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Secretion of two no...
Secretion of two novel enzymes, manganese 9S-Lipoxygenase and epoxy alcohol synthase, by the rice pathogen Magnaporthe salvinii
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- Wennman, Anneli, 1984- (författare)
- Uppsala universitet,Institutionen för farmaceutisk biovetenskap,biokemisk farmakologi
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- Oliw, Ernst H, 1948- (författare)
- Uppsala universitet,Institutionen för farmaceutisk biovetenskap,biokemisk farmakologi
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(creator_code:org_t)
- 2013
- 2013
- Engelska.
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Ingår i: Journal of Lipid Research. - 0022-2275 .- 1539-7262. ; 54:3, s. 762-775
- Relaterad länk:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- The mycelium of the rice stem pathogen, Magnaporthe salvinii, secreted linoleate 9S-lipoxygenase (9S-LOX) and epoxy alcohol synthase (EAS). The EAS rapidly transformed 9S-hydroperoxy-octadeca-10E,12Z-dienoic acid (9S-HPODE) to threo 10(11)-epoxy-9S-hydroxy-12Z-octadecenoic acid, but other hydroperoxy fatty acids were poor substrates. 9S-LOX was expressed in Pichia pastoris. Recombinant 9S-LOX oxidized 18:2n-6 directly to 9S-HPODE, the end product, and also to two intermediates, 11S-hydroperoxy-9Z,12Z-octadecenoic acid (11S-HPODE; ~5%) and 13R-hydroperoxy-9Z,11E-octadecenoic acid (13R-HPODE; ~1%). 11S- and 13R-HPODE were isomerized to 9S-HPODE, likely after oxidation to peroxyl radicals, β-fragmentation, and oxygen insertion at C-9. 18:3n-3 was oxidized at C-9, C-11, and C-13, and to 9,16-dihydroxy-10E,12,14E-octadecatrienoic acid. 9S-LOX contained catalytic manganese (Mn:protein ≥ 0.2:1; Mn/Fe, 1:0.05), and its sequence could be aligned with 77% identity to 13R-LOX with catalytic manganese (13R-MnLOX) of the Take-all fungus. The Leu350Met mutant of 9S-LOX shifted oxidation of 18:2n-6 from C-9 to C-13, and the Phe347Leu, Phe347Val, and Phe347Ala mutants of 13R-MnLOX from C-13 to C-9. In conclusion, M. salvinii secretes 9S-LOX with catalytic manganese along with a specific EAS. Alterations in the Sloane determinant of 9S-LOX and 13R-MnLOX with larger and smaller hydrophobic residues interconverted the regiospecific oxidation of 18:2n-6, presumably by altering the substrate position in relation to oxygen insertion.
Nyckelord
- Biokemisk farmakologi
- Biochemical Pharmacology
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