Information on EC 1.13.11.45 - linoleate 11-lipoxygenase

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The expected taxonomic range for this enzyme is: Eukaryota

EC NUMBER
COMMENTARY
1.13.11.45
-
RECOMMENDED NAME
GeneOntology No.
linoleate 11-lipoxygenase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
linoleate + O2 = (9Z,12Z)-(11S)-11-hydroperoxyoctadeca-9,12-dienoate
show the reaction diagram
-
-
-
-
linoleate + O2 = (9Z,12Z)-(11S)-11-hydroperoxyoctadeca-9,12-dienoate
show the reaction diagram
mechanism via suprafacial migration of O2
-
linoleate + O2 = (9Z,12Z)-(11S)-11-hydroperoxyoctadeca-9,12-dienoate
show the reaction diagram
reaction mechanism, tail-first model with rearrangement at C13 at the end of lipoxygenation, influenced by double bond configuration and the chain length of fatty acids, overview
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
oxidation
-
-
-
-
oxidation
Q8X151
possesses also 1% of hydroperoxide isomerase activity
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Linoleic acid metabolism
-
SYSTEMATIC NAME
IUBMB Comments
linoleate:oxygen 11S-oxidoreductase
The product (9Z,12Z)-(11S)-11-hydroperoxyoctadeca-9,12-dienoate, is converted, more slowly, into (9Z,11E)-(13R)-13-hydroperoxyoctadeca-9,11-dienoate. The enzyme from the fungus Gaeumannomyces graminis requires Mn2+. It also acts on alpha-linolenate, whereas gamma-linolenate is a poor substrate. Oleate and arachidonate are not substrates.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
linoleate dioxygenase
-
-
-
-
LOX-1
-
-
manganese lipoxygenase
-
-
-
-
manganese lipoxygenase
-
-
CAS REGISTRY NUMBER
COMMENTARY
9029-60-1
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
expression in Pichia pastoris
-
-
Manually annotated by BRENDA team
variant avenae
TrEMBL
Manually annotated by BRENDA team
soy bean
-
-
Manually annotated by BRENDA team
navy bean
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
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
evolution
-
9S-LOX contains catalytic manganese, and its sequence can be aligned with 77% identity to 13R-LOX with catalytic manganese lipoxygenase of the Take-all fungus. Alterations in the Sloane determinant of 9S-LOX and 13R-MnLOX with larger and smaller hydrophobic residues interconvert the regiospecific oxidation of 18:2n-6, presumably by altering the substrate position in relation to oxygen insertion
additional information
-
pentamer motif His-Val-Leu-Phe-His in MnLOX, 13R-MnLOX3 catalyzes suprafacial hydrogen abstraction and oxygenation
additional information
-
alterations in the Sloane determinant of 9S-LOX and 13R-MnLOX with larger and smaller hydrophobic residues interconvert the regiospecific oxidation of 18:2n-6, presumably by altering the substrate position in relation to oxygen insertion. The catalytic domain of 13R-MnLOX contains a pentamer motif flanked by two His metal ligands, His-Val-Leu-Phe-His, in the presumed manganese binding region. 9S-MnLOX, EC 1.13.11.58, catalyzes hydrogen abstraction at C-11 and oxygenation at C-9 and C-11 in a suprafacial manner in analogy with 13R-MnLOX
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(10Z,13Z,16Z)-nonadeca-10,13,16-trienoic acid + O2
?
show the reaction diagram
-
oxidation at the n-6 carbon and at the bis-allylic position n-8
-
-
?
(11Z,14Z)-eicosadienoic acid + O2
15-hydroxyperoxy-(9Z,11E)-eicosadienoic acid
show the reaction diagram
-
-
-
-
?
(11Z,14Z,17Z)-eicosa-11,14,17-trienoic acid + O2
15-hydroxyperoxy-11Z,13E,17Z-eicosatrienoic acid
show the reaction diagram
-
via 11-hydroperoxyoctadecadienoic acid, 13-hydroperoxyoctadecadienoic acid and 15-hydroperoxyoctadecadienoic acid
-
-
?
(11Z,14Z,17Z)-eicosa-11,14,17-trienoic acid + O2
?
show the reaction diagram
-
oxidation at the n-6 carbon and at the bis-allylic position n-8
-
-
?
