Stereoselective formation of the varietal aroma compound rose oxide during alcoholic fermentation

The potent aroma compound rose oxide was quantified in several white wines by a headspace solid-phase microextration stable isotope dilution assay (HS-SPME-SIDA) and the enantiomeric ratios of the cis diastereomers were determined by enantioselective capillary GC. The most odor-active stereoisomer (23)-cis-rose oxide was detectable in all investigated white wines ranging from 0.2 to 12 microg/L. However, its contribution to the overall aroma in some white wine varieties can be neglected as indicated by a low odor activity value (OAV). The highest concentrations were found in Gewürztraminer wines, confirming the importance of rose oxide as a varietal aroma compound in this variety. Surprisingly, the enantiomeric ratio of cis-rose oxide in all investigated wines was substantially lower than in nonfermented musts and in some wines almost racemic cis-rose oxide was detected. Fermentation studies with a model must that contained deuterated water revealed that yeast is capable of reducing the precursor 3,7-dimethyl octa-2,5-dien-1,7-diol (geranyl diol I) yielding 3,7-dimethyl-5-octen-1,7-diol (citronellyl diol I) that gives rise to cis- and trans-rose oxide after acid catalyzed cylization. The deuterium labeling pattern of the resulting rose oxide stereoisomers and a clearly detectable kinetic isotope effect indicate that at least two different reductive pathways in yeast exist that yield cis-rose oxide with different enantiomeric ratios altering the genuine enantiomeric ratio in grape musts. The presence of (+)-cis-rose oxides in wines can therefore be attributed to the reductive yeast metabolism during fermentation. This observation corroborates recent findings that the modification of terpene derived varietal aroma is an integral part of yeast metabolism and not only a simple hydrolytical process.     

Determination of authenticity, regional origin, and vintage of Slovenian wines using a combination of IRMS and SNIF-NMR analyses

The authenticity and geographical origin of wines produced in Slovenia were investigated by a combination of IRMS and SNIF-NMR methods. A total of 102 grape samples of selected wines were carefully collected in three different wine-growing regions of Slovenia in 1996, 1997, and 1998. The stable isotope data were evaluated using principal component analysis (PCA) and linear discriminant analysis (LDA). The isotopic ratios to discriminate between coastal and continental regions are the deuterium/hydrogen isotopic ratio of the methylene site in the ethanol molecule (D/H)(II) and delta(13)C values; including also delta(18)O values in the PCA and LDA made possible separation between the two continental regions Drava and Sava. It was found that delta(18)O values are modified by the meteorological events during grape ripening and harvest. The usefulness of isotopic parameters for detecting adulteration or watering and to assess the geographical origin of wines is improved only when they are used concurrently.     

Stable carbon isotopic composition of the wine and CO2 bubbles of sparkling wines: detecting C4 sugar additions

Sparkling wines have become a popular beverage in recent years, and the production of these wines is subject to adulteration during fermentation. This study investigated the stable carbon isotopic composition (expressed as delta(13)C) of the wine and of the CO(2) bubbles produced during the second fermentation for a number of sparkling wines produced in different countries around the world. Carbon isotope ratio analyses were used to estimate the addition of sugar obtained from C(4) plants (sugar cane or corn). The average delta(13)C values of the Brazilian brut, demi-sec, and doux sparkling wines were -20.5 +/- 1.2 per thousand (n = 18), -18.1 +/- 1.3 per thousand (n = 9), and -15.8 per thousand (n = 1), respectively. These values were statistically heavier (more positive carbon isotope ratio values) than the average delta(13)C of sparkling wines produced in other parts of South America (Argentina and Chile, -26.1 +/- 1.6 per thousand, n = 5) and Europe (France, Germany, Italy, Portugal, and Spain, -25.5 +/- 1.2 per thousand, n = 12), but not statistically different from sparkling wines produced in the United States or Australia. The most likely explanation for differences in the carbon isotope ratios of wines from these different regions is the addition of C(4) sugar during the production of some sparkling wines from Australia, Brazil, and the United States. The isotopic composition of the CO(2) bubbles (delta(13)C-CO(2)) followed similar trends. The average delta(13)C-CO(2) of most of the Brazilian and Argentine sparkling wines was -10.8 +/- 1.2 per thousand (n = 23), indicating that the likely source of carbon for the second fermentation was sugar cane. Conversely, the average delta(13)C-CO(2) of most of the sparkling wines produced in Chile and Europe was -22.0 +/- 1.2 per thousand (n = 13), suggesting that a different sugar (most likely sugar beet) was most used in the second fermentation. It was concluded that in many cases, the carbon isotope ratios of sparkling wine and CO(2) bubbles can provide valuable information about the sugar sources.     

