PubMed Health⌕ Search

Biomedical subjects

Denis Dubourdieu

Publications and source records attributed to Denis Dubourdieu.

15 recordsLinked to original sources

Assay of glutathione in must and wines using capillary electrophoresis and laser-induced fluorescence detection. Changes in concentration in dry white wines during alcoholic fermentation and aging.

Glutathione (GSH) was assayed in must and wine using capillary electrophoresis coupled with laser-induced fluorescence (LIF) detection. Sample preparation involved conjugating thiols with monobromobimane (MBB) in a 2-(N-cyclohexylamino)ethanesulfonic acid [CHES] buffer (179mM). The electrophoretic conditions were 30kV with a capillary length of 105cm from the inlet to the detector (120cm total length) and a 50microm inner diameter. Under these conditions, the complete separation from the other main non-volatile thiols took less than 20min. We also described the optimum conditions for derivatizing wine samples with MBB to increase eletrophoretic sensitivity. The detection limit for glutathione assay is 65nmol/L. This simple, sensitive method provides a specific assay of glutathione in reduced form, as the sample preparation technique does not modify the balance of oxidized and reduced forms. We used this method to monitor changes in the reduced glutathione content of a white wine during alcoholic fermentation and barrel aging.

Bridged Bicyclo Compounds↗

Stereoisomeric distribution of 3-mercaptohexan-1-ol and 3-mercaptohexyl acetate in dry and sweet white wines made from Vitis vinifera (Var. Sauvignon Blanc and Semillon).

The enantiomeric distribution of 3-mercaptohexan-1-ol (3MH) and 3-mercaptohexyl acetate (3MHA) in Vitis vinifera wines was determined by combining two techniques: specific purification of volatile thiols from the wines using p-hydroxymercuribenzoate and separation of the chiral molecules by gas-phase chromatography on a cyclodextrin capillary column. The R and S enantiomer ratios of these two thiols in dry white Sauvignon blanc and Semillon wines are approximately 30:70 for A3MH and 50:50 for 3MH. However, in sweet white wines made from grapes affected by "noble rot" due to the development of Botrytis cinerea on ripe grapes, the proportion of the R and S forms of 3MH is in the vicinity of 30:70. During alcoholic fermentation, a change in the ratio of the two enantiomers of 3MH in dry white wines was observed. At the beginning of fermentation (around density 1.08), the S form represented over 60%; then, at lower density, as fermentation proceeded, the enatiomeric ratio approached 50:50. The ratio of the two 3MHA enantiomers remained constant throughout fermentation. On the contrary, the distribution of the two 3MH enantiomers changed very little during fermentation of the botrytized sweet wines. The perception thresholds for the R and S forms of 3MH in hydroalcoholic model solution are similar (50 and 60 ng/L). These two enantiomers have quite different aromas: The R form is fruitier, with a zesty aroma reminiscent of grapefruit, while the S form smells more of passion fruit. The perception thresholds of the R and S enantiomers of 3MHA are slightly different (9 and 2.5 ng/L). The less odoriferous R form is reminiscent of passion fruit, while the S form has a more herbaceous odor of boxwood.

Acetates↗

PVPP-polyphenol complexes: a molecular approach.

In dry white wines, two different forms of instability occur: (i) substantial yellow or yellow-green deposits are observed principally due to flavonol quercetin; and (ii) protein instability leads to protein casse. Polyvinyl polypyrrolidone (PVPP) is used to adsorb phenols from beverages, and bentonite is used to eliminate heat instable protein. However, in both cases, their effects are still largely unknown. This study uses a multitechnique approach to gain a better molecular understanding of the association of polyphenol aglycones with PVPP compared to that of glucosides with PVPP. The work demonstrates, that with aglycones, three forces drive complex formation: hydrophobic interaction, H bonds, and van der Waals bonds. With glucosides, the sugar moiety removes or reduces these driving forces. Thus, if the interaction between proteins and polyphenols is responsible for haze and precipitates, as is classically assumed, PVPP could prevent quercetin sedimentation.

Chemical Phenomena↗

Influence of fermentation temperature on volatile thiols concentrations in Sauvignon blanc wines.