(13R)-hydroperoxylinolenic acid + O2
13-ketolinolenic acid + epoxyalcohols
show the reaction diagram
Q8X151
only mutant enzyme G316A
erythro- and threo-11-hydroxy-(12R,13R)-epoxy-(9Z,15Z)-octadecadienoic acids and one of the corresponding cis-epoxides as major products
-
?
(5E,8E,11E,14E)-icosa-5,8,11,14-tetraenoic acid + O2
(5Z,8Z,11S,12E,14Z)-11-hydroperoxyicosa-5,8,12,14-tetraenoic acid + (5Z,8Z,11Z,13E,15R)-15-hydroperoxyicosa-5,8,11,13-tetraenoic acid
show the reaction diagram
-
oxidation at the n-6 carbon and at the bis-allylic position n-8
-
-
?
(5Z,8Z,11Z,14Z,17Z)-docosa-5,8,11,14,17-pentaenoate + O2
?
show the reaction diagram
-
oxidation at the n-6 carbon and at the bis-allylic position n-8
-
-
?
(7Z,10Z,13Z,16Z,19Z)-docosa-7,10,13,16,19-pentaenoate + O2
?
show the reaction diagram
-
oxidation at the n-6 carbon and at the bis-allylic position n-8
-
-
?
(9E,12Z)-octadeca-9,12-dienoic acid + O2
?
show the reaction diagram
-
sLOX-1 oxidizes (9E,12Z)18:2 to 9-hydroperoxyoctadecadienoic acid in an R/S ratio of 60/40 and to 13-hydroperoxyoctadecadienoic acid
-
-
?
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate + O2
(9Z,11E, 13R)-13-hydroperoxyoctadeca-9,11-dienoate
show the reaction diagram
-
-
-
-
?
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate + O2
(9Z,11E,13R)-13-hydroperoxyoctadeca-9,11-dienoate
show the reaction diagram
-
-
-
-
?
(9Z,12E)-octadeca-9,12-dienoic acid + O2
?
show the reaction diagram
-
sLOX-1 oxidizes (9Z,12E)18:2 slowly to the 9-hydroperoxy metabolite with 10E,12E configuration as the main product (65%) and to 13-hydroperoxyoctadecadienoic acid
-
-
?
(9Z,12Z)-octadeca-9,12-dienoic acid + O2
(9S)-hydroperoxy-octadeca-10,12-dienoate
show the reaction diagram
-
alpha-linoleate is converted via two intermediates, (11S)-hydroperoxy-(9Z,12Z)-octadecenoate and (13R)-hydroperoxy-(9Z,11E)-octadecadienoate, which are isomerized to the end product, probably after oxidation to peroxyl radicals, beta-fragmentation, and oxygen insertion at C-9
-
-
?
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid + O2
(13R)-hydroperoxyoctadecadienoic acid
show the reaction diagram
-
via bis-allylic 11S-hydroperoxy fatty acid
-
-
?
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid + O2
?
show the reaction diagram
-
-
-
-
?
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid + O2
?
show the reaction diagram
-
oxidation at the bis-allylic n-5 carbon and at positions n-3, n-7, and n-6
-
-
?
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid + O2
?
show the reaction diagram
-
oxidation at the n-3 position
-
-
?
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid + O2
(10E,12E,14E)-9,16-dihydroxy-octadeca-10,12,14-trienoate
show the reaction diagram
-
gamma-linoleate is oxidized at C-9, C-11, and C-13
-
-
?
1,2-dilinoleoyl-sn-glycero-3-phosphocholine + O2
?
show the reaction diagram
-
-
main products with 14% and 10%, no production of 11-hydroxyoctadecanoic acid
-
?
1-linoleyl-lyso-glycerophosphatidylcholine + O2
11-hydroxyoctadecanoic acid + 13-hydroxyoctadecanoic acid
show the reaction diagram
-
via formation of the 13-hydroxyperoxy metabolite
-
-
?
11-(11R)-hydroperoxy-(9Z,12Z)-octadecadienoic acid
9-(9R)-hydroperoxy-octadecadienoic acid
show the reaction diagram
-
slow
-
-
?
