The P-OLE1 gene of Pichia angusta encodes a delta 9-fatty acid desaturase and complements the ole1 mutation of Saccharomyces cerevisiae.
Three PCR-amplified DNA fragments hybridizing with the OLE1 gene encoding delta 9-fatty acid desaturase of Saccharomyces cerevisiae were obtained using, respectively, genomic DNAs of one strain each of Kluyveromyces thermotolerans, Pichia angusta and Yarrowia lipolytica as templates. A gene designated P-OLE1 was cloned from the above fragment of P. angusta and sequenced. An open reading frame of P-OLE1 encodes a 49.6-kDa protein consisting of 451 amino acid residues, which shows high identity (62%) and similarity (89%) to that deduced from the OLE1 nucleotide sequence. Expression of P-OLE1 driven by the S. cerevisiae GAP promoter or its own promoter complemented the ole1 mutation of S.cerevisiae. Transcription of P-OLE1 in the native host was suggested to be partially repressed by oleic acid in the medium, as was that of OLE1 in S. cerevisiae and a similar gene in Y. lipolytica, but that of a similar gene in K. thermotolerans was not.
Anamnart
S
S
Department of Biotechnology, Faculty of Engineering, Osaka University, Japan.
Tomita
T
T
Fukui
F
F
Fujimori
K
K
Harashima
S
S
Yamada
Y
Y
Oshima
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004271:DNA, Fungal; D044943:Fatty Acid Desaturases / Q000235:genetics*; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D007716:Kluyveromyces / Q000201:enzymology / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D019301:Oleic Acid / Q000378:metabolism; D010843:Pichia / Q000201:enzymology / Q000235:genetics*; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D004718:Saccharomycetales / Q000201:enzymology / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013230:Stearoyl-CoA Desaturase; D014158:Transcription, Genetic
D004271:DNA, Fungal; D019301:Oleic Acid; D044943:Fatty Acid Desaturases; D013230:Stearoyl-CoA Desaturase; C029627:delta-9 fatty acid desaturase