Oleic acid desaturation in Tetrahymena pyriformis

The desaturation of oleoyl-CoA by a microsomal preparation from Tetrahymena has been studied. Desaturation of oleoyl-CoA required oxygen and NADH, and was inhibited by cyanide. HPLC analysis of fatty acid phenacyl esters, prepared from TLC-purified phospholipid, confirmed that radioactivity appeared in oleate, linoleate and gamma-linolenate. Both the time course of desaturation and the apparent desaturation of 1-palmitoyl-2-[14C]oleoylphosphatidylcholine suggested that phospholipid-bound oleate could be a substrate for desaturation. In the crude microsomal preparation, acylation of oleoyl-CoA to give oleoyl phospholipid was rapid. Therefore, preincubation in the absence of NADH was employed to create [14C]oleoyl phospholipids, and kinetic studies were carried out upon subsequent addition of NADH. When data were plotted in a double reciprocal form, a linear function was observed.