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Information on EC 5.4.4.6 - 9,12-octadecadienoate 8-hydroperoxide 8S-isomerase
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5.4.4.6 9,12-octadecadienoate 8-hydroperoxide 8S-isomeraseIUBMB Comments
The enzyme contains heme. The bifunctional enzyme from Gaeumannomyces graminis catalyses the oxidation of linoleic acid to (8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate (cf. EC 1.13.11.60, linoleate 8R-lipoxygenase), which is then isomerized to (7S,8S,9Z,12Z)-5,8-dihydroxyoctadeca-9,12-dienoate .
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The enzyme appears in viruses and cellular organisms
Reaction Schemes
Synonyms
7,8-diol synthase, 7,8-LDS, 7,8-linoleate diol synthase, 
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
7,8-diol synthase
7,8-LDS
7,8-linoleate diol synthase
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate = (7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
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-
-
-
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PATHWAY SOURCE
PATHWAYS
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SYSTEMATIC NAME
IUBMB Comments
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate hydroxymutase [(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate-forming]
The enzyme contains heme. The bifunctional enzyme from Gaeumannomyces graminis catalyses the oxidation of linoleic acid to (8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate (cf. EC 1.13.11.60, linoleate 8R-lipoxygenase), which is then isomerized to (7S,8S,9Z,12Z)-5,8-dihydroxyoctadeca-9,12-dienoate [3].
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(8R,9Z,12Z)-8-hydroperoxy-10,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxy-9,12-octadecadienoate
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
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conversion of (8R)-hydroperoxylinoleic acid into (7S,8S)-dihydroxylinoleic acid by stereospecific elimination of the pro-S hydrogen from C-7 and intramolecular suprafacial insertion of oxygen at C-7
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-
?
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
(9Z)-8-hydroperoxyoctadeca-9-enoic acid
(9Z)-7,8-dihydroxyoctadeca-9-enoic acid
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isomerase reaction of the C-terminal domain
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-
r
(9Z,12Z,15Z)-8-hydroperoxyoctadeca-9,12,15-trienoic acid
(9Z,12Z,15Z)-7,8-dihydroxyoctadeca-9,12,15-trienoic acid
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isomerase reaction of the C-terminal domain
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-
r
linoleate
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
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via (8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate, whole enzyme reaction, best substrate
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-
r
palmitoleate + O2
(9Z)-5,8-dihydroxyhexadeca-9-enoic acid
via (9Z)-8-dihydroperoxyhexadeca-9-enoate
-
-
?
(8R,9Z,12Z)-8-hydroperoxy-10,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
-
-
?
(8R,9Z,12Z)-8-hydroperoxy-10,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
-
-
?
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxy-9,12-octadecadienoate
-
-
-
?
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxy-9,12-octadecadienoate
-
-
-
-
?
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxy-9,12-octadecadienoate
bifunctional enzyme with linoleic acid (8R)-dioxygenase and hydroperoxide isomerase activities. The enzyme abstracts the 8-pro-S hydrogen from linoleic acid, which is followed by antarafacial insertion of molecular oxygen at C-8 to generate 8R-hydroperoxylinoleate. The latter is then isomerized to (7S,8S,9Z,12Z)-5,8-dihydroxy-9,12-octadecadienoate by elimination of the 7-pro-S hydrogen and intramolecular suprafacial insertion of an oxygen atom from the hydroperoxide group
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-
?
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxy-9,12-octadecadienoate
bifunctional enzyme with linoleic acid 8R-dioxygenase and hydroperoxide isomerase activities. The enzyme abstracts the 8-pro-S hydrogen from linoleic acid, which is followed by antarafacial insertion of molecular oxygen at C-8 to generate 8R-hydroperoxylinoleate. The latter is then isomerized to (7S,8S,9Z,12Z)-5,8-dihydroxy-9,12-octadecadienoate by elimination of the 7-pro-S hydrogen and intramolecular suprafacial insertion of an oxygen atom from the hydroperoxide group
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-
?
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxy-9,12-octadecadienoate
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expression in Pichia pastoris changes the position and stereospecificity of a hydroperoxide isomerase (the recombinant enzyme forms 5,8(R)-dihydroxylinoleic acid (60% 5S) and 8(R),13-dihydroxyoctadeca-(9E,11E)-dienoic acid) possibly due to N- or O-linked mannosides in the vicinity of the heme group, whereas the (8R)-dioxygenase activity is identical with native 7,8-linoleate diol synthase
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-
?
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxy-9,12-octadecadienoate
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-
-
?
