Novel fatty acid elongases and their use for the reconstitution of docosahexaenoic acid biosynthesis

Abstract

In algae, the biosynthesis of docosahexaenoic acid (22:6ω3; DHA) proceeds via the elongation of eicosapentaenoic acid (20:5ω3; EPA) to 22:5ω3, which is required as a substrate for the final Δ4 desaturation. To isolate the elongase specific for this step, we searched expressed sequence tag and genomic databases from the algae Ostreococcus tauri and Thalassiosira pseudonana, from the fish Oncorhynchus mykiss, from the frog Xenopus laevis, and from the sea squirt Ciona intestinalis using as a query the elongase sequence PpPSE1 from the moss Physcomitrella patens. The open reading frames of the identified elongase candidates were expressed in yeast for functional characterization. By this, we identified two types of elongases from O. tauri and T. pseudonana: one specific for the elongation of (Δ6-)C18-PUFAs and one specific for (Δ5-)C20-PUFAs, showing highest activity with EPA. The clones isolated from O. mykiss, X. laevis, and C. intestinalis accepted both C18- and C20-PUFAs. By coexpression of the Δ6- and Δ5-elongases from T. pseudonana and O. tauri, respectively, with the Δ5- and Δ4-desaturases from two other algae we successfully implemented DHA synthesis in stearidonic acid-fed yeast.This may be considered an encouraging first step in future efforts to implement this biosynthetic sequence into transgenic oilseed crops.

Keywords

• Ciona intestinalis
• Oncorhynchus mykiss
• Ostreococcus tauri
• polyunsaturated fatty acids
• Thalassiosira pseudonana
• Xenopus laevis