Metabolomics of Red Wines Aged Traditionally, with Chips or Staves.

Traditionally and alternatively aged wines' odour activity values (OAVs) are investigated to differentiate and highlight the differences between the selected methods. An analysis of the volatile aroma compounds of wines derived from ageing in barrels, oak chips, and staves was performed using stir bar sorptive extraction chromatography-mass spectroscopy (SBSE-GC-MS). The results showed that alcohols, esters, and oak compounds were the main contributors to aroma, and their OAVs were higher in the stave samples after 3 months than in the samples from the other two systems of ageing. Furthermore, wines aged with staves have stronger fruity, spiced, and woody aromas, while samples aged in barrels present more chemistry-driven, floral, caramelly, and creamy aromas. The staves-medium plus toast (SMPT at 3 months > 225) and chips-medium plus toast (CMPT at 3 months > 170) showed the highest levels of aromatic series, suggesting that alternative systems provided more powerful aromas than traditional systems, such as barrels-medium plus toast (BMPT at 3 months > 150). A principal component analysis (PCA), orthogonal partial least squares (OPLS) analysis, and cluster analysis allowed for a clear differentiation to be made between red wines according to ageing systems and ageing times. The odour activity values fingerprint in winemaking is a feasible approach to characterise and distinguish wines. Moreover, OAVs provide important information on the effects of production methods on wine quality and aroma profile.

Towards a better understanding of the evolution of odour-active compounds and the aroma perception of sparkling wines during ageing.

A native veil-forming yeast and a commercial yeast strain were used to elaborate sparkling wines by the Champenoise method with a grape variety traditionally used for the production of still wines. Wines aged on lees for fifteen months were sampled at five points and their physicochemical and sensory indices were analysed. Unsupervised and supervised statistical techniques were used to establish a comparison between 81 volatile compounds and eight odour descriptors (chemical, fruity, floral, fatty, balsamic, vegetal, empyreumatic and spicy). Principal component analysis of both datasets showed good separation among the samples in relation to ageing time and yeast strain. By using a partial least squares regression-based criterion, 38 odour active compounds were selected as the most influential for the ageing factor and out of them, only 27 were unique to certain aroma descriptors. These results contribute to a better understanding of the aroma perception of sparkling wines.

Using an electronic nose and volatilome analysis to differentiate sparkling wines obtained under different conditions of temperature, ageing time and yeast formats.

Effect of yeast inoculation format (F), temperature (T), and "on lees" ageing time (t) factors were evaluated on the composition of sparkling wines by a quantitative fingerprint obtained from volatile metabolites and the response of an electronic nose (E-nose). Wines elaborated according the traditional method at 10 and 14 °C, free cells and yeast biocapsules formats were monitored at 15 and 24 months of ageing time. Sixty-six volatiles identified and quantified in the eight sampling lots were subjected to a pattern recognition technique. A dual criterion based on univariate (ANOVA) and multivariate analysis (PLS-DA) through the variable importance projection (VIP) values, allowed to identify ten volatiles as potential markers for T factor, eleven for t and twelve for F factors. The discriminant models based on E-nose dataset enable a 100% correct classification of samples, in relation with t and F factors and the 83% for T factor.

Volatilome fingerprint of red wines aged with chips or staves: Influence of the aging time and toasting degree.

The influence on aroma compounds chips or staves and toasting degree have been analyzed in red wines aged for two periods of time. Ethyl propanoate, ethyl butanoate, ethyl octanoate, ethyl acetate, isoamyl acetate, isobutanol, 2-methyl-1-butanol, 2-phenylethanol, E-2-hexenol, octanal, nonanal, decanal, γ-nonalactone, furfural, 5-methylfurfural, 2-methoxy-4-vinylphenol and cis-whiskey lactone were the compounds that contribute the most to the aroma series profile. By means of principal components analysis, esters were related to the aging time; cis-whiskey lactone with the type of wood pieces and octanal, 5-methyl furfural and cis-whiskey lactone with the toasting degree. Star plot show that woody aroma compounds are dominant in wines aged with low toasting degree oak pieces, whereas medium plus toasted pieces increased the concentration of aroma compounds with fruity aroma descriptors. Wines with prominent fruity or woody aromas can be obtained depending upon the degree of toasting of wood pieces used for aging.

Evaluation of Aroma Compounds in the Process of Wine Ageing with Oak Chips.

