Paper: Development and characterization of xylose-fermenting strains of Saccharomyces cerevisiae based on structure-based engineering of key metabolic enzymes (31st Symposium on Biotechnology for Fuels and Chemicals (May 3-6, 2009))

Monday, May 4, 2009: 3:30 PM

Grand Ballroom C (InterContinental San Francisco Hotel)

Bernd Nidetzky , Institute of Biotechnology and Biochemical Engineering, Graz University of Technology, Graz, Austria

Mario Klimacek , Institute of Biotechnology and Biochemical Engineering, Graz University of Technology, Graz, Austria

Stefan Krahulec , Institute of Biotechnology and Biochemical Engineering, Graz University of Technology, Graz, Austria

Barbara Petschacher , Institute of Biotechnology and Biochemical Engineering, Graz University of Technology, Graz, Austria

Metabolic engineering of Saccharomyces cerevisiae for xylose fermentation has often relied on insertion of a heterologous pathway consisting of NAD(P)H-dependent xylose reductase (XR) and NAD-dependent xylitol dehydrogenase (XDH). Low ethanol yield and formation of fermentation by-products such as xylitol and glycerol seen for many of the strains constructed in this way have been ascribed to incomplete coenzyme recycling in the steps catalyzed by XR and XDH. We have used structure-guided engineering of Candida tenuis XR and Galactocandida mastotermitis XDH to obtain enzyme pairs that display well matched utilization of NAD(H) and NADP(H). Yeast strains producing XR and XDH variants that show altered coenzyme selectivity exhibit notably improved fermentation capabilities as compared to the reference strain expressing the genes for the wild-type enzymes.

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6-05: Development and characterization of xylose-fermenting strains of Saccharomyces cerevisiae based on structure-based engineering of key metabolic enzymes