Influence of yeast macromolecules on sweetness in dry wines: role of the saccharomyces cerevisiae protein Hsp12

Yeast autolysis during lees contact influences the organoleptic properties of wines especially by increasing their sweet taste. Although observed by winemakers, this phenomenon is poorly explained in enology. Moreover, the compounds responsible for sweetness in wine remain unidentified. This work provides new insights in this way by combining sensorial, biochemical and genetic approaches. First, we verified by sensory analysis that yeast autolysis in red wine has a significant effect on sweetness. Moderate additions of ethanol or glycerol did not have the same effect. Second, a sapid fraction was isolated from lees extracts by successive ultrafiltrations and HPLC purifications. Using nano-LC-MS/MS, peptides released by the yeast heat shock protein Hsp12p were distinctly identified in this sample. Third, we confirmed the sweet contribution of this protein by sensorial comparison of red wines incubated with two kinds of yeast strains: a wild-type strain containing the native Hsp12p and a deletion mutant strain that lacks the Hsp12p protein (Δ°HSP12 strain). Red wines incubated with wild-type strain showed a significantly higher sweetness than control wines incubated with Δ°HSP12 strains. These results demonstrated the contribution of protein Hsp12p in the sweet perception consecutive to yeast autolysis in wine.     

水蜜桃酒速酿酵母菌的筛选、鉴定及发酵条件优化

为获得快速酿造水蜜桃酒的专用酵母菌株,生产优质水蜜桃酒,以水蜜桃自然发酵醪为筛菌原料,通过TTC显色法与杜氏小管法初筛;再经耐酒精能力、发酵速度与产酒能力以及发酵液风味分析与感官评定三级复筛,获得了优质高效的酿酒酵母菌PY01(Saccharomyces cerevisiae PY01);并以初始pH值、主发酵温度、酵母接种量为自变量,以酒精度为响应值作酿酒工艺的响应面优化。结果表明,PY01菌株发酵水蜜桃酒最适发酵参数为初始pH 值3.9,发酵温度26℃,酵母接种量6.2%。在此条件下发酵可得到酒精度为12.78% vol,总糖含量为6.71 g·L^-1,风味纯正,口感醇厚,典型性的半干型水蜜桃发酵酒。本研究结果为水蜜桃发酵酿酒产业菌种的选择与工艺优化提供了基础。     

Coinoculated fermentations using saccharomyces yeasts affect the volatile composition and sensory properties of Vitis vinifera L. cv. sauvignon blanc wines

Alcoholic fermentation using Saccharomyces wine yeast is an effective means of modulating wine aroma. This study investigated the impact of coinoculating commercial yeast strains (Vin7, QA23, Vin13) on the volatile composition and sensory profile of Sauvignon Blanc wines. Small-scale replicated fermentations were conducted using single-strain and coinoculations of Vin7 with QA23 and with Vin13. The results showed that the chemical and sensory profiles of the coinoculated wines were different from both the single-strain wines and equal blends of the single-strain wines. Volatile thiol analysis indicated that the Vin7/QA23 coinoculated wines were highest in 3-mercaptohexan-1-ol (3MH) and 3-mercaptohexyl acetate (3MHA), although this pattern was not observed for the Vin7/Vin13 yeast combination. The negative white vinegar aroma and high volatile acidity measured in the Vin7 single-strain wines were not present in the coinoculated wines. This study demonstrates that coinoculations can modify the aroma profile of wines, when complementary yeasts are used.     

