La microbiologie des vins issus des raisins botrytisés au cours de l'élevage. Caractérisation des souches de "Saccharomyces cerevisiae" responsables de refermentations.

La fermentation alcoolique des vins liquoreux français issus de raisin botrytisés est arrêtée brutalement par ajout massif de dioxyde de soufre après qu'un certain équilibre est atteint entre la teneur en alcool formé et la concentration en sucres résiduels. Certaines souches de levures fermentaires survivent et parfois se multiplient provoquant une nouvelle fermentation alcoolique indésirable ; c'est la refermentation. Le suivi microbiologique de nombreux lots de vin a permis de montrer que des levures sont dans un état physiologique similaire à celui décrit chez les bactéries sous l'appellation de viable non cultivable. Cet état explique l'apparente stérilité du vin après le mutage. Au sein de l'espèce Saccharomyces cerevisiae, une sélection naturelle se produit, ne laissant souvent la place qu'à une seule souche de refermentation, tolérante au dioxyde de soufre. Une étude écologique a montré que seules certaines espèces fermentaires et oxydatives survivent. Les plus tolérantes au dioxyde de soufre forment de l'éthanal au cours de l'élevage, malgré un métabolisme ralenti, et augmente la combinaison du dioxyde de soufre libre. Cet éthanal vient progressivement combiner le dioxyde de soufre libre. La sortie de l'état viable non cultivable est probablement la clef des mécanismes engendrant les refermentations. L'utilisation du diméthyldicarbonate au moment du mutage a été étudiée en couplage avec le dioxyde de soufre. Des souches de Saccharomyces cerevisiae de refermentation ont été isolées. Elles exhibent des singularités de séquence de leur ADNr, les apparentant aux souches de voile de certains vins spéciaux. Ces souches surexpriment constitutivement le gène SSU1 et synthétisent rapidement une forte concentration d'éthanal en réponse à la présence de dioxyde de soufre. La présence de fortes concentrations de dioxyde de soufre sélectionne les souches les plus résistantes. La refermentation est donc le résultat d'une adaptation génétique et d'une sélection, fruit d'une multitude de paramètres microbiologiques, physico-chimiques et humains. Botrytis-affected wines microbiology during maturation. Characterization of Saccharomyces cerevisiae strains responsible for refermentation. ABSTRACT : The alcoholic fermentation of Botrytis-affected wines is stopped by addition of sulphur dioxide. Some fermenting yeast species can survive during maturation and grow in spite of high ethanol, sugars and sulphur dioxide concentrations. An undesirable new fermentation, named "refermentation", can sometimes occur. In this study, it was proved that some yeast species were able to survive in a viable but non-culturable-like state. This state explains the apparent sterility of wines during maturation. Within Saccharomyces cerevisiae species, an intraspecific selection was spontaneously operated. After some weeks, only one strain could often survive. An ecological study was realized. Some highly fermentative and oxidative species could survive. In spite of slower metabolism, they synthesized acetaldehyde during maturation. The exit from the VBNC state and the high sulphur dioxide binding power were the keys of refermentations. The use of dimethyldicarbonate to stop alcoholic fermentation was studied. The most efficient action was observed for the mixed sulphur dioxide and DMDC addition. Some Saccharomyces cerevisiae strains responsible for refermentations were isolated. These strains exhibited rDNA sequence singularities, showing that they were close to flor strains, responsible for velum formation in some special wines. Moreover, those strains constitutively over-expressed SSU1 gene and could rapidly synthesize high concentrations of acetaldehyde in response to sulphur dioxide addition. High sulphur dioxide concentrations had probably selected the most resistant strains. Refermentation is the result of genetic adaptation and selection, under the influence of microbiological, physical, chemical and human parameters

The cellular symphony of redox cofactor management by yeasts in wine fermentation

The original publication is available at: https://www.sciencedirect.com/Redox metabolism is pivotal in anaerobic fermentative processes such as winemaking where it results in the production of many metabolites that contribute to the aroma and flavour of wine. Key to this system are NAD+ and NADP+, which play essential roles as cofactors in maintaining cellular redox balance and regulating metabolism during fermentation. This review comprehensively explores redox metabolism under winemaking conditions, highlighting the influence of factors such as oxygen availability and vitamins including B3 and B1. Recent findings underscore the rapid assimilation and recycling dynamics of these vitamins during fermentation, reinforcing their critical role in yeast performance. Despite extensive research, the roles of diverse yeast species and specific vitamins remain insufficiently explored. By consolidating current knowledge, this review emphasises the implications of redox dynamics for metabolite synthesis and overall wine quality. Understanding these metabolic intricacies offers options to enhance fermentation efficiency and refine aroma profiles. The review also identifies gaps in studies for intracellular vitamin metabolism and underlines the need for deeper insights into non-Saccharomyces yeast metabolism. Future research directions should focus on elucidating specific metabolic responses, exploring environmental influences, and harnessing the potential of diverse yeasts to innovate and diversify wine production strategies.https://www.sciencedirect.com/science/article/pii/S0168160524004100Publisher’s versio

