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International audience; The oxygen transfer rate (OTR) of closures is a well-known parameter impacting the quality of Sauvignon blanc wines (SBw) within the first years of storage, but little research has been published on its long-term effects. The chemical changes in oxidation odor intensity in three SBw sealed with natural cork and other closures that had different known OTRs, ranging from

THE CHEMISTRY OF WINE. Organic Acids in Wine. Alcohols and Other Volatile Compounds. Carbohydrates. Dry Extract and Minerals. Nitrogen Compounds. Phenolic Compounds. Varietal Aroma. STABILIZATION AND TREATMENTS OF WINE. Chemical Nature, Origins and Consequences of the Main Organoleptic Defects. The Concept of Clarity and Colloidal Phenomena. Clarification and Stabilization Treatments: Fining Wine. Clarifying Wine by Filtration and Centrifugation. Stabilizing Wine by Physical and Physicochemical Processes. Aging Red Wines in Vat and Barrel: Phenomena Occurring During Aging. Index.

Six new triterpenoids (1-6), two known genins (7 and 8), and five known functionalized triterpenoids (9-13) were isolated from a Quercus robur heartwood extract. The purification protocol was guided by LC-HRMS by searching for structural analogues of bartogenic acid on the basis of their putative empirical formula. The structures of the new compounds were unequivocally elucidated using HRESIMS and 1D/2D NMR experiments. Sensory analyses were performed in water and in a non-oaked white wine on the pure compounds 1-13 at 5 mg/L. All molecules were perceived as bitter in water and wine, but they were mostly reported as modifying the wine taste balance. Using LC-HRMS, compounds 1-13 were observed in oaked red wine and cognac and were semiquantified in oak wood extracts. The influence of two cooperage parameters, oak species and toasting process, on compounds 1-13 content was studied. All compounds were found in quantities significantly higher in pedunculate than in sessile oak wood. Toasting is a key step in barrel manufacture and modulates the concentration of the discussed compounds. Significantly higher quantities were observed in untoasted wood compared to medium or highly toasted wood. These findings provide new insights into the molecular origin of taste changes due to oak aging.

BackgroundFermentation completion is a major prerequisite in many industrial processes involving the bakery yeastSaccharomyces cerevisiae.Stuck fermentations can be due to the combination of many environmental stresses. Among them high temperature and ethanol content are particularly deleterious especially in bioethanol and red wine production. Although the genetic causes of temperature and/or ethanol tolerance were widely investigated in laboratory conditions, few studies investigated natural genetic variations related to stuck fermentations in high gravity matrixes.ResultsIn this study, three QTLs linked to stuck fermentation in winemaking conditions were identified by using a selective genotyping strategy carried out on a backcrossed population. The precision of mapping allows the identification of two causative genesVHS1andOYE2characterized by stop-codon insertion. The phenotypic effect of these allelic variations was validated by Reciprocal Hemyzygous Assay in high gravity fermentations (>240 g/L of sugar) carried out at high temperatures (>28°C). Phenotypes impacted were related to the late stage of alcoholic fermentation during the stationary growth phase of yeast.ConclusionsThe genes identified are related to molecular functions such as Programed Cell Death, ROS metabolism and respire-fermentative switch and were never related to fermentation efficiency. Our findings open new avenues for better understanding yeast resistance mechanisms involved in high gravity fermentations.

International audience; Eight new triterpenoids (1-8), the known genin (9), and two known functionalized triterpenoids (10 and 11) were isolated from a Quercus petraea heartwood extract. The structures of the new compounds were unequivocally elucidated using HRESIMS and 1D/2D NMR experiments. Sensory analyses were performed in a non-oaked wine on the pure compounds 1-11. Except compounds 1 and 11, all molecules exhibited a sweet taste at 5 mg/L that was particularly intense for compounds 3 and 9. Using LC-HRMS, compounds 1-11 were observed in an oak wood extract and in oaked red wine and cognac. They were also semiquantified in several samples of sessile ( Q. petraea) and pedunculate ( Q. robur) oak wood extract. All compounds were found in quantities significantly higher in sessile than in pedunculate oak wood. These results support the hypothesis of their contribution to the increase in sweetness during oak aging and show that they can be used as chemical markers to identify the species of oak used for cooperage.

Chardonnay wines impart a unique complex aroma characterized by its buttery, yellow stone fruit, melon, bready, and woody notes. Among the terms used in the sensory analysis of these wines, this study investigated hazelnut-like attributes. Multidimensional gas chromatography coupled to olfactometry identified five pyrroles reminiscent of hazelnut: 1-ethylpyrrole-2-carboxaldehyde, 1H-pyrrole, 2-acetyl-1H-pyrrole (first identification in wine), 1-methylpyrrole-2-carboxaldehyde, and 1H-pyrrole-2-carboxaldehyde. Quantitative analyses demonstrated their significantly higher abundance in Chardonnay wines. However, they proved irrelevant in sensory terms, given the low amounts measured in wine compared to their olfactory detection threshold. Nevertheless, the presence of methanethiol derivatives from these pyrroles was investigated in wine. 1-Methylpyrrole-2-methanethiol and 1-ethylpyrrole-2-methanethiol were identified and exhibited hazelnut-like aroma. These compounds, which have not been observed in natural products to date, are potent volatile compounds with detection thresholds of 0.7 and 1.4 ng/L, respectively, in model wine. These findings open up promising perspectives concerning the interpretation of the typical aromatic nuances of some Chardonnay wines.

