Skip to main content
SpringerLink
Log in

Lipoxygenase-mediated metabolism of storage lipids in germinating sunflower cotyledons and β-oxidation of (9Z,11E,13S)-13-hydroxy-octadeca-9,11-dienoic acid by the cotyledonary glyoxysomes

  • Original Article
  • Published:
Planta Aims and scope Submit manuscript

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 β-oxidation. However, β-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 β-oxidation intermediates were not detected during incubation. Similar results were obtained when 13-hydroxy octadecanoic acid was used as β-oxidation substrate. On the other hand, the degradation of (9Z,11E)-octadeca-9,11-dienoic acid was accompanied by the appearance of short-chain β-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 β-oxidation by glyoxysomes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4a–c
Fig. 5
Fig. 6
Fig 7
Fig. 8a,b

Similar content being viewed by others

Abbreviations

RP-HPLC :

Reverse-phase HPLC

SP-HPLC :

Straight-phase HPLC

13-HPODE :

(9Z,11E,13S)-13-Hydroperoxy-octadeca-9,11-dienoic acid

13-HODE :

(9Z,11E,13S)-13-Hydroxy-octadeca-9,11-dienoic acid

13-KODE :

(9Z,11E)-13-Oxo-octadeca-9,11-dienoic acid

9-KTDE :

(5Z,7E)-9-Oxo-tetradeca-5,7-dienoic acid

LOX :

Lipoxygenase

TAG :

Triacylglycerol

RT :

Retention time

References

  • Balkenhohl T, Kühn H, Wasternack C, Feussner I (1998) A lipase specific for esterified oxygenated polyenoic fatty acids in lipid bodies of cucumber cotyledons. In: Sánchez J, Cerdá-Olmedo E, Martínez-Force E (eds.) Advances in plant lipid research. Secretariado de Publicaciones de la Universidad de Sevilla, Sevilla, pp 320–322

  • Behrends W, Thieringer R, Engeland K, Kunau W-H, Kindl H (1988) The glyoxysomal β-oxidation system in cucumber seedlings: identification of enzymes required for the degradation of unsaturated fatty acids. Arch Biochem Biophys 263:170–177

    CAS  PubMed  Google Scholar 

  • Bronstein JC, Bull AW (1997) Substrate specificity and characterization of partially purified rat liver 13-hydroxyoctadecadienoic acid (13-HODE) dehydrogenase. Arch Biochem Biophys 348:219–225

    Article  CAS  PubMed  Google Scholar 

  • Engeland K, Kindl H (1991) Evidence for a peroxisomal fatty acid beta-oxidation involving d-3-hydroxyacyl-CoAs. Characterization of two forms of hydro-lyase that convert d-(−)-3-hydroxyacyl-CoA into 2-trans-enoyl-CoA. Eur J Biochem 200:171–178

    CAS  PubMed  Google Scholar 

  • Fang X, Kaduce TL, Spector AA (1999) 13-(S)-Hydroxyoctadecadienoic acid (13-HODE) incorporation and conversion to novel products by endothelial cells. J Lipid Res 40:699–707

    CAS  PubMed  Google Scholar 

  • Feussner I, Kindl H (1994) Particulate and soluble lipoxygenase isoenzymes—comparison of molecular and enzymatic properties. Planta 194:22–28

    Article  CAS  Google Scholar 

  • Feussner I, Wasternack C, Kindl H, Kühn H (1995) Lipoxygenase-catalyzed oxygenation of storage lipids is implicated in lipid mobilization during germination. Proc Natl Acad Sci USA 92:11849–11853

    CAS  PubMed  Google Scholar 

  • Feussner I, Bachmann A, Höhne M, Kindl H (1998a) All three acyl moieties of trilinolein are efficiently oxygenated by recombinant His-tagged lipid body lipoxygenase in vitro. FEBS Lett 431:433–436

    Article  CAS  PubMed  Google Scholar 

  • Feussner I, Blée E, Weichert H, Rousset C, Wasternack C (1998b) Fatty acid catabolism at the lipid body membrane of germinating cucumber cotyledons. In: Sánchez J, Cerdá-Olmedo E, Martínez-Force E (eds.) Advances in plant lipid research. Secretariado de Publicaciones de la Universidad de Sevilla, Sevilla, pp 311–313

  • Feussner I, Kühn H, Wasternack C (2001) The lipoxygenase dependent degradation of storage lipids. Trends Plant Sci 6:268–273

    Article  CAS  PubMed  Google Scholar 

  • Gerbling H, Gerhardt B (1988) Peroxisomal degradation of branched-chain 2-oxo acids. Plant Physiol 91:1387–1392

    Google Scholar 

  • Gerhardt B (1983) Localization of β-oxidation enzymes in peroxisomes isolated from non fatty plant tissues. Planta 159:238–246

    CAS  Google Scholar 

  • Gerhardt B, Fischer K, Deittert M, Wenzel B (1999) Substrate inhibition and affinities of the glyoxysomal β-oxidation of sunflower cotyledons. Planta 209:355–363

    Article  CAS  PubMed  Google Scholar 

  • Graham IA, Eastmond PJ (2002) Pathways of straight and branched chain fatty acid catabolism in higher plants. Prog Lipid Res 41:156–181

    Article  CAS  PubMed  Google Scholar 

  • Gurvitz A, Hamilton B, Ruis H, Hartig A (2001) Peroxisomal degradation of trans-unsaturated fatty acids in the yeast Saccharomyces cerevisiae. J Biol Chem 276:895–903

    Article  CAS  PubMed  Google Scholar 

  • Hadjiagapiou C, Travers J, Fertel R, Sprecher H (1990) Metabolism of 15-hydroxy-5,8,11,13-eicosatetraenoic acid by MOLT-4 cells and blood T-lymphocytes. J Biol Chem 265:4369–4373

