title: Cloning of the xylose reductase gene of Candida milleri.
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.
authors.affiliation: Department of Food Science and Biotechnology, Kangwon National University, Chuncheon 200-701, Republic of Korea.
authors.forename: Hyoun-Soo
authors.initials: HS
authors.lastname: Sim
authors.forename: Eun-Hee
authors.initials: EH
authors.lastname: Park
authors.forename: Se-Young
authors.initials: SY
authors.lastname: Kwon
authors.forename: Sang-Ki
authors.initials: SK
authors.lastname: Choi
authors.forename: Su-Han
authors.initials: SH
authors.lastname: Lee
authors.forename: Myoung-Dong
authors.initials: MD
authors.lastname: Kim
mesh_terms: D000449:Aldehyde Reductase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D002175:Candida / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D016000:Cluster Analysis; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D005516:Food Microbiology; D015870:Gene Expression; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D007716:Kluyveromyces / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014993:Xylitol / Q000378:metabolism; D014994:Xylose / Q000378:metabolism
chemical_list: D004271:DNA, Fungal; D014994:Xylose; D000449:Aldehyde Reductase; D014993:Xylitol