Transcriptomics unravels the adaptive molecular mechanisms of Brettanomyces bruxellensis under SO2 stress in wine condition

CITATION: Valdetara, F. et al. 2020. Transcriptomics unravels the adaptive molecular mechanisms of Brettanomyces bruxellensis under SO2 stress in wine condition. Food Microbiology, 90. doi:10.1016/j.fm.2020.103483.The original publication is available at https://www.sciencedirect.com/journal/food-microbiologySulfur dioxide is generally used as an antimicrobial in wine to counteract the activity of spoilage yeasts, including Brettanomyces bruxellensis. However, this chemical does not exert the same effectiveness on different B. bruxellensis yeasts since some strains can proliferate in the final product leading to a negative sensory profile due to 4-ethylguaiacol and 4-ethylphenol. Thus, the capability of deciphering the general molecular mechanisms characterizing this yeast species’ response in presence of SO2 stress could be considered strategic for a better management of SO2 in winemaking. A RNA-Seq approach was used to investigate the gene expression of two strains of B. bruxellensis, AWRI 1499 and CBS 2499 having different genetic backgrounds, when exposed to a SO2 pulse. Results revealed that sulphites affected yeast culturability and metabolism, but not volatile phenol production suggesting that a phenotypical heterogeneity could be involved for the SO2 cell adaptation. The transcriptomics variation in response to SO2 stress confirmed the strain-related response in B. bruxellensis and the GO analysis of common differentially expressed genes showed that the detoxification process carried out by SSU1 gene can be considered as the principal specific adaptive response to counteract the SO2 presence. However, nonspecific mechanisms can be exploited by cells to assist the SO2 tolerance; namely, the metabolisms related to sugar alcohol (polyols) and oxidative stress, and structural compounds.https://www.sciencedirect.com/science/article/pii/S0740002020300721?via%3DihubPublishers versio

The production of reduced-alcohol wines using Gluzyme Mono® 10.000 BG-treated grape juice

The original publication is available at http://www.sasev.org/.High alcohol wines have become a major challenge in the international wine trade. Several physical processes are used to produce wines with reduced-alcohol content, all of which involve the selective extraction of ethanol based on volatility or diffusion. In this study, the possibility of Gluzyme Mono® 10.000 BG (Gluzyme) (Novozymes, South Africa) to reduce the glucose content of synthetic grape juice before fermentation was investigated in order to produce wine with reduced-alcohol content. Gluzyme is a glucose oxidase preparation from Aspergillus oryzae, currently used in the baking industry. Glucose oxidase catalyses the oxidation of glucose to gluconic acid and hydrogen peroxide(H2O2) in the presence of molecular oxygen. Gluzyme was initially used in synthetic grape juice, where different enzyme concentrations and factors influencing its efficiency were investigated under winemaking conditions. The results showed up to 0.5% v/v less alcohol at an enzyme concentration of 20 kU compared to the control samples. This reduction in alcohol was increased to 1 and 1.3% v/v alcohol at pH 3.5 and pH 5.5 respectively in aerated (8 mg/L O2) synthetic grape juice using 30 kU enzyme. Secondly, Gluzyme was used to treat Pinotage grape must before fermentation. Gluzyme-treated wines at 30 kU enzyme concentration after fermentation contained 0.68% v/v less alcohol than the control wines. A decrease in acetic acid concentration of the treated compared to control wines was also observed.Publishers' versio

Comparative characterization of endo-polygalacturonase (Pgu1) from Saccharomyces cerevisiae and Saccharomyces paradoxus under winemaking conditions