Many modern alcoholic beverages are subjected to ageing processes during which compounds extracted from wood contribute decisively to the overall beverage character. Wines represent a perfect example of beverage in which ageing is a crucial technological manufacturing step. During winemaking, producers accelerate chemical changes in wine composition by traditional and alternative methods, such as the use of oak wood barrels and/or oak wood chips. Our research aimed to investigate the overall volatile composition and sensory quality of red wines aged for two timeframes, namely, 1.5 and 3 months, and with two technological variants, i.e., American and French oak wood chips. Red grapes from the Feteasca neagra (Vitis vinifera) variety were harvested from a vineyard in the North-East region of Romania. Stir bar sorptive extraction and gas chromatography coupled with mass spectrometry (SBSE-GC-MS) was used to extract minor aromas present in wine samples. The results showed clear differences between wines treated with American and French oak chips. Furthermore, ageing for 3 months increased the concentration of cis-whiskey lactone and guaiacol in American oak-treated wine samples. For wines aged with French oak chips, we observed higher concentrations of furfural, 5-methylfurfural, 4-vinylguaiacol, and trans-whiskey lactone. The increased presence of chemical compounds in wine aged with French oak chips generated prominent smoky, licorice, and toasty aromas, whereas in wines aged with American oak chips, notes of vanilla, toasty, and cacao aromas were noticed. Moreover, red wines aged with American and French oak chips were discriminated by chemometric analysis, which confirmed the evolution of aroma compounds.

Effects of ADH2 overexpression in Saccharomyces bayanus during alcoholic fermentation.

The effect of overexpression of the gene ADH2 on metabolic and biological activity in Saccharomyces bayanus V5 during alcoholic fermentation has been evaluated. This gene is known to encode alcohol dehydrogenase II (ADH II). During the biological aging of sherry wines, where yeasts have to grow on ethanol owing to the absence of glucose, this isoenzyme plays a prominent role by converting the ethanol into acetaldehyde and producing NADH in the process. Overexpression of the gene ADH2 during alcoholic fermentation has no effect on the proteomic profile or the net production of some metabolites associated with glycolysis and alcoholic fermentation such as ethanol, acetaldehyde, and glycerol. However, it affects indirectly glucose and ammonium uptakes, cell growth, and intracellular redox potential, which lead to an altered metabolome. The increased contents in acetoin, acetic acid, and L-proline present in the fermentation medium under these conditions can be ascribed to detoxification by removal of excess acetaldehyde and the need to restore and maintain the intracellular redox potential balance.

Study of the potential use of mesoporous nanomaterials as fining agent to prevent protein haze in white wines and its impact in major volatile aroma compounds and polyols.

Mesoporous nanomaterials were used to prevent protein haze in Muscat Ottonel and Pedro Ximénez wines. Major volatile compounds, polyols and turbidity were analyzed in fined wines and the results were compared with untreated and fined with bentonite wines. Two commercial mesoporous nanomaterial labelled as SBA-15 and MCM-41 provided a notable turbidity reduction and prevent protein haze as was demonstrated by the heat stability tests. Only, the siliceous material labelled as KIT-6 was not suitable for wine fining. All the chemical families were affected by the fining agents, although the loss of aroma compounds was lower with nanomaterials than with bentonite. The impact on volatile compounds concentration depends on tested wine, independently on the fining agent used. Wines were distinguished by principal components analysis, which was carried out using the volatile compounds as classifying variables. Wines treated with SBA-15 were closest to the control wine in terms of volatile compounds concentration.

Influence of two yeast strains in free, bioimmobilized or immobilized with alginate forms on the aromatic profile of long aged sparkling wines.

Production of sparkling wines involve a second alcoholic fermentation and contact with yeast less over an extended period of time, which influences the aroma composition and sensory quality of the resulting wines. Sparkling wines obtained with two yeast strains inoculated as free cells, immobilized in alginate bed and bioimmobilized as biocapsules, were aged during 32 months. Among the volatile compounds, high Odor Activity Values were obtained with isoamyl acetate, ethyl propanoate, ethyl butanoate, ethyl 3-methylbutanoate, ethyl hexanoate, ethyl octanoate, hexanol, 2-methoxy-4-vinylphenol, decanal, octanoic acid, decanoic acid and TDN. Taken together these contribute more than 70% of the overall aromatic series value. Although some results rely more on the yeast strain than the inoculation format, specific aroma compounds were associated with the immobilization format, allowing the classification of sparkling wines by PCA. As a result the aroma quality of sparkling wines could be improved using immobilized yeasts.