The effect of Saccharomyces cerevisiae strains on the amount of 4-mercapto-4-methylpentan-2-one, a major varietal aroma of Sauvignon blanc wines, was demonstrated by previous research work. However, the influence of different alcoholic fermentation parameters on the levels of volatile thiols (4-mercapto-4-methylpentan-2-one, 3-mercaptohexan-1-ol and 3-mercaptohexyl acetate) in wines has not yet been investigated. The impact of fermentation temperature on the final amount of volatiles thiols and on some other analytical parameters (ethanol, total acidity, residual sugars, volatile acidity) was determined in a model medium and in grape juice. Interaction between fermentation temperature and yeast strain was also tested. The fermentation temperature influenced the amount of volatile thiols irrespective of the yeast strain used. The final levels of 4MMP and 3MH in model medium and in wines were higher when the alcoholic fermentation is conducted at 20 degrees C than at 13 degrees C. The 3MHA, which was correlated with the amount of 3MH determined in wines, was also higher when the alcoholic fermentation was conducted at 20 degrees C. From a technological point of view, the choice of yeast strain and fermentation temperature has a decisive influence on the concentrations of the varietal aromas of Sauvignon blanc wines.

Fermentation↗

A novel method for quantification of 2-methyl-3-furanthiol and 2-furanmethanethiol in wines made from Vitis vinifera grape varieties.

A rapid, easy method has been developed for isolating and quantifying 2-methyl-3-furanthiol (2M3F) in wines. Until now, it was not possible to quantify this highly odoriferous compound, with a smell reminiscent of cooked meat, in wine. The original aspect of this method is the specific release of volatile thiols using a cysteamine solution applied in reverse flow to sample percolation on the basis of a p-hydroxymercuribenzoate (pHMB)-volatile thiol conjugate formed by the direct addition of pHMB to 50 mL of wine. Purification of volatile thiols in wines is much faster and easier than our previous method. This method may also be used to assay 2-furanmethanethiol in wine. This thiol's strong aroma of roasted coffee has been shown to contribute to the "roast coffee" aroma of certain wines. Assaying 2M3F by this method showed that it was present in the wines analyzed (red and white Bordeaux, Loire Valley Sauvignon blanc, white Burgundy, and Champagne) at concentrations up to 100 ng/L, i.e., significantly above the olfactory perception threshold for this compound in model dilute alcohol solution.

Fruit↗

Breeding strategies for combining fermentative qualities and reducing off-flavor production in a wine yeast model.

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.

DNA Shuffling↗

Molecular genetic study of introgression between Saccharomyces bayanus and S. cerevisiae.

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.

Beer↗

Origin of (-)-geosmin on grapes: on the complementary action of two fungi, botrytis cinerea and penicillium expansum.

One of the consequences of rot on grapes is the development of volatile compounds giving fungal, mouldy or earthy odours. Among these compounds, (-)-geosmin (trans-1,10-dimethyl-trans-9-decalol), a powerful aromatic compound with an earthy smell is a persistent defect in grape juice and wines made with at least partially rotten grapes. A microbiota analysis of rotten grapes containing (-)-geosmin was carried out on sites from four French regions from 1999 to 2002, to clarify the involvement in geosmin appearance of Streptomyces spp. and Penicillium spp., two types of microorganisms present on grape, that are known for their ability to produce geosmin. In earthy grapes, Botrytis cinerea was largely present. Different species of Streptomyces were also isolated, but their pH sensitivity was an extremely limiting parameter for their development on grape juice, grapes or stem, and consequently for their potentiality to generate geosmin in the vineyard. Penicillium expansum, producing geosmin on a model medium, was omnipresent. Penicillium carneum, which is also a geosmin producer, was represented by a single colony during the 4 years of this study. P. expansum alone was able to produce geosmin on a model medium but not on grapes. However, after 7 days' pre-culture of some B. cinerea strains on grape juice, this juice became favourable to geosmin production by P. expansum. We demonstrated the necessary and complementary action of B. cinerea and P. expansum in geosmin production in grape juice and in crushed grape berries.

Beverages↗

Characterization of Penicillium species isolated from grape berries by their internal transcribed spacer (ITS1) sequences and by gas chromatography-mass spectrometry analysis of geosmin production.

