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Applied and Environmental Microbiology, December 2002, p. 5981-5989, Vol. 68, No. 12
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.12.5981-5989.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Aquaporin Expression Correlates with Freeze Tolerance in Baker's Yeast, and Overexpression Improves Freeze Tolerance in Industrial Strains

An Tanghe,1 Patrick Van Dijck,1,2 Françoise Dumortier,1 Aloys Teunissen,1,{dagger} Stefan Hohmann,3 and Johan M. Thevelein1*

Laboratorium voor Moleculaire Celbiologie,1 Vlaams Interuniversitair Instituut voor Biotechnologie (VIB), Institute of Botany and Microbiology, Katholieke Universiteit Leuven, B-3001 Leuven-Heverlee, Flanders, Belgium,2 Department of Cell and Molecular Biology/Microbiology, Lundberg Laboratory, Göteborg University, S-405 30 Göteborg, Sweden3

Received 16 April 2002/ Accepted 23 August 2002

Little information is available about the precise mechanisms and determinants of freeze resistance in baker's yeast, Saccharomyces cerevisiae. Genomewide gene expression analysis and Northern analysis of different freeze-resistant and freeze-sensitive strains have now revealed a correlation between freeze resistance and the aquaporin genes AQY1 and AQY2. Deletion of these genes in a laboratory strain rendered yeast cells more sensitive to freezing, while overexpression of the respective genes, as well as heterologous expression of the human aquaporin gene hAQP1, improved freeze tolerance. These findings support a role for plasma membrane water transport activity in determination of freeze tolerance in yeast. This appears to be the first clear physiological function identified for microbial aquaporins. We suggest that a rapid, osmotically driven efflux of water during the freezing process reduces intracellular ice crystal formation and resulting cell damage. Aquaporin overexpression also improved maintenance of the viability of industrial yeast strains, both in cell suspensions and in small doughs stored frozen or submitted to freeze-thaw cycles. Furthermore, an aquaporin overexpression transformant could be selected based on its improved freeze-thaw resistance without the need for a selectable marker gene. Since aquaporin overexpression does not seem to affect the growth and fermentation characteristics of yeast, these results open new perspectives for the successful development of freeze-resistant baker's yeast strains for use in frozen dough applications.

* Corresponding author. Mailing address: Laboratorium voor Moleculaire Celbiologie, Institute of Botany and Microbiology, Katholieke Universiteit Leuven, Kasteelpark Arenberg 31, B-3001 Leuven-Heverlee, Flanders, Belgium. Phone: 32-16-32 15 07. Fax: 32-16-32 19 79. E-mail: Johan.Thevelein{at}bio.kuleuven.ac.be

{dagger} Present address: Department of Pharmacochemistry, Vrije Universiteit Amsterdam, 1081HV Amsterdam, The Netherlands.

Applied and Environmental Microbiology, December 2002, p. 5981-5989, Vol. 68, No. 12
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.12.5981-5989.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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