BRENDA - Enzyme Database show

Catalytic convergence of manganese and iron lipoxygenases by replacement of a single amino acid

Wennman, A.; Jerneren, F.; Hamberg, M.; Oliw, E.H.; J. Biol. Chem. 287, 31757-31765 (2012)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.13.11.45
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
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.13.11.45
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
1.13.11.45
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
1.13.11.45
K52N
site-directed mutagenesis
Gaeumannomyces graminis
1.13.11.45
L176K
site-directed mutagenesis
Gaeumannomyces graminis
1.13.11.45
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
1.13.11.45
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
1.13.11.45
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
1.13.11.45
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
1.13.11.45
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
1.13.11.45
Y158C
site-directed mutagenesis
Gaeumannomyces graminis
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.13.11.45
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
1.13.11.45
0.0027
-
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid
pH 9.0, temperature not specified in the publication, mutant L336V
Gaeumannomyces graminis
1.13.11.45
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
1.13.11.45
0.0065
-
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid
pH 9.0, temperature not specified in the publication, mutant L336V
Gaeumannomyces graminis
1.13.11.45
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
EC Number
Metals/Ions
Commentary
Organism
Structure
1.13.11.45
Mn2+
manganese-containing enzyme
Gaeumannomyces graminis
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.13.11.45
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate + O2
Gaeumannomyces graminis
-
(9Z,11E, 13R)-13-hydroperoxyoctadeca-9,11-dienoate
-
-
?
1.13.11.45
linoleate + O2
Gaeumannomyces graminis
-
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate
-
-
?
1.13.11.45
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
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.13.11.45
Gaeumannomyces graminis
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.13.11.45
recombinant mini 13R-MnLOX from Pichia pastoris by hydrophobic interaction chromatography and gel filtration
Gaeumannomyces graminis
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.13.11.45
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate + O2
-
725486
Gaeumannomyces graminis
(9Z,11E, 13R)-13-hydroperoxyoctadeca-9,11-dienoate
-
-
-
?
1.13.11.45
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate + O2
-
725486
Gaeumannomyces graminis
(9Z,11E,13R)-13-hydroperoxyoctadeca-9,11-dienoate
-
-
-
?
1.13.11.45
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid + O2
-
725486
Gaeumannomyces graminis
?
-
-
-
?
1.13.11.45
linoleate + O2
-
725486
Gaeumannomyces graminis
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate
-
-
-
?
1.13.11.45
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
725486
Gaeumannomyces graminis
?
-
-
-
-
1.13.11.45
additional information
oxidation of 18:1 n-6 is performed by 13R-MnLOX
725486
Gaeumannomyces graminis
?
-
-
-
-
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.13.11.45
9
-
assay at
Gaeumannomyces graminis
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.13.11.45
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
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.13.11.45
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
1.13.11.45
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
1.13.11.45
K52N
site-directed mutagenesis
Gaeumannomyces graminis
1.13.11.45
L176K
site-directed mutagenesis
Gaeumannomyces graminis
1.13.11.45
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
1.13.11.45
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
1.13.11.45
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
1.13.11.45
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
1.13.11.45
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
1.13.11.45
Y158C
site-directed mutagenesis
Gaeumannomyces graminis
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.13.11.45
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
1.13.11.45
0.0027
-
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid
pH 9.0, temperature not specified in the publication, mutant L336V
Gaeumannomyces graminis
1.13.11.45
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
1.13.11.45
0.0065
-
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid
pH 9.0, temperature not specified in the publication, mutant L336V
Gaeumannomyces graminis
1.13.11.45
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 (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.13.11.45
Mn2+
manganese-containing enzyme
Gaeumannomyces graminis
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.13.11.45
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate + O2
Gaeumannomyces graminis
-
(9Z,11E, 13R)-13-hydroperoxyoctadeca-9,11-dienoate
-
-
?
1.13.11.45
linoleate + O2
Gaeumannomyces graminis
-
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate
-
-
?
1.13.11.45
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
?
-
-
-
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.13.11.45
recombinant mini 13R-MnLOX from Pichia pastoris by hydrophobic interaction chromatography and gel filtration
Gaeumannomyces graminis
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.13.11.45
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate + O2
-
725486
Gaeumannomyces graminis
(9Z,11E, 13R)-13-hydroperoxyoctadeca-9,11-dienoate
-
-
-
?
1.13.11.45
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate + O2
-
725486
Gaeumannomyces graminis
(9Z,11E,13R)-13-hydroperoxyoctadeca-9,11-dienoate
-
-
-
?
1.13.11.45
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid + O2
-
725486
Gaeumannomyces graminis
?
-
-
-
?
1.13.11.45
linoleate + O2
-
725486
Gaeumannomyces graminis
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate
-
-
-
?
1.13.11.45
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
725486
Gaeumannomyces graminis
?
-
-
-
-
1.13.11.45
additional information
oxidation of 18:1 n-6 is performed by 13R-MnLOX
725486
Gaeumannomyces graminis
?
-
-
-
-
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.13.11.45
9
-
assay at
Gaeumannomyces graminis
General Information
EC Number
General Information
Commentary
Organism
1.13.11.45
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
1.13.11.45
additional information
pentamer motif His-Val-Leu-Phe-His in MnLOX, 13R-MnLOX3 catalyzes suprafacial hydrogen abstraction and oxygenation
Gaeumannomyces graminis
General Information (protein specific)
EC Number
General Information
Commentary
Organism
1.13.11.45
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
1.13.11.45
additional information
pentamer motif His-Val-Leu-Phe-His in MnLOX, 13R-MnLOX3 catalyzes suprafacial hydrogen abstraction and oxygenation
Gaeumannomyces graminis