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxy-9,12-octadecadienoate
expression in Pichia pastoris changes the position and stereospecificity of a hydroperoxide isomerase (the recombinant enzyme forms 5,8(R)-dihydroxylinoleic acid (60% 5S) and 8(R),13-dihydroxyoctadeca-(9E,11E)-dienoic acid) possibly due to N- or O-linked mannosides in the vicinity of the heme group, whereas the (8R)-dioxygenase activity is identical with native 7,8-linoleate diol synthase
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-
?
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxy-9,12-octadecadienoate
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-
-
-
?
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
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-
-
r
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
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-
-
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r
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
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isomerase reaction of the C-terminal domain
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r
alpha-linolenate + O2
(9Z,12Z,15Z)-7,8-dihydroperoxyoctadeca-9,12,15-trienoic acid
via (9Z,12Z,15Z)-8-hydroperoxyoctadeca-9,12,15-trienoic acid, best substrate with respect to whole enzyme activity
-
-
?
alpha-linolenate + O2
(9Z,12Z,15Z)-7,8-dihydroperoxyoctadeca-9,12,15-trienoic acid
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via (9Z,12Z,15Z)-8-hydroperoxyoctadeca-9,12,15-trienoic acid, whole enzyme reaction
-
-
?
oleate + O2
(9Z)-7,8-dihydroxyoctadeca-9-enoic acid
via (9Z)-8-hydroperoxyoctadeca-9-enoic acid
-
-
?
oleate + O2
(9Z)-7,8-dihydroxyoctadeca-9-enoic acid
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via (9Z)-8-hydroperoxyoctadeca-9-enoic acid, whole enzyme reaction
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-
?
additional information
?
-
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the bifunctional enzyme converts linoleic acid to a product, identified as (9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoic acid (7,8-DiHODE) by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) and nuclear magnetic resonance (NMR) spectroscopy. The specific activity and catalytic efficiency (kcat/Km) of 7,8-diol synthase for the conversion of fatty acid to dihydroxy fatty acid follows the descending order linoleic acid, alpha-linolenic acid, and oleic acid, indicating that the enzyme is a 7,8-linoleate diol synthase (7,8-LDS). The reaction via (9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoic acid (8-HPODE) as an intermediate and the accumulation of 8-HPODE is due to a higher 8-dioxygenase activity in the N-terminal domain than hydroperoxide isomerase activity in the C-terminal domain, EC 5.4.4.6. 8-HPODE is subsequently isomerized to (7S,8S,9Z,12Z)-dihydroxyoctadeca-9,12-dienoic acid (7,8-DiHODE) or (5S,8R,9Z,12Z)-dihydroxyoctadeca-9,12-dienoic acid (5,8-DiHODE) by the C-terminal hydroperoxide isomerase domain of diol synthase
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-
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additional information
?
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the bifunctional enzyme converts linoleic acid to a product, identified as (9Z,12Z)7,8-dihydroxyoctadeca-9,12-dienoic acid (7,8-DiHODE) by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) and nuclear magnetic resonance (NMR) spectroscopy. The specific activity and catalytic efficiency (kcat/Km) of 7,8-diol synthase for the conversion of fatty acid to dihydroxy fatty acid follows the descending order linoleic acid, alpha-linolenic acid, oleic acid, and palmitoleic acid, indicating that the enzyme is a 7,8-linoleate diol synthase (7,8-LDS). Substrate specificity, overview. The reaction via 8-hydroperoxy-9,12(Z,Z)-octadecadienoic acid (8-HPODE) as an intermediate and the accumulation of 8-HPODE is due to a higher 8-dioxygenase activity in the N-terminal domain than hydroperoxide isomerase activity in the C-terminal domain, EC 5.4.4.6. 8-HPODE is subsequently isomerized to (7S,8S,9Z,12Z)-dihydroxyoctadeca-9,12-dienoic acid (7,8-DiHODE) or (5S,8R,9Z,12Z)-dihydroxyoctadeca-9,12-dienoic acid (5,8-DiHODE) by the C-terminal hydroperoxide isomerase domain of diol synthase. No activity with conjugated linoleic acid, docosapentaenoic acid, and docosahexaenoic acid, and no isomerase activity and no whole enzyme acitivity, but dioxygenase activity with eicosenoic acid, eicosadienoic acid, dihomo-gamma-linolenic acid, and arachidonic acid
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(8R,9Z,12Z)-8-hydroperoxy-10,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxy-9,12-octadecadienoate
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
(8R,9Z,12Z)-8-hydroperoxy-10,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
-
-
?
(8R,9Z,12Z)-8-hydroperoxy-10,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
-
-
?
