Isolation of xylose reductase gene of Pichia stipitis and its expression in Saccharomyces cerevisiae.
A NADPH/NADH-dependent xylose reductase gene was isolated from the xylose-assimilating yeast, Pichia stipitis. DNA sequence analysis showed that the gene consists of 951 bp. The gene introduced in Saccharomyces cerevisiae was transcribed to mRNA, and a considerable amount of enzyme activity was observed constitutively, whereas transcription and translation in P stipitis were inducible. S. cerevisiae carrying the xylose reductase gene could not, however, grow on xylose medium, and could not produce ethanol from xylose. Since xylose uptake and accumulation of xylitol by S. cerevisiae were observed, the conversion of xylitol to xylulose seemed to be limited.
International Center of Cooperative Research in Biotechnology, Faculty of Engineering, Osaka University, Japan.
S
S
Takuma
N
N
Nakashima
M
M
Tantirungkij
S
S
Kinoshita
H
H
Okada
T
T
Seki
T
T
Yoshida
D000449:Aldehyde Reductase; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D005798:Genes, Bacterial; D007700:Kinetics; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010843:Pichia / Q000201:enzymology / Q000235:genetics*; D010957:Plasmids; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013401:Sugar Alcohol Dehydrogenases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D014158:Transcription, Genetic; D014994:Xylose / Q000378:metabolism
D004269:DNA, Bacterial; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014994:Xylose; D013401:Sugar Alcohol Dehydrogenases; D000449:Aldehyde Reductase; C023142:polyol dehydrogenase (NADP)