Lipid Composition Analysis Reveals Mechanisms of Ethanol Tolerance in the Model Yeast Saccharomyces cerevisiae.

Saccharomyces cerevisiae is an important unicellular yeast species within the biotechnological and the food and beverage industries. A significant application of this species is the production of ethanol, where concentrations are limited by cellular toxicity, often at the level of the cell membrane. Here, we characterize 61 S. cerevisiae strains for ethanol tolerance and further analyze five representatives with various ethanol tolerances. The most tolerant strain, AJ4, was dominant in coculture at 0 and 10% ethanol. Unexpectedly, although it does not have the highest noninhibitory concentration or MIC, MY29 was the dominant strain in coculture at 6% ethanol, which may be linked to differences in its basal lipidome. Although relatively few lipidomic differences were observed between strains, a significantly higher phosphatidylethanolamine concentration was observed in the least tolerant strain, MY26, at 0 and 6% ethanol compared to the other strains that became more similar at 10%, indicating potential involvement of this lipid with ethanol sensitivity. Our findings reveal that AJ4 is best able to adapt its membrane to become more fluid in the presence of ethanol and that lipid extracts from AJ4 also form the most permeable membranes. Furthermore, MY26 is least able to modulate fluidity in response to ethanol, and membranes formed from extracted lipids are least leaky at physiological ethanol concentrations. Overall, these results reveal a potential mechanism of ethanol tolerance and suggest a limited set of membrane compositions that diverse yeast species use to achieve this. IMPORTANCE Many microbial processes are not implemented at the industrial level because the product yield is poorer and more expensive than can be achieved by chemical synthesis. It is well established that microbes show stress responses during bioprocessing, and one reason for poor product output from cell factories is production conditions that are ultimately toxic to the cells. During fermentative processes, yeast cells encounter culture media with a high sugar content, which is later transformed into high ethanol concentrations. Thus, ethanol toxicity is one of the major stresses in traditional and more recent biotechnological processes. We have performed a multilayer phenotypic and lipidomic characterization of a large number of industrial and environmental strains of Saccharomyces to identify key resistant and nonresistant isolates for future applications.

Effect of transient thermal shocks on alcoholic fermentation performance.

Stuck and sluggish fermentations are among the main problems in winemaking industry leading to important economic losses. Several factors have been described as causes of stuck and sluggish fermentations, being exposure to extreme temperatures barely studied. The objective of this study was to identify thermal conditions leading to stuck and sluggish fermentations, focusing on the impact of an abrupt and transient decrease/increase of temperature on fermentation performance and yeast viability/vitality. Different strains of Saccharomyces cerevisiae, SBB11, T73, and PDM were evaluated in synthetic grape must fermentations. Cold shocks (9 °C and 1.5 °C for 16 h) carried out on different days during the fermentation process were unable to alter fermentation performance. Conversely, shock temperatures higher than 32 °C, applied in early stages of the process, lead to sluggish fermentation showing a delay directly related to the temperature increase. Fermentation delay was associated with a decrease in cell vitality. The impact of the heat shock on fermentation performance was different depending on the strain evaluated and nitrogen supplementation (with or without diammonium phosphate addition). None of the conditions evaluated produced a stuck fermentation and importantly, in all cases must nutrition improved fermentation performance after a heat shock.