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Characterization and complementation of a Pichia stipitis mutant unable to grow on D-xylose or L-arabinose.

N Q Shi1, K Prahl, J Hendrick

  • 1Department of Bacteriology, University of Wisconsin, Madison 53706, USA.

Applied Biochemistry and Biotechnology
|June 13, 2000
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Summary
This summary is machine-generated.

Pichia stipitis yeast can metabolize L-arabinose, a key sugar in maize. A mutant deficient in xylitol dehydrogenase (XDH) revealed that XDH is crucial for L-arabinose utilization.

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Area of Science:

  • Biochemistry
  • Microbiology
  • Biotechnology

Background:

  • Pichia stipitis yeast metabolizes various pentose sugars, including D-xylose, D-arabinose, and L-arabinose.
  • Efficient utilization of these sugars, particularly L-arabinose found in maize residues, is vital for biomass processing.
  • Understanding the metabolic pathways is crucial for optimizing industrial applications.

Purpose of the Study:

  • To elucidate the L-arabinose degradation pathway in Pichia stipitis.
  • To identify the specific enzyme deficiency in a mutant strain (FPL-MY30) unable to utilize L-arabinose.
  • To confirm the role of xylitol dehydrogenase (XDH) in L-arabinose metabolism.

Main Methods:

  • Generation and characterization of a Pichia stipitis mutant (FPL-MY30) with impaired pentose utilization.
  • Enzyme activity assays for oxidoreductases and pentulokinases involved in pentose metabolism.
  • Genetic complementation by transforming the mutant with the PsXYL2 gene encoding XDH.

Main Results:

  • The mutant FPL-MY30 exhibited deficiencies in D-xylitol dehydrogenase (D-XDH), D- and L-arabinitol dehydrogenases, and D-ribitol dehydrogenase.
  • Restoration of D-XDH activity and L-arabinose growth capacity in FPL-MY30 upon transformation with the PsXYL2 gene.
  • Identification of xylitol as a common intermediate in both D-xylose and L-arabinose metabolic pathways.

Conclusions:

  • The PsXYL2 gene, encoding D-xylitol dehydrogenase, is essential for L-arabinose utilization in Pichia stipitis.
  • The D-xylose and L-arabinose metabolic pathways converge at the xylitol intermediate.
  • This study clarifies a critical step in pentose sugar assimilation, relevant for lignocellulosic biomass conversion.