Foam aptitude of trepat and monastrell red varieties in cava elaboration. 2. Second fermentation and aging

The foam properties of sparkling wines (Cava) made from two red autochthonous grape varieties, Trepat and Monastrell, and coupages, including different percentages of them, were studied during second fermentation and aging. The effect of second fermentation on foam gave the highest decreases when the base wines had the highest foam values, while gave the lowest decreases or even increases for the base wines with the lowest foam characteristics. However, the greater the HM and Sigma of the base wine, the greater the foam values of the sparkling wine. Base wine determinations for quality control in cellars could provide information about future sparkling wine foaming. Acidity parameters, ethanol, sulfur, and polysaccharides contents were correlated to foam characteristics in the sparkling wines. In terms of color and foaming, wines made from the red varieties Trepat and Monastrell blended with white variety wines could be appropriate for elaborating "blanc de noirs" sparkling wines.     

Natural abundance hydrogen isotope affiliation between the reactants and the products in glucose fermentation with yeast

In glucose fermentation, the hydrogen source of products such as ethanol and glycerol is the medium and the sugar. The site-specific natural isotope ratios of the products, (D/H)(i), and that of the medium and sugar, (D/H)(k), may be related by a matrix, A, of redistribution coefficients, a(ik), that characterizes the specific genealogies of the hydrogen atoms. (D/H)(i) = [A](D/H)(k), where (D/H)(i) and (D/H)(k) are the column vectors of the isotope ratios of sites i and k that can be measured by (2)H NMR. The complete redistribution matrix was determined in a set of isotope labeling experiments. Thus, we obtained a mathematical model representing the hydrogen isotope affiliation during alcoholic fermentation. It not only provides information about the biochemical reaction mechanism but also can be used to estimate the isotopic data of the products, based on those of the substrate and the medium. The results prove, in a quantitative way, that the metabolites contain isotopic information about the precursor in a biotransformation and can be used to identify its origin. The method established for the study of the hydrogen-transfer mechanism can be applied to other chemical and biochemical reactions.     

Sensory and analytical study of rose sparkling wines manufactured by second fermentation in the bottle

The sensory and analytical characteristics of five rose sparkling wines manufactured by the traditional method have been determined. Moreover, the changes that take place in the nitrogen and volatile fraction of the wines during the second fermentation and the aging with the yeasts have been studied. Each of these wines was made from a single industrial rose base wine of the Garnacha Tinta variety, with five selected yeasts strains. The base wine had a low content in free amino acids, 16 mg/L, and the yeast consumed more peptides than free amino acids during second fermentation. From the application of the two-way analysis of variance, yeast strain, and aging time factors to the data of volatile compounds, it has been found that most of the differences between these sparkling wines are due to the aging time. It has been verified that these rose sparkling wines have foam of good quality and that the grape variety Garnacha Tinta is suitable for the production of rose sparkling wines.     

Accurate determination of 2,4,6-trichloroanisole in wines at low parts per trillion by solid-phase microextraction followed by GC-ECD

A headspace solid-phase microextraction (HS-SPME) procedure at 30 degrees C with a 100 microm PDMS fiber of a saturated NaCl solution stirred at 1100 rpm combined to GC-ECD for the 2,4,6-trichloroanisol (TCA) determination in wines has been developed. Due to the matrix complexity and ethanol absorption into the fiber, the internal standard selection was crucial to obtain unbiased results. Thus, matrix effects were observed when analyzing different types of Spanish wines (white, early, and vintage red wines) spiked with TCA at low concentration levels (i.e., <40 ng L(-)(1)). In contrast, the use of 2,4,6-tribromoanisole (TBA) as internal standard overcame these matrix effects, whereas the use of 2,4,6-trichlorophenyl ethyl ether led to inconsistent results. The developed HS-SPME-GC-ECD methodology reaches a limit of quantitation for TCA in wine within 2.9-18 ng L(-)(1), with a relative standard deviation of 2.5-13.4%, depending on the TCA concentration level and wine characteristics. This analytical method is comparable to the existing methodologies based on HS-SPME followed by GC-MS in terms of accuracy, precision, length of determination, and length of quantification; however, analysis cost is reduced.     