Background and Aims Winemakers observe a sensory effect of oak ageing in wines, but the phenomenon has never been fully described. The aim of this study was to determine the influence of the ageing container on the sweetness of dry wines and to evaluate the contribution of oak volatile compounds. Methods and Results Both white and red dry wines were produced in various containers, such as stainless steel and oak tanks, and new and used oak barrels. Sensory analysis (with or without use of nose clip) by a panel of experts showed an increase in sweetness when wine was in contact with wood. Oak volatile compounds (vanillin, β-methyl-γ-octalactone and eugenol) were quantified by gas chromatography-mass spectrometry and then added to the wines to obtain the same concentration in all samples. To determine the contribution of the volatiles to the increase in sweetness, the treatments were tasted again after such an ‘oak aroma’ homogenisation. The ranking of the wines remained unchanged. Conclusion These results demonstrate that the sweet perception of dry wines depends on the nature of the ageing container, and that these oak volatiles are not involved in this phenomenon. Significance of the Study This work confirms an empirical observation and establishes for the first time the sweetening effect of oak ageing, strongly suggesting the existence of sweet non-volatile compounds in oak wood.

Quercoresinosides A and B (1 and 2), two new lignans, were isolated from a toasted Quercus petraea heartwood extract along with a known compound, 3-methoxy-4-hydroxyphenol 1-O-β-d-(6′-O-galloyl)glucopyranoside (3). The purification protocol was based on a taste-guided approach that sought to reveal new bitter compounds released from oak wood into wines and spirits. HRMS and NMR data were used to establish that compounds 1 and 2 are lignan derivatives bearing a glucosyl unit and a galloyl unit at the same positions. Hydrolysis of these compounds showed that they could be distinguished by the absolute configuration of their respective lyoniresinol genin as determined by chiral LC-HRMS in comparison with (+)- and (−)-lyoniresinol standards. Sensory analyses were performed in a non-oaked wine on the pure compounds 1–3. The three molecules exhibited a bitter taste at 2 mg/L that was particularly intense for compounds 2 and 3. Finally, LC-HRMS demonstrated the occurrence of compounds 1–3 in oaked wine and brandy, which supports the hypothesis of their contributions to the increase in bitterness during oak aging.

International audience; Oak aging is a crucial step in winemaking during which the organoleptic properties of wine are modified. Various parameters affect the chemical composition of oak wood including botanical origin, which has been previously shown to be a determinant factor. This study focused on the development of a LC-HRMS method to assay four recently discovered taste-active triterpenes (three sweet and one bitter). The method was applied to evaluate the effect of oak species (Quercus petraea and Quercus robur) on the concentration of these molecules in wood. The results showed that sessile oak was richer in sweet triterpenes and poorer in the bitter one than pedunculate oak, with high interindividual variations within species. Furthermore, a triterpenoid index was calculated to reveal the triterpenoid composition of oak wood. This index appears to be a promising tool for the unambiguous discrimination of oak species and could offer new insights into oak wood selection by coopers and the monitoring of oak aging by winemakers.

International audience; The most famous sweet wine in France is made from botrytised grapes in the Sauternes region. Its distinctive, complex aromas are undoubtedly due to noble rot as a result of Botrytis cinerea development on perfectly ripe Sauvignon blanc and Semillon grapes. The contribution of volatile thiols in botrytised wine aroma was recently reported. The objective of this work was to investigate the impact of B. cinerea development on the aromatic potential of grape juice. The level of S-3-(hexan-1-ol)cysteine (P-3SH), the cysteinylated precursor of the most abundant volatile thiol in wine (3-sulfanylhexanol, 3SH), was determined during grape botrytisation and several grape overripening processes. Production was considerably amplified when B. cinerea had developed on the grapes in both on- and off-vine experiments. The P-3SH levels were increased about 100-fold between the healthy and botrytised stages in one week. We demonstrated that the grape metabolism produced aroma precursor under post-harvest conditions. When grapes were left to overrippen, the vine also produced P-3SH, but levels increased more moderately (about 10-fold). Moreover, a determination of P-3SH distribution demonstrated that B. cinerea was not directly responsible for precursor formation but probably stimulated the grape metabolic pathway involved in this formation.

International audience; A preparative HPLC method, which preserves wine aromas and isolates fruity characteristics in specific fractions, was applied to red wine aroma extracts. Various odor-active zones were detected in typical fractions by GC-O analysis of their extracts. Through further GC-MS analyses, the aromatic compounds responsible for 15 of these odoriferous zones were identified as various ethyl esters and alkyl acetates. In view of their olfactory thresholds, the concentrations of these compounds had no direct impact on the fruity aroma of red wines. Nevertheless, an overall sensory effect of “red-” or “black-berry” nuances was clearly established. Higher than average levels of ethyl propanoate, ethyl 2-methylpropanoate, and ethyl 2-methylbutanoate were involved in black-berry aromas, whereas ethyl butanoate, ethyl hexanoate, ethyl octanoate, and ethyl 3-hydroxybutanoate conferred red-berry aromas.

International audience; 2-Furanmethanethiol, a compound contained in many kinds of food, was identified for the first time in five types of miso fermented soybean paste (red salty rice miso, thin-colored salty rice miso, weak salty rice miso, barley miso, and soy miso) by specific extraction of volatile thiols using p-hydroxymercuribenzoate. In the triangle test with red salty rice miso, which included a higher concentration of 2-furanmethanethiol, and thin-colored salty rice miso, which included a lower concentration of 2-furanmethanethiol, it was shown that the aroma of thin-colored salty rice miso was similar to that of red salty rice miso by adding 2-furanmethanethiol into thin-colored salty rice miso. In addition, a quantitative descriptive analysis (QDA) clearly shows that 2-furanmethanethiol contributed to the intensity of three odor qualities “thick, complex”, “sweet”, and “pleasant aroma like coffee beans” in six odor qualities and was a very important component of miso aroma.