Wine strains of Saccharomyces cerevisiae have no to weak natural pectinase activity, despite their genetic ability to secrete an endo-polygalacturonase. The addition of external pectinase of fungal origin has therefore become a common step of winemaking in order to enhance the extraction of compounds located in the grape berry skins during maceration and to ease wine clarification after maturation. Recently, the strong pectinase activity of a wine strain of Saccharomyces paradoxus has been reported. In this study, the endo-polygalacturonase-encoding gene of S. paradoxus was sequenced and its activity was characterised, compared with that of S. cerevisiae and tested under winemaking conditions through overexpression of both genes individually in S. cerevisiae. A few differences in the amino acids sequences between the two proteins were revealed and the activity of the Pgu1 enzyme of S. paradoxus was shown to be weaker under winemaking conditions. Clear indicators of extracellular activity were observed in the wines made with both recombinant strains (i.e. enzyme activity in cell-free wine, higher methanol concentration and higher free-run wine), but the actual composition of the wines fermented with the mutants was only sparingly altered. Although unexpectedly found in lower concentrations in the latter wines, phenolic compounds were shown to be the most discriminatory components. Overexpressing the PGU1 gene of S. paradoxus or that of S. cerevisiae did not make much difference, showing that the higher activity of S. paradoxus strains under laboratory conditions could be due to a different regulation mechanism rather than to a different sequence of PGU1. © 2011 Springer-Verlag.Articl

La microbiologie des vins issus de raisins botrytisés au cours de l'élevage. Caractérisation des souches de Saccharomyces cerevisiae responsables de refermentation

La fermentation alcoolique des vins liquoreux issus de raisin botrytisés est arrêtee brutalement par ajout massif de dioxyde de soufre. Certaines sousches de levures survivent et parfois provoquent une refermentation. Ces levures survivent dans un état comparable à celui qualifié de viable non cultivable chez les bactéries. Seules certaines espèces fermentaires et oxydatives survivent. L'utilisation potentielle du diméthyldicarbonate en complément du sioxyde de soufre a été étudiée. Les souches de saccharomyces cerevisiae de refermentation isolées exhibent des singularités de séquence de leur ADNr, les apparentant aux souches de voile de certains vins spéciaux. Ces souches surexpriment constitutivement le gène ssu1 et synthétisent rapidement une forte concentration d'éthanal en réponse à la présence de dioxyde de soufre. La refermentation est le résultat d'une adaptation génétique et d'une sélection, fruit d'une multitude de paramètres microbiologiques physico-chimiques et humains.TOULOUSE-ENSIACET (315552325) / SudocSudocFranceF

Quantitative PCR: An appropriate tool to detect viable but not culturable <I>Brettanomyces bruxellensis </I>in wine

AgriwetenskappeInstituut Vir WynbiotegnologiePlease help us populate SUNScholar with the post print version of this article. It can be e-mailed to: [email protected]

Exploring the phenotypic diversity of oenological traits in<i>Kluyveromyces marxianus</i>strains

AbstractThe use of non-Saccharomyces yeasts in the winemaking process may have several positive outcomes. Kluyveromyces marxianus has recently been revealed as a promising species for this industry. While the majority of studies mention the use of K. marxianus in various industries including food production (e.g. dairy and cocoa), recent studies have also shown that its aroma and pectinase production make it a suitable yeast for the wine industry. Nevertheless, only particular strain, IWBT Y885, was investigated. In this study, five different K. marxianus strains as well as one protoplast fusant (BF2020) were compared to strain Y885. These comparisons focused on various oenological traits such as fermentation performance, fermentation metabolites, hydrogen sulfide, and pectinase production. Throughout the study, variations were found between the K. marxianus strains investigated. Indeed, although common traits such as high pectinase activity appeared conserved among K. marxianus strains, a fairly large phenotypic diversity was also evident. Using cluster analysis, strain groupings emerged with strains L01, L05, Y885, and BF2020 grouping together. Similarly, strains L02 and L04 grouped together while strain L03 appearing to show the most variation between the strains investigated. Variation between strains was observed regardless of the original source of isolation.</jats:p

Metabolic engineering of wine yeast and advances in yeast selection methods for improved wine quality and safety.

Please help us populate SUNScholar with the post print version of this article. It can be e-mailed to: [email protected] Vir Wynbiotegnologi

PGU1 gene natural deletion is responsible for the absence of endo-polygalacturonase activity in some wine strains of Saccharomyces cerevisiae

The PGU1 gene encodes an endo-polygalacturonase enzyme in Saccharomyces cerevisiae. The literature reports that most S. cerevisiae strains possess this gene, despite a wide range of enzyme activity levels. Nevertheless, a few wine strains lack the PGU1 gene. We investigated the PGU1 locus sequence in these strains. The results indicated that the gene had been replaced by a partial Ty mobile element, whereas the gene promoter was still at the expected location. As all the strains lacking the PGU1 gene experienced the same phenomenon, it was tempting to hypothesize a common phylogenetic origin. However, fingerprints only allowed grouping of a few of them within one cluster. © 2007 Federation of European Microbiological Societies.Articl