Influence of fatty acids on wine foaming

The influence of fatty acids (free and bound as ethyl esters) on wine foaming was studied in different white wines and the corresponding sparkling wines. Moreover, from three of these wines the foam formed by CO(2) injection was separated, and two fractions were then obtained: foam wine (FW) and remainder wine (RW). In these fractions and the sparkling wines produced from them, foam properties and fatty acids were also determined. The free fatty acids C8, C10, and C12 were negatively correlated with foamability (HM), whereas the ethyl esters of hexanoic, octanoic, and decanoic acids were positively related to HM. The value of HM was directly proportional to the ratio of esterified to unesterified fatty acids. This was confirmed by the changes that occur in the esterification ratio during the second fermentation and aging. No influence was observed on either the Bikerman coefficient or the stability time of foam.     

Grape contribution to wine aroma: production of hexyl acetate, octyl acetate, and benzyl acetate during yeast fermentation is dependent upon precursors in the must

Wine is a complex consumer product produced predominately by the action of yeast upon grape juice musts. Model must systems have proven ideal for studies of the effects of fermentation conditions on the production of certain wine volatiles. To identify grape-derived precursors to acetate esters, model fermentation systems were developed by spiking precursors into model must at different concentrations. Solid-phase microextraction-gas chromatgraphy mass spectrometry analysis of the fermented wines showed that a variety of grape-derived aliphatic alcohols and aldehydes are precursors to acetate esters. The C6 compounds hexan-1-ol, hexenal, (E)-2-hexen-1-ol, and (E)-2-hexenal are all precursors to hexyl acetate, and octanol and benzyl alcohol are precursors to octyl acetate and benzyl acetate, respectively. In these cases, the postfermentation concentration of an acetate ester increased proportionally with the prefermentation concentration of the respective precursor in the model must. Determining viticultural or winemaking methods to alter the prefermentation concentration of precursor compounds or change the precursor-to-acetate ester ratio will have implications upon the final flavor and aroma of wines.     

Influence of prefermentative treatments to the major volatile compounds of Assyrtiko wines

A study of the volatile fraction of Assyrtiko wines, using gas chromatography coupled with olfactometry, was realized. Twenty-seven volatile compounds were identified as potent odorants, most of them originating from the fermentation process. Quantification of the major volatile compounds was realized developing a rapid analytical method based on fractionation of a 50 mL wine aliquot using C 18-reversed phase adsorbent. After elution of the volatile compounds with pentane-diethyl ether and concentration under nitrogen, the final wine extract was injected in a gas chromatography-flame ionization detection system. The method allows satisfactory determination of more than 15 volatile compounds of wine. The linearity of the method gave a typical r (2) between 0.990 and 0.999, while reproducibility ranged from 5.1 to 12.2% (as relative standard deviation) with 9.5% as the average. The method was applied to wines produced by Assyrtiko grapes (AOC Santorini), for two consecutive years, to compare the effect of skin contact prior to fermentation and the must clarification process. Direct press and skin contact wines were differentiated analytically; however, highly significant differences were not. Inversely, the differences found between direct press/clarified and nonclarified wines were significant. Wines produced by direct press and clarified must presented significantly higher levels of ethylic esters and fusel alcohol acetates but lower fusel alcohol levels, leading probably to more fruity wines. This difference, between clarified and nonclarified grape musts, was not significant in the case of the wines produced by skin contact of Assyrtiko berries. These findings were validated by preference sensory analysis tests.     