International audience; The premature aging of red Vitis vinifera L. wines is mainly associated with the formation of an intense off-flavor reminiscent of prunes. The compounds responsible for this deterioration in red wine flavor have not previously been identified. Sensory descriptive analysis associated with a gas chromatography-olfactometry (GC-O) technique was first performed to find characteristic odoriferous zones of 15 aged red wines with or without a marked prune aroma. Afterward, high-pressure liquid chromatography, gas chromatography, and multidimensional gas chromatography coupled with mass spectrometry (MDGC-MS) were used to identify the odorants reminiscent of prunes in prematurely aged red wines and in the dried fruit. Three compounds were detected with a strong odor of prunes: gamma-nonalactone, beta-damascenone, and 3-methyl-2,4-nonanedione. The perception threshold of the latter beta-diketone in a model hydroalcoholic solution is 16 ng/L. Identified for the first time in aged red wines, this very powerful volatile compound was also suggested to produce the characteristic prune aroma of prematurely aged red wines. The presence of 3-methyl-2,4-nonanedione was also detected in prunes for the first time.

International audience; Botrytized wines (BW) are famous for their distinctive, complex aromas. To date, only a few studies have analysed the volatile compounds involved in their typical flavours. In this paper, GC-O was applied to BW and dry white wines (DW) made from the same grape varieties to characterize the main odorants responsible for their sensory differences. Surprisingly, only two odorous zones, with grapefruit or curry nuances, were apparently specific to BW. However, GC-AEDA revealed important differences in the FD values between BW and DW, making it possible to screen potent odorants of BW, such as 3-mercaptohexan-1-ol, homofuraneol®, furaneol®, sotolon, methional, and phenylacetaldehyde. GC-MS quantification of homofuraneol®, furaneol®, norfuraneol®, phenylacetaldehyde, and methional in 14 BW, mostly at levels above their perception thresholds, confirmed their contribution to the aroma of BW. Increased concentrations of some of these odorants in BW were shown to be associated with grape botrytization, partially through the desiccation process.

The Saccharomyces bayanus var. uvarum yeasts are associated with spontaneous fermentation of must. Some strains were shown to be enological yeasts of interest in different winemaking processes. The molecular typing of S. bayanus var. uvarum at the strain level has become significant for wine microbiologists. Four microsatellite loci were defined from the exploration of genomic DNA sequence of S. bayanus var. uvarum. The 40 strains studied were homozygote for the locus considered. The discriminating capacity of the microsatellite method was found to be equal to that of karyotypes analysis. Links between 37 indigenous strains with the same geographic origin could be established through the analysis of microsatellite patterns. The analysis of microsatellite polymorphism is a reliable method for wine S. bayanus var. uvarum strains and their hybrids with Saccharomyces cerevisiae identification in taxonomic, ecological studies and winemaking applications.

In agricultural sciences, breeding strategies have historically been used to select new, optimized plant varieties or animal breeds. Similar strategies are possible for genetic improvement of wine yeasts. We optimized 11 relevant enological traits in a single clone using successive hybridization and segregation steps. A hybrid obtained by crossing two parent strains derived from commercial wine yeasts showed that some of the traits were readily optimized. Dominance/recessivity, heterosis and transgression were observed among 51 segregating progeny. On the basis of this information, all the optimal characters from both parents were combined in a single strain following two targeted sexual crosses. This article presents a powerful methodology for obtaining a single wine strain with numerous fermentative qualities that does not produce off-flavors.

The genomic constitution of different S. bayanus strains and natural interspecific Saccharomyces hybrids has been studied by genetic and molecular methods. Unlike S. bayanus var. uvarum, some S. bayanus var. bayanus strains (the type culture CBS 380, CBS 378, CBS 425, CBS 1548) harbour a number of S. cerevisiae subtelomeric sequences: Y', pEL50, SUC, RTM and MAL. The two varieties, having 86-100% nDNA-nDNA reassociation, are partly genetically isolated from one another but completely isolated from S. cerevisiae. Genetic and molecular data support the maintaining of var. bayanus and var. uvarum strains in the species S. bayanus. Using Southern hybridization with species-specific molecular markers, RFLP of the MET2 gene and flow cytometry analysis, we showed that the non-S. cerevisiae parents are different in lager brewing yeasts and in wine hybrid strains. Our results suggest that S. pastorianus is a hybrid between S. cerevisiae and S. bayanus var. bayanus, while S. bayanus var. uvarum contributed to the formation of the wine hybrids S6U and CID1. According to the partial sequence of ACT1 gene and flow cytometry analysis, strain CID1 is a triple hybrid between S. cerevisiae, S. kudriavzevii and S. bayanus var. uvarum.

S-Cysteine conjugate precursors of three volatile thiols were monitored in Vitis vinifera L cv Sauvignon blanc grapes during fruit ripening to assess the influence of vine water and nitrogen status on the grape aroma potential in field conditions. Four dry farmed plots were studied in the Pessac-Leognan and Graves appellations (Bordeaux area) in 1998, which was a very dry vintage, and in 1999, when regular summer rainfall occurred. Soil water-holding capacity ranged from very low to high. Soil total nitrogen content was related to soil organic matter content, which was highly variable on the four plots. Vine vigour was enhanced by both high water and nitrogen status. Major compounds in grapes depended mainly on vine water status. Water deficit-stressed vines produced small berries with low sugar and low total acidity. Grape aroma potential was highest in vines under mild water deficit and moderate nitrogen supply. Severe water deficit stress seemed to limit aroma potential, as did nitrogen deficiency. Consequences for site selection and irrigation management for Sauvignon blanc are discussed. Copyright © 2004 Society of Chemical Industry

We analyzed the variability of volatile acidity and glycerol production by Saccharomyces cerevisiae on a large sample of high sugar musts. The production of volatile acidity was inversely correlated with the maximum cell population and the assimilable nitrogen concentration. The higher the nitrogen concentration, the less volatile acidity was produced. An approach to minimize volatile acidity production during high sugar fermentations by adjustment of assimilable nitrogen in musts was investigated in terms of both quantity and addition time. It was found that the optimal nitrogen concentration in the must is 190 mgN· l −1 . The best moment for nitrogen addition was at the beginning of fermentation. Addition at the end of the growth phase had less effect on volatile acidity reduction. We suggest that by stimulating cell growth, nitrogen addition provides NADH in the redox-equilibrating process, which in turn reduces volatile acidity formation.

