BRENDA - Enzyme Database
show all sequences of 1.5.99.B4

Characterization of flavin-containing opine dehydrogenase from bacteria

Watanabe, S.; Sueda, R.; Fukumori, F.; Watanabe, Y.; PLoS ONE 10, e0138434 (2015)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
PpOdhB, PpOdhC, and PpOdhA genes in the PpOdhABC gene cluster, the genes encoding the beta-, gamma-, and alpha-subunits are arranged in tandem on the bacterial genome, genetic structure overview, and amino acid sequence determination, analysis, and comparisons, genetic structure, phylogenetic analysis, recombinant expression as N-terminally His6-tagged proteins in Escherichia coli strain BL21(DE3), recombinant expression of the PpOdhABC gene cluster in Pseudomonas putida
Pseudomonas putida
the gene cluster from Bradyrhizobium japonicum consists of genes OdhB1-C-A-B2, from which two proteins, OdhAB1C and OdhAB2C, appear through the assembly of each beta-subunit together with common alpha- and gamma-subunits, and amino acid sequence determination, analysis, and comparisons, genetic structure, phylogenetic analysis, recombinant expression as N-terminally His6-tagged proteins in Escherichia coli strain BL21(DE3), recombinant expression of the PpOdhABC gene cluster in Pseudomonas putida
Bradyrhizobium japonicum
Inhibitors
Inhibitors
Commentary
Organism
Structure
L-arginine
substrate inhibition
Bradyrhizobium japonicum
L-lysine
-
Bradyrhizobium japonicum
L-ornithine
-
Bradyrhizobium japonicum
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0078
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
0.077
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with nitroblue tetrazolium (NBT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Pseudomonas putida
0.127
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with 4-iodonitrotetrazolium violet (INT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Pseudomonas putida
0.379
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme; pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Pseudomonas putida
0.476
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
1.29
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with nitroblue tetrazolium (NBT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
3.54
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
4.81
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
6.52
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
12.9
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 4-iodonitrotetrazolium violet (INT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Fe2+
in iron-sufur clusters, [4Fe-4S] and [2Fe-2S], overview
Bradyrhizobium japonicum
Fe2+
in iron-sufur clusters, [4Fe-4S] and [2Fe-2S], overview
Pseudomonas putida
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O
Pseudomonas putida
i.e. nopaline
L-arginine + 2-oxoglutarate + FADH2
-
-
?
N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O
Pseudomonas putida KT 2240
i.e. nopaline
L-arginine + 2-oxoglutarate + FADH2
-
-
?
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
Pseudomonas putida
i.e. D-octopine
L-arginine + pyruvate + FADH2
-
-
?
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
Bradyrhizobium japonicum
i.e. D-octopine
L-arginine + pyruvate + FADH2
-
-
?
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
Bradyrhizobium japonicum USDA110
i.e. D-octopine
L-arginine + pyruvate + FADH2
-
-
?
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
Pseudomonas putida KT 2240
i.e. D-octopine
L-arginine + pyruvate + FADH2
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Bradyrhizobium japonicum
Q89E96 AND Q89E94 AND Q89E97 AND
subunits BjOdhA (bll7191 or ooxA), BjOdhB1 (bll7193), and BjOdhB2 (bll7190 or soxB)
-
Bradyrhizobium japonicum USDA110
Q89E96 AND Q89E94 AND Q89E97 AND
subunits BjOdhA (bll7191 or ooxA), BjOdhB1 (bll7193), and BjOdhB2 (bll7190 or soxB)
-
Pseudomonas putida
Q88EK6 AND Q88EK5
subunits alpha and beta; isolated from soil
-
Pseudomonas putida KT 2240
Q88EK6 AND Q88EK5
subunits alpha and beta; isolated from soil
-
Purification (Commentary)
Commentary
Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, dialysis, and gel filtration
Bradyrhizobium japonicum
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, dialysis, and gel filtration
Pseudomonas putida
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
additional information
analysis of specificity of electron acceptors using 2,6-dichloroindophenol, 4-iodonitrotetrazolium violet (INT) or nitroblue tetrazolium (NBT) together with phenazine methosulfate (PMS) (electron-transfer intermediate), ferricyanide, horse heart cytochrome c, and NAD(P)+
743588
Pseudomonas putida
?
-
-
-
-
additional information
analysis of specificity of electron acceptors using 2,6-dichloroindophenol, 4-iodonitrotetrazolium violet (INT) or nitroblue tetrazolium (NBT) together with phenazine methosulfate (PMS) (electron-transfer intermediate), ferricyanide, horse heart cytochrome c, and NAD(P)+. Enzyme BjOdhAB2C and BjOdhAB1C both function as octopine-specific OpnDH, poor activity with nopaline
743588
Bradyrhizobium japonicum
?
