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Background  The understanding of the molecular basis of yeast tolerance to ethanol may guide the design of rational strategies to increase process performance in industrial alcoholic fermentations. A set of 21 genes encoding multidrug transporters from the ATP-Binding Cassette (ABC) Superfamily and Major Facilitator Superfamily (MFS) in  S. cerevisiae  were scrutinized for a role in ethanol stress resistance.    Results  A yeast multidrug resistance ABC transporter encoded by the  PDR18  gene, proposed to play a role in the incorporation of ergosterol in the yeast plasma membrane, was found to confer resistance to growth inhibitory concentrations of ethanol.  PDR18  expression was seen to contribute to decreased  3  H-ethanol intracellular concentrations and decreased plasma membrane permeabilization of yeast cells challenged with inhibitory ethanol concentrations. Given the increased tolerance to ethanol of cells expressing  PDR18 , the final concentration of ethanol produced during high gravity alcoholic fermentation by yeast cells devoid of  PDR18  was lower than the final ethanol concentration produced by the corresponding parental strain. Moreover, an engineered yeast strain in which the  PDR18  promoter was replaced in the genome by the stronger  PDR5  promoter, leading to increased  PDR18  mRNA levels during alcoholic fermentation, was able to attain a 6 % higher ethanol concentration and a 17 % higher ethanol production yield than the parental strain. The improved fermentative performance of yeast cells over-expressing  PDR18  was found to correlate with their increased ethanol tolerance and ability to restrain plasma membrane permeabilization induced throughout high gravity fermentation.    Conclusions   PDR18  gene over-expression increases yeast ethanol tolerance and fermentation performance leading to the production of highly inhibitory concentrations of ethanol.  PDR18  overexpression in industrial yeast strains appears to be a promising approach to improve alcoholic fermentation performance for sustainable bio-ethanol production.

microbial cell factories

Increased expression of the yeast multidrug resistance ABC transporter Pdr18 leads to increased ethanol tolerance and ethanol production in high gravity alcoholic fermentation