    CAS  PubMed  Google Scholar 

  • Hadjiagapiou C, Travers J, Fertel R, Sprecher H (1992) β-oxidation of 12(S)-hydroxy-5,8,10,14-eicosatetraenoic acid by MOLT-4 lymphocytes. Arch Biochem Biophys 292:112–120

    CAS  PubMed  Google Scholar 

  • Hooks MA (2002) Molecular biology, enzymology, and physiology of β-oxidation. In: Baker A, Graham IA (eds.) Plant peroxisomes. Kluwer, Dordrecht, pp 19–55

  • Huang AHC (1993) Lipases. In: Moore JTS (ed) Lipid metabolism in plants. CRC Press, London, pp 473–503

  • Huang AHC (1996) Oleosins and oil bodies in seeds and other organs. Plant Physiol 110:1055–1061

    Article  CAS  PubMed  Google Scholar 

  • Kleiter AE, Gerhardt B (1998) Glyoxysomal β-oxidation of long-chain fatty acids: completeness of degradation. Planta 206:125–130

    Article  CAS  Google Scholar 

  • Kobayashi Y, Shimazaki T, Taguchi H, Sato F (1990) Highly stereocontrolled total synthesis of leukotriene-B4, 20-hydroxyleukotriene-B4, leukotriene-B3, and their analogs. J Org Chem 55:5324–5335

    CAS  Google Scholar 

  • Liang XQ, Zhu D, Schulz H (1999) Δ3,5,7,Δ2,4,6-Trienoyl-CoA isomerase, a novel enzyme that functions in the β-oxidation of polyunsaturated fatty acids with conjugated double bonds. J Biol Chem 274:13830–13835

    Article  CAS  PubMed  Google Scholar 

  • Pantke-Böcker S, Pohnert G, Fischerlui I, Boland W, Peters AF (1995) Synthesis and absolute-configuration of desmarestene, the gamete-releasing and gamete-attracting pheromone of the brown-algae Desmarestia-Aculeata and D-Firma (Phaeophyceae). Tetrahedron 51:7927–7936

    Article  Google Scholar 

  • Sattler W, Mohr D, Stocker R (1994) Rapid isolation of lipoproteins and assessment of their peroxidation by high-performance liquid chromatography postcolumn chemiluminescence. Methods Enzymol 233:469–489

    Article  CAS  PubMed  Google Scholar 

  • Shoukry K, Schulz H (1998) Significance of the reductase-dependent pathway for the β-oxidation of unsaturated fatty acids with odd-numbered double bonds—mitochondrial metabolism of 2-trans-5-cis-octadienoyl-CoA. J Biol Chem 273:6892–6899

    Article  CAS  PubMed  Google Scholar 

  • Trelease RN, Doman DC (1984) Mobilization of oil and wax reserves. In: Murray DR (ed) Seed physiology. Academic Press, Sydney, pp 202–245

  • Vollenweider S, Weber H, Stolz S, Chetelat A, Farmer EE (2000) Fatty acid ketodienes and fatty acid ketotrienes: michael addition acceptors that accumulate in wounded and diseased Arabidopsis leaves. Plant J 24:467–476

    Article  CAS  PubMed  Google Scholar 

  • Weichert H, Kolbe A, Kraus A, Wasternack C, Feussner I (2002) Metabolic profiling of oxylipins in germinating cucumber seedlings—lipoxygenase-dependent degradation of triacylglycerols and biosynthesis of volatile aldehydes. Planta 215:612–619

    Article  CAS  PubMed  Google Scholar 

  • Woldegiorgis G, Spennetta T, Corkey BE, Williamson JR, Shrago E (1985) Extraction of tissue long-chain acyl-CoA esters and measurement by reverse-phase high-performance liquid chromatography. Anal Biochem 150:8-12

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the Deutsche Forschungsgemeinschaft SFB 363/B23 and C5 (I.F. and C.W.) and by a grant from the Deutsche Forschungsgemeinschaft to H.K. (Ku 961/7–2).

Author information

Authors and Affiliations

  1. Abteilung Biochemie der Pflanze, Georg-August-Universität Göttingen, Justus-von-Liebig-Weg 11, 37077, Göttingen, Germany

    Ivo Feussner

  2. Institut für Botanik, Universität Münster, Schlossgarten 3, 48149, Münster, Germany

    Bernt Gerhardt & Kerstin Fischer

  3. Leibniz-Institut für Pflanzenbiochemie, Weinberg 3, 06120, Halle, Germany

    Thomas J. Balkenhohl & Claus Wasternack

  4. Abteilung Bioorganische Chemie, Max-Planck-Institut für Chemische Ökologie, Hans Knöll Strasse 8, 07745, Jena, Germany

    Georg Pohnert

  5. Universitätsklinikum Charité, Institut für Biochemie, Humboldt Universität Berlin, Monbijoustrasse 2, 10115, Berlin, Germany

    Hartmut Kühn

Authors
  1. Bernt Gerhardt
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kerstin Fischer
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Thomas J. Balkenhohl
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Georg Pohnert
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hartmut Kühn
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Claus Wasternack
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Ivo Feussner
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ivo Feussner.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gerhardt, B., Fischer, K., Balkenhohl, T.J. et al. Lipoxygenase-mediated metabolism of storage lipids in germinating sunflower cotyledons and β-oxidation of (9Z,11E,13S)-13-hydroxy-octadeca-9,11-dienoic acid by the cotyledonary glyoxysomes. Planta 220, 919–930 (2005). https://doi.org/10.1007/s00425-004-1408-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-004-1408-1

Keywords

Search

Navigation