-
-
-
-
additional information
analysis of specificity of electron acceptors using 2,6-dichloroindophenol, 4-iodonitrotetrazolium violet (INT) or nitroblue tetrazolium (NBT) together with phenazine methosulfate (PMS) (electron-transfer intermediate), ferricyanide, horse heart cytochrome c, and NAD(P)+. Enzyme BjOdhAB2C and BjOdhAB1C both function as octopine-specific OpnDH, poor activity with nopaline
743588
Bradyrhizobium japonicum USDA110
?
-
-
-
-
additional information
analysis of specificity of electron acceptors using 2,6-dichloroindophenol, 4-iodonitrotetrazolium violet (INT) or nitroblue tetrazolium (NBT) together with phenazine methosulfate (PMS) (electron-transfer intermediate), ferricyanide, horse heart cytochrome c, and NAD(P)+
743588
Pseudomonas putida KT 2240
?
-
-
-
-
N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O
i.e. nopaline
743588
Pseudomonas putida
L-arginine + 2-oxoglutarate + FADH2
-
-
-
?
N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O
i.e. nopaline
743588
Pseudomonas putida KT 2240
L-arginine + 2-oxoglutarate + FADH2
-
-
-
?
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
i.e. D-octopine
743588
Pseudomonas putida
L-arginine + pyruvate + FADH2
-
-
-
?
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
i.e. D-octopine
743588
Bradyrhizobium japonicum
L-arginine + pyruvate + FADH2
-
-
-
?
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
i.e. D-octopine
743588
Bradyrhizobium japonicum USDA110
L-arginine + pyruvate + FADH2
-
-
-
?
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
i.e. D-octopine
743588
Pseudomonas putida KT 2240
L-arginine + pyruvate + FADH2
-
-
-
?
Subunits
Subunits
Commentary
Organism
heterododecamer
alpha4beta4gamma4
Bradyrhizobium japonicum
heterododecamer
alpha4beta4gamma4, 4 * 42000, beta-subunit, + 4 * 9000, gamma-subunit, + 4 * 45000, alpha-subunit, SDS-PAGE
Pseudomonas putida
More
BjOdhB1 and BjOdhB2 both assemble with common alpha- and gamma-subunits, and function as the same octopine dehydrogenase. Most of the purified BjOdhAB2C exists as alpha4beta4gamma4 as determined by gel-filtration, while BjOdhB1 is clearly dominant in purified BjOdhAB1C, thereby confirming that the molar ratio of FAD:FMN of the former (1.8:1.0) is similar to that of PpOdhABC. The gene cluster from Bradyrhizobium japonicum consists of genes OdhB1-C-A-B2, from which two proteins, OdhAB1C and OdhAB2C, appear through the assembly of each beta-subunit together with common alpha- and gamma-subunits, overview. A poor phylogenetic relationship exists between OdhB1 and OdhB2 in spite of them both functioning as octopine dehydrogenases, which provided clear evidence for the acquisition of novel functions by subunit exchange
Bradyrhizobium japonicum
More
PpOpnDH (alpha4beta4gamma4) contained 2 FAD (alpha- and beta-subunits), 1 FMN (between the alpha- and beta-subunits), and 1 [2Fe-2S] iron-sulfur cluster (gamma-subunit) within the structural unit of alphabetagamma. The beta- and alpha-subunit are both necessary for substrate binding, but the beta-subunit functions as a catalytic subunit by itself
Pseudomonas putida
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Bradyrhizobium japonicum
30
-
assay at
Pseudomonas putida
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.093
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with nitroblue tetrazolium (NBT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
0.098
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
0.173
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
0.21
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 4-iodonitrotetrazolium violet (INT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
0.252
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
2.3
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
17.33
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Pseudomonas putida
23.17
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with nitroblue tetrazolium (NBT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Pseudomonas putida
91.5
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with 4-iodonitrotetrazolium violet (INT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Pseudomonas putida
298.67
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Pseudomonas putida
315
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
additional information
-
pH optimum of BjOdhAB2C and BjOdhAB1C, overview
Bradyrhizobium japonicum
9
-
assay at
Bradyrhizobium japonicum
9
-
assay at
Pseudomonas putida
Cofactor
Cofactor
Commentary
Organism
Structure
FAD
-
Bradyrhizobium japonicum
FAD
-
Pseudomonas putida
FMN
-
Bradyrhizobium japonicum
FMN
required for activity, the gamma-subunit has no effect on the binding of FMN
Pseudomonas putida
additional information
electron acceptor specificities of BjOdhAB2C and BjOdhAB1C, overview
Bradyrhizobium japonicum
[2Fe-2S]-center
