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Literature summary for 1.13.11.45 extracted from

  • Wennman, A.; Jerneren, F.; Hamberg, M.; Oliw, E.H.
    Catalytic convergence of manganese and iron lipoxygenases by replacement of a single amino acid (2012), J. Biol. Chem., 287, 31757-31765.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
construction, expression, and secretion of truncated 13R-MnLOX, from mini DNA coding for the sequence Glu-Phe-His6-Leu-Gln-Thr39-Val-Leu-Pro to Val618, in Pichia pastoris Gaeumannomyces graminis

Protein Variants

Protein Variants Comment Organism
F337I site-directed mutagenesis, replacement of Phe337 with Ile changes the stereochemistry of the 13-hydroperoxy metabolites of 18:2n-6 and 18:3n-3 (from 100% R to 69-74% S) with little effect on regiospecificity. The abstraction of the pro-S hydrogen of 18:2n-6 is retained, suggesting antarafacial hydrogen abstraction and oxygenation. The mutant shows highly reduced activity compared to the wild-type enzyme Gaeumannomyces graminis
G332A site-directed mutagenesis, replacement of Gly332 with the larger hydrophobic residue selectively augments dehydration of (9Z,11E,13R,15Z)-13-hydroperoxyoctadeca-9,12,15-trienoic acid and increases the oxidation at C-13 of 18:1n-6 Gaeumannomyces graminis
K52N site-directed mutagenesis Gaeumannomyces graminis
L176K site-directed mutagenesis Gaeumannomyces graminis
L336A site-directed mutagenesis, replacement of Leu336 with the smaller hydrophobic residue shifts the oxygenation from C-13 toward C-9 with formation of 9S- and 9R-hydroperoxy metabolites of 18:2n-6 and 18:3n-3, the mutant shows highly reduced activity compared to the wild-type enzyme Gaeumannomyces graminis
L336F site-directed mutagenesis, replacement of Leu336 with the larger hydrophobic residue selectively augments dehydration of (9Z,11E,13R,15Z)-13-hydroperoxyoctadeca-9,12,15-trienoic acid and increases the oxidation at C-13 of 18:1n-6, the mutant shows highly reduced activity compared to the wild-type enzyme Gaeumannomyces graminis
L336G site-directed mutagenesis, replacement of Leu336 with the smaller hydrophobic residue shifts the oxygenation from C-13 toward C-9 with formation of 9S- and 9R-hydroperoxy metabolites of 18:2n-6 and 18:3n-3, the mutant shows highly reduced activity compared to the wild-type enzyme Gaeumannomyces graminis
L336V site-directed mutagenesis, replacement of Leu336 with the smaller hydrophobic residue shifts the oxygenation from C-13 toward C-9 with formation of 9S- and 9R-hydroperoxy metabolites of 18:2n-6 and 18:3n-3, the mutant shows highly reduced activity compared to the wild-type enzyme Gaeumannomyces graminis
additional information hydrophobic replacements of Leu336 can modify the hydroperoxide configurations at C-9 with little effect on the R configuration at C-13 of the 18:2n-6 and 18:3n-3 metabolites Gaeumannomyces graminis
Y158C site-directed mutagenesis Gaeumannomyces graminis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0005
-
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid pH 9.0, temperature not specified in the publication, wild-type enzyme Gaeumannomyces graminis
0.0027
-
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid pH 9.0, temperature not specified in the publication, mutant L336V Gaeumannomyces graminis
0.0036
-
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid pH 9.0, temperature not specified in the publication, mutants L336A and L336F Gaeumannomyces graminis
0.0065
-
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid pH 9.0, temperature not specified in the publication, mutant L336V Gaeumannomyces graminis
0.0073
-
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid pH 9.0, temperature not specified in the publication, mutant F337I Gaeumannomyces graminis

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ manganese-containing enzyme Gaeumannomyces graminis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate + O2 Gaeumannomyces graminis
-
(9Z,11E, 13R)-13-hydroperoxyoctadeca-9,11-dienoate
-
?
linoleate + O2 Gaeumannomyces graminis
-
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate
-
?
additional information Gaeumannomyces graminis 13R-MnLOX forms (11S)-hydroperoxyoctadecadienoic acid and (11R)-hydroperoxyoctadecatrienoic acid as intermediates during the linear phase of oxidation and the (13R)-hydroperoxides as main end products ?
-
?

Organism

Organism UniProt Comment Textmining
Gaeumannomyces graminis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant mini 13R-MnLOX from Pichia pastoris by hydrophobic interaction chromatography and gel filtration Gaeumannomyces graminis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate + O2
-
Gaeumannomyces graminis (9Z,11E, 13R)-13-hydroperoxyoctadeca-9,11-dienoate
-
?
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate + O2
-
Gaeumannomyces graminis (9Z,11E,13R)-13-hydroperoxyoctadeca-9,11-dienoate
-
?
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid + O2
-
Gaeumannomyces graminis ?
-
?
linoleate + O2
-
Gaeumannomyces graminis (9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate
-
?
additional information 13R-MnLOX forms (11S)-hydroperoxyoctadecadienoic acid and (11R)-hydroperoxyoctadecatrienoic acid as intermediates during the linear phase of oxidation and the (13R)-hydroperoxides as main end products Gaeumannomyces graminis ?
-
?
additional information oxidation of 18:1 n-6 is performed by 13R-MnLOX Gaeumannomyces graminis ?
-
?

Synonyms

Synonyms Comment Organism
13R-MnLOX
-
Gaeumannomyces graminis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
assay at Gaeumannomyces graminis

General Information

General Information Comment Organism
evolution overview of possible lipoxygenation positions of linoleic acid and partial alignment of 13R-MnLOX, sLOX-1, 8R-LOX, human 5-LOX, and rabbit 15-LOX covering an important region for regio- and stereospecificity. A few LOXs, including 9S-MnLOX, deviate from this rule of R/S stereospecificity Gaeumannomyces graminis
additional information pentamer motif His-Val-Leu-Phe-His in MnLOX, 13R-MnLOX3 catalyzes suprafacial hydrogen abstraction and oxygenation Gaeumannomyces graminis