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Identification, Separation, and Characterization of Acyl-Coenzyme A Dehydrogenases Involved in Mitochondrial beta -Oxidation in Higher Plants1

Kornelia Bode, Mark A. Hooks, and Ivan Couée2, *

Station de Physiologie Végétale, Institut National de la Recherche Agronomique, Centre de Recherches de Bordeaux, Boîte Postale 81, 33883 Villenave d'Ornon cedex, France

The existence in higher plants of an additional beta -oxidation system in mitochondria, besides the well-characterized peroxisomal system, is often considered controversial. Unequivocal demonstration of beta -oxidation activity in mitochondria should rely on identification of the enzymes specific to mitochondrial beta -oxidation. Acyl-coenzyme A dehydrogenase (ACAD) (EC 1.3.99.2,3) activity was detected in purified mitochondria from maize (Zea mays L.) root tips and from embryonic axes of early-germinating sunflower (Helianthus annuus L.) seeds, using as the enzyme assay the reduction of 2,6-dichlorophenolindophenol, with phenazine methosulfate as the intermediate electron carrier. Subcellular fractionation showed that this ACAD activity was associated with mitochondrial fractions. Comparison of ACAD activity in mitochondria and acyl-coenzyme A oxidase activity in peroxisomes showed differences of substrate specificities. Embryonic axes of sunflower seeds were used as starting material for the purification of ACADs. Two distinct ACADs, with medium-chain and long-chain substrate specificities, respectively, were separated by their chromatographic behavior, which was similar to that of mammalian ACADs. The characterization of these ACADs is discussed in relation to the identification of expressed sequenced tags corresponding to ACADs in cDNA sequence analysis projects and with the potential roles of mitochondrial beta -oxidation in higher plants.

1   This work was partly funded by the Aquitaine (France) Regional Council.
2   Present address: Centre National de la Recherche Scientifique UMR 6553, Université de Rennes I, Campus Scientifique de Beaulieu, Bâtiment 14, 263 Avenue du Général Leclerc, 35042 Rennes cedex, France.
*   Corresponding author; e-mail ivan.couee{at}univ-rennes1.fr; fax 33-299286915.

Plant Physiol. (1999) 119: 1305-1314
Copyright Clearance Center:   0032-0889/99/119//10
© 1999 American Society of Plant Physiologists




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