BRENDA - Enzyme Database show

Expression of 5,8-LDS of Aspergillus fumigatus and its dioxygenase domain. A comparison with 7,8-LDS, 10-dioxygenase, and cyclooxygenase

Hoffmann, I.; Jerneren, F.; Garscha, U.; Oliw, E.H.; Arch. Biochem. Biophys. 506, 216-222 (2010)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.13.11.60
expression in Escherichia coli and in insect cells
Aspergillus fumigatus
1.13.11.60
expression in Escherichia coli and in insect cells
Gaeumannomyces graminis
5.4.4.5
expression in Escherichia coli and in insect cells
Aspergillus fumigatus
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.13.11.60
C1006S
the hydroperoxide isomerase activity of 5,8-LDS mutant is abolished
Aspergillus fumigatus
1.13.11.60
C1006S/V328F
the mutant forms after reduction of hydroperoxides to alcohols, (8R)-hydroperoxy-(9Z,12Z)-octadecadienoic acid and (10R)-hydroxy-(8E,12Z)-octadecadienoic acid in the same relative amounts
Aspergillus fumigatus
1.13.11.60
C1006S/V328L
the mutant forms after reduction of hydroperoxides to alcohols, (8R)-hydroperoxy-(9Z,12Z)-octadecadienoic acid and (10R)-hydroxy-(8E,12Z)-octadecadienoic acid in the same relative amounts
Aspergillus fumigatus
1.13.11.60
C1006S/V328S
the mutant forms after reduction of hydroperoxides to alcohols, (8R)-hydroperoxy-(9Z,12Z)-octadecadienoic acid and (10R)-hydroxy-(8E,12Z)-octadecadienoic acid in the same relative amounts
Aspergillus fumigatus
1.13.11.60
additional information
replacements of Tyr and Cys in the conserved YRWH and FXXGPHXCLG sequences abolished 8R-dioxygenase and hydroperoxide isomerase activities, respectively. N-terminal expression constructs of 5,8- and 7,8-LDS (674 of 1079, and 673 of 1165 residues), containing one additional alpha-helix compared to cyclooxygenase-1, yields prominent 8R-DOX activities with apparently unchanged or slightly lower substrate affinities, respectively; replacements of Tyr and Cys in the conserved YRWH and FXXGPHXCLG sequences abolishes 8R-dioxygenase (8-DOX) and hydroperoxide isomerase activities, respectively. Val328 of 5,8-LDS does not influence the position of oxygenation
Aspergillus fumigatus
1.13.11.60
Y374F
mutation in the conserved sequence YRWH results in loss of linoleate 8R-lipoxygenase activity, whereas the hydroperoxide isomerase activity is retained
Aspergillus fumigatus
5.4.4.5
C1006S
the hydroperoxide isomerase activity of 5,8-LDS mutant is abolished
Aspergillus fumigatus
5.4.4.5
additional information
replacements of Tyr and Cys in the conserved YRWH and FXXGPHXCLG sequences abolishes 8R-dioxygenase (8-DOX) and hydroperoxide isomerase activities, respectively. Val328 of 5,8-LDS does not influence the position of oxygenation
Aspergillus fumigatus
5.4.4.5
Y374F
mutation in the conserved sequence YRWH results in loss of linoleate 8R-lipoxygenase activity, whereas the hydroperoxide isomerase activity is retained
Aspergillus fumigatus
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.13.11.60
additional information
-
additional information
kinetic data of truncated enzymes
Aspergillus fumigatus
1.13.11.60
additional information
-
additional information
kinetic data of truncated enzymes
Gaeumannomyces graminis
5.4.4.5
additional information
-
additional information
kinetic data of truncated enzymes
Aspergillus fumigatus
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.13.11.60
linoleate + O2
Gaeumannomyces graminis
-
(8R,10E,12Z)-8-hydroperoxy-10,12-octadecadienoate
-
-
?
1.13.11.60
linoleate + O2
Aspergillus fumigatus
i.e. (9Z,12Z)-octadeca-9,12-dienoate
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
-
-
?
1.13.11.60
linoleate + O2
Aspergillus fumigatus
all-cis-9,12-octadecadienoic acid
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
-
-
?
5.4.4.5
(8R,9Z,12Z)-8-hydroperoxy-10,12-octadecadienoate
Aspergillus fumigatus
-
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.13.11.60
Aspergillus fumigatus
-
-
-
1.13.11.60
Aspergillus fumigatus
C1KH66
-
-
1.13.11.60
Gaeumannomyces graminis
-
-
-
5.4.4.5
Aspergillus fumigatus
C1KH66
-
-
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
1.13.11.60
mycelium
-
Aspergillus fumigatus
-
1.13.11.60
mycelium
-
Gaeumannomyces graminis
-
5.4.4.5
mycelium
-
Aspergillus fumigatus
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.13.11.60
(11Z)-icosa-11-enoate + O2
-
711024
Aspergillus fumigatus
?
-
-
-
?
1.13.11.60
(11Z,14Z)-icosa-11,14-dienoic acid + O2
-
711024
Aspergillus fumigatus
?
