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J. Lipid Res.
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A more recent version of this article appeared on October 1, 2004

Papers In Press, published online ahead of print August 1, 2004
J. Lipid Res., doi:10.1194/jlr.M400181-JLR200
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Submitted on May 11, 2004
Revised on July 20, 2004
Accepted on July 21, 2004

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

Astrid Meyer, Helene Kirsch, Frédéric Domergue, Amine Abbadi, Petra Sperling, Jörg Bauer, Petra Cirpus, Thorsten K. Zank, Hervé Moreau, Thomas J. ;Roscoe, Ulrich Zähringer, and Ernst Heinz

Biozentrum Klein Flottbek, Universität Hamburg, Hamburg, Hamburg 22609

Corresponding Author: astrid_meyer{at}gmx.net

In algae the biosynthesis of docosahexaenoic acid (22:6omega 3, DHA) proceeds via the elongation of eicosapentaenoic acid (20:5omega 3, EPA) to 22:5omega 3, which is required as substrate for the final 4-desaturation. In order to isolate the elongase specific for this step, we searched EST 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 (6-)C18-PUFAs and one specific for (5-)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 6- and 5-elongases from T. pseudonana and O. tauri, respectively, with the 5- and 4-desaturases from two other algae, we successfully implemented DHA synthesis in stearidonic acid-fed yeast. This may be considered as encouraging first step in future efforts to implement this biosynthetic sequence into transgenic oilseed crops.


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