A891G
site-directed mutagenesis, the mutant converts linoleic acid to 7,8-DiHODE as major product and 8,11-DiHODE as minor product, which is the similar pattern as the wild-type enzyme
Colletotrichum gloeosporioides
C1038S
site-directed mutagenesis, the mutant produces a low amount of 7,8-DiHODE with the accumulation of 8-HPODE
Colletotrichum gloeosporioides
H1036A
site-directed mutagenesis, the mutant produces a low amount of 7,8-DiHODE with the accumulation of 8-HPODE
Colletotrichum gloeosporioides
H1036A/C1038S
site-directed mutagenesis, the double mutant converts linoleic acid to only 8-HPODE
Colletotrichum gloeosporioides
additional information
comparison of substrate specificities of wild-type an d mutant enzymes, overview
Colletotrichum gloeosporioides
N895D
site-directed mutagenesis, the mutant produces a reduced amount of 8-HPODE, compared to wild-type, without the accumulation of DiHODE
Colletotrichum gloeosporioides
N895L
site-directed mutagenesis, the mutant converts linoleic acid to 7,8-DiHODE as major product and 8,11-DiHODE as minor product, which is the similar pattern as the wild-type enzyme
Colletotrichum gloeosporioides
N895Q
site-directed mutagenesis, the mutant converts linoleic acid to 7,8-DiHODE as major product and 8,11-DiHODE as minor product, which is the similar pattern as the wild-type enzyme
Colletotrichum gloeosporioides
Q898E
site-directed mutagenesis, the mutant converts linoleic acid to 8,11-DiHODE as major product and 7,8-DiHODE as minor product
Colletotrichum gloeosporioides
Q898L
site-directed mutagenesis, the mutant converts linoleic acid to 8,11-DiHODE as major product and 7,8-DiHODE as minor product
Colletotrichum gloeosporioides
Y380F
site-directed mutagenesis, the mutant enzyme produces trace amount of 7,8-DiHODE without the accumulation of 8-HPODE when incubated with linoleic acid as substrate
Colletotrichum gloeosporioides
0.0193
linoleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0193
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.022
linoleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0264
alpha-linolenate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0264
alpha-linolenate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0293
oleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0293
oleate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0305
palmitoleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0305
palmitoleate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0319
eicosadienoic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0319
eicosadienoate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0324
eicosenoic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0324
eicosenoate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0333
oleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0358
arachidonic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0358
arachidonate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0368
dihomo-gamma-linolenic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0369
dihomo-gamma-linolenate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0371
alpha-linolenate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.048
palmitoleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.39
purified recombinant enzyme, whole enzyme activity, substrate palmitoleate, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.48
purified recombinant enzyme, whole enzyme activity, substrate oleate, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.75
purified recombinant enzyme, whole enzyme activity, substrate alpha-linolenate, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.87
pH 6.5, 40°C
Colletotrichum gloeosporioides
0.87
purified recombinant enzyme, whole enzyme activity, substrate (8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate, pH 6.5, 40°C
Colletotrichum gloeosporioides
1.17
purified recombinant enzyme, N-terminal domain activity, substrate arachidate, pH 6.5, 40°C
Colletotrichum gloeosporioides
1.18
purified recombinant enzyme, N-terminal domain activity, substrate dihomo-gamma-linolenate, pH 6.5, 40°C
Colletotrichum gloeosporioides
1.2
purified recombinant enzyme, N-terminal domain activity, substrate eicosadienoate, pH 6.5, 40°C
Colletotrichum gloeosporioides
1.39
purified recombinant enzyme, N-terminal domain activity, substrate eicosenoate, pH 6.5, 40°C
Colletotrichum gloeosporioides
alpha-linolenate + O2
741712
Colletotrichum gloeosporioides
(8R,9Z,12Z)-8-hydroperoxy-9,12,15-octadecatrienoate
?
alpha-linolenate + O2
via (9Z,12Z,15Z)-8-hydroperoxyoctadeca-9,12,15-trienoic acid, best substrate with respect to whole enzyme activity
741712
Colletotrichum gloeosporioides
(9Z,12Z,15Z)-7,8-dihydroxyoctadeca-9,12,15-trienoic acid
?