-
-
-
?
1.13.11.60
alpha-linolenate + O2
-
711024
Aspergillus fumigatus
?
-
-
-
?
1.13.11.60
elaidate + O2
-
711024
Aspergillus fumigatus
?
-
-
-
?
1.13.11.60
linoleate + O2
-
711024
Gaeumannomyces graminis
(8R,10E,12Z)-8-hydroperoxy-10,12-octadecadienoate
-
-
-
?
1.13.11.60
linoleate + O2
i.e. (9Z,12Z)-octadeca-9,12-dienoate
711024
Gaeumannomyces graminis
(8R,10E,12Z)-8-hydroperoxy-10,12-octadecadienoate
-
-
-
?
1.13.11.60
linoleate + O2
i.e. (9Z,12Z)-octadeca-9,12-dienoate
711024
Aspergillus fumigatus
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
-
-
-
?
1.13.11.60
linoleate + O2
all-cis-9,12-octadecadienoic acid
711024
Aspergillus fumigatus
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
-
-
-
?
1.13.11.60
additional information
the recombinant enzyme expressed in insect cells, oxygenates 16:1n-7, 18:1n-7, 18:2n-6, 18:3n-3, 20:1n-9, 20:1n-11, and 20:2n-6 at the allylic carbon closest to the carboxyl group
711024
Gaeumannomyces graminis
?
-
-
-
-
1.13.11.60
palmitoleate + O2
-
711024
Aspergillus fumigatus
?
-
-
-
?
5.4.4.5
(8R,9Z,12Z)-8-hydroperoxy-10,12-octadecadienoate
-
711024
Aspergillus fumigatus
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
-
?
5.4.4.5
(8R,9Z,12Z)-9-hydroperoxy-10,12-octadecadienoate
-
711024
Aspergillus fumigatus
(5S,8R,9Z,12Z)-5,8-dihydroxyoctadeca-9,12-dienoate
-
-
-
?
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.13.11.60
expression in Escherichia coli and in insect cells
Aspergillus fumigatus
1.13.11.60
expression in Escherichia coli and in insect cells
Gaeumannomyces graminis
5.4.4.5
expression in Escherichia coli and in insect cells
Aspergillus fumigatus
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.13.11.60
C1006S
the hydroperoxide isomerase activity of 5,8-LDS mutant is abolished
Aspergillus fumigatus
1.13.11.60
C1006S/V328F
the mutant forms after reduction of hydroperoxides to alcohols, (8R)-hydroperoxy-(9Z,12Z)-octadecadienoic acid and (10R)-hydroxy-(8E,12Z)-octadecadienoic acid in the same relative amounts
Aspergillus fumigatus
1.13.11.60
C1006S/V328L
the mutant forms after reduction of hydroperoxides to alcohols, (8R)-hydroperoxy-(9Z,12Z)-octadecadienoic acid and (10R)-hydroxy-(8E,12Z)-octadecadienoic acid in the same relative amounts
Aspergillus fumigatus
1.13.11.60
C1006S/V328S
the mutant forms after reduction of hydroperoxides to alcohols, (8R)-hydroperoxy-(9Z,12Z)-octadecadienoic acid and (10R)-hydroxy-(8E,12Z)-octadecadienoic acid in the same relative amounts
Aspergillus fumigatus
1.13.11.60
additional information
replacements of Tyr and Cys in the conserved YRWH and FXXGPHXCLG sequences abolished 8R-dioxygenase and hydroperoxide isomerase activities, respectively. N-terminal expression constructs of 5,8- and 7,8-LDS (674 of 1079, and 673 of 1165 residues), containing one additional alpha-helix compared to cyclooxygenase-1, yields prominent 8R-DOX activities with apparently unchanged or slightly lower substrate affinities, respectively
Aspergillus fumigatus
1.13.11.60
additional information
replacements of Tyr and Cys in the conserved YRWH and FXXGPHXCLG sequences abolishes 8R-dioxygenase (8-DOX) and hydroperoxide isomerase activities, respectively. Val328 of 5,8-LDS does not influence the position of oxygenation
Aspergillus fumigatus
1.13.11.60
Y374F
mutation in the conserved sequence YRWH results in loss of linoleate 8R-lipoxygenase activity, whereas the hydroperoxide isomerase activity is retained
Aspergillus fumigatus
5.4.4.5
C1006S
the hydroperoxide isomerase activity of 5,8-LDS mutant is abolished
Aspergillus fumigatus
5.4.4.5
additional information
replacements of Tyr and Cys in the conserved YRWH and FXXGPHXCLG sequences abolishes 8R-dioxygenase (8-DOX) and hydroperoxide isomerase activities, respectively. Val328 of 5,8-LDS does not influence the position of oxygenation
Aspergillus fumigatus
5.4.4.5
Y374F
mutation in the conserved sequence YRWH results in loss of linoleate 8R-lipoxygenase activity, whereas the hydroperoxide isomerase activity is retained
Aspergillus fumigatus
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.13.11.60
additional information
-
additional information
kinetic data of truncated enzymes
Aspergillus fumigatus
1.13.11.60
additional information
-
additional information
kinetic data of truncated enzymes
Gaeumannomyces graminis
5.4.4.5
additional information
-
additional information
kinetic data of truncated enzymes
Aspergillus fumigatus
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.13.11.60
linoleate + O2
Gaeumannomyces graminis
-
(8R,10E,12Z)-8-hydroperoxy-10,12-octadecadienoate
-
-
?
