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The genome of Bacillus subtilis 168 was modified by replacing the gene encoding the essential aldose 1-epimerase (gerK gene) with the functional gene encoding xylitol dehydrogenase (xylA gene) from Candida tropicalis. A mutant, obtained by the suicide plasmid system, showed high xylitol dehydrogenase activity and was able to utilize xylitol as the sole carbon source. After 48 h cultivation of the mutant in a 250-ml flask containing 50 g/l xylitol, it produced xylitol at a concentration of 25.3 g/l with a yield of 84% with respect to the consumed xylitol and a specific activity of 302.1 U/mg protein. Xylitol production was further improved by disruption of a gene (xylB gene) encoding xylose reductase. B. subtilis 158 produced xylitol at a yield of 46% with respect to consumed xylitol and a specific activity of 553.6 U/mg protein after 48 h cultivation in 50 g/l xylitol, followed by scaling up to 1 l fermentor with a fermentation time of 144 h.
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