arachidonate + O2
741712
Colletotrichum gloeosporioides
(5Z,8Z,11Z,14Z)-8-hydroxyeicosa-5,8,11,14-tetraenoic acid
?
arachidonic acid + O2
741712
Colletotrichum gloeosporioides
8-hydroxyarachidonic acid
?
dihomo-gamma-linolenate + O2
741712
Colletotrichum gloeosporioides
(8Z,11Z,14Z)-8-hydroxyeicosa-8,11,14-trienoic acid
?
dihomo-gamma-linolenic acid + O2
741712
Colletotrichum gloeosporioides
8-hydroxydihomo-gamma-linolenic acid
?
eicosadienoate + O2
741712
Colletotrichum gloeosporioides
(11Z,14Z)-8-hydroxyeicosa-11,14-dienoic acid
?
eicosadienoic acid + O2
741712
Colletotrichum gloeosporioides
8-hydroxyeicosadienoic acid
?
eicosenoate + O2
741712
Colletotrichum gloeosporioides
(11Z)-8-hydroxyeicosa-11-enoic acid
?
eicosenoic acid + O2
741712
Colletotrichum gloeosporioides
8-hydroxyeicosenoic acid
?
linoleate + O2
741712
Colletotrichum gloeosporioides
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
?
linoleate + O2
741712
Colletotrichum gloeosporioides KACC 40961
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
?
linoleate + O2
741712
Colletotrichum gloeosporioides
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
?
linoleate + O2
741712
Colletotrichum gloeosporioides
(7S,8S)-7,8-dihydroxy-(9Z,12Z)-octadecadienoic acid
?
linoleate + O2
741712
Colletotrichum gloeosporioides
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoic acid
?
additional information
bifunctional enzyme, the second domain isomerizes the product (8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate to (7S,8S,9Z,12Z)-5,8-dihydroxyoctadeca-9,12-dienoate, cf. EC 5.4.4.6
741712
Colletotrichum gloeosporioides
?
?
additional information
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 (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 (9Z,12Z)-7S,8S-dihydroxyoctadeca-9,12-dienoic acid (7,8-DiHODE) or (5R,8R,9Z,12Z)-5,8-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
741712
Colletotrichum gloeosporioides
?
?
additional information
bifunctional enzyme, the second domain isomerizes the product (8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate to (7S,8S,9Z,12Z)-5,8-dihydroxyoctadeca-9,12-dienoate, cf. EC 5.4.4.6
741712
Colletotrichum gloeosporioides KACC 40961
?
?
additional information
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 (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 (9Z,12Z)-7S,8S-dihydroxyoctadeca-9,12-dienoic acid (7,8-DiHODE) or (5R,8R,9Z,12Z)-5,8-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
741712
Colletotrichum gloeosporioides KACC 40961
?
?
oleate + O2
741712
Colletotrichum gloeosporioides
(9Z)-8-hydroperoxyoctadec-9-enoate
?
oleate + O2
741712
Colletotrichum gloeosporioides KACC 40961
(9Z)-8-hydroperoxyoctadec-9-enoate
?
oleate + O2
via (9Z)-8-hydroperoxyoctadeca-9-enoic acid
741712
Colletotrichum gloeosporioides
(9Z)-7,8-dihydroxyoctadeca-9-enoic acid
?
palmitoleate + O2
741712
Colletotrichum gloeosporioides
(9Z)-8-hydroperoxyhexadec-9-enoate
?
palmitoleate + O2
741712
Colletotrichum gloeosporioides KACC 40961
(9Z)-8-hydroperoxyhexadec-9-enoate
?
palmitoleate + O2
via (9Z)-dihydroperoxyhexadeca-9-enoate
741712
Colletotrichum gloeosporioides
(9Z)-5,8-dihydroxyhexadeca-9-enoic acid
?