[4Fe-4S] and [2Fe-2S] clusters bind to two different types of [Fe-S] binding sites in the gamma- and alpha-subunits, respectively. The [2Fe-2S] cluster is important for structural folding and enzyme catalysis
Bradyrhizobium japonicum
[2Fe-2S]-center
[4Fe-4S] and [2Fe-2S] clusters bind to two different types of [Fe-S] binding sites in the gamma- and alpha-subunits, respectively. Cysteine residues C380, C382, C415, and C420 coordinate with the [Fe-S] clusters. The [2Fe-2S] cluster is important for structural folding and enzyme catalysis
Pseudomonas putida
[4Fe-4S]-center
[4Fe-4S] and [2Fe-2S] clusters bind to two different types of [Fe-S] binding sites in the gamma- and alpha-subunits, respectively
Bradyrhizobium japonicum
[4Fe-4S]-center
[4Fe-4S] and [2Fe-2S] clusters bind to two different types of [Fe-S] binding sites in the gamma- and alpha-subunits, respectively. Cysteine residues C56, C61, C64, and C77 coordinate with the [Fe-S] clusters
Pseudomonas putida
Cloned(Commentary) (protein specific)
Commentary
Organism
PpOdhB, PpOdhC, and PpOdhA genes in the PpOdhABC gene cluster, the genes encoding the beta-, gamma-, and alpha-subunits are arranged in tandem on the bacterial genome, genetic structure overview, and amino acid sequence determination, analysis, and comparisons, genetic structure, phylogenetic analysis, recombinant expression as N-terminally His6-tagged proteins in Escherichia coli strain BL21(DE3), recombinant expression of the PpOdhABC gene cluster in Pseudomonas putida
Pseudomonas putida
the gene cluster from Bradyrhizobium japonicum consists of genes OdhB1-C-A-B2, from which two proteins, OdhAB1C and OdhAB2C, appear through the assembly of each beta-subunit together with common alpha- and gamma-subunits, and amino acid sequence determination, analysis, and comparisons, genetic structure, phylogenetic analysis, recombinant expression as N-terminally His6-tagged proteins in Escherichia coli strain BL21(DE3), recombinant expression of the PpOdhABC gene cluster in Pseudomonas putida
Bradyrhizobium japonicum
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FAD
-
Bradyrhizobium japonicum
FAD
-
Pseudomonas putida
FMN
-
Bradyrhizobium japonicum
FMN
required for activity, the gamma-subunit has no effect on the binding of FMN
Pseudomonas putida
additional information
electron acceptor specificities of BjOdhAB2C and BjOdhAB1C, overview
Bradyrhizobium japonicum
[2Fe-2S]-center
[4Fe-4S] and [2Fe-2S] clusters bind to two different types of [Fe-S] binding sites in the gamma- and alpha-subunits, respectively. The [2Fe-2S] cluster is important for structural folding and enzyme catalysis
Bradyrhizobium japonicum
[2Fe-2S]-center
[4Fe-4S] and [2Fe-2S] clusters bind to two different types of [Fe-S] binding sites in the gamma- and alpha-subunits, respectively. Cysteine residues C380, C382, C415, and C420 coordinate with the [Fe-S] clusters. The [2Fe-2S] cluster is important for structural folding and enzyme catalysis
Pseudomonas putida
[4Fe-4S]-center
[4Fe-4S] and [2Fe-2S] clusters bind to two different types of [Fe-S] binding sites in the gamma- and alpha-subunits, respectively
Bradyrhizobium japonicum
[4Fe-4S]-center
[4Fe-4S] and [2Fe-2S] clusters bind to two different types of [Fe-S] binding sites in the gamma- and alpha-subunits, respectively. Cysteine residues C56, C61, C64, and C77 coordinate with the [Fe-S] clusters
Pseudomonas putida
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
L-arginine
substrate inhibition
Bradyrhizobium japonicum
L-lysine
-
Bradyrhizobium japonicum
L-ornithine
-
Bradyrhizobium japonicum
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0078
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
0.077
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with nitroblue tetrazolium (NBT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Pseudomonas putida
0.127
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with 4-iodonitrotetrazolium violet (INT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Pseudomonas putida
0.379
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme; pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Pseudomonas putida
0.476
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
1.29
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with nitroblue tetrazolium (NBT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
3.54
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
4.81
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
6.52
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
12.9
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 4-iodonitrotetrazolium violet (INT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Fe2+
in iron-sufur clusters, [4Fe-4S] and [2Fe-2S], overview
Bradyrhizobium japonicum
Fe2+
in iron-sufur clusters, [4Fe-4S] and [2Fe-2S], overview
Pseudomonas putida
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O
Pseudomonas putida
i.e. nopaline
L-arginine + 2-oxoglutarate + FADH2
-
-
?