When Sauvignon blanc or Gros Manseng grape must was percolated through an immobilized γ-glutamyltranspeptidase column, there was a significant increase in the concentration of S-3-(hexan-1-ol)-l-cysteine, the precursor of 3-mercaptohexan-1-ol, a compound that contributes to the varietal aroma of wines made from these grapes. Low- and high-resolution liquid secondary ion mass sepectrometry (LSIMS) analyses established the presence of S-3-(hexan-1-ol)-glutathione in Sauvignon blanc must. The identification of this compound suggests that the S-3-(hexan-1-ol)-l-cysteine in grapes is produced by the catabolism of S-3-(hexan-1-ol)-glutathione. As is the case in other plant or animal organisms, S-glutathione conjugates may be involved in certain detoxification systems in vines. Keywords: Vitis vinifera; Sauvignon blanc; varietal aroma precursors; S-cysteine conjugate; S-glutathione conjugate; aroma potential; tryptophanase; γ-glutamyltranspeptidase; LSIMS

Powdery mildew due to the fungus Uncinula necator is an important disease for the vineyard. The development of the fungus at the surface of the berries leads to the occurrence of a very characteristic and sometimes intense mushroom-type odor cited as an important default for grapes quality. Gas chromatography/olfactometry, gas chromatography, and multidimensional gas chromatogaphy/mass spectrometry techniques were used to investigate the most important odorants of grapes diseased by powdery mildew. Among 22 odorants detected, strongly odorant compounds were identified or tentatively identified in purified extracts obtained from grapes diseased by powdery mildew. Aroma extraction dilution analysis (AEDA) analysis revealed that 1-octen-3-one (mushroom odor), (Z)-1,5-octadien-3-one (geranium-leaf odor), and an unidentified odorous zone (fishy-mushroom like odor) were the most potent volatiles of the diseased grapes. In the presence of nonproliferating Saccharomyces cerevisiae yeast cells, and consequently during alcoholic fermentation, the enzymatic reduction of 1-octen-3-one and (Z)-1,5-octadien-3-one to much less odorant compounds, namely 3-octanone and (Z)-5-octen-3-one, was shown. Those results explain to some extent the disappearance of the fungal aroma specific to powdery mildew grapes during alcoholic fermentation.

International audience; Yeast lees influence the organoleptic properties of wines by increasing their sweet taste. This effect is in part due to the protein Hsp12p, which is regulated by different stress response pathways in Saccharomyces cerevisiae. This work investigated the genetics and environmental factors influencing the expression level of the HSP12 gene in an enological context. RT-qPCR confirmed that the HSP12 expression level is regulated by temperature change and ethanol content during the alcoholic fermentation but not by the sugar content. Moreover, this gene shows an important variation according to the yeast strain used. For the first time yeast strain is demonstrated to play an important role in the perception of sweetness in red wine due to post-fermentation lees autolysis. Interestingly, a correlation between the expression level of HSP12 and the sweetness perception was found using yeast strains of different origins. All of the findings provide new insights on the contribution of yeast to wine taste.

International audience; The modification of wine taste during oak ageing is due to the felease of non-volatile compounds from wood. Among these molecules, some lignans have been previously described as exhibiting bitterness. However, the lack of knowledge concerning this class of compounds in oak wood led us to explore both their structural diversity and their sensory properties. Nine lignans were isolated from extracts of Quercus petraea oak heartwood. Among them, one new compound called quercoresinol was identified and four other molecules were described for the first time in Quercus genus. The presence of these lignans in oaked wine was then established and their gustatory properties were evaluated. Lyoniresinol was the bitterest compound with a detection threshold of 1.5 mg/l. An LC HRMS quantitative method was performed to study the influence of oenological practices on lyoniresinol concentration in wine.

Wines from four Galician white varieties (Albariño, Loureira, Treixadura and Dona Branca) with 1992 to 1995 vintages have been analysed, in order to obtain their characterisation by the aromatic index. Monoterpenes, higher alcohols, ethyl esters, fatty acids, acetates, volatile phenols and heavy sulphur compounds were analysed by gas chromatography alone and together with mass spectrometry. Flavour compositions were similar for each wine obtained from grapes harvested in different years. Comparisons between odour unit values (OUV) from each wine variety and from these vintages were similar. However, odour profiles of Galician wines from these years were clearly differentiated. A correct differentiation could be achieved between samples elaborated with different varieties, when principal component and linear discriminant analysis were applied to the OUV data.

Summary Using genetic and flow cytometric analyses, we showed that wine strain S6U is an allotetraploid of S. cerevisiae × S. bayanus . Hybrid constitution of the strain and its meiotic segregants was confirmed by Southern hybridization analysis of their chromosomal DNAs using four S. cerevisiae cloned genes: LYS2 (chr. II), TRK 1 (chr. X), ARG 4 (chr. VIII), ACT 1 (chr. VI) and PCR/RFLP analysis of the MET 2 gene (chr. XIV). Monosporic progeny of strain S6U was highly viable in first generation but completely non-viable in the second one. According to the genetic analysis, sherry strain S. cerevisiae SBY 2592 was found to be an autotetraploid heterozygous for homo-heterothallism.