Potential application of a glucose-transport-deficient mutant of Schizosaccharomyces pombe for removing gluconic acid from grape must.

Musts from rotten grapes typically contain high levels of gluconic acid, which can raise severe problems in winemaking processes. In this work, the ability of the glucose-transport-deficient mutant YGS-5 of Schizosaccharomyces pombe to completely or partly remove gluconic acid from a synthetic glucose-containing medium and the potential use of this yeast strain for the same purpose in musts and wines were examined. Surprisingly, the S. pombe YGS-5 strain successfully removed 93% of the initial gluconic acid (2.5 gL(-1)) and 80% of the initial malic acid (1.0 gL(-1)) within 30 h after inoculation. Also, the yeast strain produced no volatile compounds other than those obtained in fermentations conducted with the wine yeast Saccharomyces cerevisiae. S. pombe YGS-5 could thus be used to remove gluconic acid present in musts from rotten grapes. On the basis of these results, various ways of using S. pombe YGS-5 to treat musts containing gluconic acid in order to solve the problems due to the high gluconic acid concentrations in botrytized grape must are proposed.

Biological aging of sherry wines using pure cultures of two flor yeast strains under controlled microaeration.

Sherry wines obtained after biological aging for an average of 0, 2, and 4 years were inoculated separately with the flor yeast strains Saccharomyces cerevisiae var. capensis and Saccharomyces bayanus and subjected to short, periodic microaeration to a dissolved oxygen concentration of 4 mg L(-1) after formation of the yeast film. A principal component analysis with the acetaldehyde, ethanol, volatile acidity, and glycerol concentrations obtained was performed. The first principal component was found to account for 49.5% of the overall variance and to be defined mainly by glycerol and ethanol. The second component accounted for 38.8% of the variance and was defined by volatile acidity and acetaldehyde. The conditions used in the tests allowed the biological aging of the wines to be substantially shortened. Thus, 42 days after flor-film formation by S. cerevisiae var. capensis, 0- and 2-year-old wines exhibited parameter values similar to those obtained for the wine aged for 4 years. The wines inoculated with S. bayanus exhibited high acetaldehyde concentrations and ethanol levels above 15% (v/v)-sherry wines with alcohol concentrations below 14.5% are undesirable-, so one need not exclude the sequential or simultaneous inoculation of S. bayanus together with S. cerevisiae var. capensis in order to improve the biological aging process.

Effect of Schizosaccharomyces pombe on aromatic compounds in dry sherry wines containing high levels of gluconic acid.

Volatile compounds have been determined in control dry sherry wines and those supplemented with gluconic acid, which were inoculated with the Schizosaccharomyces pombe 1379 (ATCC 26760) yeast strain. These compounds were grouped, according to volatiles exhibiting the identical odor quality, into nine groups of the same odor character (aromatic series) as a way of establishing the aroma profile for the studied wines. Control and supplemented wines showed changes in the balsamic, spicy, roasty, and fruity aromatic series, and tasters judged the aroma as typical of wines subjected to biological aging. This fission yeast may be used as a treatment to reduce gluconic acid contents in wines obtained from rotten grapes, making feasible the incorporation of these wines into the biological aging process. In addition, this procedure may also help to accelerate the traditional biological aging in sherry winemaking due to the contribution of some specific compounds by S. pombe to the wine.

Gluconic acid consumption in wines by Schizosaccharomyces pombe and its effect on the concentrations of major volatile compounds and polyols.

Schizosaccharomyces pombe 1379 (ATCC 26760) yeast strain in wine substantially increases acetaldehyde and 1,1-diethoxyethane concentrations and to decreases levo-2,3-butanediol, glycerol, acetoin, and gluconic acid concentrations. In this study, S. pombe has been used for the first time to reduce gluconic acid in wine under aerobic conditions. Only acetaldehyde and acetoin exhibited significantly higher levels in the wines containing gluconic acid. The high in vitro specific activity of alcohol dehydrogenase observed may be directly related to the high production of acetaldehyde by the studied fission yeast.

Gas chromatographic quantification of major volatile compounds and polyols in wine by direct injection.