4.86
palmitoleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
5.62
oleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
6.58
linoleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
10.5
alpha-linolenate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
17.2
dihomo-gamma-linolenic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
17.2
dihomo-gamma-linolenate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
17.7
eicosadienoic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
17.7
eicosadienoate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
18.2
arachidonic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
18.2
arachidonate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
21.2
palmitoleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
21.2
palmitoleate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
22
eicosenoic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
22
eicosenoate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
24.7
oleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
24.7
oleate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
25.1
alpha-linolenate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
25.2
alpha-linolenate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
35
linoleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
35
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
A891G
site-directed mutagenesis, the mutant converts linoleic acid to 7,8-DiHODE as major product and 8,11-DiHODE as minor product, which is the similar pattern as the wild-type enzyme
Colletotrichum gloeosporioides
C1038S
site-directed mutagenesis, the mutant produces a low amount of 7,8-DiHODE with the accumulation of 8-HPODE
Colletotrichum gloeosporioides
H1036A
site-directed mutagenesis, the mutant produces a low amount of 7,8-DiHODE with the accumulation of 8-HPODE
Colletotrichum gloeosporioides
H1036A/C1038S
site-directed mutagenesis, the double mutant converts linoleic acid to only 8-HPODE
Colletotrichum gloeosporioides
additional information
comparison of substrate specificities of wild-type an d mutant enzymes, overview
Colletotrichum gloeosporioides
N895D
site-directed mutagenesis, the mutant produces a reduced amount of 8-HPODE, compared to wild-type, without the accumulation of DiHODE
Colletotrichum gloeosporioides
N895L
site-directed mutagenesis, the mutant converts linoleic acid to 7,8-DiHODE as major product and 8,11-DiHODE as minor product, which is the similar pattern as the wild-type enzyme
Colletotrichum gloeosporioides
N895Q
site-directed mutagenesis, the mutant converts linoleic acid to 7,8-DiHODE as major product and 8,11-DiHODE as minor product, which is the similar pattern as the wild-type enzyme
Colletotrichum gloeosporioides
Q898E
site-directed mutagenesis, the mutant converts linoleic acid to 8,11-DiHODE as major product and 7,8-DiHODE as minor product
Colletotrichum gloeosporioides
Q898L
site-directed mutagenesis, the mutant converts linoleic acid to 8,11-DiHODE as major product and 7,8-DiHODE as minor product
Colletotrichum gloeosporioides
Y380F
site-directed mutagenesis, the mutant enzyme produces trace amount of 7,8-DiHODE without the accumulation of 8-HPODE when incubated with linoleic acid as substrate
Colletotrichum gloeosporioides
0.0193
linoleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0193
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.022
linoleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0264
alpha-linolenate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0264
alpha-linolenate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0293
oleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0293
oleate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0305
palmitoleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0305
palmitoleate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0319
eicosadienoic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0319
eicosadienoate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0324
eicosenoic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0324
eicosenoate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0333
oleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0358
arachidonic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0358
arachidonate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0368
dihomo-gamma-linolenic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0369
dihomo-gamma-linolenate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.0371
alpha-linolenate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.048
palmitoleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.39
purified recombinant enzyme, whole enzyme activity, substrate palmitoleate, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.48
purified recombinant enzyme, whole enzyme activity, substrate oleate, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.75
purified recombinant enzyme, whole enzyme activity, substrate alpha-linolenate, pH 6.5, 40°C
Colletotrichum gloeosporioides
0.87
pH 6.5, 40°C
Colletotrichum gloeosporioides
0.87
purified recombinant enzyme, whole enzyme activity, substrate (8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate, pH 6.5, 40°C
Colletotrichum gloeosporioides
1.17
purified recombinant enzyme, N-terminal domain activity, substrate arachidate, pH 6.5, 40°C
Colletotrichum gloeosporioides
1.18
purified recombinant enzyme, N-terminal domain activity, substrate dihomo-gamma-linolenate, pH 6.5, 40°C
Colletotrichum gloeosporioides
1.2
purified recombinant enzyme, N-terminal domain activity, substrate eicosadienoate, pH 6.5, 40°C
Colletotrichum gloeosporioides
1.39
purified recombinant enzyme, N-terminal domain activity, substrate eicosenoate, pH 6.5, 40°C
Colletotrichum gloeosporioides
alpha-linolenate + O2
741712
Colletotrichum gloeosporioides
(8R,9Z,12Z)-8-hydroperoxy-9,12,15-octadecatrienoate
?
alpha-linolenate + O2
via (9Z,12Z,15Z)-8-hydroperoxyoctadeca-9,12,15-trienoic acid, best substrate with respect to whole enzyme activity
741712
Colletotrichum gloeosporioides
(9Z,12Z,15Z)-7,8-dihydroxyoctadeca-9,12,15-trienoic acid
?