Geosmin (trans-1,10-dimethyl-trans-9-decalol), an earthy-musty compound, has been identified in wines and in grape juice, in which its presence is highly detrimental to the aromatic quality. Geosmin has a biological origin, and the analysis of rotten grape microflora has been done on two grape varieties (Semillon, Cabernet Sauvignon) from six parcels of the Bordeaux region over 3 years (1999, 2000, 2001). Forty-three Penicillium-related species have been analyzed by gas chromatography-mass spectrometry (GC-MS) for their geosmin production. GC-MS analysis has demonstrated that the earthy odor was always correlated with the presence of geosmin. Phenotypic characterization of Penicillium spp. being ambiguous, a molecular characterization by rDNA internal transcribed spacer (ITS1) sequencing was performed for all strains. The results evidenced that all strains producing geosmin belonged to only one species, P. expansum, and that the other strains, not producing geosmin, belonged to three species: P. purpurogenum, P. thomii, and Talaromyces wortmanii.

DNA, Fungal↗

Contribution of benzenemethanethiol to smoky aroma of certain Vitis vinifera L. wines.

Benzenemethanethiol, a volatile thiol with a strong empyreumatic aroma reminiscent of smoke, has been identified in boxwood (Buxus sempervirens L.) as well as in red and white Vitis vinifera L. wines. The perception threshold in a model hydroalcoholic solution is approximately 0.3 ng/L. All of the wines analyzed for this study contained this compound in concentrations of several dozen nanograms per liter. The Chardonnay wines had 30-40 ng/L. Sensory discrimination between a wine containing 7 ng/L benzenemethanethiol and the same wine with an additional 4 ng/L is very significant; the difference in smell is described as "empyreumatic". This compound can therefore significantly contribute to the aroma of certain wines (Sauvignon Blanc, Semillon, Chardonnay, etc.) containing concentrations as high as 30-100 times higher than their perception threshold.

Benzaldehydes↗

Role of certain volatile thiols in the bouquet of aged champagne wines.

A method for the specific extraction of volatile thiols by use of p-hydroxymercuribenzoate has made it possible to identify certain flavor-active volatile thiols in Champagne wines. Benzenemethanethiol, 2-furanmethanethiol, and ethyl 3-mercaptopropionate were present in these wines at concentrations considerably higher than their perception thresholds. Their concentrations increased gradually in proportion to the bottle aging period and sharply as a result of disgorging. The contribution of these volatile thiols to the empyreumatic nuances of the bouquet of aged Champagne wines was demonstrated for the first time.

3-Mercaptopropionic Acid↗

Influence of assimilable nitrogen on volatile acidity production by Saccharomyces cerevisiae during high sugar fermentation.

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.

Journal Article↗

Sulfur aroma precursor present in S-glutathione conjugate form: identification of S-3-(hexan-1-ol)-glutathione in must from Vitis vinifera L. cv. Sauvignon blanc.

When Sauvignon blanc or Gros Manseng grape must was percolated through an immobilized gamma-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.

Chromatography, Liquid↗

Impact odorants contributing to the fungus type aroma from grape berries contaminated by powdery mildew (Uncinula necator); incidence of enzymatic activities of the yeast Saccharomyces cerevisiae.

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.

Ascomycota↗

Volatile components of Loureira, Dona Branca, and Treixadura wines.

White wines experimentally produced from the white grape varieties Loureira, Dona Branca, and Treixadura have been analyzed over four consecutive harvests. The contents of monoterpenes, volatile phenols, alcohols, sulfur components, acetates, fatty acids, and ethyl esters were determined by gas chromatography (FID) and gas chromatography-mass spectrometry. The mean values from four vintages confirmed that these wines have characteristic profiles. Loureira wines are characterized by a high content of free terpenes, 1-hexanol, ethyl acetate, and fatty acids. Dona Branca wines present the highest concentrations of methanol and sulfur constituents, and the lowest concentrations of higher alcohols, acetates, diethyl succinate, and ethyl esters. The levels of monoterpenes in Treixadura wines are very low, but they have the highest concentrations of volatile phenols, principally due to the vanillin, diethyl succinate, ethyl lactate, and ethyl esters. These results were confirmed by principal component and linear discriminant analysis, which show a clear differentiation among these wines as a function of the varietal origin.

Chromatography, Gas↗