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Applied and Environmental Microbiology, October 2005, p. 6185-6192, Vol. 71, No. 10
0099-2240/05/$08.00+0 doi:10.1128/AEM.71.10.6185-6192.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.
Engineering of a Novel Saccharomyces cerevisiae Wine Strain with a Respiratory Phenotype at High External Glucose Concentrations
C. Henricsson,1 M. C. de Jesus Ferreira,2 K. Hedfalk,1 K. Elbing,1,
C. Larsson,1
R. M. Bill,3
J. Norbeck,1
S. Hohmann,4 and
L. Gustafsson1*
Department of Chemistry and Bioscience—Molecular Biotechnology, Chalmers University of Technology, Box 462, SE-405 30 Göteborg, Sweden,1 Université de Montpellier II, Laboratoire de Stress et Plasticité Cérébrale, FRE2693, bat24 C090, place Eugene Bataillon, 34095 Montpellier, France,2 The School of Life and Health Sciences, Aston University, Aston Triangle, Birmingham B4 7ET, United Kingdom,3 Department of Cell and Molecular Biology—Microbiology, Göteborg University, Box 462, SE-405 30 Göteborg, Sweden4
Received 17 February 2005/ Accepted 3 June 2005
The recently described respiratory strain Saccharomyces cerevisiae KOY.TM6*P is, to our knowledge, the only reported strain of S. cerevisiae which completely redirects the flux of glucose from ethanol fermentation to respiration, even at high external glucose concentrations (27). In the KOY.TM6*P strain, portions of the genes encoding the predominant hexose transporter proteins, Hxt1 and Hxt7, were fused within the regions encoding transmembrane (TM) domain 6. The resulting chimeric gene, TM6*, encoded a chimera composed of the amino-terminal half of Hxt1 and the carboxy-terminal half of Hxt7. It was subsequently integrated into the genome of an hxt null strain. In this study, we have demonstrated the transferability of this respiratory phenotype to the V5 hxt1-7
strain, a derivative of a strain used in enology. We also show by using this mutant that it is not necessary to transform a complete hxt null strain with the TM6* construct to obtain a non-ethanol-producing phenotype. The resulting V5.TM6*P strain, obtained by transformation of the V5 hxt1-7 strain with the TM6* chimeric gene, produced only minor amounts of ethanol when cultured on external glucose concentrations as high as 5%. Despite the fact that glucose flux was reduced to 30% in the V5.TM6*P strain compared with that of its parental strain, the V5.TM6*P strain produced biomass at a specific rate as high as 85% that of the V5 wild-type strain. Even more relevant for the potential use of such a strain for the production of heterologous proteins and also of low-alcohol beverages is the observation that the biomass yield increased 50% with the mutant compared to its parental strain.
* Corresponding author. Mailing address: Department of Chemistry and Bioscience—Molecular Biotechnology, Chalmers University of Technology, Box 462, SE-405 30 Göteborg, Sweden. Phone: 46 31 773 25 97. Fax: 46 31 773 25 99. E-mail: lena.gustafsson{at}chembio.chalmers.se.
Present address: W1212 Biomolecular Science Tower, Department of Molecular Genetics & Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261.
Applied and Environmental Microbiology, October 2005, p. 6185-6192, Vol. 71, No. 10
0099-2240/05/$08.00+0 doi:10.1128/AEM.71.10.6185-6192.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.
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