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxy-9,12-octadecadienoate
-
-
-
?
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxy-9,12-octadecadienoate
-
-
-
-
?
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxy-9,12-octadecadienoate
-
-
-
?
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
(7S,8S,9Z,12Z)-7,8-dihydroxy-9,12-octadecadienoate
-
-
-
-
?
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
-
r
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
-
-
r
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
heme
-
ferric hemeprotein. The proximal heme ligand of linoleate diol synthase is tentatively identified as His379 and the important tyrosine for catalysis as residue376 (apparent consensus EFNXXXYXWH). The distal heme ligand is tentatively identified as His203 (apparent consensus THXXFXT)
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
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catalysis declines due to suicide inactivation; catalysis declines due to suicide inactivation. Following column chromatography, e.g. ion exchange or zinc-affinity chromatography, hydroperoxide isomerase activity is often decreased and sometimes even undetectable by TLC, whereas 8-dioxygenase activity is always present. However, storage of the enzyme for a few days on ice often restores the hydroperoxide isomerase and appeares to increase the LDS activity as well
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0371
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
-
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
0.0223
alpha-linolenate
-
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
0.0333
oleate
-
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
0.048
palmitoleate
-
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
10.5
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
-
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
6.58
alpha-linolenate
-
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
5.62
oleate
-
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
4.86
palmitoleate
-
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
23.45
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
-
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
33.62
alpha-linolenate
-
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
14.41
oleate
-
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
8.125
palmitoleate
-
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.39
-
purified recombinant enzyme, whole enzyme activity, substrate palmitoleate, pH 6.5, 40°C
0.48
-
purified recombinant enzyme, whole enzyme activity, substrate oleate, pH 6.5, 40°C
0.75
-
purified recombinant enzyme, whole enzyme activity, substrate alpha-linolenate, pH 6.5, 40°C
0.87
-
purified recombinant enzyme, whole enzyme activity, substrate (8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate, pH 6.5, 40°C
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
the maximum activity for the conversion of linoleic acid to (9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoic acid (8-HPODE) by 8-dioxygenase in the N-terminal domain of 7,8-LDS is observed at pH 7.0 and 20°C, whereas that for the conversion of linoleic acid to (9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoic acid (7,8-DiHODE) by the whole enzyme is observed at pH 6.5 and 40°C
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
i.e. Colletotrichum gloeosporioides
UniProt
brenda
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
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UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
Sequence
LIDS_GAEGR
1165
0
128322
Swiss-Prot
A0A0S2SWE4_COLGL
1127
0
126591
TrEMBL
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homotetramer
tetramer
additional information
homotetramer
-
4 * 127000, recombinant His-tagged enzyme, SDS-PAGE, 4 * 127413, sequence calculation
homotetramer
Colletotrichum gloeosporioides KACC 40961
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4 * 127000, recombinant His-tagged enzyme, SDS-PAGE, 4 * 127413, sequence calculation
-
additional information
-
the enzyme diol synthase is a fusion protein, consisting of an N-terminal dioxygenase domain and a C-terminal cytochrome P450/hydroperoxide isomerase domain
additional information
Colletotrichum gloeosporioides KACC 40961
-
the enzyme diol synthase is a fusion protein, consisting of an N-terminal dioxygenase domain and a C-terminal cytochrome P450/hydroperoxide isomerase domain
-
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POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
glycoprotein
glycoprotein
-
7,8-LDS contains five tentative N-linked glycosylation sites, one of which has high probability of glycosylation. This N-glycosylation site, Asn216, is located near the distal heme-binding region along with tentative sites for O-linked glycosylation. Asn216Gln shows no detectable enzyme activity, although expression of Asn216Gln is confirmed by Western blot analysis. alpha-Mannosidase treatment does not change the 8R-dioxygenase activity but abolishes the hydroperoxide isomerase activity. Proper N- and O-linked glycosylation could be important for the hydroperoxide isomerase. Expression of 7,8-LDS in insect cells yields recombinant 7,8-LDS with the native hydroperoxide isomerase activity; 7,8-LDS contains five tentative N-linked glycosylation sites, one of which has high probability of glycosylation. This N-glycosylation site, Asn216, is located near the distal heme-binding region along with tentative sites for O-linked glycosylation. Asn216Gln shows no detectable enzyme activity, although expression of Asn216Gln is confirmed by Western blot analysis. alpha-Mannosidase treatment does not change the 8R-dioxygenase activity but abolishes the hydroperoxide isomerase activity. Proper N- and O-linked glycosylation could be important for the hydroperoxide isomerase. Expression of 7,8-LDS in insect cells yields recombinant 7,8-LDS with the native hydroperoxide isomerase activity