Methanol, propanol, isobutanol, isoamyl alcohol, 2-phenylethanol, acetaldehyde, 1,1-diethoxyethane, acetoin, ethyl acetate, ethyl lactate, and ethyl succinate and the polyols 2,3-butanediol (levo and meso forms) and glycerol were quantified by direct injection of wine samples. Linear responses over the usual concentration ranges for these compounds and r2 values from 0.9932 to 0.9998 were obtained. The confidence limits for the mean values ranged from 2.34% for diethyl succinate to 8.52% for 1,1-diethoxyethane, both at a probability level of 0.05. Relative errors ranged from 8 to 10% for the polyols and 1,1-diethoxyethane and were all less than 5% for alcohols and acetaldehyde. The proposed method is useful with a view to identifying relationships between alcoholic fermentation byproducts and controlling biological or chemical aging in wines.

Effect of gluconic acid consumption during simulation of biological aging of sherry wines by a flor yeast strain on the final volatile compounds.

Flor yeast Saccharomyces cerevisiae (capensis G1) strain assimilates gluconic acid during the aerobic biological aging process of sherry wines and exerts significant changes on the final volatile compounds of wines, especially a decrease in volatile acidity and butanoic, isobutanoic, 2-methylbutanoic, and 3-methylbutanoic acids. This decrease may have a favorable effect on the quality of sherry wines.

Sweet wine production by two osmotolerant Saccharomyces cerevisiae strains.

The use of Saccharomyces cerevisiae to produce sweet wine is difficult because yeast is affected by a hyperosmotic stress due to the high sugar concentrations in the fermenting must. One possible alternative could be the coimmobilization of the osmotolerant yeast strains S. cerevisiae X4 and X5 on Penicillium chrysogenum strain H3 (GRAS) for the partial fermentation of raisin musts. This immobilized has been, namely, as yeast biocapsules. Traditional sweet wine (that is, without fermentation of the must) and must partially fermented by free yeast cells were also used for comparison. Partially fermented sweet wines showed higher concentration of the volatile compounds than traditionally produced wines. The wines obtained by immobilized yeast cells reached minor concentrations of major alcohols than wines by free cells. The consumption of specific nitrogen compounds was dependent on yeast strain and the cellular immobilization. A principal component analysis shows that the compounds related to the response to osmotic stress (glycerol, acetaldehyde, acetoin, and butanediol) clearly differentiate the wines obtained with free yeasts but not the wines obtained with immobilized yeasts.

Use of a Schizosaccharomyces pombe mutant to reduce the content in gluconic acid of must obtained from rotten grapes.

Schizosaccharomyces pombe YGS-5 and Saccharomyces cerevisiae G1 strains were used in order to develop an effective method for reducing the gluconic acid content of musts without altering the development of alcoholic fermentation or detracting from quality in the resulting wines. The best results in synthetic media were obtained by using a temperature of 24 degrees C and a sulfur dioxide rate below 100 mg/L under semiaerobic conditions. Sequential inoculation of the musts with YGS-5 first and fermentative G1 yeasts then reduced their gluconic acid content by 85% within 43 h; by contrast, simultaneous inoculation with YGS-5 and G1 provided a reduction of only 40%. The wines with the best sensory and analytical properties were obtained by sequentially inoculating the musts with YGS-5 and, once gluconic acid was removed, G1. The wine obtained by sequential inoculation without removing YGS-5 was that exhibiting the highest odorant activity value (OAV) for the volatile compounds in the floral odor series. A protocol for treating musts containing gluconic acid was developed and tested at the pilot plant scale. The treatment reduced the gluconic acid content by 70% within 46 h with no adverse effect on the analytical or sensory quality of the resulting wines.

Use of a novel immobilization yeast system for winemaking.

Penicillium was used to immobilize Saccharomyces cerevisiae, without using physico-chemical external supports, to form yeast biocapsules. The biocapsules, once the Penicillium was killed by the ethanol produced, were used in a grape must fermentation. Must fermentation was carried out for 160 h with the biocapsules and for 300 h with free yeast cells. Acetaldehyde (84 vs. 63 mg/l), isobutanol (217 vs. 194 mg/l), L: -proline (7.7 vs. 6.5 mM: ) and aspartic acid (0.42 vs. 0 mM: ) in final wine were higher with the biocapsules than with free cells.