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Appl. Environ. Microbiol. doi:10.1128/AEM.02707-06
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Metabolic engineering of Saccharomyces cerevisiae for conversion of D-glucose to xylitol and other five-carbon sugars and sugar alcohols

Mervi H. Toivari*, Laura Ruohonen, Andrei N. Miasnikov, Peter Richard, and Merja Penttilä

VTT Technical Research Centre of Finland, P.O. Box 1000, FI-02044 VTT, Espoo, Finland, Danisco Global Innovation, Sokeritehtaantie 20, FI-02460 Kantvik, Finland

* To whom correspondence should be addressed. Email: Mervi.Toivari{at}vtt.fi.


  Abstract

Recombinant Saccharomyces cerevisiae strains that produce sugar alcohols xylitol and ribitol and the pentose sugar D-ribose, from D-glucose, in a single fermentation step are described. A transketolase-deficient S. cerevisiae strain accumulated D-xylulose 5-phosphate intracellularly and released ribitol and pentose sugars (D-ribose, D-ribulose, D-xylulose) into the growth medium. Expression of the xylitol dehydrogenase encoding gene XYL2 from Pichia stipitis in the transketolase-deficient strain resulted in 8.5-fold enhancement in the total amount of the excreted sugar alcohols ribitol and xylitol. The additional introduction of the 2-deoxy-glucose 6-phosphate phosphatase encoding gene, DOG1, into the transketolase-deficient strain expressing XYL2 resulted in a further 1.6-fold increase in ribitol production. Finally, the deletion of the endogenous xylulokinase encoding gene, XKS1, was necessary to increase the amount of xylitol to 50% of the 5-carbon sugar alcohols excreted.




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