Cloning and expression of the Delta(9) fatty acid desaturase gene from Cryptococcus curvatus ATCC 20509 containing histidine boxes and a cytochrome b(5) domain

To allow genetic modification of the fatty acid biosynthesis routes in the lipid-accumulating yeast Cryptococcus curvatus the Delta(9) fatty acid desaturase gene was cloned and characterized. The 1668-bp gene encodes a protein of 556 amino acids with a calculated molecular mass of 62 kDa. The gene shows strong homology to previously cloned Delta(9) fatty acid desaturase genes from yeast and rat. Homology includes three histidine boxes characteristic for membrane-bound desaturases and a cytochrome b(5) domain responsible for electron transport. The Delta(9) desaturase gene has a high G +/- C content of 61% and displays a codon usage different from that of Saccharomyces cerevisiae, but similar to that of the basidiomycete Schizophyllum commune. Expression of the Delta(9) desaturase gene of C. curvatus ATCC 20509 was studied in the presence of different fatty acids in the growth medium. Repression of desaturase mRNA signals was found if fatty acids with a double bond at the Delta(9) position were present Fatty acids with a double bond at another position (Delta(10) or Delta(6) or saturated fatty acids had no effect on the transcription of the cloned gene.