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Applied and Environmental Microbiology, December 2001, p. 5668-5674, Vol. 67, No. 12
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.12.5668-5674.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Deletion of the GRE3 Aldose Reductase Gene and Its Influence on Xylose Metabolism in Recombinant Strains of Saccharomyces cerevisiae Expressing the xylA and XKS1 Genes

K. L. Träff,1,2 R. R. Otero Cordero,2,dagger W. H. van Zyl,2 and B. Hahn-Hägerdal1,*

Department of Applied Microbiology, Lund University, 221 00 Lund, Sweden,1 and Department of Microbiology, University of Stellenbosch, 7602 Matieland, South Africa2

Received 8 June 2001/Accepted 27 September 2001

Saccharomyces cerevisiae ferments hexoses efficiently but is unable to ferment xylose. When the bacterial enzyme xylose isomerase (XI) from Thermus thermophilus was produced in S. cerevisiae, xylose utilization and ethanol formation were demonstrated. In addition, xylitol and acetate were formed. An unspecific aldose reductase (AR) capable of reducing xylose to xylitol has been identified in S. cerevisiae. The GRE3 gene, encoding the AR enzyme, was deleted in S. cerevisiae CEN.PK2-1C, yielding YUSM1009a. XI from T. thermophilus was produced, and endogenous xylulokinase from S. cerevisiae was overproduced in S. cerevisiae CEN.PK2-1C and YUSM1009a. In recombinant strains from which the GRE3 gene was deleted, xylitol formation decreased twofold. Deletion of the GRE3 gene combined with expression of the xylA gene from T. thermophilus on a replicative plasmid generated recombinant xylose utilizing S. cerevisiae strain TMB3102, which produced ethanol from xylose with a yield of 0.28 mmol of C from ethanol/mmol of C from xylose. None of the recombinant strains grew on xylose.

* Corresponding author. Mailing address: Department of Applied Microbiology, Lund University, P.O. Box 124, 221 00 Lund, Sweden. Phone: 46 46 222 8428. Fax: 46 46 222 4203. E-mail: Barbel.Hahn-Hagerdal{at}tmb.lth.se.

dagger Present address: Institute for Wine Biotechnology, University of Stellenbosch, 7602 Matieland, South Africa.

Applied and Environmental Microbiology, December 2001, p. 5668-5674, Vol. 67, No. 12
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.12.5668-5674.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Brat, D., Boles, E., Wiedemann, B. (2009). Functional Expression of a Bacterial Xylose Isomerase in Saccharomyces cerevisiae. Appl. Environ. Microbiol. 75: 2304-2311 [Abstract] [Full Text]  
  • Jin, Y.-S., Laplaza, J. M., Jeffries, T. W. (2004). Saccharomyces cerevisiae Engineered for Xylose Metabolism Exhibits a Respiratory Response. Appl. Environ. Microbiol. 70: 6816-6825 [Abstract] [Full Text]  
  • Toivari, M. H., Salusjarvi, L., Ruohonen, L., Penttila, M. (2004). Endogenous Xylose Pathway in Saccharomyces cerevisiae. Appl. Environ. Microbiol. 70: 3681-3686 [Abstract] [Full Text]  


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