arachidonate + O2
741712
Colletotrichum gloeosporioides
(5Z,8Z,11Z,14Z)-8-hydroxyeicosa-5,8,11,14-tetraenoic acid
?
arachidonic acid + O2
741712
Colletotrichum gloeosporioides
8-hydroxyarachidonic acid
?
dihomo-gamma-linolenate + O2
741712
Colletotrichum gloeosporioides
(8Z,11Z,14Z)-8-hydroxyeicosa-8,11,14-trienoic acid
?
dihomo-gamma-linolenic acid + O2
741712
Colletotrichum gloeosporioides
8-hydroxydihomo-gamma-linolenic acid
?
eicosadienoate + O2
741712
Colletotrichum gloeosporioides
(11Z,14Z)-8-hydroxyeicosa-11,14-dienoic acid
?
eicosadienoic acid + O2
741712
Colletotrichum gloeosporioides
8-hydroxyeicosadienoic acid
?
eicosenoate + O2
741712
Colletotrichum gloeosporioides
(11Z)-8-hydroxyeicosa-11-enoic acid
?
eicosenoic acid + O2
741712
Colletotrichum gloeosporioides
8-hydroxyeicosenoic acid
?
linoleate + O2
741712
Colletotrichum gloeosporioides
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
?
linoleate + O2
741712
Colletotrichum gloeosporioides KACC 40961
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
?
linoleate + O2
741712
Colletotrichum gloeosporioides
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
?
linoleate + O2
741712
Colletotrichum gloeosporioides
(7S,8S)-7,8-dihydroxy-(9Z,12Z)-octadecadienoic acid
?
linoleate + O2
741712
Colletotrichum gloeosporioides
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoic acid
?
additional information
bifunctional enzyme, the second domain isomerizes the product (8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate to (7S,8S,9Z,12Z)-5,8-dihydroxyoctadeca-9,12-dienoate, cf. EC 5.4.4.6
741712
Colletotrichum gloeosporioides
?
?
additional information
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 (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 (9Z,12Z)-7S,8S-dihydroxyoctadeca-9,12-dienoic acid (7,8-DiHODE) or (5R,8R,9Z,12Z)-5,8-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
741712
Colletotrichum gloeosporioides
?
?
additional information
bifunctional enzyme, the second domain isomerizes the product (8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate to (7S,8S,9Z,12Z)-5,8-dihydroxyoctadeca-9,12-dienoate, cf. EC 5.4.4.6
741712
Colletotrichum gloeosporioides KACC 40961
?
?
additional information
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 (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 (9Z,12Z)-7S,8S-dihydroxyoctadeca-9,12-dienoic acid (7,8-DiHODE) or (5R,8R,9Z,12Z)-5,8-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
741712
Colletotrichum gloeosporioides KACC 40961
?
?
oleate + O2
741712
Colletotrichum gloeosporioides
(9Z)-8-hydroperoxyoctadec-9-enoate
?
oleate + O2
741712
Colletotrichum gloeosporioides KACC 40961
(9Z)-8-hydroperoxyoctadec-9-enoate
?
oleate + O2
via (9Z)-8-hydroperoxyoctadeca-9-enoic acid
741712
Colletotrichum gloeosporioides
(9Z)-7,8-dihydroxyoctadeca-9-enoic acid
?
palmitoleate + O2
741712
Colletotrichum gloeosporioides
(9Z)-8-hydroperoxyhexadec-9-enoate
?
palmitoleate + O2
741712
Colletotrichum gloeosporioides KACC 40961
(9Z)-8-hydroperoxyhexadec-9-enoate
?
palmitoleate + O2
via (9Z)-dihydroperoxyhexadeca-9-enoate
741712
Colletotrichum gloeosporioides
(9Z)-5,8-dihydroxyhexadeca-9-enoic acid
?
