Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Cloning of the Xylose Reductase Gene of Candida milleri

  • Sim, Hyoun-Soo (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Park, Eun-Hee (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Kwon, Se-Young (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Choi, Sang-Ki (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Lee, Su-Han (Department of Food Technology and Service, Eulji University) ;
  • Kim, Myoung-Dong (Department of Food Science and Biotechnology, Kangwon National University)
  • Received : 2013.05.07
  • Accepted : 2013.05.12
  • Published : 2013.07.28
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Abstract

The entire nucleotide sequence of the xylose reductase (XR) gene in Candida milleri CBS8195 sourdough yeast was determined by degenerate polymerase chain reaction (PCR) and genome walking. The sequence analysis revealed an open-reading frame of 981 bp that encoded 326 amino acids with a predicted molecular mass of 36.7 kDa. The deduced amino acid sequence of XR of C. milleri was 64.7% homologous to that of Kluyveromyces lactis. The cloned XR gene was expressed in Saccharomyces cerevisiae, and the resulting recombinant S. cerevisiae strain produced xylitol from xylose, indicating that the C. milleri XR introduced into S. cerevisiae is functional. An enzymatic activity assay and semiquantitative reverse transcription-PCR revealed that the expression of CmXR was induced by xylose. The GenBank Accession No. for CmXR is KC599203.

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References

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