Metabolic influence of Botrytis cinerea infection in champagne base wine

Botrytis cinerea infection of grape berries leads to changes in the chemical composition of grape and the corresponding wine and, thus, affects wine quality. The metabolic effect of Botrytis infection in Champagne base wine was investigated through a (1)H NMR-based metabolomic approach. Isoleucine, leucine, threonine, valine, arginine, proline, glutamine, γ-aminobutyric acid (GABA), succinate, malate, citrate, tartarate, fructose, glucose, oligosaccharides, amino acid derivatives, 2,3-butanediol, acetate, glycerol, tyrosine, 2-phenylethanol, trigonelline, and phenylpropanoids in a grape must and wine were identified by (1)H NMR spectroscopy and contributed to metabolic differentiations between healthy and botrytized wines by using multivariate statistical analysis such as principal component analysis (PCA). Lowered levels of glycerol, 2,3-butanediol, succinate, tyrosine, valine derivative, and phenylpropanoids but higher levels of oligosaccharides in the botrytized wines were main discriminant metabolites, demonstrating that Botrytis infection of grape caused the fermentative retardation during alcoholic fermentation because the main metabolites responsible for the differentiation are fermentative products. Moreover, higher levels of several oligosaccharides in the botrytized wines also indicated the less fermentative behavior of yeast in the botrytized wines. This study highlights a metabolomic approach for better understanding of the comprehensive metabolic influences of Botrytis infection of grape berries in Champagne wines.     

Effect of addition of commercial grape seed tannins on phenolic composition, chromatic characteristics, and antioxidant activity of red wine

The effect of addition of grape seed tannins on the phenolic composition, chromatic characteristics, and antioxidant activity of red wine was studied. Two highly pure commercial grape seed tannins (GSE100 and GSE300) were selected, and their phenolic compositions were determined. Two types of red wines were made with Castela?o/Tinta Miu?da (3/2, w/w) grapevine varieties by fermentation on skin using two different maceration times, which correspond to the wines rich and poor in polyphenols, respectively. Each of these wines was used for experimentation with the addition of GSE100 and GSE300 before and immediately after alcoholic fermentation. Phenolic composition, chromatic characteristics, and antioxidant activity of the finished red wines were analyzed by HPLC-DAD, CIElab 76 convention, and DPPH radical test, respectively. The results showed that the addition of grape seed tannins had obvious effects of increasing color intensity and antioxidant activity only in the wines poor in polyphenols. Although GSE300 contained much higher amounts of di- and trimer procyanidins and a lower amount of polymeric proanthocyanidins, it provided effects of increasing the color intensity and antioxidant activity of the wines poor in polyphenols similar to those of GSE100. Furthermore, GSE100 released more gallic acid to wines than GSE300, although no gallic acid was detected in GSE100. Tannins added after alcoholic fermentation had a better effect on phenolic composition of red wine than tannins added before alcoholic fermentation.     

Influence of a Saccharomyces cerevisiae selected strain in the volatile composition of rosé wines. Evolution during fermentation

There has been considerable controversy about the use of selected pure strains in wine fermentation. For that reason it is important to determine the influence of this vinification technique in the composition of wine because it arises from the type of yeast and the subsequent evolution during fermentation. This study researches the volatile composition of rosé wines from the Garnacha must, inoculated with one selected NA33 strain of Saccharomyces cerevisiae. The inoculated yeast did not predominate in all of the samples. These samples showed a behavior intermediate between those of the control and samples in which NA33 did predominate. The greatest concentration of higher alcohols was in the control wine, and its evolution was similar in all fermentations. The esters formed at the end of the fermentation and their concentrations were higher in the control than in the inoculated samples. In the control, acids were produced above all, in the first half of fermentation, and decreased from then onward. In the sample in which the yeast predominated, the synthesis occurred later and to a lesser extent than in the control.     