4.86
palmitoleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
5.62
oleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
6.58
linoleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
10.5
alpha-linolenate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
17.2
dihomo-gamma-linolenic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
17.2
dihomo-gamma-linolenate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
17.7
eicosadienoic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
17.7
eicosadienoate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
18.2
arachidonic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
18.2
arachidonate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
21.2
palmitoleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
21.2
palmitoleate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
22
eicosenoic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
22
eicosenoate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
24.7
oleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
24.7
oleate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
25.1
alpha-linolenate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
25.2
alpha-linolenate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
35
linoleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
35
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
100
palmitoleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
170
oleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
340
alpha-linolenate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
370
linoleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
466.1
dihomo-gamma-linolenate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
470
dihomo-gamma-linolenic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
508.4
arachidonate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
510
arachidonic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
554.9
eicosadienoate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
560
eicosadienoic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
679
eicosenoate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
680
eicosenoic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
690
palmitoleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
695.1
palmitoleate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
840
oleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
843
oleate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
950
alpha-linolenate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
954.54
alpha-linolenate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
1813.5
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
1820
linoleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
100
palmitoleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
170
oleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
340
alpha-linolenate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
370
linoleate
bifunctional enzyme, pH 6.5, 40°C
Colletotrichum gloeosporioides
466.1
dihomo-gamma-linolenate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
470
dihomo-gamma-linolenic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
508.4
arachidonate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
510
arachidonic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
554.9
eicosadienoate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
560
eicosadienoic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
679
eicosenoate
recombinant enzyme, N-terminal domain activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
680
eicosenoic acid
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
690
palmitoleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
695.1
palmitoleate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
840
oleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
843
oleate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
950
alpha-linolenate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
954.54
alpha-linolenate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
1813.5
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
recombinant enzyme, whole enzyme activity, pH 6.5, 40°C
Colletotrichum gloeosporioides
1820
linoleate
N-terminal 8-dioxygenase domain, pH 6.5, 40°C
Colletotrichum gloeosporioides
741712
Seo
Characterization of a recombi ...
Colletotrichum gloeosporioides, Colletotrichum gloeosporioides KACC 40961
Appl. Microbiol. Biotechnol.
100
3087-3099
2016
1
-
2
-
11
-
-
20
-
-
2
1
1
7
-
-
1
-
-
1
9
-
25
3
1
4
-
1
20
5
-
-
-
-
-
-
1
-
2
-
-
11
-
-
-
-
20
-
-
2
1
1
-
-
1
-
1
9
-
25
3
4
-
1
20
5
-
-
-
-
-
-
-
20
20
742730
Seo
Production of 7,8-dihydroxy u ...
Colletotrichum gloeosporioides, Colletotrichum gloeosporioides KACC 40961
J. Agric. Food Chem.
64
8555-8562
2016
1
1
2
-
1
-
-
-
-
-
-
2
-
11
-
-
1
-
-
1
-
-
14
-
3
1
-
-
-
1
-
-
-
-
-
-
1
1
2
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
1
-
1
-
-
14
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
743200
Sooman
Discovery of a novel linoleat ...
Colletotrichum graminicola, Fusarium oxysporum, Colletotrichum graminicola M1.001, Colletotrichum graminicola M1.001 / M2 / FGSC 10212, Fusarium oxysporum Fo5176
Lipids
50
1243-1252
2015
-
-
1
-
-
-
-
-
-
-
-
5
-
18
-
-
-
-
-
-
-
-
29
-
8
2
2
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
-
29
-
2
2
-
-
-
-
-
-
-
2
2
-
-
-
724132
Hoffmann
7,8- and 5,8-linoleate diol sy ...
Aspergillus fumigatus, Gaeumannomyces graminis
Arch. Biochem. Biophys.
539
87-91
2013
-
-
2
-
7
-
-
-
-
-
-
-
-
6
-
-
-
-
-
-
-
-
5
2
4
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
7
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
5
2
-
-
-
-
-
-
-
-
-
2
2
-
-
-
711024
Hoffmann
Expression of 5,8-LDS of Asper ...
Aspergillus fumigatus, Gaeumannomyces graminis
Arch. Biochem. Biophys.
506
216-222
2010
-
-
2
-
7
-
-
2
-
-
-
3
-
6
-
-
-
-
-
2
-
-
10
-
10
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
7
-
-
-
-
2
-
-
-
3
-
-
-
-
-
2
-
-
10
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-
714317
Jernerén
Reaction mechanism of 5,8-lino ...
Aspergillus clavatus
Biochim. Biophys. Acta
1801
503-507
2010
-
-
-
-
-
-
-
-
-
-
-
1
-
7
-
-
-
-
-
-
-
-
2
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
704466
Brodhun
Identification of PpoA from As ...
