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Applied and Environmental Microbiology, April 2002, p. 1604-1609, Vol. 68, No. 4
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.4.1604-1609.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Reduced Oxidative Pentose Phosphate Pathway Flux in Recombinant Xylose-Utilizing Saccharomyces cerevisiae Strains Improves the Ethanol Yield from Xylose

Marie Jeppsson, Björn Johansson, Bärbel Hahn-Hägerdal, and Marie F. Gorwa-Grauslund*

Department of Applied Microbiology, Lund University, 221 00 Lund, Sweden

Received 18 September 2001/ Accepted 7 January 2002

In recombinant, xylose-fermenting Saccharomyces cerevisiae, about 30% of the consumed xylose is converted to xylitol. Xylitol production results from a cofactor imbalance, since xylose reductase uses both NADPH and NADH, while xylitol dehydrogenase uses only NAD+. In this study we increased the ethanol yield and decreased the xylitol yield by lowering the flux through the NADPH-producing pentose phosphate pathway. The pentose phosphate pathway was blocked either by disruption of the GND1 gene, one of the isogenes of 6-phosphogluconate dehydrogenase, or by disruption of the ZWF1 gene, which encodes glucose 6-phosphate dehydrogenase. Decreasing the phosphoglucose isomerase activity by 90% also lowered the pentose phosphate pathway flux. These modifications all resulted in lower xylitol yield and higher ethanol yield than in the control strains. TMB3255, carrying a disruption of ZWF1, gave the highest ethanol yield (0.41 g g-1) and the lowest xylitol yield (0.05 g g-1) reported for a xylose-fermenting recombinant S. cerevisiae strain, but also an 84% lower xylose consumption rate. The low xylose fermentation rate is probably due to limited NADPH-mediated xylose reduction. Metabolic flux modeling of TMB3255 confirmed that the NADPH-producing pentose phosphate pathway was blocked and that xylose reduction was mediated only by NADH, leading to a lower rate of xylose consumption. These results indicate that xylitol production is strongly connected to the flux through the oxidative part of the pentose phosphate pathway.

* Corresponding author. Mailing address: Department of Applied Microbiology, Lund University, P.O. Box 124, 221 00 Lund, Sweden. Phone: 46 46 222 0619. Fax: 46 46 222 4203. E-mail: Marie-Francoise.Gorwa{at}tmb.lth.se.

Applied and Environmental Microbiology, April 2002, p. 1604-1609, Vol. 68, No. 4
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.4.1604-1609.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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