannins appeared to be better copigments than condensed tannins as the copigmentation effectiveness (Cp) was found to be between two to four times higher. The effects of these tannins were dose-dependent because a higher addition resulted in a greater impact on copigmentation. In general, an increase in pH and ethanol content resulted in a decrease of the effect of tannins on color. Independent of intrinsic wine conditions, hydrolysable tannins, more specifically gallotannin, remain the most effective in increasing red wine color. These results prove that supplementation with oenological tannins, especially hydrolysable tannins, could be an interesting tool for the improvement of the red wine color.

Simultaneous Analysis of Organic Acids, Glycerol and Phenolic Acids in Wines Using Gas Chromatography-Mass Spectrometry.

Fermented beverages, particularly wines, exhibit variable concentrations of organic and phenolic acids, posing challenges in their accurate determination. Traditionally, enzymatic methods or chromatographic analyses, mainly high-performance liquid chromatography (HPLC), have been employed to quantify these compounds individually in the grape must or wine. However, chromatographic analyses face limitations due to the high sugar content in the grape must. Meanwhile, phenolic acids, found in higher quantities in red wines than in white wines, are typically analyzed using HPLC. This study presents a novel method for the quantification of organic acids (OAs), glycerol, and phenolic acids in grape musts and wines. The approach involves liquid-liquid extraction with ethyl acetate, followed by sample derivatization and analysis using gas chromatography-mass spectrometry (GC-MS) in selected ion monitoring (SIM) detection mode. The results indicated successful detection and quantification of all analyzed compounds without the need for sample dilution. However, our results showed that the method of adding external standards was more suitable for quantifying wine compounds, owing to the matrix effect. Furthermore, this method is promising for quantifying other metabolites present in wines, depending on their extractability with ethyl acetate. Fermented beverages, particularly wines, exhibit variable concentrations of organic and phenolic acids, posing challenges in their accurate determination. Traditionally, enzymatic methods or chromatographic analyses, mainly high-performance liquid chromatography (HPLC), have been employed to quantify these compounds individually in the grape must or wine. The approach of this proposed method involves (i) methoximation of wine compounds in a basic medium, (ii) acidification with HCl, (iii) liquid-liquid extraction with ethyl acetate, and (iv) silyl derivatization to analyze samples with gas chromatography-mass spectrometry (GC-MS) in ion monitoring detection mode (SIM). The results indicated successful detection and quantification of all analyzed compounds without the need for sample dilution. However, our results showed that the method of adding external standards was more suitable for quantifying wine compounds, owing to the matrix effect. Furthermore, this method is promising for quantifying other metabolites present in wines, depending on their extractability with ethyl acetate. In other words, the proposed method may be suitable for profiling (targeted) or fingerprinting (untargeted) strategies to quantify wine metabolites or to classify wines according to the type of winemaking process, grape, or fermentation.

Comparison of ancestral and traditional methods for elaborating sparkling wines.

This work compares the ancestral method for elaborating sparkling wines with the most widely used traditional method. Ancestral method is a single fermentation procedure in which the fermenting grape must is bottled before the end of alcoholic fermentation whereas traditional method involves a second fermentation of a base wine inside a bottle. Macabeo grapes were used to elaborate a traditional sparkling wine and two ancestral sparkling wines, one with a low yeast population and one with a high yeast population. The findings indicate that ancestral sparkling wines have lower ethanol content and can be elaborated using lower sulphur dioxide levels. In general, ancestral sparkling wines showed similar protein concentration, higher polysaccharide content, similar or better foamability (HM) than the traditional sparkling wine., No differences were found in the foam stability (HS). In addition, the sensory analysis indicated that ancestral sparkling wines have smaller bubble size, lower CO2 aggressivity, they seemed to have longer ageing time and were scored better than the traditional sparkling wine. These results therefore indicate that the ancestral method is of great interest for the elaboration of high-quality sparkling wines.

Use of Glutathione, Pure or as a Specific Inactivated Yeast, as an Alternative to Sulphur Dioxide for Protecting White Grape Must from Browning.

One of the problems that most seriously affects oenology today is enzymatic browning, especially when grapes are infected by grey rot. We studied the capacity of glutathione (GSH) and a specific inactivated dry yeast rich in glutathione (IDY-GSH) to protect white grape must from browning compared to that of sulphur dioxide (SO2). The results indicate that SO2 drastically reduces the oxygen consumption rate (by around 72%), protects hydroxycinnamic acids from oxidation and prevents grape must against browning even in the presence of laccase. Specifically, the presence of SO2 reduced the colour's blue-yellow component (b*) by around 91% in control conditions and around 76% in the presence of laccase. GSH, pure or in the form of IDY-GSH, also reduces the oxygen consumption rate (by 23% and 36%, respectively) but to a lesser extent than SO2. GSH also favours the formation of grape reaction product (GRP) from hydroxycinnamic acids and effectively protects grape must against browning in healthy grape conditions. Specifically, the presence of GSH reduced b* by around 81% in control conditions. Nevertheless, in the presence of laccase, it was not effective enough, reducing b* by around 39% in the case of pure GSH and 24% in the case of IDY-GSH. Therefore, both forms of GSH can be considered as interesting alternative tools to SO2 for preventing browning in white grape must, but only when the grapes are healthy.