glycoprotein
-
7,8-LDS contains five tentative N-linked glycosylation sites, one of which has high probability of glycosylation. This N-glycosylation site, Asn216, is located near the distal heme-binding region along with tentative sites for O-linked glycosylation. Asn216Gln shows no detectable enzyme activity, although expression of Asn216Gln is confirmed by Western blot analysis. alpha-Mannosidase treatment does not change the 8R-dioxygenase activity but abolishes the hydroperoxide isomerase activity. Proper N- and O-linked glycosylation could be important for the hydroperoxide isomerase. Expression of 7,8-LDS in insect cells yields recombinant 7,8-LDS with the native hydroperoxide isomerase activity; 7,8-LDS contains five tentative N-linked glycosylation sites, one of which has high probability of glycosylation. This N-glycosylation site, Asn216, is located near the distal heme-binding region along with tentative sites for O-linked glycosylation. Asn216Gln shows no detectable enzyme activity, although expression of Asn216Gln is confirmed by Western blot analysis. alpha-Mannosidase treatment does not change the 8R-dioxygenase activity but abolishes the hydroperoxide isomerase activity. Proper N- and O-linked glycosylation could be important for the hydroperoxide isomerase. Expression of 7,8-LDS in insect cells yields recombinant 7,8-LDS with the native hydroperoxide isomerase activity
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
-
comparison of substrate specificities of wild-type and mutant enzymes, overview
additional information
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reaction conditions for the production of (7S,8S,9Z,12Z)-dihydroxyoctadeca-9,12-dienoic acid from linoleic acid by recombinant Escherichia coli expressing 7,8-linoleate diol synthase from Glomerella cingulata are optimized using response surface methodology, overview. The optimal reaction conditions are pH 7.0, 18.6°C, 10.8% v/v dimethyl sulfoxide, 44.9 g/l cells, and 14.3 g/l linoleic acid, with agitation at 256 rpm. Under these conditions, recombinant cells produce 7,8-dihydroxy unsaturated fatty acids in the range of 7.0-9.8 g/l from 14.3 g/l linoleic acid, 14.3 g/l oleic acid, and plant oil hydrolysates such as waste oil and olive oil containing 14.3 g/l linoleic acid or oleic acid. Comparisons of quantitative production of dihydroxy unsaturated fatty acids by microorganisms
additional information
Colletotrichum gloeosporioides KACC 40961
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reaction conditions for the production of (7S,8S,9Z,12Z)-dihydroxyoctadeca-9,12-dienoic acid from linoleic acid by recombinant Escherichia coli expressing 7,8-linoleate diol synthase from Glomerella cingulata are optimized using response surface methodology, overview. The optimal reaction conditions are pH 7.0, 18.6°C, 10.8% v/v dimethyl sulfoxide, 44.9 g/l cells, and 14.3 g/l linoleic acid, with agitation at 256 rpm. Under these conditions, recombinant cells produce 7,8-dihydroxy unsaturated fatty acids in the range of 7.0-9.8 g/l from 14.3 g/l linoleic acid, 14.3 g/l oleic acid, and plant oil hydrolysates such as waste oil and olive oil containing 14.3 g/l linoleic acid or oleic acid. Comparisons of quantitative production of dihydroxy unsaturated fatty acids by microorganisms
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additional information
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treatment with alpha-mannosidase to shorten N- and O-linked mannosides inhibis the hydroperoxide isomerase but not the 8(R)-dioxygenase; treatment with alpha-mannosidase to shorten N- and O-linked mannosides inhibis the hydroperoxide isomerase but not the 8(R)-dioxygenase
additional information
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treatment with alpha-mannosidase to shorten N- and O-linked mannosides inhibis the hydroperoxide isomerase but not the 8(R)-dioxygenase; treatment with alpha-mannosidase to shorten N- and O-linked mannosides inhibis the hydroperoxide isomerase but not the 8(R)-dioxygenase
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged wild-enzyme from Escherichia coli strain ER2566 by nickel affinity chromatography
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recombinant His-tagged wild-type and mutant enzymes 8fold from Escherichia coli strain ER2566 by nickel affinity chromatography
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Spodoptera frugiperda (Sf21) insect cells with native 8R-dioxygenase and hydroperoxide isomerase activities
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expression in Pichia pastoris changes the position and stereospecificity of a hydroperoxide isomerase possibly due to N- or O-linked mannosides in the vicinity of the heme group, whereas the 8(R)-dioxygenase activity is identical with native 7,8-linoleate diol synthase; expression in Pichia pastoris changes the position and stereospecificity of a hydroperoxide isomerase possibly due to N- or O-linked mannosides in the vicinity of the heme group, whereas the 8(R)-dioxygenase activity is identical with native 7,8-linoleate diol synthase
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phylogenetic tree, recombinant expression of wild-type and mutant His-tagged enzymes in Escherichia coli strain ER2566
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recombinant expression of His-tagged enzyme in Escherichia coli strain ER2566
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Su, C.; Sahlin, M.; Oliw, E.H.