Musty, earthy odors are highly detrimental to the aromatic quality of wines. A characteristic aroma of freshly tilled earth, damp cellar was studied in some red and white wines of different origins. The extraction and purification of the wines marked by this odor have shown after analysis by gas chromatography-olfactometry a unique strong odorous zone having the same odor as the one perceived at tasting. The compound responsible for this odorous zone was identified by gas chromatography-mass spectrometry as geosmin (trans-1,10-dimethyl-trans-9-decalol), which possesses a distinctive earthy odor. Geosmin may be present in wines at levels higher than the racemic geosmin olfactory perception threshold, thus suggesting its contribution to their off-aroma. Moreover, the presence of this compound in juice taken from freshly crushed grapes suggests that microorganisms that develop on the grapes may contribute to the presence of this compound in wines.

Sweet fortified wines, traditionally aged under strong oxidation conditions, have a characteristic aroma. An experimental study investigated the aging of white sweet fortified wines under various conditions. The development of various molecules, previously identified as characteristic of the aroma of this type of wine, was monitored by analysis. The development of these compounds during accelerated aging was affected by oxidation and the color of the wine. Aging in oak containers, variable storage temperatures, and variable modes of oxidation affect the formation of many of the volatile compounds responsible for the aging aroma of vins doux naturels (fortified wines). Furfural (1), 5-ethoxymethylfurfural (2), and sotolon (3) always reach concentrations above their perception thresholds in wood-aged wines, especially when new oak is used. The isomers of methyl-gamma-octalactone (4 and 5) are reliable indicators of aging in oak. Among the many identifiable volatile phenols, only the vanillin (7) content increases with aging, particularly if the container is made of wood and there is a high degree of oxidation. This molecule frequently reaches and even exceeds its perception threshold. We show here that vanillin and isomers of methyl-gamma-octalactone have a major impact on the aromas of fortified wines.

3-Mercapto-2-methylpropanol, an odorous compound recently identified in red wines, was extracted from a red wine variety, using a low-temperature vacuum distillation coupled with a specific reversible thiol capture. The analysis of the diacetylated wine extract by enantioselective multidimensional gas chromatography with mass spectrometry detection revealed the presence of only one enantiomer, (R)-3-mercapto-2-methylpropanol. Although both of the optical isomers are characterized by the same broth and sweat odour, they have very different odour thresholds. The amounts of (R)-3-mercapto-2-methylpropanol in young Cabernet-Sauvignon and Merlot wines can exceed its perception threshold, thus suggesting its contribution to the aroma of these wines.

Toasting changes both the quantity and the quality of the extractable substances in the oak wood of barrels used for the aging of fine wines and spirits. Mastery and repeatability of toasting are vital in the production of quality barrels to be used for the aging of wines and spirits. In this study, we show that it is possible for a given cooperage to differentiate barrel toasting levels by analyzing a certain number of volatile and semivolatile compounds resulting from the thermal degradation of oak. Toasted wood components, which can be extracted by the wines or spirits during the aging process, are normally analyzed after the wood has soaked in standard alcoholic solutions and can be used to control the intensity of the wood toasting. The results of the comparative analysis presented in this work show that headspace analysis with a microextractive method using a stationary polydimethylsiloxane type phase is a promising technique for analyzing toasted oak wood from barrels. It is easier to use than the traditional maceration and extraction method and provides similar information.

The thermoprotective effect of yeast parietal mannoproteins improves the protein stability of white wines aged on their lees (sur lie). The substance responsible for this is an N-glycosylated, 31.8 kDa mannoprotein that corresponds to a parietal invertase fragment of Saccharomyces cerevisiae. This mannoprotein is released into the wine during autolysis of the lees by the combined action of β-glucanases from the cell wall and the yeast's vacuolar protease. This mannoprotein may be obtained industrially by extracting yeast mannoproteins using enzymatic digestion of the cell wall with commercially prepared β-glucanases (GlucanexTM -Novo-Nordisk). A wine's susceptibility to protein breakdown may be considerably reduced by adding this mannoprotein extract. As a result, less bentonite is needed to stabilise the wine. © 1999 Society of Chemical Industry

Pectolytic enzymatic preparations used in enology come from Aspergillus niger. They have a desirable principal activity and several secondary activities. Among these secondary activities, three classes of esterases were detected: an esterase specific for ester linkage between cinnamic acid and alcohol (cinnamate esterase); an esterase acting on ester linkage between two phenolic rings (depsidase); and an esterase hydrolysing ester linkage between phenolic acids and straight-chain alcohols (phenyl esterase). Only the first class of enzyme is able to hydrolyse the cinnamic acid tartrate ester of the white must. The authors tried to purify the cinnamate esterase from the other esterase of an A niger industrial pectolytic preparation. This purification was done in two steps. A gel filtration on S200 HR Sephacryl was first used. This step eluted two fractions: esterases and pectin esterase. The second step was affinity chromatography on a 4B Sepharose EAH grafted with chlorogenic acid. This step eluted two fractions: an unretained fraction containing the phenyl esterase and the depsidase; and a retained fraction with the cinnamate esterase. This enzyme has a molecular weight of 240 kDa and contains two submits of 120 kDa. A new classification for these three esterases is suggested. © 1998 Society of Chemical Industry.

This study is dedicated to quantification, by gas chromatography combined with mass spectrometry, of some volatile compounds founded during ageing of sweet natural wines. Besides determination of the thresholds olfactory perception enable us knowing better the more active compound in sweet natural wines aroma.The results enable to know better volatile compounds which are able to produce ageing aromas characteristic of sweet natural wines. These products are always rich in sugar and grown up in a more or less oxidising environment. Logically, many of the compounds identified in this work (derived furanic compounds and many others) are Iinked with chemical like Maillard reactions between sugars and amino-acids. Others compounds as a sotolon need oxidising phenomena to increase in the environment.Comparatively, oldest Port wines have been analysed. The compounds measured in these Port wines also exist in French sweet natural wines of type Banyuls and Rivesaltes, keeped under oxidising environment. But the dillerences of concentrations in some compounds discriminate them from the Port wines.

