Project description:Our study involves a transcriptomic approach to the analysis of industrial yeast metabolism. Historically, among the hundreds of yeast species, Saccharomyces cerevisiae has played an important role in scientific investigations and industrial applications, and it is universally acknowledged as one of the model systems for eukaryotic organisms. Yeast is also an important component of the wine fermentation process and determines various attributes of the final product. Our research takes a holistic approach to the improvement of industrial yeast strains by integrating large data sets from various yeast strains during fermentation. This means that analysis can be done in such a way as to co-evaluate several parameters simultaneously to identify points of interest and target genes for metabolic engineering. Eventually we hope to construct an accurate information matrix and a more complete cellular map for the fermenting yeast. This will enable accurate model-building for industrial yeast and facilitated the design of intelligent yeast improvement strategies which can be applied via traditional avenues of molecular biology. Experiment Overall Design: Five different Saccharomyces cerevisiae strains used in industrial winemaking processes were used in synthetic must (MS300) fermentations. All fermentations were carried out in triplicate, so each sample is represented by three completely independent biological repeats. Samples for microarray analysis were taken at three different time points during fermentation, representative of the exponential (day2), early stationary (day5) and late stationary (day14) growth stages.
Project description:Comparative gene expression analysis of two wine yeast strains at three time points (days 2, 5 and 14) during fermentation of colombar must. In our study we conducted parallel fermentations with the VIN13 and BM45 wine yeast strains in two different media, namely MS300 (syntheticmust) and Colombar must. The intersection of transcriptome datasets from both MS300 (simulated wine must;GSE11651) and Colombar fermentations should help to delineate relevant and ânoisyâ changes in gene expression in response to experimental factors such as fermentation stage and strain identity. Experiment Overall Design: Two industrial wine yeast strains (BM45 and VIN13) grown micro-aerobically in Colombar must. Microarray analysis was performed at three time points during fermentation (days 2, 5 and 14), representing the exponential, early and late stationary growth phases respectively.
Project description:Second fermentation in a bottle supposes such specific conditions that undergo yeasts to a set of stress situations like high ethanol, low nitrogen, low pH or sub-optimal temperature. Also, yeast have to grow until 1 or 2 generations and ferment all sugar available while they resist increasing CO2 pressure produced along with fermentation. Because of this, yeast for second fermentation must be selected depending on different technological criteria such as resistance to ethanol, pressure, high flocculation capacity, and good autolytic and foaming properties. All of these stress factors appear sequentially or simultaneously, and their superposition could amplify their inhibitory effects over yeast growth. Considering all of the above, it has supposed interesting to characterize the adaptive response of commercial yeast strain EC1118 during second-fermentation experiments under oenological/industrial conditions by transcriptomic profiling. We have pointed ethanol as the most relevant environmental condition in the induction of genes involved in respiratory metabolism, oxidative stress, autophagy, vacuolar and peroxisomal function, after comparison between time-course transcriptomic analysis in alcoholic fermentation and transcriptomic profiling in second fermentation. Other examples of parallelism include overexpression of cellular homeostasis and sugar metabolism genes. Finally, this study brings out the role of low-temperature on yeast physiology during second-fermentation. S. cerevisiae EC1118 pre-adapted to ethanol cells and sucrose (20 g/L) were added to 20 L of base wine (Cavas Freixenet, Sant Sadurní D’Anoia, Spain). Complete volume was bottled with 350 mL each one. All were sealed and incubated in static conditions at 16ºC for approximately 40 days after tirage. Three samples were taken during the process for transcriptional study of the physiological adaptation of yeast cells to industrial second fermentation conditions. A sample corresponding to exponential-growth phase under unstressed conditions (in YPD at 28ºC) was used as an external reference. Three timepoints from second-fermentation were monitored and three biological replicates from each timepoint were analyzed.
Project description:We used genome-wide expression analyses to study the response of Saccharomyces cerevisiae to stress throughout a 15-day wine fermentation. Forty percent of the yeast genome significantly changed expression levels to mediate long-term adaptation to an environment in which ethanol is both a stressor and a carbon source. Within this set, we identify a group of 223 genes, designated as the Fermentation Stress Response (FSR), that are dramatically and permanently induced; FSR genes exhibited changes ranging from four-to eighty-fold. The FSR is novel; 62% of the genes involved have not been implicated in global stress responses and 28% of the genes have no functional annotation. Genes involved in respiratory metabolism and gluconeogenesis were expressed during fermentation despite the presence of high concentrations of glucose. Ethanol, rather than nutrient depletion, was responsible for entry of yeast cells into stationary phase. Ethanol seems to regulate yeast metabolism through hitherto undiscovered regulatory networks during wine fermentation. Keywords: time course, stress response, fermentation
Project description:Gene expression analysis of a time course experiment of a synthetic must (nitrogen-poor) fermentation by a natural wine yeast. Three replicates of five time points taken at 24, 48, 80, 96 and 144 hours after yeast inoculation
Project description:Gene expression analysis of a time course experiment of a synthetic must (nitrogen-poor) fermentation by a natural wine yeast, supplemented at 72 hours with 200 mg/l of nitrogen Three replicates of five time points taken at 24, 48, 80, 96 and 144 hours after yeast inoculation. Time points 24 and 48 hours are common to Sluggish fermentation. Time points at 80, 96 and 144 hours are exclusive of this experiment.
Project description:Transcriptomic analyses of fermenting yeast are increasingly being carried out under small scale simulated winemaking conditions. It is not known to what degree data generated from such experiments are a reflection of transcriptional processes in large-scale commercial fermentation tanks. In this experiment we set out to determine the effect of scale, or fermentation volume, on the transcriptional respone of wine yeast strains. Parallel fermentations were carried out in laboratory fermentation vials and commercial fermentation tanks using the same wine media and inoculated yeast strain. Comparative transcriptomic analyses were carried out at three time points during alcoholic fermentation.
Project description:Comparative gene expression analysis of two wine yeast strains at three time points (days 2, 5 and 14) during fermentation of colombar must. In our study we conducted parallel fermentations with the VIN13 and BM45 wine yeast strains in two different media, namely MS300 (syntheticmust) and Colombar must. The intersection of transcriptome datasets from both MS300 (simulated wine must;GSE11651) and Colombar fermentations should help to delineate relevant and ‘noisy’ changes in gene expression in response to experimental factors such as fermentation stage and strain identity.
Project description:In wine fermentation, the blending of non-Saccharomyces yeast with Saccharomyces cerevisiae to improve the complexity of wine has become common practice, but data regarding the impact on yeast physiology and on genetic and metabolic regulation remain limited. Here we describe a transcriptomic analysis of single species and mixed species fermentations.
Project description:The main objectives of this study were to expand our understanding of NSF1 gene function in industrial S. cerevisiae M2 strain during fermentation by finding the largest maximal clique of co-expressed genes (i.e. Interdependent Correlation Cluster), and to establish the impact of Nsf1p on genome-wide gene expression during the fermentation process with possible implications related to wine quality and S. cerevisiae adapation to stressful fermentation conditions The Affymetrix Yeast 2.0 microarrays were used to capture the global gene expression profile of M2 and M2 nsf1∆ grown under fermentation conditions in Riesling grape must at 18°C with no shaking at various time points. The analysis of this microarray dataset expanded our understanding of the mechanism of action and the roles of NSF1 under fermentation stress conditions.