title: Lipoxygenase-mediated metabolism of storage lipids in germinating sunflower cotyledons and beta-oxidation of (9Z,11E,13S)-13-hydroxy-octadeca-9,11-dienoic acid by the cotyledonary glyoxysomes.
abstract: During the early stages of germination, a lipid-body lipoxygenase is expressed in the cotyledons of sunflowers (Helianthus annuus L.). In order to obtain evidence for the in vivo activity of this enzyme during germination, we analyzed the lipoxygenase-dependent metabolism of polyunsaturated fatty acids esterified in the storage lipids. For this purpose, lipid bodies were isolated from etiolated sunflower cotyledons at different stages of germination, and the storage triacylglycerols were analyzed for oxygenated derivatives. During the time course of germination the amount of oxygenated storage lipids was strongly augmented, and we detected triacylglycerols containing one, two or three residues of (9Z,11E,13S)-13-hydro(pero)xy-octadeca-9,11-dienoic acid. Glyoxysomes from etiolated sunflower cotyledons converted (9Z,11E,13S)-13-hydroxy-octadeca-9,11-dienoic acid to (9Z,11E)-13-oxo-octadeca-9,11-dienoic acid via an NADH-dependent dehydrogenase reaction. Both oxygenated fatty acid derivatives were activated to the corresponding CoA esters and subsequently metabolized to compounds of shorter chain length. Cofactor requirement and formation of acetyl-CoA indicate degradation via beta-oxidation. However, beta-oxidation only proceeded for two consecutive cycles, leading to accumulation of a medium-chain metabolite carrying an oxo group at C-9, equivalent to C-13 of the parent (9Z,11E,13S)-13-hydroxy-octadeca-9,11-dienoic acid. Short-chain beta-oxidation intermediates were not detected during incubation. Similar results were obtained when 13-hydroxy octadecanoic acid was used as beta-oxidation substrate. On the other hand, the degradation of (9Z,11E)-octadeca-9,11-dienoic acid was accompanied by the appearance of short-chain beta-oxidation intermediates in the reaction mixture. The results suggest that the hydroxyl/oxo group at C-13 of lipoxygenase-derived fatty acids forms a barrier to continuous beta-oxidation by glyoxysomes.
authors.affiliation: Institut für Botanik, Universität Münster, Schlossgarten 3, 48149, Münster, Germany.
authors.forename: Bernt
authors.initials: B
authors.lastname: Gerhardt
authors.forename: Kerstin
authors.initials: K
authors.lastname: Fischer
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Balkenhohl
authors.forename: Georg
authors.initials: G
authors.lastname: Pohnert
authors.forename: Hartmut
authors.initials: H
authors.lastname: Kühn
authors.forename: Claus
authors.initials: C
authors.lastname: Wasternack
authors.forename: Ivo
authors.initials: I
authors.lastname: Feussner
mesh_terms: D002851:Chromatography, High Pressure Liquid; D018548:Cotyledon / Q000254:growth & development* / Q000378:metabolism; D018525:Germination / Q000502:physiology; D020676:Glyoxysomes / Q000378:metabolism*; D006368:Helianthus / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism*; D008041:Linoleic Acids / Q000378:metabolism; D050356:Lipid Metabolism; D008054:Lipid Peroxides / Q000378:metabolism*; D008084:Lipoxygenase / Q000378:metabolism*; D010084:Oxidation-Reduction; D013997:Time Factors
chemical_list: D008041:Linoleic Acids; D008054:Lipid Peroxides; C038649:13-hydroperoxy-9,11-octadecadienoic acid; C024348:13-hydroxy-9,11-octadecadienoic acid; D008084:Lipoxygenase