11-(11S)-hydroperoxy-(9Z,12Z)-octadecadienoic acid
13-(13R)-hydroperoxy-(9Z,11E)-octadecadienoic acid
show the reaction diagram
-
47% the rate of conversion of linolenic acid
-
-
?
11-(11S)-hydroperoxy-(9Z,12Z)-octadecadienoic acid
13-(13R)-hydroperoxy-(9Z,11E)-octadecadienoic acid
show the reaction diagram
-
rapid
-
-
?
11-(11S)-hydroperoxy-(9Z,12Z)-octadecadienoic acid methyl ester
13-(13R)-hydroperoxy-(9Z,11E)-octadecadienoic acid methyl ester
show the reaction diagram
-
-
-
-
?
11-hydroperoxyoctadecadienoic acid + O2
11,18-dihydroxy-12E,14Z,16E-eicosatrienoic acid
show the reaction diagram
-
-
two diastereoisomers
-
?
alpha-linolenic acid + O2
?
show the reaction diagram
-
-
-
-
?
alpha-linolenic acid + O2
(9Z,11E,15Z)-(13)-hydroperoxyoctadecatrienoic acid
show the reaction diagram
-
-
-
-
?
alpha-linolenic acid + O2
(13R)-hydroperoxy octadecadienoic acid + 9-hydroperoxyoctadecadienoic acid
show the reaction diagram
-
-
-
-
?
gamma-linolenic acid + O2
(6Z,9Z,11E)-(13)-hydroperoxyoctadecatrienoic acid + (6Z,9Z,12Z)-(11)-hydroperoxyoctadecatrienoic acid
show the reaction diagram
-
-
-
-
?
linoleate + O2
(9Z,12Z)-(11S)-11-hydroperoxyoctadeca-9,12-dienoate
show the reaction diagram
-
-
-
-
?
linoleate + O2
(9Z,12Z)-(11S)-11-hydroperoxyoctadeca-9,12-dienoate
show the reaction diagram
-
via bis-allylic 11S-hydroperoxy fatty acid
-
-
?
linoleate + O2
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate
show the reaction diagram
-
-
-
-
?
linoleic acid + O2
(9Z,11E)-(13R)-hydroperoxyoctadecadienoic acid
show the reaction diagram
-
-
-
-
?
linoleic acid + O2
(9Z,11E)-(13R)-hydroperoxyoctadecadienoic acid
show the reaction diagram
-
-
-
-
?
linoleic acid + O2
(9Z,11E)-(13R)-hydroperoxyoctadecadienoic acid
show the reaction diagram
Q8X151
-
-
-
?
linoleic acid + O2
(9Z,11E)-(13R)-hydroperoxyoctadecadienoic acid + (9Z,12Z)-(11S)-hydroperoxyoctadecadienoic acid
show the reaction diagram
-
-
-
ir
linoleic acid + O2
?
show the reaction diagram
-
-
-
-
?
linolenic acid + O2
11-(11S)-hydroperoxy-(9Z,12Z)-octadecadienoic acid
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
Mn-LO likely binds fatty acids tail-first and oxygenates many C16, C18, C20, and C22 fatty acids to significant amounts of bis-allylic hydroperoxides. Unsaturated C16-C22 fatty acids, other than 18:3n-3, 18:2n-6, or 17:3n-3, are poor substrates, possibly because of ineffective enzyme activation through Mn2+ to Mn3+ by the produced hydroperoxides, substrate specificities of wild-type and mutant enzymes, overview
-
-
-
additional information
?
-
-
the enzyme catalyzes the rearrangement of bis-allylic S-hydroxyperoxides to allylic R-hydroperoxides, and the oxygenation of 18:2n-6 by suprafacial hydrogen abstraction at C11 and O2 insertion at the bis-allylic position C11 and, with double bond migation, at the allylic position C13, overview, 20:4n-6 is a poor substrate, substrate specificities of wild-type and mutant enzymes, overview
-
-
-
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
-
-
-
additional information
?
-
-
(9Z,12E)18:2 is not oxidized at a detectable rate. sLOX-1 oxidizes (9E,12Z)18:2 more efficiently than (9Z,12E)18:2
-
-
-
additional information
?
