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Applied and Environmental Microbiology, March 2003, p. 1499-1503, Vol. 69, No. 3
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.3.1499-1503.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Ethanol Tolerance in the Yeast Saccharomyces cerevisiae Is Dependent on Cellular Oleic Acid Content

Kyung Man You,{dagger} Claire-Lise Rosenfield, and Douglas C. Knipple*

Department of Entomology, Cornell University, New York State Agricultural Experiment Station, Geneva, New York 14456

Received 26 July 2002/ Accepted 19 November 2002

In this investigation, we examined the effects of different unsaturated fatty acid compositions of Saccharomyces cerevisiae on the growth-inhibiting effects of ethanol. The unsaturated fatty acid (UFA) composition of S. cerevisiae is relatively simple, consisting almost exclusively of the mono-UFAs palmitoleic acid ({Delta}9Z-C16:1) and oleic acid ({Delta}9Z-C18:1), with the former predominating. Both UFAs are formed in S. cerevisiae by the oxygen- and NADH-dependent desaturation of palmitic acid (C16:0) and stearic acid (C18:0), respectively, catalyzed by a single integral membrane desaturase encoded by the OLE1 gene. We systematically altered the UFA composition of yeast cells in a uniform genetic background (i) by genetic complementation of a desaturase-deficient ole1 knockout strain with cDNA expression constructs encoding insect desaturases with distinct regioselectivities (i.e., {Delta}9 and {Delta}11) and substrate chain-length preferences (i.e., C16:0 and C18:0); and, (ii) by supplementation of the same strain with synthetic mono-UFAs. Both experimental approaches demonstrated that oleic acid is the most efficacious UFA in overcoming the toxic effects of ethanol in growing yeast cells. Furthermore, the only other UFA tested that conferred a nominal degree of ethanol tolerance is cis-vaccenic acid ({Delta}11Z-C18:1), whereas neither {Delta}11Z-C16:1 nor palmitoleic acid ({Delta}9Z-C16:1) conferred any ethanol tolerance. We also showed that the most ethanol-tolerant transformant, which expresses the insect desaturase TniNPVE, produces twice as much oleic acid as palmitoleic acid in the absence of ethanol and undergoes a fourfold increase in the ratio of oleic acid to palmitoleic acid in response to exposure to 5% ethanol. These findings are consistent with the hypothesis that ethanol tolerance in yeast results from incorporation of oleic acid into lipid membranes, effecting a compensatory decrease in membrane fluidity that counteracts the fluidizing effects of ethanol.

* Corresponding author. Mailing address: Department of Entomology, Cornell University, New York State Agricultural Experiment Station, P.O. Box 462, Geneva, NY 14456. Phone: (315) 787-2363. Fax: (315) 787-2326. E-mail: dck2{at}cornell.edu.

{dagger} Present address: Department of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang 790-784, South Korea.

Applied and Environmental Microbiology, March 2003, p. 1499-1503, Vol. 69, No. 3
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.3.1499-1503.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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