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First published online March 13, 2003; 10.1104/pp.102.018317

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Plant Physiol, April 2003, Vol. 131, pp. 1648-1660

New Insight into Phaeodactylum tricornutum Fatty Acid Metabolism. Cloning and Functional Characterization of Plastidial and Microsomal Delta 12-Fatty Acid Desaturases1,[w]

Frédéric Domergue,* Patricia Spiekermann, Jens Lerchl,2 Christoph Beckmann, Oliver Kilian, Peter G. Kroth, Wilhem Boland, Ulrich Zähringer, and Ernst Heinz

Institut für Allgemeine Botanik, Universität Hamburg, Ohnhorststrasse 18, 22609 Hamburg, Germany (F.D., P.S., E.H.); BASF Plant Science GmbH, BPS-A30, D-67056 Ludwigshafen, Germany (J.L.); Fachbereich Biologie, Universität Konstanz, D-78457 Konstanz, Germany (O.K., P.K.); Max-Planck-Institut für Chemische Ökologie, Carl-Zeiss-Promenade 10, D-07745 Jena, Germany (C.B., W.B.); and Forschungszentrum Borstel, Parkallee 22, D-23845 Borstel, Germany (U.Z.)

In contrast to 16:3 plants like rapeseed (Brassica napus), which contain alpha -linolenic acid (18:3Delta 9,12,15) and hexadecatrienoic acid (16:3Delta 7,10,13) as major polyunsaturated fatty acids in leaves, the silica-less diatom Phaeodactylum tricornutum contains eicosapentaenoic acid (EPA; 20:5Delta 5,8,11,14,17) and a different isomer of hexadecatrienoic acid (16:3Delta 6,9,12). In this report, we describe the characterization of two cDNAs having sequence homology to Delta 12-fatty acid desaturases from higher plants. These cDNAs were shown to code for a microsomal and a plastidial Delta 12-desaturase (PtFAD2 and PtFAD6, respectively) by heterologous expression in yeast (Saccharomyces cerevisiae) and Synechococcus, respectively. Using these systems in the presence of exogenously supplied fatty acids, the substrate specificities of the two desaturases were determined and compared with those of the corresponding rapeseed enzymes (BnFAD2 and BnFAD6). The microsomal desaturases were similarly specific for oleic acid (18:1Delta 9), suggesting that PtFAD2 is involved in the biosynthesis of EPA. In contrast, the plastidial desaturase from the higher plant and the diatom clearly differed. Although the rapeseed plastidial desaturase showed high activity toward the omega 9-fatty acids 18:1Delta 9 and 16:1Delta 7, in line with the fatty acid composition of rapeseed leaves, the enzyme of P. tricornutum was highly specific for 16:1Delta 9. Our results indicate that in contrast to EPA, which is synthesized in the microsomes, the hexadecatrienoic acid isomer found in P. tricornutum (16:3Delta 6,9,12) is of plastidial origin.

1 This work was supported in part by the European Community program Human Potential (Marie Curie Fellowship under contract no. HPMF-CT-1999-00148 to F.D.), by the Fachagentur für Nachwachsende Rohstoffe (grant to P.S.), by the Deutsche Forschungsgemeinschaft (grant no. Transregio-SFB TR1, TP A1 to P.G.K. and grant no. SFB 436 to W.B.), by the Fonds der Chemischen Industrie, and by BASF Plant Science GmbH.

2 Present address: Plant Science Sweden AB, SE-26831 Svalöv, Sweden.

[w] The online version of this article contains Web-only data. The supplemental material is available at www.plantphysiol.org.

* Corresponding author; e-mail fredDo{at}botanik.uni-hamburg.de; fax 49-40-42816-254.

© 2003 American Society of Plant Biologists


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