Identification of amino acids in wines by one- and two-dimensional nuclear magnetic resonance spectroscopy

Amino acids are minor compounds in wines, but they have a profound influence on wine quality, and amino acids composition can be used to differentiate wines according to the vine variety, geographical origin, and year of production. The NMR signals of amino acids in NMR spectra are overlapped by the signals of other compounds present and especially by the signals of dominant compounds such as water, ethanol, and glycerol. In this work we used 1D (1)H and (13)C, 2D homonuclear COSY, TOCSY, and 2D heteronuclear HSQC and HMQC pulse sequences, also with an incorporated WET pulse sequence element that allows the simultaneous suppression of several frequencies. Complete (1)H and (13)C NMR assignments for 17 amino acids commonly present in wine and of gamma-aminobutyric acid at pH 3 have been achieved in wine sample of Sauvignon from the Coastal wine-growing region of Slovenia, vintage 1994.     

Formation of furfurylthiol exhibiting a strong coffee aroma during oak barrel fermentation from furfural released by toasted staves

Furfurylthiol (FFT) is formed in white wines during alcoholic fermentation in the barrel from the furfural released by toasted staves. The quantity of furfural released into the must has a decisive effect on the quantity of FFT in the finished wine. Wines fermented in new barrels thus contain larger quantities of FFT than those fermented in used barrels. Fermentation conditions favorable to an excess production of H(2)S (hydrogen sulfide) by the yeast promote the formation of this volatile thiol. The presence of this volatile thiol in white wines is, therefore, closely related to the yeast's sulfur metabolism.     

Influence of ethanol content on the extent of copigmentation in a cencibel young red wine

The effect of ethanol content on the copigmentation of a Cencibel young red wine was studied, by means of ethanol elimination and reconstitution of the initial volume with different ethanol proportions. The reference wine (14.0% ethanol, 0.40 g/L volatile acidity) showed a bathochromic shift of 4 nm and a color enhancement (DeltaColor) of 41%, lower than that found for the reconstituted wine with the same ethanol content (53%). This discrepancy could be attributable to the loss of acetic acid during the ethanol elimination step. DeltaColor was 95% for the reconstituted wine without ethanol and decreased until 18% for the reconstituted wine with 22% ethanol. Copigmentation was important for reconstituted wines with ethanol contents typical for table red wine, showing DeltaColor between 53 and 57%. An increase in ethanol content in reconstituted wines was accompanied by an increase in the pH value. Perceivable changes in color (DeltaE > 1) followed every increase in ethanol content.     

Optical sensor for sulfur dioxide determination in wines

A method for the determination of free and total sulfur dioxide in wines, based on the use of an optical sensor that employs a dichlorobis(diphenylphosphino)methane dipalladium I complex [Pd(2)(dppm)(2)Cl(2)] immobilized in a PVC membrane plasticized with o-nitrophenyloctylether (o-NPOE) is described. A sensing membrane [4.2% Pd(2)(dppm)(2)Cl(2), 20.8% PVC, and 75% o-NPOE] was adapted to the tip of a bifurcated optical fiber bundle to perform reflectance measurements at 550 nm. The detection system consisted of two cells (40 mL), which hold the sample solution (plus reagents) and the optical sensor, respectively. For the determination of free SO(2), a wine sample was mixed with H(2)SO(4) solution in the sample cell, into which N(2) was bubbled, providing mixing of the solutions and conducting the SO(2) formed toward the detection cell. For determination of total SO(2), a KOH solution was mixed with the wine in the sample cell. Afterward, an H(2)SO(4) solution was added to the cell, and then N(2) was bubbled to conclude the measurement. Linear responses up to 50 and 150 mg L(-1) were obtained for free and total SO(2), with detection limits of 0.37 and 0.70 mg L(-1), respectively. The repeatability of the method was evaluated by carrying out 10 measurements using a single wine sample, providing relative standard deviation values of 2.2 and 2.5% for free and total SO(2), respectively. The sensing membrane prepared from 10 muL of the cocktail solution lasted for 80 measurements, whereas those prepared from 200 muL can be used for 250 measurements. The method was applied to free and total SO(2) determination in wines, and the results did not show significant difference from those obtained with the Ripper reference method at a confidence level of 95%.