Isolation and characterization of a delta-9 fatty acid desaturase gene from the oleaginous yeast Cryptococcus curvatus CBS 570.
The oleaginous yeast Cryptococcus curvatus is of industrial interest because it can accumulate triacylglycerols up to 60% of the cell dry weight. We are aiming at genetic modification of fatty acid biosynthesis for the production of tailor-made triacylglycerols in C. curvatus. As a first step in the development of a transformation and expression system a gene encoding the delta-9 fatty acid desaturase of C. curvatus (CBS 570) was cloned. The 1470 bp gene encodes a protein of 493 amino acids with a calculated molecular mass of 55 kDa. The gene shows strong similarity to previous cloned delta-9 desaturase genes from rat and Saccharomyces cerevisiae, 62 and 72%, respectively. Expression of the delta-9 desaturase gene was studied. Supplementation of the growth medium with oleic acid (C18:1(c9)) showed a strong repression (90%) on the mRNA level, while supplementation with petroselinic acid (C18:1(c6)) had no effect on the amount of mRNA.
Meesters
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Agrotechnical Research Institute (ATO-DLO), Wageningen, The Netherlands.
Eggink
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D003454:Cryptococcus / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics*; D004271:DNA, Fungal / Q000235:genetics; D044943:Fatty Acid Desaturases / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D005800:Genes, Fungal; D007536:Isomerism; D008969:Molecular Sequence Data; D019301:Oleic Acid / Q000494:pharmacology; D009829:Oleic Acids / Q000494:pharmacology; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013230:Stearoyl-CoA Desaturase
D003062:Codon; D004271:DNA, Fungal; D009829:Oleic Acids; D012331:RNA, Fungal; D012333:RNA, Messenger; D019301:Oleic Acid; C008820:petroselinic acid; D044943:Fatty Acid Desaturases; D013230:Stearoyl-CoA Desaturase; C029627:delta-9 fatty acid desaturase