-
-
cf. EC 1.13.11.58, LC- and GC-MS analysis product analysis, overview
-
-
-
additional information
?
-
-
oxidation of 18:1 n-6 is performed by 13R-MnLOX
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate + O2
(9Z,11E, 13R)-13-hydroperoxyoctadeca-9,11-dienoate
show the reaction diagram
-
-
-
-
?
linoleate + O2
(9Z,11S,12Z)-11-hydroperoxyoctadeca-9,12-dienoate
show the reaction diagram
-
-
-
-
?
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
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
maximum activity between 0.24 and 0.49 mM in phosphate buffer
Ca2+
-
maximum activity between 0.24 and 0.92 mM in phosphate buffer
Fe2+
-
non-heme iron
Manganese
-
mononuclear Mn center, 0.94 mol Mn per mol of protein
Mg2+
-
activation less effective than with Ca2+ in phosphate buffer
Mn2+
-
activation less effective than with Ca2+ in phosphate buffer
Mn2+
-
contains 0.5-1.0 atom per enzyme molecule
Mn2+
-
1 mol per mol of protein
Mn2+
-
a Mn-lipoxygenase
Mn2+
-
required for activity
Mn2+
-
manganese-containing enzyme
Mn2+
-
manganese 13R-lipoxygenase contains catalytic manganese, the catalytic domain of 13R-MnLOX contains a pentamer motif flanked by two His metal ligands, His-Val-Leu-Phe-His, in the presumed manganese binding region
additional information
-
no activity with Fe3+
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
arachidonic acid
-
inhibits at 0.01 mM
eicosatetraynoic acid
-
50% of control activity at 0.01 mM
N-(3-phenoxycinnamyl) acetohydroxamic acid
Q8X151
0.3 mM, complete inhibition
N-(3-phenoxycinnamyl)acetohydroxamic acid
-
50% of control activity at 0.05 mM
Tween 20
-
inhibition of Ca2+-activated enzyme at pH 7.5 and 6.4, no inhibition at pH 6.4 in the absence of Ca2+
eicosatetraynoic acid
Q8X151
0.1 mM, 75% inhibition
additional information
-
no inhibition by lipoxygenase inhibitors zileuton, esculetin and ICI 230487
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.01
-
(13R)-hydroperoxylinolenic acid
Q8X151
mutant enzyme G316A
0.0005
-
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid
-
pH 9.0, temperature not specified in the publication, wild-type enzyme
0.0027
-
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid
-
pH 9.0, temperature not specified in the publication, mutant L336V
0.0036
-
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid
-
pH 9.0, temperature not specified in the publication, mutants L336A and L336F
0.0065
-
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid
-
pH 9.0, temperature not specified in the publication, mutant L336V
0.0073
-
(9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid
-
pH 9.0, temperature not specified in the publication, mutant F337I
0.0024
-
alpha-linolenic acid
-
-
0.0071
-
alpha-linolenic acid
-
pH 9.0, 21C
0.0024
-
linoleate
-
pH 7.5, 18:3n-3 substrate
0.0044
-
linoleic acid
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
40
-
alpha-linolenic acid
-
-
40
-
linoleate
-
pH 7.5, 18:3n-3 substrate
18.3
-
linoleic acid
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
18
-
-
pH 9.0, 21C
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.4
-
-
in the absence of Ca2+
7
-
-
broad pH optimum
7.2
-
-
in the presence of 0.37 mM Ca2+
7.5
-
-
assay at
9
-
-
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
7
-
in the absence of Ca2+
4
9
-
in the presence of 0.37 mM Ca2+
5
11
-
at pH 5.0 64% of maximum activity, at pH 11 69% of maximum activity
7
-
-
93% of maximum activity
8
-
-
98% of maximum activity
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
23
-
-
assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
12
60
-
8.8% of maximal activity at 12C and 100% of maximal activity at 60C, activity decreases sharply at 63C
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
73000
-
-
SDS-PAGE after deglycosylation
135000
-
-
gel filtration with Superdex 200 in 6 M urea
160000
-
-
gel filtration with Superdex 200
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 80-100000, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
glycoprotein
-
-
glycoprotein
-
N-and O-glycosilation
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
63
-
-
incubation for 5 min over 63C decreases activity, no activity at 75C
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
9S-LOX activity is less robust during expression and purification, possibly due to proteolysis, stability is not improved by addition of 0.001 mM pepstatin A, 0.001 mM leupeptin, or 1 mM EDTA to the growth medium
