A novel Delta12-fatty acid desaturase gene from methylotrophic yeast Pichia pastoris GS115.
The methylotrophic yeast Pichia pastoris GS115, a widely used strain in production of various heterologous proteins, especially membrane-bound enzymes, can also produce linoleic and linolenic acids, which indicates the existence of membrane-bound Delta12 and Delta15-fatty acid desaturases. This paper describes the cloning and functional characterization of a novel Delta12-fatty acid desaturase gene from this methylotrophic yeast. The open reading frame of the gene (named Pp-FAD12) is 1263 bp in size and encodes a 420-amino-acid peptide. The deduced Pp-FAD12 protein shows high identity (50-67%) with Delta12-fatty acid desaturases from other fungi. It also shows a high identity (57%) with Delta15-fatty acid desaturase (named Sk-FAD15) from Saccharomyces kluyveri. Expression of Pp-FAD12 in polyunsaturated fatty acids non-producing yeast Saccharomyces cerevisiae demonstrated that its product converted oleic acid (18 : 1) to linoleic acid (18 : 2). This result suggests that Pp-FAD12 encodes a novel Delta12-fatty acid desaturase in P. pastoris GS115. This is the first report about the cloning and functional characterization of Delta12-fatty acid desaturase gene in methylotrophic yeast.
Wei
D Sh
DSh
Tianjin Key Laboratory of Microbial Functional Genomics, Department of Microbiology, Nankai University, Tianjin 300071, PR China.
Li
M Ch
MCh
Zhang
X X
XX
Zhou
H
H
Xing
L J
LJ
D001483:Base Sequence; D044943:Fatty Acid Desaturases / Q000235:genetics* / Q000378:metabolism; D005800:Genes, Fungal; D019787:Linoleic Acid / Q000378:metabolism; D008969:Molecular Sequence Data; D019301:Oleic Acid / Q000378:metabolism; D016366:Open Reading Frames; D010843:Pichia / Q000201:enzymology / Q000235:genetics*; D017385:Sequence Homology
D019301:Oleic Acid; D019787:Linoleic Acid; D044943:Fatty Acid Desaturases; C040448:delta-12 fatty acid desaturase