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Applied and Environmental Microbiology, September 2009, p. 5761-5772, Vol. 75, No. 18
0099-2240/09/$08.00+0     doi:10.1128/AEM.00845-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Genome-Wide Identification of Saccharomyces cerevisiae Genes Required for Maximal Tolerance to Ethanol{triangledown} ,{dagger}

Miguel C. Teixeira, Luís R. Raposo, Nuno P. Mira, Artur B. Lourenço, and Isabel Sá-Correia*

Institute for Biotechnology and BioEngineering, Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001 Lisbon, Portugal

Received 14 April 2009/ Accepted 20 July 2009

The understanding of the molecular basis of yeast resistance to ethanol may guide the design of rational strategies to increase process performance in industrial alcoholic fermentations. In this study, the yeast disruptome was screened for mutants with differential susceptibility to stress induced by high ethanol concentrations in minimal growth medium. Over 250 determinants of resistance to ethanol were identified. The most significant gene ontology terms enriched in this data set are those associated with intracellular organization, biogenesis, and transport, in particular, regarding the vacuole, the peroxisome, the endosome, and the cytoskeleton, and those associated with the transcriptional machinery. Clustering the proteins encoded by the identified determinants of ethanol resistance by their known physical and genetic interactions highlighted the importance of the vacuolar protein sorting machinery, the vacuolar H+-ATPase complex, and the peroxisome protein import machinery. Evidence showing that vacuolar acidification and increased resistance to the cell wall lytic enzyme β-glucanase occur in response to ethanol-induced stress was obtained. Based on the genome-wide results, the particular role of the FPS1 gene, encoding a plasma membrane aquaglyceroporin which mediates controlled glycerol efflux, in ethanol stress resistance was further investigated. FPS1 expression contributes to decreased [3H]ethanol accumulation in yeast cells, suggesting that Fps1p may also play a role in maintaining the intracellular ethanol level during active fermentation. The increased expression of FPS1 confirmed the important role of this gene in alcoholic fermentation, leading to increased final ethanol concentration under conditions that lead to high ethanol production.

* Corresponding author. Mailing address: Institute for Biotechnology and BioEngineering, Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001 Lisbon, Portugal. Phone: 351-218417682. Fax: 351-218419199. E-mail: isacorreia{at}ist.utl.pt

{triangledown} Published ahead of print on 24 July 2009.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, September 2009, p. 5761-5772, Vol. 75, No. 18
0099-2240/09/$08.00+0     doi:10.1128/AEM.00845-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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