Information on EC 1.13.11.77 - oleate 10S-lipoxygenase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.13.11.77
-
RECOMMENDED NAME
GeneOntology No.
oleate 10S-lipoxygenase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
alpha-linolenate + O2 = (8E,10S,12Z,15Z)-10-hydroperoxyoctadeca-8,12,15-trienoate
show the reaction diagram
(3)
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-
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linoleate + O2 = (8E,10S,12Z)-10-hydroperoxyoctadeca-8,12-dienoate
show the reaction diagram
(2)
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-
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oleate + O2 = (8E,10S)-10-hydroperoxyoctadeca-8-enoate
show the reaction diagram
(1)
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-
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SYSTEMATIC NAME
IUBMB Comments
oleate:oxygen (10S)-oxidoreductase
Binds Fe2+. The enzyme isolated from the bacterium Pseudomonas sp. 42A2 has similar activity with all the three Delta9 fatty acids. cf. EC 1.13.11.62, linoleate 10R-lipoxygenase.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
linoleate 10S-dioxygenase is an ancient relative of cyclooxygenase in cyanobacteria, linoleate 10S-dioxygenase works in tandem with a catalase-related protein with specific 10S-hydroperoxide lyase activity. Neighboring cyanobacterial genes, dioxygenase and catalase, are identified as linoleate 10S-dioxygenase and 10Shydroperoxide lyase, respectively. The Nostoc linoleate 10S-dioxygenase, the sequence of which contains the signature catalytic sequence of cyclooxygenases and fungal linoleate dioxygenases (YRWH), appears to be a heme dioxygenase ancestor
metabolism
the immediate downstream gene, Np-cat, encodes a specific linoleate 10S-hydroperoxide lyase (10S-hydroperoxyoleate being metabolized at only 3% of the rate) converting the product of Np-diox and working in tandem with it
physiological function
enzyme Np-diox is an oleate and linoleate heme 10S-dioxygenase-alpha-linolenate is also efficiently metabolized at lower concentrations, but it much more rapidly leads to enzyme inactivation
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
alpha-linolenate + O2
(8E,10S,12Z,15Z)-10-hydroperoxyoctadeca-8,12,15-trienoate
show the reaction diagram
arachidonate + O2
?
show the reaction diagram
cis-12-octadecenoate + O2
?
show the reaction diagram
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8.9% activity compared to linoleate
-
-
?
linoleate + O2
(8E,10S,12Z)-10-hydroperoxyoctadeca-8,12-dienoate
show the reaction diagram
oleate + O2
(8E,10S)-10-hydroperoxyoctadeca-8-enoate
show the reaction diagram
petroselinoate + O2
?
show the reaction diagram
-
26.4% activity compared to linoleate
-
-
?
triolein + O2
?
show the reaction diagram
-
0.4% activity compared to linoleate
-
-
?
vaccenic acid + O2
?