Aspergillus nidulans
J. Biol. Chem.
284
11792-11805
2009
-
-
1
-
2
-
1
4
-
-
2
1
-
1
-
-
1
-
-
-
1
-
7
1
1
1
-
-
4
1
-
-
2
-
-
-
-
-
1
2
-
2
-
-
1
-
4
-
-
2
1
-
-
-
1
-
-
1
-
7
1
1
-
-
4
1
-
-
-
-
-
-
-
4
4
704475
Garscha
Leucine/valine residues direct ...
Gaeumannomyces graminis
J. Biol. Chem.
284
13755-13765
2009
-
-
1
-
4
-
-
-
-
-
-
1
-
2
-
-
-
-
-
-
-
-
2
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
4
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
702018
Garscha
Pichia expression and mutagene ...
Gaeumannomyces graminis, Gaeumannomyces graminis var. avenae
Biochem. Biophys. Res. Commun.
373
579-583
2008
-
-
1
-
3
-
-
-
-
-
-
2
-
6
-
1
1
-
-
-
-
-
4
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
3
-
-
-
-
-
-
-
-
2
-
-
1
1
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
703700
Garscha
Critical amino acids for the 8 ...
Gaeumannomyces graminis
FEBS Lett.
582
3547-3551
2008
-
-
1
-
11
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
3
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
-
11
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
285246
Hörnsten
Cloning of linoleate diol synt ...
Gaeumannomyces graminis
J. Biol. Chem.
274
28219-28224
1999
-
-
1
-
-
-
-
-
-
1
1
-
-
3
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
285251
Oliw
Catalytic properties of linole ...
Gaeumannomyces graminis
Adv. Exp. Med. Biol.
469
679-685
1999
2
-
-
-
-
-
2
-
-
-
1
1
-
3
-
-
-
-
-
1
-
-
1
1
-
-
-
-
-
-
-
-
1
-
-
-
2
-
-
1
-
-
-
-
2
-
-
-
-
1
1
-
-
-
-
-
1
-
-
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
285244
Su
A protein radical and ferryl i ...
Gaeumannomyces graminis
J. Biol. Chem.
273
20744-20751
1998
-
-
-
-
-
-
1
-
-
-
-
2
-
5
-
-
2
-
-
2
-
-
5
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
2
-
-
-
-
2
-
-
-
-
-
2
-
-
-
2
-
2
-
-
5
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
285243
Su
Purification and characterizat ...
Gaeumannomyces graminis
J. Biol. Chem.
271
14112-14118
1996
-
-
-
-
-
2
2
2
-
-
2
1
1
1
-
-
1
-
-
1
1
1
2
1
-
-
-
-
1
1
-
-
1
-
1
-
-
-
-
1
-
-
2
-
2
-
2
-
-
2
1
1
-
-
1
-
1
1
1
2
1
-
-
-
1
1
-
-
1
-
-
-
-
-
-
285249
Su
Studies on linoleic acid 8R-di ...
Gaeumannomyces graminis
Lipids
30
43-50
1995
-
-
-
-
-
1
16
-
-
-
-
1
-
6
-
-
1
-
-
-
-
-
1
-
1
-
-
-
-
1
-
-
-
-
1
-
-
-
-
-
-
-
1
-
16
-
-
-
-
-
1
-
-
-
1
-
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-
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1
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1
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1
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285247
Brodowsky
BW A4C and other hydroxamic ac ...
Gaeumannomyces graminis
Eur. J. Pharmacol.
254
43-47
1994
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12
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1
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4
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1
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1
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1
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1
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6
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6
12
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1
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1
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1
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1
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1
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285248
Brodowsky
Linoleic acid 8R-dioxygenase a ...
Gaeumannomyces graminis
Ann. N. Y. Acad. Sci.
744
314-316
1994
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-
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5
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1
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3
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-
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1
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2
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5
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5
5
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1
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1
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285250
Hamberg
Sequential oxygenation of lino ...
Gaeumannomyces graminis
Arch. Biochem. Biophys.
309
77-80
1994
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2
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1
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4
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2
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4
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285245
Brodowsky
A linoleic acid (8R)-dioxygena ...
Gaeumannomyces graminis
J. Biol. Chem.
267
14738-14745
1992
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1
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1
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1
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1
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1
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