N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O
Pseudomonas putida KT 2240
i.e. nopaline
L-arginine + 2-oxoglutarate + FADH2
-
-
?
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
Pseudomonas putida
i.e. D-octopine
L-arginine + pyruvate + FADH2
-
-
?
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
Bradyrhizobium japonicum
i.e. D-octopine
L-arginine + pyruvate + FADH2
-
-
?
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
Bradyrhizobium japonicum USDA110
i.e. D-octopine
L-arginine + pyruvate + FADH2
-
-
?
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
Pseudomonas putida KT 2240
i.e. D-octopine
L-arginine + pyruvate + FADH2
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, dialysis, and gel filtration
Bradyrhizobium japonicum
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, dialysis, and gel filtration
Pseudomonas putida
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
additional information
analysis of specificity of electron acceptors using 2,6-dichloroindophenol, 4-iodonitrotetrazolium violet (INT) or nitroblue tetrazolium (NBT) together with phenazine methosulfate (PMS) (electron-transfer intermediate), ferricyanide, horse heart cytochrome c, and NAD(P)+
743588
Pseudomonas putida
?
-
-
-
-
additional information
analysis of specificity of electron acceptors using 2,6-dichloroindophenol, 4-iodonitrotetrazolium violet (INT) or nitroblue tetrazolium (NBT) together with phenazine methosulfate (PMS) (electron-transfer intermediate), ferricyanide, horse heart cytochrome c, and NAD(P)+. Enzyme BjOdhAB2C and BjOdhAB1C both function as octopine-specific OpnDH, poor activity with nopaline
743588
Bradyrhizobium japonicum
?
-
-
-
-
additional information
analysis of specificity of electron acceptors using 2,6-dichloroindophenol, 4-iodonitrotetrazolium violet (INT) or nitroblue tetrazolium (NBT) together with phenazine methosulfate (PMS) (electron-transfer intermediate), ferricyanide, horse heart cytochrome c, and NAD(P)+. Enzyme BjOdhAB2C and BjOdhAB1C both function as octopine-specific OpnDH, poor activity with nopaline
743588
Bradyrhizobium japonicum USDA110
?
-
-
-
-
additional information
analysis of specificity of electron acceptors using 2,6-dichloroindophenol, 4-iodonitrotetrazolium violet (INT) or nitroblue tetrazolium (NBT) together with phenazine methosulfate (PMS) (electron-transfer intermediate), ferricyanide, horse heart cytochrome c, and NAD(P)+
743588
Pseudomonas putida KT 2240
?
-
-
-
-
N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O
i.e. nopaline
743588
Pseudomonas putida
L-arginine + 2-oxoglutarate + FADH2
-
-
-
?
N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O
i.e. nopaline
743588
Pseudomonas putida KT 2240
L-arginine + 2-oxoglutarate + FADH2
-
-
-
?
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
i.e. D-octopine
743588
Pseudomonas putida
L-arginine + pyruvate + FADH2
-
-
-
?
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
i.e. D-octopine
743588
Bradyrhizobium japonicum
L-arginine + pyruvate + FADH2
-
-
-
?
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
i.e. D-octopine
743588
Bradyrhizobium japonicum USDA110
L-arginine + pyruvate + FADH2
-
-
-
?
