Current Genetics

, Volume 18, Issue 6, pp 493–500

Isolation and characterization of the Pichia stipitis xylitol dehydrogenase gene, XYL2, and construction of a xylose-utilizing Saccharomyces cerevisiae transformant

  • Peter Kötter
  • René Amore
  • Cornelis P. Hollenberg
  • Michael Ciriacy
Original Articles

DOI: 10.1007/BF00327019

Cite this article as:
Kötter, P., Amore, R., Hollenberg, C.P. et al. Curr Genet (1990) 18: 493. doi:10.1007/BF00327019

Summary

A P. stipitis cDNA library in λgt11 was screened using antisera against P. stipitis xylose reductase and xylitol dehydrogenase, respectively. The resulting cDNA clones served as probes for screening a P. stipitis genomic library. The genomic XYL2 gene was isolated and the nucleotide sequence of the 1089 bp structural gene, and of adjacent non-coding regions, was determined. The XYL2 open-reading frame codes for a protein of 363 amino acids with a predicted molecular mass of 38.5 kDa. The XYL2 gene is actively expressed in S. cerevisiae transformants. S. cerevisiae cells transformed with a plasmid, pRD1, containing both the xylose reductase gene (XYL1) and the xylitol dehydrogenase gene (XYL2), were able to grow on xylose as a sole carbon source. In contrast to aerobic glucose metabolism, S. cerevisiae XYL1-XYL2 transformants utilize xylose almost entirely oxidatively.

Key words

Xylitol dehydrogenase gene Pichia stipitis Saccharomyces cerevisiae Xylose utilization 

Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • Peter Kötter
    • 1
  • René Amore
    • 1
  • Cornelis P. Hollenberg
    • 1
  • Michael Ciriacy
    • 1
  1. 1.Institut für MikrobiologieHeinrich-Heine-UniversitätDüsseldorfGermany