Characterization and complementation of a Pichia stipitis mutant unable to grow on d-xylose or l-arabinose |
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| Authors: | Shi Nian-Qing Prahl Kristine Hendrick Jim Cruz Jose Lu Ping Cho Jae-Yong Jones Sharon Jeffries Thomas |
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| Institution: | (1) Department of Bacteriology, University of Wisconsin, 1550 Linden Drive, 53706 Madison, WI;(2) Department of Biochemistry, University of Wisconsin, 420 Henry Mall, 53706 Madison, WI;(3) Forest Service, Forest Products Laboratory, USDA, 1 Gifford Pinchot Drive, 53705 Madison, WI;(4) Present address: University of Wissonsin-Marathon County, 518 S. 7th Avenue, 54401 Wausau, WI;(5) Present address: Ophidian Pharmaceuticals Inc., 5445 E. Cheryl Parkway, 53711 Madison, WI;(6) Present address: Department of Chemical Engineering, University of Vigo, AsLagoas, 32004 Ourense, Spain;(7) Present address: Merck, P.O. Box 2000, RY80Y-125, 07065 Rahway, NJ;(8) Present address: NCl-Frederick Cancer Research and Development Center, 21701 Frederick, MD;(9) Present address: Department of Anatomy, University of Wisconsin, 53706 Madison, WI |
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| Abstract: | Pichia stipitis CBS 6054 will grow on d-xylose, d-arabinose, and l-arabinose. d-Xylose and l-arabinose are abundant in seed hulls of maize, and their utilization is important in processing grain residues. To elucidate
the degradation pathway for l-arabinose, we obtained a mutant, FPL-MY30, that was unable to grow on d-xylose and l-arabinose but that could grow on d-arabinitol. Activity assays of oxidoreductase and pentulokinase enzymes involved in d-xylose, d-arabinose, and l-arabinose pathways indicated that FPL-MY30 is deficient in d-xylitol dehydrogenase (D-XDH), d- and l-arabinitol dehydrogenases, and d-ribitol dehydrogenase. Transforming FPL-MY30 with a gene for xylitol dehydrogenase (PsXYL2), which was cloned from CBS 6054 (Gen Bank AF127801), restored the D-XDH activity and the capacity for FPL-MY30 to grow on
l-arabinose. This suggested that FPL-MY30 is critically deficient in XYL2 and that the d-xylose and l-arabinose metabolic pathways have xylitolas a common intermediate. The capacity for FPL-MY30 to grow on d-arabinitol could proceed through d-ribulose. |
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| Keywords: | d-Xylose" target="_blank">d-Xylose l-arabinose" target="_blank">l-arabinose Pichia stipitis fermentation metabolism mutant complementation |
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