Molecular cloning and characterization of the Delta9 fatty acid desaturase gene and its promoter region from Saccharomyces kluyveri.
A gene encoding Delta9 fatty acid desaturase, which catalyzes the first reaction of fatty acid desaturation, has been cloned from Saccharomyces kluyveri, a species capable of producing polyunsaturated fatty acids (poly-UFA). The open reading frame of this gene (named Sk-OLE1) consists of 1443 bp, encoding 480 amino acids. The deduced Sk-OLE1 protein shared 63-74% identity with those from other yeasts. Expression of the Sk-OLE1 gene in Saccharomyces cerevisiae complemented genetic disruption of the S. cerevisiae OLE1 gene (Sc-OLE1). Fatty acid analysis showed that the ratio of palmitoleic acid to oleic acid in the Sk-OLE1-expressing S. cerevisiae strain was lower than that of the S. cerevisiae strain expressing the Sc-OLE1 gene. The promoter region of the Sk-OLE1 gene was shorter than that of the Sc-OLE1 gene and appeared to lack the fatty acid-regulated sequences present in the Sc-OLE1 promoter region. Analysis of transcriptional expression revealed that the Sk-OLE1 gene was not actually repressed in either S. kluyveri or S. cerevisiae by exogenous UFAs. The DDBJ accession number for the sequence reported in this paper is AB071696.
Kajiwara
Susumu
S
Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan. skajiwar@bio.titech.ac.jp
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D044943:Fatty Acid Desaturases / Q000235:genetics* / Q000378:metabolism; D005231:Fatty Acids, Unsaturated / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012440:Saccharomyces / Q000201:enzymology / Q000235:genetics*; D013230:Stearoyl-CoA Desaturase
D005231:Fatty Acids, Unsaturated; D044943:Fatty Acid Desaturases; D013230:Stearoyl-CoA Desaturase; C029627:delta-9 fatty acid desaturase