Three flavor-active volatile thiols (4-mercapto-4-methylpentan-2-one, 4-mercapto-4-methylpentan-2-ol and 3-mercaptohexan-1-ol) involved in Vitis vinifera L. var. Sauvignon blanc wine aroma can be generated in vitro from nonvolatile must extracts by the enzyme action of a cell-free extract obtained from the gastrointestinal bacterium Eubacterium limosum. The specificity of a cysteine conjugate β-lyase (EC 4.4.1.13) which is contained in the cell-free extract, strongly suggested the existence of precursor for these volatile thiols having a structure of S-cysteine conjugate. Gas-phase chromatography analysis coupled with mass spectrometry of must extract in the form of trimethylsilylated derivatives, verified this hypothesis. The release of flavor-active volatile thiols during alcoholic fermentation of the must is shown to be due to the degradation, by yeast, of the corresponding S-cysteine conjugates. This mechanism explains the amplification of the typical Sauvignon blanc grape aroma during alcoholic ferme...

Three new flavour-active mercapto-alcohols have been identified in Sauvignon blanc wines: 4-mercapto-4-methylpentan-2-ol, 3-mercaptohexan-1-ol and 3-mercapto-3-methylbutan-1-ol. The first two have a perception threshold of the order of 60 ng/l in an aqueous alcohol solution and their respective odours are reminiscent of citrus zest and grapefruit. The perception threshold of 3-mercapto-3-methylbutan-1-ol, with its odour of cooked leeks, is higher (1500 ng/l in the same solution). These three compounds may contribute towards the typical aroma of Sauvignon blanc wines.

Volatile thiols were purified from a dichloromethane extract of Sauvignon blanc wine by the reversible combination of the thiols with p-hydroxymercuribenzoate, fixation of the resulting complex in an anion exchange column, and finally release of the thiols with cysteine. By analyzing the purified thiol extract from 500 mL of wine, using gas chromatography coupled with mass spectrometry, it was possible to measure the concentrations of five volatile compounds at once: 4-mercapto-4-methylpentan-2-one, 3-mercaptohexyl acetate, 3-mercaptohexan-1-ol, 4-mercapto-4-methylpentan-2-ol, and 3-mercapto-3-methylbutan-1-ol. Analysis of 10 typical Sauvignon Blanc wines (Bordeaux, Sancerre) showed that the first three compounds listed above are expected to be involved in varietal aroma, as they may be present at concentrations greatly in excess of their respective perception thresholds. Keywords: Volatile thiol; aroma; Sauvignon blanc wine; Vitis vinifera; p-hydroxymercuribenzoate

The unpleasant "sawdustI odour sometimes found in wines aged in new barrels was studied by gas chromatography olfactory detection and mass spec- trometry. A particular aromatic zone has several chromatographic peaks, corre- sponding to certain characteristic odours which can be detected by smelling the gas effluent. These include the subtle rancid, weedy odour of "sawdustI. Various carbonyl components have been identi-ed in this particular zone. These include (E)-2-nonenal, 3-octen-1-one, (E)-2-octenal and 1-decanal. These molecules are the primary cause of disagreeable oak wood odours. By measuring (E)-2-nonenal in wines, after derivation by O-(2,3,4,5-pentaNuobenzyl)-hydroxilamine (PFBOA), it was possible to correlate the concentration of this unsaturated aldehyde with the intensity of the "sawdustI character noted in tasting. The possible explana- tions for the presence of these carbonyl compounds are debatable. The carbonyl content varied from one wood sample to another. However, controling the toast- ing of the wood, both in the laboratory and in the cooperage, produced a major reduction in the extractable (E)-2-nonenal content and eradicated the "sawdustI character of the wine. 1998 SCI. (

A very odorous compound, 4-mercapto-4-methylpentan-2-one has been identified in the box tree, Buxus sempervirens L., and in broom, Sarothamnus scoparius (L.) Koch by a selective extraction of volatile thiol compounds. Although the concentration varied depending on weather and the places where samples were collected, the maximum concentration of the thiol from these plants reached ca. 22 ng/g fresh leaves (box tree) and 16 ng/g fresh stem (broom), respectively. Considering the very low perception threshold of this thiol, the characteristic aroma of these plants is due to 4-mercapto-4-methylpentan-2-one. A plant that contains this thiol abundantly has not been reported before. © 1997 John Wiley & Sons, Ltd.

Toasting wood to be used in barrels for aging wine and brandy produces a great number of volatile and odiferous compounds. Some of these have a "toasty caramel" aroma. Analysis by high-performance gas chromatography of toasted oak extracts, combined with olfactory detection, enabled various Chromatographic peaks with these specific aromas to be isolated. These same odors were simultaneously studied by heating glucose both with and without proline. Aromatic compounds of interest were identified thanks to a combination of gas chromatography and both mass and infrared spectrometry. In addition to already-known substances such as 2-hydroxy-3-methyl-2-cyclopenten-1-one (cyclotene) (1) and 3-hydroxy-2-methyl-4H-pyran-4-one (maltol) (2), we identified, for the first time, 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP) (3), 4-hydroxy-2,5-dimethyl-3(2H)-furan-3-one (furaneol) (4), and 2,3-dihydro-5-hydroxy-6-methyl-4H-pyran-4-one (dihydromaltol) (5) as compounds which actively contribute to the "toasty caramel" aroma of heated oak. Their role in the aroma of wines and spirits is thus well worth studying. These molecules were absent from the glucose pyrolysis but were relatively plentiful after heating in the presence of proline. Therefore, Maillard reactions were seen to be the cause of these substances in the heated oak wood.

