BRENDA - Enzyme Database show
show all sequences of 5.4.4.6

Production of 7,8-dihydroxy unsaturated fatty acids from plant oils by whole recombinant cells expressing 7,8-linoleate diol synthase from Glomerella cingulata

Seo, M.J.; Kang, W.R.; Shin, K.C.; Oh, D.K.; J. Agric. Food Chem. 64, 8555-8562 (2016)

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
DMSO
activates the whole enzyme activity
Colletotrichum gloeosporioides
Cloned(Commentary)
Commentary
Organism
recombinant expression of His-tagged enzyme in Escherichia coli strain ER2566
Colletotrichum gloeosporioides
Engineering
Amino acid exchange
Commentary
Organism
additional information
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, 18.6C, 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
Colletotrichum gloeosporioides
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
Colletotrichum gloeosporioides
-
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
r
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Colletotrichum gloeosporioides
A0A0S2SWE4
i.e. Colletotrichum gloeosporioides
-
Colletotrichum gloeosporioides KACC 40961
A0A0S2SWE4
i.e. Colletotrichum gloeosporioides
-
Purification (Commentary)
Commentary
Organism
recombinant His-tagged wild-enzyme from Escherichia coli strain ER2566 by nickel affinity chromatography
Colletotrichum gloeosporioides
Source Tissue
Source Tissue
Commentary
Organism
Textmining
mycelium
-
Colletotrichum gloeosporioides
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
-
742730
Colletotrichum gloeosporioides
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
-
r
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
isomerase reaction of the C-terminal domain
742730
Colletotrichum gloeosporioides
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
-
r
(9Z)-8-hydroperoxyoctadeca-9-enoic acid
isomerase reaction of the C-terminal domain
742730
Colletotrichum gloeosporioides
(9Z)-7,8-dihydroxyoctadeca-9-enoic acid
-
-
-
r
(9Z,12Z,15Z)-8-hydroperoxyoctadeca-9,12,15-trienoic acid
isomerase reaction of the C-terminal domain
742730
Colletotrichum gloeosporioides
(9Z,12Z,15Z)-7,8-dihydroxyoctadeca-9,12,15-trienoic acid
-
-
-
r
alpha-linolenate + O2
via (9Z,12Z,15Z)-8-hydroperoxyoctadeca-9,12,15-trienoic acid, whole enzyme reaction
742730
Colletotrichum gloeosporioides
(9Z,12Z,15Z)-7,8-dihydroperoxyoctadeca-9,12,15-trienoic acid
-
-
-
?
linoleate
via (8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate, whole enzyme reaction, best substrate
742730
Colletotrichum gloeosporioides
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
-
r
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, 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
742730
Colletotrichum gloeosporioides
?
-
-
-
-
oleate + O2
via (9Z)-8-hydroperoxyoctadeca-9-enoic acid, whole enzyme reaction
742730
Colletotrichum gloeosporioides
(9Z)-7,8-dihydroxyoctadeca-9-enoic acid
-
-
-
?
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
37
-
assay at
Colletotrichum gloeosporioides
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Colletotrichum gloeosporioides
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
DMSO
activates the whole enzyme activity
Colletotrichum gloeosporioides
Cloned(Commentary) (protein specific)
Commentary
Organism
recombinant expression of His-tagged enzyme in Escherichia coli strain ER2566
Colletotrichum gloeosporioides
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
additional information
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, 18.6C, 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
Colletotrichum gloeosporioides
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
Colletotrichum gloeosporioides
-
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
r
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant His-tagged wild-enzyme from Escherichia coli strain ER2566 by nickel affinity chromatography
Colletotrichum gloeosporioides
Source Tissue (protein specific)
Source Tissue
Commentary
Organism
Textmining
mycelium
-
Colletotrichum gloeosporioides
-
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
-
742730
Colletotrichum gloeosporioides
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
-
r
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
isomerase reaction of the C-terminal domain
742730
Colletotrichum gloeosporioides
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
-
r
(9Z)-8-hydroperoxyoctadeca-9-enoic acid
isomerase reaction of the C-terminal domain
742730
Colletotrichum gloeosporioides
(9Z)-7,8-dihydroxyoctadeca-9-enoic acid
-
-
-
r
(9Z,12Z,15Z)-8-hydroperoxyoctadeca-9,12,15-trienoic acid
isomerase reaction of the C-terminal domain
742730
Colletotrichum gloeosporioides
(9Z,12Z,15Z)-7,8-dihydroxyoctadeca-9,12,15-trienoic acid
-
-
-
r
alpha-linolenate + O2
via (9Z,12Z,15Z)-8-hydroperoxyoctadeca-9,12,15-trienoic acid, whole enzyme reaction
742730
Colletotrichum gloeosporioides
(9Z,12Z,15Z)-7,8-dihydroperoxyoctadeca-9,12,15-trienoic acid
-
-
-
?
linoleate
via (8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate, whole enzyme reaction, best substrate
742730
Colletotrichum gloeosporioides
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
-
r
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, 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
742730
Colletotrichum gloeosporioides
?
-
-
-
-
oleate + O2
via (9Z)-8-hydroperoxyoctadeca-9-enoic acid, whole enzyme reaction
742730
Colletotrichum gloeosporioides
(9Z)-7,8-dihydroxyoctadeca-9-enoic acid
-
-
-
?
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
37
-
assay at
Colletotrichum gloeosporioides
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Colletotrichum gloeosporioides
Other publictions for EC 5.4.4.6
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
741712
Seo
Characterization of a recombi ...
Colletotrichum gloeosporioides, Colletotrichum gloeosporioides KACC 40961
Appl. Microbiol. Biotechnol.
100
3087-3099
2016
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1
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11
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4
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1
1
