Christian Schlechtriem

Highly unsaturated fatty acid synthesis in marine fish: Cloning, functional characterization, and nutritional regulation of fatty acyl Δ6 desaturase of Atlantic cod (Gadus morhua L.)

This study reports the cloning, functional characterization, tissue expression, and nutritional regulation of a Δ6 fatty acyl desaturase of Atlantic cod (Gadus morhua). PCR primers were designed based on the sequences of conserved motifs in available fish desaturases and used to isolate a cDNA fragment from cod liver, with full-length cDNA obtained by rapid amplification of cDNA ends. The cDNA for the putative desaturase was shown to comprise 1980 bp, including a 261-bp 5′-UTR, a 375-bp 3′-UTR, and an ORF of 1344 bp that specified a protein of 447 amino acids. The protein sequence included three histidine boxes, two transmembrane regions, and an N-terminal cytochrome b5 domain containing the heme-binding motif HPGG, all characteristic of microsomal fatty acyl desaturases. The cDNA displayed Δ6 desaturase activity in a yeast expression system. Quantitative real-time PCR assay of gene expression in cod showed that the Δ6 desaturase gene was expressed highly in brain, to a slightly lesser extent in liver, kidney, intestine, red muscle, and gill, and at much lower levels in white muscle, spleen, and heart. The expression of the Δ6 desaturase gene did not appear to be under significant nutritional regulation, with levels in liver and intestine being barely altered in fish fed a vegetable oil blend, in comparison with levels in fish fed fish oil. This was reflected in enzyme activity, as hepatocytes or enterocytes showed very little highly unsaturated FA biosynthesis activity irrespective of diet. Further studies are required to determine why the Δ6 desaturase appears to be barely functional in cod under the conditions tested.

Incorporation and metabolism of fatty acids by desaturation and elongation in the nematode, Panagrellus redivivus

The free-living nematode Panagrellus redivivus can be mass produced in monoxenic solid culture on Saccharomyces cerevisiae and therefore could be useful as a live food for marine fish or crustacean larvae in the rapidly expanding aquaculture industry. However, this will depend on their lipid and fatty acid composition and so this was investigated in mass produced P. redivivus grown on S. cerevisiae in three different media. Live nematodes were also incubated with [1-14C]-labelled fatty acids and the desaturation and elongation of the fatty acids were determined. The combined results from the growth trials on different media and the metabolic studies with labelled fatty acids indicated the presence of Δ9, Δ12, Δ6 and Δ5 fatty acid desaturase activities, and elongase activities active towards C18, C16 and shorter chain fatty acids. The presence of Δ15, and therefore the ability to produce n-3 polyunsaturated fatty acids, was suggested by the compositional data, but could not be conclusively established from metabolic studies.