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Rare earth element (REE), dissolved organic carbon (DOC) and trace-element (Al, Mn, Fe, Sr, Ba, U and Th) concentrations were measured in fourteen well water samples (

Aspergillus oryzae was found to secrete two distinct β-glucosidases when it was grown in liquid culture on various substrates. The major form had a molecular mass of 130 kDa and was highly inhibited by glucose. The minor form, which was induced most effectively on quercetin (3,3′,4′,5,7-pentahydroxyflavone)-rich medium, represented no more than 18% of total β-glucosidase activity but exhibited a high tolerance to glucose inhibition. This highly glucose-tolerant β-glucosidase (designated HGT-BG) was purified to homogeneity by ammonium sulfate precipitation, gel filtration, and anion-exchange chromatography. HGT-BG is a monomeric protein with an apparent molecular mass of 43 kDa and a pI of 4.2 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and isoelectric focusing polyacrylamide gel electrophoresis, respectively. Using p -nitrophenyl-β- d -glucoside as the substrate, we found that the enzyme was optimally active at 50°C and pH 5.0 and had a specific activity of 1,066 μmol min −1 mg of protein −1 and a K m of 0.55 mM under these conditions. The enzyme is particularly resistant to inhibition by glucose ( K i , 1.36 M) or glucono-δ-lactone ( K i , 12.5 mM), another powerful β-glucosidase inhibitor present in wine. A comparison of the enzyme activities on various glycosidic substrates indicated that HGT-BG is a broad-specificity type of fungal β-glucosidase. It exhibits exoglucanase activity and hydrolyzes (1→3)- and (1→6)-β-glucosidic linkages most effectively. This enzyme was able to release flavor compounds, such as geraniol, nerol, and linalol, from the corresponding monoterpenyl-β- d -glucosides in a grape must (pH 2.9, 90 g of glucose liter −1 ). Other flavor precursors (benzyl- and 2-phenylethyl-β- d -glucosides) and prunin (4′,5,7-trihydroxyflavanone-7-glucoside), which contribute to the bitterness of citrus juices, are also substrates of the enzyme. Thus, this novel β-glucosidase is of great potential interest in wine and fruit juice processing because it releases aromatic compounds from flavorless glucosidic precursors.

Sclerotinia sclerotiorum secretes β-galactosidase, giving very low activity when cultivated in a medium with glucose as the carbon source. This production is greatly enhanced during growth on different pectin related polysaccharides. An extracellular β-galactosidase was purified 26-fold by ammonium sulfate precipitation, gel filtration and ion exchange chromatography. This enzyme has a Km of 0.15 mg/ml, a Mr of 120 000 and a pI of 6.0. Enzyme activity was optimal at pH 4.0 and 50°C.

Cultures of Sclerotinia sclerotiorum grown on different pectin-related polysaccharides (citrus pectin, apple pectin, sodium polygalacturonate), carboxymethylcellulose (CMC) or glucose as the only carbon source were examined daily for polygalacturonase and pectinase activities. Electrophoretic forms of polygalacturonase and pectin methylesterase activities were revealed using analytical IEF and sodium polygalacturonate and citrus pectin as substrates in overlay gels. A sequence in the production of pectic enzymes and isoenzyme synthesis was found in pectic-polymer cultures corresponding to the induction of several isoenzymes. Enzyme activities in glucose media were associated with three polygalacturonase and two pectinmethylesterase isoforms which were produced constitutively. Sodium-dodecyl-sulphate polyacrylamide-gel electrophoresis followed by immunoblotting with polyclonal antibodies against an exo-polymethylgalacturonase and an exo-polygalacturonase revealed that these exo-enzymes were secreted from the beginning of cultivation in the different culture media showing characteristics of constitutive enzymes.

Aims: Grenache noir and Syrah are the predominant grape varieties in the French Rhone valley vineyard. This study aimed at identifying the odorants generated from glycoconjugates extracted from wines made with Grenache noir and Syrah.Methods and results: Synthetic model wines enriched with glycoconjugates, treated or not with enzymes, were stored at 45 °C for 3 weeks, or at 13 °C for 18 months. Aromas generated were extracted and analyzed by GC-Olfactometry (only samples from accelerated aging) and were further quantitatively determined by GC-MS. Analysis of the extracts allowed to identify 49 odorants, including 27 that could be aglycons, or related compounds, of glycoconjugates from the grapes. In addition, the active compounds were quantified in similar experiments led in conditions of natural aging for 18 months.Conclusion: The two varieties, Grenache noir and Syrah, were distinguishable by 14 odorant zones. Multivariate analyses (PCA) performed with the amounts of aroma compounds formed during both model and natural aging confirmed the effect of the glycosidase treatment on the acceleration of the aroma compounds formation and on the increase of the varietal differences of the wines.Significance and impact of study: GC-Olfactometry coupled with GCMS were good techniques to indentify and apreciate the odorants generated from glycoconjugates in the wines of Syrah and Grenache Noir, but in the context of a blend of odors, these techniques showed their limits and did not permit to determine the real impact of a molecule in the global aroma of the wine perceived by the taster. Other methods as additive techniques should be used to complete this study.