1.13.11.60
linoleate + O2
Aspergillus fumigatus
i.e. (9Z,12Z)-octadeca-9,12-dienoate
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
-
-
?
1.13.11.60
linoleate + O2
Aspergillus fumigatus
all-cis-9,12-octadecadienoic acid
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
-
-
?
5.4.4.5
(8R,9Z,12Z)-8-hydroperoxy-10,12-octadecadienoate
Aspergillus fumigatus
-
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
?
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
1.13.11.60
mycelium
-
Aspergillus fumigatus
-
1.13.11.60
mycelium
-
Gaeumannomyces graminis
-
5.4.4.5
mycelium
-
Aspergillus fumigatus
-
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.13.11.60
(11Z)-icosa-11-enoate + O2
-
711024
Aspergillus fumigatus
?
-
-
-
?
1.13.11.60
(11Z,14Z)-icosa-11,14-dienoic acid + O2
-
711024
Aspergillus fumigatus
?
-
-
-
?
1.13.11.60
alpha-linolenate + O2
-
711024
Aspergillus fumigatus
?
-
-
-
?
1.13.11.60
elaidate + O2
-
711024
Aspergillus fumigatus
?
-
-
-
?
1.13.11.60
linoleate + O2
-
711024
Gaeumannomyces graminis
(8R,10E,12Z)-8-hydroperoxy-10,12-octadecadienoate
-
-
-
?
1.13.11.60
linoleate + O2
i.e. (9Z,12Z)-octadeca-9,12-dienoate
711024
Gaeumannomyces graminis
(8R,10E,12Z)-8-hydroperoxy-10,12-octadecadienoate
-
-
-
?
1.13.11.60
linoleate + O2
i.e. (9Z,12Z)-octadeca-9,12-dienoate
711024
Aspergillus fumigatus
(8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
-
-
-
?
1.13.11.60
linoleate + O2
all-cis-9,12-octadecadienoic acid
711024
Aspergillus fumigatus
(8R,9Z,12Z)-8-hydroperoxyoctadeca-9,12-dienoate
-
-
-
?
1.13.11.60
additional information
the recombinant enzyme expressed in insect cells, oxygenates 16:1n-7, 18:1n-7, 18:2n-6, 18:3n-3, 20:1n-9, 20:1n-11, and 20:2n-6 at the allylic carbon closest to the carboxyl group
711024
Gaeumannomyces graminis
?
-
-
-
-
1.13.11.60
palmitoleate + O2
-
711024
Aspergillus fumigatus
?
-
-
-
?
5.4.4.5
(8R,9Z,12Z)-8-hydroperoxy-10,12-octadecadienoate
-
711024
Aspergillus fumigatus
(7S,8S,9Z,12Z)-7,8-dihydroxyoctadeca-9,12-dienoate
-
-
-
?
5.4.4.5
(8R,9Z,12Z)-9-hydroperoxy-10,12-octadecadienoate
-
711024
Aspergillus fumigatus
(5S,8R,9Z,12Z)-5,8-dihydroxyoctadeca-9,12-dienoate
-
-
-
?
General Information
EC Number
General Information
Commentary
Organism
1.13.11.44
additional information
expression of enzyme residues 1-673, 7,8-LDS_1-673, results in only 8-DOX activity, and of residues 1-643, 7,8-LDS_1-643, in undetectable activity
Aspergillus fumigatus
1.13.11.60
malfunction
replacements of Tyr and Cys in the conserved YRWH and FXXGPHXCLG sequences abolished 8R-dioxygenase and hydroperoxide isomerase activities, respectively
Aspergillus fumigatus
1.13.11.60
additional information
Val-328 of 5,8-LDS does not influence the position of oxygenation in contrast to the homologous residues Val-349 of COX-1 and Leu-384 of 10R-dioxygenase. About 675 amino acids are sufficient to support 8-DOX activity
Aspergillus fumigatus
General Information (protein specific)
EC Number
General Information
Commentary
Organism
1.13.11.44
additional information
expression of enzyme residues 1-673, 7,8-LDS_1-673, results in only 8-DOX activity, and of residues 1-643, 7,8-LDS_1-643, in undetectable activity
Aspergillus fumigatus
1.13.11.60
malfunction
replacements of Tyr and Cys in the conserved YRWH and FXXGPHXCLG sequences abolished 8R-dioxygenase and hydroperoxide isomerase activities, respectively
Aspergillus fumigatus
1.13.11.60
additional information
Val-328 of 5,8-LDS does not influence the position of oxygenation in contrast to the homologous residues Val-349 of COX-1 and Leu-384 of 10R-dioxygenase. About 675 amino acids are sufficient to support 8-DOX activity
Aspergillus fumigatus