4-mercapto-4-methylpentan-2-one (4-MMP), a strongly odorant compound responsible for the « boxtree » or « broom plant » odour of the Sauvignon wines, can be enzymaticaly released in vitro from an odourless must extract. The enzyme source used is a cell-free extract of the gastrointestinal bacterium Eubacterium limosum. This crude preparation exhibits a cysteine β-lyase activity which requires the presence of pyridoxal phosphate. The release of 4-MMP is inhibited when the substrate is previously treated with N-hydroxysuccimide acetate which reacts with a primary amine. The same bacterial extract is also able to release 4-MMP, pyruvic acid and ammonium, from S-(4-méthylpentan-2-one)-L-cysteine. On the other hand, the cleavage of S-(4-méthylpentan-2-one)D,L-homocysteine and S-(4-méthylpentan-2-one)- glutathione is very limited. These results suggest that the precursor of 4-MMP in Sauvignon must is a S-cysteine conjugate. Such an aroma precursor in grapes or in other fruits has never been round berore.

p-Hydroxymercuribenzoic acid (pHMB), reacts reversibly with thiols. Using this principle, an extraction method of the thiols present in wines has been established. Injecting the extracts from Vitis vinifera L. var. Sauvignon wines on to a gas-chromatographic column using the purge and trap technique and a flame photometric detector enabled us to discover some sulphur compounds present in wines from this cultivar. On several capillary columns of different polarities, one of the peaks possesses the same retention time as the potent odourant, 4-mercapto-4-methylpentan-2-one, which evokes odours of box tree and cat urine. The identification of this compound in Sauvignon wines was confirmed by mass spectrometry. These results confirm previous work obtained by coupling gas chromatography and olfactometry, and show that this thiol is present in Sauvignon wines. The perception threshold of this compound in water and in wine is very low (close to 0.1 ng/1 and 3 ng/1s respectively) and despite its low concentration in Sauvignon wines, 4-mercapto-4-methylpentan-2-one seems to play a major role in the varietal aroma of the wines from this cultivar.

International audience; Grapes (Vitis spp.) are economically significant fruit species. Many scientific advances have been achieved in understanding physiological, biochemical, and molecular aspects of grape berry maturation. Some of these advances have led to the improvement of wine quality through better grape growing practices; but this avenue of research is still wide open due to the complexity of the biochemical mechanisms involved in the development and ripening of grapes and their response to environment. The Biochemistry of the Grape Berry is a pioneer offering for readers - giving a focused and integrated coverage of the key biochemical and molecular aspects of grape berry development and ripening. Readers will benefit from a comprehensive and updated coverage on key steps, mechanisms and regulation in grapevine biochemistry and molecular biology, provided by an international team of leading scientists. It provides useful references on this research area and original data from laboratories worldwide. The comprehensive range of topics covered in this e-book is valuable to a multidisciplinary readership including plant physiologists, enologists, microbiologists, wine makers, biochemists and students. Thus The Biochemistry of the Grape Berry represents a step forward in the comprehension of grape berry biochemistry, wine quality and health benefits.

International audience; Several studies have reported the beneficial influence of non-Saccharomyces yeasts and their potential applications in the wine industry, mainly in mixed-culture fermentation with S. cerevisiae. The potential impact of 15 non-Saccharomyces strains from 7 species on 4-methyl-4-sulfanylpentan-2-one (4MSP) and 3-sulfanylhexan-1-ol (3SH) release in model medium and Sauvignon Blanc must was evaluated after partial fermentation. Whereas the impact of non-Saccharomyces on 4MSP release in both media was low, some M. pulcherrima, T. delbrueckii and K. thermotolerans strains had a high capacity to release 3SH, despite their minimal fermentation activity. As previously demonstrated for Saccharomyces yeast, this contribution is strain dependant. Taking into account their dynamic and quantitative presence during the whole process, the real impact of non-Saccharomyces yeast on 4MSP and 3SH release was evaluated using a recreated community simulating the yeast ecosystem. Our results revealed a positive impact on 3SH release in Sauvignon Blanc wines by promoting non-Saccharomyces yeast activity and delaying the growth of S. cerevisiae. Some non-Saccharomyces yeast strains are capable of making a positive contribution to volatile thiol release in wines, essentially during the pre-fermentation stage in winemaking, when this microbiological sub-population is dominant.

International audience; Volatile thiols, compounds that contribute strongly to the varietal aroma, are present in much higher concentrations in sweet wines than in dry wines. This positive effect, due to the presence of Botrytis cinerea on the berries, in fact results from a strong enrichment of cysteine S-conjugate precursors in botrytized berries. In the present study, a convenient model was investigated to reproduce and therefore study this phenomenon. A Vitis vinifera cell culture was used as a simple model, and we focused on S-3-(hexan-1-ol)-l-cysteine (P-3SH), the cysteinylated precursor of 3-sulfanylhexanol. We demonstrated that grapevine cells were able to produce P-3SH and that the presence of B. cinerea considerably increased the precursor level (up to 1000-fold). This positive result was determined to be due to metabolites secreted by the fungus. These molecules were temperature sensitive, unstable over time, and their production was activated in the presence of grapevine cells. Moreover, part of the pathway leading to P-3SH was deciphered: it was directly derived from the cleavage of S-3-(hexan-1-ol)-l-glutathione, which itself was generated after a conjugation of glutathione on (E)-2-hexenal.

International audience; 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.