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
4C, 3 mM NaN3, 0.2 mM GSH, several months, no loss of activity
-
4C, stable for months
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
phenyl-Sepharose column chromatography
Q8X151
recombinant mini 13R-MnLOX from Pichia pastoris by hydrophobic interaction chromatography and gel filtration
-
recombinant wild-type and mutant enzymes from Pichia pastoris by hydrophobic interaction chromatography, dialfiltration, and gel filtration
-
recombinant wild-type and mutant enzymes from Pichia pastoris cell medium by hydrophobic interaction and ion exchange chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
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
-
expressed in Pichia pastoris
Q8X151
expression in Pichia pastoris
-
expression of wild-type and G316A mutant enzymes in Pichia pastoris strain X-33, secretion to the cell medium
-
recombinant expression of wild-type and mutant enzymes in Pichia pastoris
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
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
F347A
-
site-directed mutagenesis, the mutant oxidizes octadeca-9,11-dienoic acid to 11-hydroperoxyoctadecadienoate/(13R)-hydroperoxyoctadecadienoate/(9S)-hydroperoxyoctadecadienoate in an initial ratio of 50/42/8 and with almost complete consumption of 11-hydroperoxyoctadecadienoate to a ratio of 2/84/14. The (9E,11Z,15E)-octadeca-9,11,15-trienoic acid is transformed to 11- and (13R)-hydroperoxyoctadecatrienoic acid and to traces of (9S)-hydroperoxyoctadecatrienoic acid, essentially as native 13R-MnLOX
F347L
-
site-directed mutagenesis, (9E,11Z,15E)-octadeca-9,11,15-trienoic acid is oxidized to a 2:3 mixture of (9S)- and (13R)-dihydroxyoctadecatrienoic acid
F347V
-
site-directed mutagenesis
G316A
Q8X151
catalytically active, 7-8% of hydroperoxide isomerase activity
G316A
-
site-directed mutagenesis, the mutant MnIII-LO G316A oxygenates mainly 16:3n-3 at positions n-7 and n-6, 19:3n-3 at n-10, n-8, and n-6, and 20:3n-3 at n-10 and n-8
G316A
-
site-directed mutagenesis, the mutant changes the the position of lipoxygenation toward the carboxyl group of 20:2n-6 and 20:3n-3 and prevents the bis-allylic hydroperoxide of 20:3n-3 but not of 20:2n-6to interact with the catalytic metall
G316T
Q8X151
catalytically inactive
G316V
Q8X151
catalytically inactive
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
H274Q
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no enzymic activity, loss of more than 95% of manganese content
H278E
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no enzymic activity, loss of more than 95% of manganese content
H462E
-
no enzymic activity, loss of more than 95% of manganese content
H463Q
-
no significant change in activity
K52N
-
site-directed mutagenesis
L176K
-
site-directed mutagenesis
L336A
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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
L336F
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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
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
N466L
-
no significant change in activity
N466L
-
site-directed mutagenesis, the mutation hardly affects the bis-allylic hydroperoxide rearrangement
N466Q
-
no significant change in activity
Q467N
-
no significant change in activity
S469A
-
site-directed mutagenesis, the mutation hardly affects the bis-allylic hydroperoxide rearrangement, the mutant shows the same substrate profile as the wild-type enzyme
Y158C
-
site-directed mutagenesis
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
additional information
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V602 deletion mutant, no enzymic activity, loss of more than 95% of manganese content
additional information
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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
additional information
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replacement Phe347 in 13R-MnLOX and changes the regiospecific oxidation of 18:2 n-6 in a consistent way, but the n-3 double bond of 18:3n-3 can reduce this effect. Mutations are designed to convert the pentamer motif to a hexamer motif to mimic FeLOX, but the mutants with the His-Val-Leu-Phe-Thr-His and His-Val-Leu-Phe-Gly-His motives are inactive, overview