Yeast Delta 12 fatty acid desaturase: gene cloning, expression, and function.
In an effort to elucidate the molecular mechanisms of fatty acid desaturation in yeast, a complete gene encoding Delta 12 fatty acid desaturase of Saccharomyces kluyveri was cloned. The open reading frame of this gene (named Sk-FAD2) consists of 1,251 bp, encoding 416 amino acids. The deduced Sk-FAD2 protein had 37-55% identity with those from other filamentous fungi. Unlike the genes of these other fungi, S. cerevisiae expressing Sk-FAD2 was found to be capable of synthesizing the dienoic fatty acid hexadecadienoic acid as well as linoleic acid. Moreover, the Sk-FAD2-disrupted strain of S. kluyveri was unable to produce polyunsaturated fatty acids. These results suggested that Sk-FAD2 protein is a unique Delta 12 fatty acid desaturase in S. kluyveri. Analysis of transcriptional expression revealed that Sk-FAD2 was not repressed by exogenous unsaturated fatty acids but responded to low-temperature stress.
Watanabe
Kyoko
K
Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Kanagawa, Japan.
Oura
Takahiro
T
Sakai
Hiromichi
H
Kajiwara
Susumu
S
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D044943:Fatty Acid Desaturases / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D005227:Fatty Acids / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D020868:Gene Silencing; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis; D012440:Saccharomyces / Q000201:enzymology* / Q000235:genetics; D016415:Sequence Alignment; D013696:Temperature
D005227:Fatty Acids; D012333:RNA, Messenger; D044943:Fatty Acid Desaturases; C040448:delta-12 fatty acid desaturase