The thermotolerant yeast Kluyveromyces marxianus displays a potential to be used for ethanol production from both whey and lignocellulosic biomass at elevated temperatures, which is highly alluring to reduce the cost of the bioprocess. Nevertheless, contrary to Saccharomyces cerevisiae, K. marxianus cannot tolerate high ethanol concentrations. We report the transcriptional profile alterations in K. marxianus under ethanol stress in order to gain insights about mechanisms involved with ethanol response. Time-dependent changes have been characterized under the exposure of 6% ethanol and compared with the unstressed cells prior to the ethanol addition. Our results reveal that the metabolic flow through the central metabolic pathways is impaired under the applied ethanol stress. Consistent with these results, we also observe that genes involved with ribosome biogenesis are downregulated and gene-encoding heat shock proteins are upregulated. Remarkably, the expression of some gene-encoding enzymes related to unsaturated fatty acid and ergosterol biosynthesis decreases upon ethanol exposure, and free fatty acid and ergosterol measurements demonstrate that their content in K. marxianus does not change under this stress. These results are in contrast to the increase previously reported with S. cerevisiae subjected to ethanol stress and suggest that the restructuration of K. marxianus membrane composition differs in the two yeasts which gives important clues to understand the low ethanol tolerance of K. marxianus compared to S. cerevisiae.

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Fermentation is one of the oldest forms of food preservation in the world. In South "merica, most fermented beverages are nondairy products featuring several other food raw materials such as cereals, fruits, and vegetables. Generally, natural fermentations are carried out by yeast and lactic acid bacteria forming a complex microbiota that acts in cooperation. Yeast have a prominent role in the production of beverages, due to the ability to accumulate high levels of ethanol and to produce highly desirable aroma compounds, but lactic acid bacteria are particularly important in fermentation because they produce desirable acids, flavor compounds, and peptides that inhibit the growth of undesirable organisms. "mong the South "merica beverages based on cereals and vegetables, the fermented beverages chicha, caxiri, cauim and champús, and cachaça, a fermented and distilled beverage, could be cited. Genetic and physiological analyses of Saccharomyces cerevisiae strains isolated from cachaça have been shown to present interesting traits for beer production, such as flocculation and production of aroma compounds, fundamental to high-quality beer. The study of these traditional beverages allows the identification of new microorganism strains displaying enhanced resistance or new flavor and aroma profiles that could lead to applications in several industries and ultimately new products.

This study identified phenotypic traits appropriate for biotechnological applications of 118 yeasts isolated from cachaça distilleries. Different properties were verified: capacity to use alternative carbon sources; ability to tolerate high concentrations of sucrose, ethanol, methanol, aluminum and zinc as well as different pH values and foam production. Pichia guilliermondii and Pichia anomala strains were identified as the most promising ones for application in the second-generation biofuel industry, showing ability to grow on high glycerol concentrations. Other isolates, identified as Saccharomyces cerevisiae, produced bioethanol comparable to the industrial strains, and were therefore ideal for use in the first-generation ethanol industry. Some of these strains also showed high resistance to aluminum, as observed in sugarcane juice, and to inter-cycle washings with diluted sulphuric acid, as performed in the industrial bioethanol production process. In summary, yeast isolates from cachaça distilleries displayed robustness and phenotypic plasticity, which makes them interesting for biotechnological applications.

The simultaneous saccharification and fermentation (SSF) process is a promising strategy to obtain ethanol from cellulosic biomass. In this study, sugarcane bagasse was supplemented with ricotta whey to increase the sugar, vitamin, and trace metal concentrations in the fermentation medium. The optimum conditions for SSF ethanol production from a mixture of sugarcane bagasse and ricotta whey produced by Kluyveromyces marxianus CCT 7735 were evaluated considering five factors: cellulase concentration, cellulosic biomass concentration, pH, temperature, and agitation. The highest ethanol yield was 49.65 g/L with a cellulosic biomass of 80 g/L, pH value of 5.05, agitation at 65 rpm and temperature of 39.2ºC. The results demonstrated that a mixture of the cellulosic residue of sugarcane bagasse and ricotta whey is promising for ethanol production because the ethanol yield in the mixture was higher than that in single substrate of sugarcane bagasse.

This work was performed to verify the potential of yeast strains isolated from cachaça distilleries for two specific biotechnological applications: beer and bioethanol production. In the beer production, the strains were tested for characteristics required in brewery practices, such as: capacity to ferment maltose and maltotriose, ability to grow at lowest temperatures, low HS production, and flocculation profile. Among the strains tested, two of them showed appropriate characteristics to produce two different beer styles: lager and ale. Moreover, both strains were tested for cachaça production and the results confirmed the capacity of these strains to improve the quality of cachaça. In the bioethanol production, the fermentation process was performed similarly to that used by bioethanol industries: recycling of yeast biomass in the fermentative process with sulfuric acid washings (pH 2.0). The production of ethanol, glycerol, organic acids, dry cell weight, carbohydrate consumption, and cellular viability were analyzed. One strain presented fermentative parameters similar to PE2, industrial/commercial strain, with equivalent ethanol yields and cellular viability during all fermentative cycles. This work demonstrates that cachaça distilleries seem to be an interesting environment to select new yeast strains to be used in biotechnology applications as beer and bioethanol production.

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This study suggests that the use of selected yeast strains could contribute to obtain a cachaça similar to those produced traditionally, while getting wide acceptation in the market, yet presenting more homogeneous organoleptic characteristics, and thus contributing to the PDO implementation.