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3
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1
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1
4
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2
1
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2
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11
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4
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1
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1
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1
4
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5
2
1
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4
2
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-
-
-
-
-
-
4
4
742730
Seo
Production of 7,8-dihydroxy u ...
Colletotrichum gloeosporioides, Colletotrichum gloeosporioides KACC 40961
J. Agric. Food Chem.
64
8555-8562
2016
1
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1
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1
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-
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1
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3
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1
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1
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8
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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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1
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1
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1
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8
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1
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1
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-
-
-
-
-
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-
702018
Garscha
Pichia expression and mutagene ...
Gaeumannomyces graminis, Gaeumannomyces graminis var. avenae
Biochem. Biophys. Res. Commun.
373
579-583
2008
-
-
1
-
3
-
-
-
-
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4
-
4
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1
1
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8
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2
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4
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4
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2
2
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8
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703700
Garscha
Critical amino acids for the 8 ...
Gaeumannomyces graminis
FEBS Lett.
582
3547-3551
2008
-
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1
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11
-
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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
1
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11
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1
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-
1
-
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-
-
-
-
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-
-
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285246
Hrnsten
Cloning of linoleate diol synt ...
Gaeumannomyces graminis
J. Biol. Chem.
274
28219-28224
1999
-
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1
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1
1
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1
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1
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1
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285251
Oliw
Catalytic properties of linole ...
Gaeumannomyces graminis
Adv. Exp. Med. Biol.
469
679-685
1999
2
-
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2
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1
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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1
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1
1
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285244
Su
A protein radical and ferryl i ...
Gaeumannomyces graminis
J. Biol. Chem.
273
20744-20751
1998
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1
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2
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2
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2
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2
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2
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4
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-
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-
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285249
Su
Studies on linoleic acid 8R-di ...
Gaeumannomyces graminis
Lipids
30
43-50
1995
-
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-
-
-
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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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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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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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1
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1
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-
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285250
Hamberg
Sequential oxygenation of lino ...
Gaeumannomyces graminis, Gaeumannomyces graminis var.graminis
Arch. Biochem. Biophys.
309
77-80
1994
-
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4
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2
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5
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-
-
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285245
Brodowsky
A linoleic acid (8R)-dioxygena ...
Gaeumannomyces graminis
J. Biol. Chem.
267
14738-14745
1992
-
-
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-
-
-
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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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