Vinylphenols (4-vinylphenol and 4-vinylguaiacol) are natural constituents of wine and can play a role in wine aroma. However, only white wines contain important quantities of these volatile substances which, beyond a certain concentration (limit threshold = 725 μg litre−1 of 4-vinylguaiacol+4-vinylphenol (1:1)), can be responsible for a depreciating ‘phenolic’ or ‘pharmaceutic’ characteristic, Saccharomyces cerevisiae possesses a type-(E) enzymic activity, substituted cinnamate carboxy-lyase (SCD), which is capable of transforming, by non-oxidative decarboxylation, the phenolic acids in the must, (E) p-coumaric and (E) ferulic acids, into corresponding vinylphenols. This endocellular activity is constitutive, it is only expressed during alcoholic fermentation and with a variable intensity depending on the yeast strain. Furthermore, the enzyme is rapidly inhibited by catechic tannins, which explains why, in comparison with white wines, red and rose wines contain low levels of vinylphenols despite having more precursors. A yeast strain with a weak SCD activity has been selected and its use in vinification should eliminate the appearance of the phenolic taint in white wines coming from grapes rich in decarboxylable hydroxycinnamic acids.

Commonly used techniques for the identification of industrial yeast strains are usually time-consuming and cumbersome. Moreover, some of these methods may give ambiguous results. A novel strategy has been developed for identifying yeast strain employing polymerase chain reaction technology. Using customised oligonucleotides, some regions of the yeast genome between δ elements are amplified to give an ‘amplified’ sequence polymorphisml (Skolnick and Wallace 1988) characteristic of the strains. With this technique it is possible to identify individual strains of Saccharomyces cerevisiae.

Among Saccharomyces yeast, S. cerevisiae and S. bayanus var. uvarum are related species, sharing the same ecosystem in sympatry. The physiological and technological properties of a large collection of genetically-identified S. bayanus var. uvarum wine strains were investigated in a biometric study and their fermentation behavior was compared at 24 degrees C and 13 degrees C. The variability of the phenotypic traits was considered at both intraspecific and interspecific levels. Low ethanol tolerance at 24 degrees C and production of high levels of 2-phenylethanol and its acetate were clearly revealed as discriminative technological traits, distinguishing the S. bayanus var. uvarum strains from S. cerevisiae. Although some S. bayanus var. uvarum strains produced very small amounts of acetic acid, this was not a species-specific trait, as the distribution of values was similar in both species. Fermentation kinetics at 24 degrees C showed that S. bayanus var. uvarum maintained a high fermentation rate after Vmax, with low nitrogen requirements, but stuck fermentations were observed at later stages. In contrast, a shorter lag phase compared with S.cerevisiae, higher cell viability, and the ability to complete alcoholic fermentation at 13 degrees C confirmed the low-temperature adaptation trait of S.bayanus var. uvarum. This study produced a phenotypic characterization data set for a collection of S. bayanus var. uvarum strains, thus paving the way for industrial developments using this species as a new genetic resource.

International audience; Wine has very rich and complex aromas but is paradoxically obtained from grape juice that has very little odour itself. This aromatic transformation is mainly due to the action of yeasts during the alcoholic fermentation. First, yeasts produce secondary metabolites that give to wines its basic 'winy' characteristic. Second, some enzymatic activities may transform non-odorous compounds of certain grape varieties into varietal aromas, giving the specific characteristics of 'cepage'. Depending on the yeast strain, the concentration of aromatic molecules is determined, modifying the organoleptic perception of wine. The enzymatic activities of yeast metabolism and their genetic control have been widely studied, allowing genetic selection approaches as well as molecular engineering for selecting more specialized commercial starters.

International audience; During red wine fermentation, high temperatures may cause stuck fermentation by affecting the physiology of fermenting yeast. This deleterious effect is the result of the complex interaction of temperature with other physicochemical parameters of grape juice, such as sugar and lipid content. The genetic background of fermenting yeast also interacts with this complex matrix and some strains are more resistant to high temperatures than others. Here, the temperature tolerance of nine commercial starters was evaluated, demonstrating that, at high sugar concentrations, half of them are sensitive to temperature. Using a classical backcross approach, one thermo-sensitive commercial starter was genetically improved by introducing quantitative trait loci conferring resistance to temperature. With this breeding program it is possible to obtain a thermo-resistant strain sharing most of its genome with the initial commercial starter. The parental and improved strains were compared for population growth and fermentation ability in various conditions. Despite their common genetic background, these two strains showed slight physiological differences in response to environmental changes that enable identification of the key physiological parameters influencing stuck fermentation.

A Selection of papers presented at the 6th Symposium in Vino Analytica Scientia - In Vino 2009 ; International audience; Sweet wines made from botrytized grapes contain much higher concentrations of volatile thiols, especially 3-sulfanylhexan-1-ol (3SH), than dry white wines. Three new specific volatile thiols (3-sulfanylpentan-1-ol (3SP), 3-sulfanylheptan-1-ol (3SHp), and 2-methyl-3-sulfanylbutan-1-ol (2M3SB) were recently identified in Sauternes wines. Like most volatile thiols, these compounds were almost totally absent from must, mainly being formed during alcoholic fermentation. In this work, we describe the identification and quantification of three new cysteine-S-conjugate precursors in must made from Botrytis-infected grapes. S-3-(pentan-1-ol)-l-cysteine (P-3SP), S-3-(heptan-1-ol)-l-cysteine (P-3SHp), and S-3-(2-methylbutan-1-ol)-l-cysteine (P-2M3SB) were identified by direct GC–MS analysis of their derivative forms obtained by silylation of an enriched fraction, isolated from must by affinity chromatography. Concentrations were considerably higher when Botrytis cinerea had developed on the grapes. In botrytized must, the mean levels of P-3SP, P-3SHp, and P-2M3SB were in the vicinity of 700, 50, and 500 nM, respectively, whereas concentrations in healthy must ranged from 0 to 50 nM. This indicated that these three new sulfanyl alcohols, responsible for the characteristic aroma of botrytized wines, were formed by the yeast metabolism during alcoholic fermentation from the corresponding non-volatile cysteine-S-conjugate precursors. Moreover, these results highlighted the predominant role of botrytization in developing grape aroma potential.