The range of polysaccharide-degrading enzymes and glycosidases formed by the phytopathogenic fungus Sclerotinia sclerotiorum was monitored following growth on 16 carbohydrate substrates. Endo- and exoenzymes capable of degrading cellulosic, hemicellulosic, and pectinolytic polysaccharides were secreted. Pectinolytic activities were produced constitutively on all of the substrates tested. Cellulolytic enzymes were not induced in simple sugar (i.e., glucose or xylose) media. Polysaccharide growth substrates and cellulase inducers increased all of the enzyme activities tested. Gel filtration analysis revealed the appearance of new molecular forms of pectinase, β-xylosidase, and cellobiosidase during induction on pectin and carboxymethyl cellulose media.

High-precision oxygen isotope analyses were carried out on dissolved phosphate extracted from discharge waters from three wastewater treatment plants (WTP) located in western France, as well as on the different phosphate-based fertilizers applied by farmers in the same region. Measured delta(18)O values of phosphate from chemical fertilizers range from 19.6 to 23.1 per thousand, while those of phosphate from WTP discharge waters are more tightly grouped between 17.7 and 18.1 per thousand. The variability in delta(18)O values of phosphate fertilizers is attributed to oxygen isotope variations of the phosphorite deposits from which France's fertilizers are manufactured. The significance of the delta(18)O values of phosphate from WTP discharge waters is less straightforward. At present, it is not clear whether these values are primary isotopic compositions corresponding, e.g., to the oxygen isotope composition of phosphate builders included in detergents (delta(18)O(P)=17.9 per thousand), or represent secondary values reflecting biological recycling of the phosphate in equilibrium with ambient WTP water The restricted difference in isotopic composition obtained between phosphate from fertilizers and phosphate from WTP discharge waters (2 per thousand), as well as the fairly large internal isotopic variability observed in both end-members (/=1.5 per thousand), cast doubt about the possibility that the oxygen isotope composition could serve as a tracer for the source of anthropogenic phosphates in waters.

We investigated the role of hexose transporters in a Saccharomyces cerevisiae strain derived from an industrial wine strain by carrying out a functional analysis of HXT genes 1–7 under enological conditions. A strain in which the sugar carrier genes HXT1–HXT7 were deleted was constructed and the HXT genes were expressed individually or in combination to evaluate their role under wine alcoholic fermentation conditions. No growth or fermentation was observed in winemaking conditions for the hxt1-7Δ strain. The low-affinity carriers Hxt1 and Hxt3 were the only carriers giving complete fermentation of sugars when expressed alone, indicating that these carriers play a predominant role in wine fermentation. However, these two carriers have different functions. The Hxt3 transporter is thought to play a major role, as it was the only carrier that gave an almost normal fermentation profile when produced alone. The hxt1 carrier was much less effective during the stationary phase and its role is thought to be restricted to the beginning of fermentation. The high-affinity carriers Hxt2, Hxt6 and/or Hxt7 were also required for normal fermentation. These high-affinity transporters have different functions: hxt2 is involved in growth initiation, whereas Hxt6 and/or Hxt7 are required at the end of alcoholic fermentation. This work shows that the successful alcoholic fermentation of wine involves at least four or five hexose carriers, playing different roles at various stages in the fermentation cycle. Copyright © 2002 John Wiley & Sons, Ltd.

Dissolved concentrations were determined for Fe, Mn, Al, Cu, Zn, La, U, Th, Cd and As in a wetland and its recipient stream to reveal the effect of seasonal changes in environmental conditions on the cycling and transfer of trace elements at the transition between terrestrial and aquatic ecosystems. These preliminary results from the wetland show marked seasonal changes in dissolved concentration for all elements except Zn and Cu. Concentrations are found to be low until about mid-February and then increase abruptly. The onset of trace element release appears to coincide with a marked decline in redox potential and increase of organic carbon content. Because this decline is itself correlated with a pronounced increase in temperature and dissolved Fe, Mn and organic carbon content, we suggest that the microorganisms which use soil iron and manganese oxy-hydroxides as electron acceptors catalyzed the change in redox conditions and induced an increase of DOC. Temporal changes were also observed in the recipient stream which showed marked positive concentration peaks during stormflow events (except Zn). The seasonal processes occurring in the wetland appear to play a major role in determining the amount of trace elements which are transferred from the wetland to the river.

Aims: The aim of this study was to demonstrate the impact of the volatile compounds, arising from glycosidic precursors contained in the berries of Vitis vinifera L.cv Syrah and Grenache noir varieties, on wine aromas from these varieties.Methods and results: The sensory analysis was used to compare Grenache noir and Syrah wines. The role played by the glycosidic precursors on the future odorant profile of the wines from both varieties was demonstrated in an experiment by increasing their natural content in glycosides. Then, odorant compounds were generated by aging treatments, heating at 45 °C for 3 weeks, preceded or not by enzyme addition, or natural aging for 18 months. The wines were then submitted to a selected and trained panel. Samples were compared using triangular test. Furthermore, a quantitative descriptive analysis was carried out to determine the aroma attributes describing and discriminating the wines from the two varieties.Conclusion: The glycoconjugates increased the global aromatic complexity, and enhanced the fruity aromas in Grenache wines and the leather or olive aromas in Syrah wines. The use of glycosidase enzymes led to a stewed fruit character in Grenache wines, whereas in Syrah wines, the samples enriched with glycosides differed according to the « terroir ».Significance and impact of study: This study showed the impact of the glycosidic fraction of the grapes on the varietal aroma of wines. Furthermore, comparisons of the results obtained by both aging techniques highlight the experimental interest of the aging model but also its limits.