A protein radical and ferryl intermediates are generated by linoleate diol synthase, a ferric hemeprotein with dioxygenase and hydroperoxide isomerase activities
J. Biol. Chem.
273
20744-20751
1998
Gaeumannomyces graminis, Gaeumannomyces graminis (Q9UUS2)
brenda
Brodowsky, I.D.; Hamberg, M.; Oliw, E.H.
A linoleic acid (8R)-dioxygenase and hydroperoxide isomerase of the fungus Gaeumannomyces graminis. Biosynthesis of (8R)-Hydroxylinoleic acid and (7S,8S)-dihydroxylinoleic acid from (8R)-Hydroperoxylinoleic acid
J. Biol. Chem.
267
14738-14745
1992
Gaeumannomyces graminis
brenda
Hörnsten, L.; Su, C.; Osbourn, A.E.; Garosi, P.; Hellman, U.; Wernstedt, C.; Oliw, E.H.
Cloning of linoleate diol synthase reveals homology with prostaglandin H synthase
J. Biol. Chem.
274
28219-28224
1999
Gaeumannomyces graminis (Q9UUS2)
brenda
Brodowsky, I.D.; Hamberg, M.; Oliw, E.H.
BW A4C and other hydroxamic acids are potent inhibitors of linoleic acid 8R-dioxygenase of the fungus Gaeumannomyces graminis
Eur. J. Pharmacol.
254
43-47
1994
Gaeumannomyces graminis
brenda
Su, C.; Brodowsky, I.D.; Oliw, E.H.
Studies on linoleic acid 8R-dioxygenase and hydroperoxide isomerase of the fungus Gaeumannomyces graminis
Lipids
30
43-50
1995
Gaeumannomyces graminis
brenda
Hamberg, M.; Zhang, L.Y.; Brodowsky, I.D.; Oliw, E.H.
Sequential oxygenation of linoleic acid in the fungus Gaeumannomyces graminis: Stereochemistry of dioxygenase and hydroperoxide isomerase reactions
Arch. Biochem. Biophys.
309
77-80
1994
Gaeumannomyces graminis, Gaeumannomyces graminis var.graminis
brenda
Oliw, E.H.; Su, C.; Sahlin, M.
Catalytic properties of linoleate diol synthase of the fungus Gaeumannomyces graminis: A comparison with PGH synthases
Adv. Exp. Med. Biol.
469
679-685
1999
Gaeumannomyces graminis
brenda
Garscha, U.; Oliw, E.
Pichia expression and mutagenesis of 7,8-linoleate diol synthase change the dioxygenase and hydroperoxide isomerase
Biochem. Biophys. Res. Commun.
373
579-583
2008
Gaeumannomyces graminis, Gaeumannomyces graminis (Q9UUS2), Gaeumannomyces graminis var. avenae, Gaeumannomyces graminis var. avenae (Q9UUS2)
brenda
Garscha, U.; Oliw, E.H.
Critical amino acids for the 8(R)-dioxygenase activity of linoleate diol synthase. A comparison with cyclooxygenases
FEBS Lett.
582
3547-3551
2008
Gaeumannomyces graminis (Q9UUS2)
brenda
Seo, M.J.; Shin, K.C.; An, J.U.; Kang, W.R.; Ko, Y.J.; Oh, D.K.
Characterization of a recombinant 7,8-linoleate diol synthase from Glomerella cingulate
Appl. Microbiol. Biotechnol.
100
3087-3099
2016
Colletotrichum gloeosporioides (A0A0S2SWE4), Colletotrichum gloeosporioides KACC 40961 (A0A0S2SWE4)
brenda
Seo, M.J.; Kang, W.R.; Shin, K.C.; Oh, D.K.
Production of 7,8-dihydroxy unsaturated fatty acids from plant oils by whole recombinant cells expressing 7,8-linoleate diol synthase from Glomerella cingulata
J. Agric. Food Chem.
64
8555-8562
2016
Colletotrichum gloeosporioides (A0A0S2SWE4), Colletotrichum gloeosporioides KACC 40961 (A0A0S2SWE4)
brenda
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