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Linolenate 9R-Dioxygenase and Allene Oxide Synthase Activities of Lasiodiplodia theobromae

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Lipids

Abstract

Jasmonic acid (JA) is synthesized from linolenic acid (18:3n-3) by sequential action of 13-lipoxygenase, allene oxide synthase (AOS), and allene oxide cyclase. The fungus Lasiodiplodia theobromae can produce large amounts of JA and was recently reported to form the JA precursor 12-oxophytodienoic acid. The objective of our study was to characterize the fatty acid dioxygenase activities of this fungus. Two strains of L. theobromae with low JA secretion (~0.2 mg/L medium) oxygenated 18:3n-3 to 5,8-dihydroxy-9Z,12Z,15Z-octadecatrienoic acid as well as 9R-hydroperoxy-10E,12Z,15Z-octadecatrienoic acid, which was metabolized by an AOS activity into 9-hydroxy-10-oxo-12Z,15Z-octadecadienoic acid. Analogous conversions were observed with linoleic acid (18:2n-6). Studies using [11S-2H]18:2n-6 revealed that the putative 9R-dioxygenase catalyzed stereospecific removal of the 11R hydrogen followed by suprafacial attack of dioxygen at C-9. Mycelia from these strains of L. theobromae contained 18:2n-6 as the major polyunsaturated acid but lacked 18:3n-3. A third strain with a high secretion of JA (~200 mg/L) contained 18:3n-3 as a major fatty acid and produced 5,8-dihydroxy-9Z,12Z,15Z-octadecatrienoic acid from added 18:3n-3. This strain also lacked the JA biosynthetic enzymes present in higher plants.

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Abbreviations

AOS:

Allene oxide synthase

CP:

Chiral phase

P450:

Cytochrome P450

DiHODE:

Dihydroxyoctadecadienoic acid

DOX:

Dioxygenase

HHDTrE:

Hydroxyhexadecatrienoic acid

HPHDTrE:

Hydroperoxyhexadecatrienoic acid

HOME:

Hydroxyoctadecenoic acid

HPODE:

Hydroperoxyoctadecadienoic acid

HPOTrE:

Hydroperoxyoctadecatrienoic acid

JA:

(−)-Jasmonic acid

JAs:

Jasmonates

(+)-7-iso-JA:

3R,7S-JA

LDS:

Linoleate diol synthase

LOX:

Lipoxygenase

MO:

Methyloxime

NP:

Normal phase

OPDA:

Oxophytodienoic acid

RP:

Reversed phase

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Acknowledgments

Supported by VR Medicine (03X-06523) and by The Knut and Alice Wallenberg Foundation (KAW 2004.123). F Eng was supported by a project of Dr Mariano Gutierrez-Rojas from UAM Campus Iztapalapa (Mexico). Presented at the Third European Workshop on Lipid Mediators, Pasteur Institute, Paris June 3–4, 2010.

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Authors and Affiliations

  1. Department of Pharmaceutical Biosciences, Uppsala Biomedical Center, Uppsala University, P.O. Box 591, 751 24, Uppsala, Sweden

    Fredrik Jernerén & Ernst H. Oliw

  2. Cuban Research Institute on Sugar Cane Byproducts, Via Blanca and Carretera Central 804, P.O. Box 4026, Havana, Cuba

    Felipe Eng

  3. Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 77, Stockholm, Sweden

    Mats Hamberg

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  1. Fredrik Jernerén
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  2. Felipe Eng
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  3. Mats Hamberg
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  4. Ernst H. Oliw
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Correspondence to Ernst H. Oliw.

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Jernerén, F., Eng, F., Hamberg, M. et al. Linolenate 9R-Dioxygenase and Allene Oxide Synthase Activities of Lasiodiplodia theobromae . Lipids 47, 65–73 (2012). https://doi.org/10.1007/s11745-011-3622-5

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  • DOI: https://doi.org/10.1007/s11745-011-3622-5

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