Oxygen consumption rate of lees during sparkling wine (Cava) aging; influence of the aging time.

Sparkling wines elaborated with a traditional method need to age in the bottle in contact with wine lees because yeast autolysis enriches the wines in colloids and improves their effervescence, foam and aromatic complexity. It is generally considered that lees protect the wine against oxidation because they consume small amounts of oxygen that can permeate the crown cap. However, to our knowledge there is no specific study on this subject using lees from real sparkling wine. Therefore, the oxygen consumption rate (OCR) of the lees of sparkling wines from the first to the ninth year of aging time was measured using a noninvasive fluorescence measurement method. The results indicate that lees really consume oxygen and that their OCR tended to decrease with the wine aging time. These data suggest that the lees' capacity to protect against oxidation decreases over time, which could affect the ability of sparkling wines to age properly.

Oenological tannins to prevent Botrytis cinerea damage in grapes and musts: Kinetics and electrophoresis characterization of laccase.

Enzymatic parameters (KM and Vmax), residual activity, effect of bentonite and electrophoresis characterization of laccase in the presence of different oenological tannins (OT) were investigated in relation to B. cinerea negative effects in grapes and musts. Five OT were tested (gallotannin, ellagitannin, quebracho, grape-skin and grape-seed) in comparison with ascorbic acid (AA), sulfur dioxide (SO2) and bentonite. We added OT, AA, SO2 and bentonite to botrytized must obtained by inoculation of grapes with B. cinerea strain 213. Laccase activity was measured by the syringaldazine method at different concentrations of substrate. Enzymatic parameters were determined using Michaelis-Menten and Lineweaver-Burk plots. The B. cinerea strain was also grown in a liquid medium for laccase production. Molecular weight of laccases and effect of OT upon these laccases were studied by SDS-PAGE. Results confirm that bentonite, contrary to OT, did not permit to reduce laccase activity. Regardless the tannin considered, Vmax, KM and laccase activity were reduced and gallotannin, grape-skin and grape-seed tannin presented the greatest ability. Efficiency of grape-seed tannin addition in order to reduce the laccase activity, was comparable to that of AA or SO2 at the typical doses employed in oenology for each one. Oenological tannins appear to be excellent processing aids to prevent laccase effects and contribute to reduce the use of SO2 in grapes and musts.

Influence of grape seeds on wine composition and astringency of Tempranillo, Garnacha, Merlot and Cabernet Sauvignon wines.

BACKGROUND: The aim of this work was to study the influence of grape berry morphology, especially the seed weight percentage, on the tannin concentration and astringency of red wine. Clusters of Tempranillo, Garnacha, Merlot, and Cabernet Sauvignon were characterized and their seeds were extracted and macerated in a model wine solution. In parallel, we elaborated three types of wines of each cultivar. One wine was made with only grape juice, one wine was made adding the appropriate proportion of seeds to the grape juice, and the last wine was elaborated with the complete destemmed and crushed berries. RESULTS: Merlot and Cabernet Sauvignon grapes, which have higher percentage of seed weight with respect to the berry weight than Tempranillo and Garnacha grapes originated wines with higher tannin concentration and astringency than Tempranillo and Garnacha wines. CONCLUSION: The main conclusion of this study is that the seed weight percentage with respect to the berry weight is one of the main determinants of the final tannin concentration and astringency of red wines.

Chemical characterization, antioxidant properties and oxygen consumption rate of 36 commercial oenological tannins in a model wine solution.

The chemical composition (CC), antioxidant capacity (AC) and oxygen consumption rate (OCR) of 36 different commercial tannins were measured. The CC was analyzed by total polyphenol index, Bate-Smith, methyl-cellulose, Folin-Ciocalteu, OIV official method and phloroglucinolisis. The AC was measured by different methods (ABTS, CUPRAC, DPPH, FRAP, ORAC) using Trolox as standard. The OCR was measured using a non-invasive method based on luminescence. The results indicate that it is possible to obtain differentiation between procyanidins/prodelphinidins, profisetinidins/prorobinetidins, gallotannins and ellagitannins by PCA based on their CC data. It is also possible to separate condensed from hydrolysable tannins by PCA based on their AC data. The results show that ellagitannins are the fastest oxygen consumers of the various oenological tannins, followed in descending order by condensed tannins and finally gallotannins. The combination of CC, AC and OCR analyses enable to classify tannins according to their effectiveness in protecting wines against oxidation.

Oxygen consumption rates by different oenological tannins in a model wine solution.

The kinetics of oxygen consumption by different oenological tannins were measured in a model wine solution using the non-invasive method based on luminiscence. The results indicate that the oxygen consumption rate follows second-order kinetics depending on tannin and oxygen concentrations. They also confirm that the oxygen consumption rate is influenced by temperature in accordance with Arrhenius law. The indications are that ellagitannins are the fastest oxygen consumers of the different oenological tannins, followed in decreasing order by quebracho tannins, skin tannins, seed tannins and finally gallotannins. This methodology can therefore be proposed as an index for determining the effectiveness of different commercial tannins in protecting wines against oxidation.