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
alpha-linolenate + O2
(8E,10S,12Z,15Z)-10-hydroperoxyoctadeca-8,12,15-trienoate
show the reaction diagram
linoleate + O2
(8E,10S,12Z)-10-hydroperoxyoctadeca-8,12-dienoate
show the reaction diagram
oleate + O2
(8E,10S)-10-hydroperoxyoctadeca-8-enoate
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Fe2+
-
the enzyme contains 0.55 mol of Fe2+ per mol of protein
Mg2+
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the presence of 1 mM Mg2+ increases enzyme activity 1.5fold
Mn2+
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the enzyme contains 25 mmol Mn2+ per mol of protein
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(8,10,12-EEZ)-conjugated linoleic acid derivative
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strong inhibition
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alpha-calendic acid
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(8,10,12-ZEE)-conjugated linoleic acid derivative, strong inhibition
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alpha-eleostearic acid acid
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(9,11,13-ZEE)-conjugated linoleic acid derivative, strong inhibition
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alpha-linolenic acid
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13% inhibition
Ba2+
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complete inhibition at 1 mM
beta-calendic acid
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(8,10,12-EEE)-conjugated linoleic acid derivative, complete inhibition
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beta-eleostearic acid acid
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(9,11,13-EEE)-conjugated linoleic acid derivative, complete inhibition
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catalpic acid
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(9,11,13-EEZ)-conjugated linoleic acid derivative
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EDTA
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99% inhibition at 5 mM
Fe3+
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complete inhibition at 1 mM
jacaric acid
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(8,10,12-ZEZ)-conjugated linoleic acid derivative, low inhibition
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Mg2+
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concentrations of Mg2+ higher than 1.25 mM inhibit activity strongly (90% of activity lost at 1.75 mM)
oleate
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at concentrations of oleate higher than 2.1 mM, the enzyme activity is inhibited by excess of substrate
oxidized linoleic acid
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6% inhibition
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punicic acid
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(9,11,13-ZEZ)-conjugated linoleic acid derivative
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Zn2+
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complete inhibition at 1 mM
additional information
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
chaperone Gro7
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activates the enzyme about 200%
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.013 - 0.73
alpha-linolenate
0.063 - 0.66
linoleate
0.113 - 0.74
oleate
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
31
alpha-linolenate
recombinant enzyme, pH and temperature not specified in the publication
135
linoleate
recombinant enzyme, pH and temperature not specified in the publication
178
oleate
recombinant enzyme, pH and temperature not specified in the publication
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2400
alpha-linolenate
recombinant enzyme, pH and temperature not specified in the publication
2100
linoleate
recombinant enzyme, pH and temperature not specified in the publication
1600
oleate
recombinant enzyme, pH and temperature not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.096
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purified recombinant enzyme, substrate alpha-linolenate, pH 9.0, 35°C
0.236
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purified recombinant enzyme, substrate linoleate, pH 9.0, 35°C
0.345
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purified recombinant enzyme, substrate oleate, pH 9.0, 35°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.5 - 9
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9
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recombinant His-tagged enzyme
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 30
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35
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recombinant His-tagged enzyme
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45000
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1 * 45000, SDS-PAGE
50000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 63000, recombinant His-tagged enzyme, SDS-PAGE
monomer
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45 - 70
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44% of activity still remains at 55°C. The enzyme is stable up to 45 °C, but sharply decreases after 50°C. Activity is completely lost at 70°C
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, 1 week, no loss of activity (thawing and freezing decreases enzyme activity by 30%)
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Q Sepharose column chromatography and Macro-Prep SE 100/40 gel filtration
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recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
gene Npun_R5469, sequence comparisons and phylogenetic tree, recombinant expression of His-tagged enzyme in Escherichia coli
recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3), coexpression with chaperone Gro7
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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optimization of conditions for 10S-hydroxy-8(E)-octadecenoic acid production from oleic acid by recombinant Escherichia coli cells expressing Nostoc punctiforme 10S-dioxygenase with the aid of a chaperone plasmid. Optimal conditions are pH 9.0, 35°C, 15 % v/v dimethyl sulfoxide, 40 g cells/l, and 10 g oleic acid/l. The activity of recombinant cells expressing 10S-dioxygenase is increased by 200% with the aid of chaperone Gro7. Production of 10S-hydroxy-8(E)-octadecenoic acid, 10S-hydroxy-8,12(E,Z)-octadecadienoic acid, and 10S-hydroxy-8,12,15(E,Z,Z)-octadecatrienoic acid from unsaturated fatty acids including oleic acid, linoleic acid, and a-linolenic acid, respectively. The soluble 10S-dioxygenase content in the crude extract of recombinant cells co-expressing the 10S-dioxygenase gene with pGro7 is higher than that of recombinant cells co-expressing pKJE7 or pG-KJE8. Production of 360 mg/g/h 10S-hydroxy-8(E)-octadecenoic acid from oleic acid