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
i.e. D-octopine
743588
Pseudomonas putida KT 2240
L-arginine + pyruvate + FADH2
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
heterododecamer
alpha4beta4gamma4
Bradyrhizobium japonicum
heterododecamer
alpha4beta4gamma4, 4 * 42000, beta-subunit, + 4 * 9000, gamma-subunit, + 4 * 45000, alpha-subunit, SDS-PAGE
Pseudomonas putida
More
BjOdhB1 and BjOdhB2 both assemble with common alpha- and gamma-subunits, and function as the same octopine dehydrogenase. Most of the purified BjOdhAB2C exists as alpha4beta4gamma4 as determined by gel-filtration, while BjOdhB1 is clearly dominant in purified BjOdhAB1C, thereby confirming that the molar ratio of FAD:FMN of the former (1.8:1.0) is similar to that of PpOdhABC. The gene cluster from Bradyrhizobium japonicum consists of genes OdhB1-C-A-B2, from which two proteins, OdhAB1C and OdhAB2C, appear through the assembly of each beta-subunit together with common alpha- and gamma-subunits, overview. A poor phylogenetic relationship exists between OdhB1 and OdhB2 in spite of them both functioning as octopine dehydrogenases, which provided clear evidence for the acquisition of novel functions by subunit exchange
Bradyrhizobium japonicum
More
PpOpnDH (alpha4beta4gamma4) contained 2 FAD (alpha- and beta-subunits), 1 FMN (between the alpha- and beta-subunits), and 1 [2Fe-2S] iron-sulfur cluster (gamma-subunit) within the structural unit of alphabetagamma. The beta- and alpha-subunit are both necessary for substrate binding, but the beta-subunit functions as a catalytic subunit by itself
Pseudomonas putida
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Bradyrhizobium japonicum
30
-
assay at
Pseudomonas putida
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.093
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with nitroblue tetrazolium (NBT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
0.098
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
0.173
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
0.21
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 4-iodonitrotetrazolium violet (INT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
0.252
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
2.3
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
17.33
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Pseudomonas putida
23.17
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with nitroblue tetrazolium (NBT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Pseudomonas putida
91.5
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with 4-iodonitrotetrazolium violet (INT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Pseudomonas putida
298.67
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Pseudomonas putida
315
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
additional information
-
pH optimum of BjOdhAB2C and BjOdhAB1C, overview
Bradyrhizobium japonicum
9
-
assay at
Bradyrhizobium japonicum
9
-
assay at
Pseudomonas putida
General Information
General Information
Commentary
Organism
evolution
opine dehydrogenase (OpnDH) belongs to a group of so-called dye-linked dehydrogenases that catalyze the oxidation of various organic acids, amino acids, and alcohols in the presence of an artificial electron acceptor, such as 2,6-dichloroindophenol, in which FAD and/or FMN is commonly contained as a prosthetic group(s). This enzyme is phylogenetically related to hydrogen cyanide synthase, EC 1.4.99.5, from bacteria, D-hydroxyproline dehydrogenase from bacteria, and L-proline dehydrogenase, EC 1.5.5.2, from archaea. In contrast to Agrobacterium species, PpOpnDH and BjOpnDH genes are located on the chromosome. Flavin-containing OpnDH (the beta-subunit) belongs to the D-amino acid oxidase (DAD) superfamily (pfam01266). An ancestor of this protein family may inherently possess FMN between the alpha- and beta-subunits, and OpnDHtype-1 and OpnDHtype-2 may have acquired the same substrate specificity independently, convergent evolution of OpnDH
Bradyrhizobium japonicum
evolution
opine dehydrogenase (OpnDH) belongs to a group of so-called dye-linked dehydrogenases that catalyze the oxidation of various organic acids, amino acids, and alcohols in the presence of an artificial electron acceptor, such as 2,6-dichloroindophenol, in which FAD and/or FMN is commonly contained as a prosthetic group(s). This enzyme is phylogenetically related to hydrogen cyanide synthase, EC 1.4.99.5, from bacteria, D-hydroxyproline dehydrogenase from bacteria, and L-proline dehydrogenase, EC 1.5.5.2, from archaea. In contrast to Agrobacterium species, PpOpnDH and BjOpnDH, genes are located on the chromosome. Pseudomonas putida strain KT2440 very recently acquired this ability by horizontal gene transfer (not plasmid transfer). Convergent evolution of OpnDH
Pseudomonas putida
additional information
a poor phylogenetic relationship exists between OdhB1 and OdhB2 in spite of them both functioning as octopine dehydrogenases, which provided clear evidence for the acquisition of novel functions by subunit exchange
Bradyrhizobium japonicum
additional information
the beta-subunit functions as a catalytic subunit by itself
Pseudomonas putida
physiological function
a potential physiological role in opine catabolism may be limited to (active) BjOpnDH2 because the OdhA and OdhC proteins preferentially associate with OdhB2 over OdhB1
Bradyrhizobium japonicum
General Information (protein specific)
General Information
Commentary
Organism
evolution
opine dehydrogenase (OpnDH) belongs to a group of so-called dye-linked dehydrogenases that catalyze the oxidation of various organic acids, amino acids, and alcohols in the presence of an artificial electron acceptor, such as 2,6-dichloroindophenol, in which FAD and/or FMN is commonly contained as a prosthetic group(s). This enzyme is phylogenetically related to hydrogen cyanide synthase, EC 1.4.99.5, from bacteria, D-hydroxyproline dehydrogenase from bacteria, and L-proline dehydrogenase, EC 1.5.5.2, from archaea. In contrast to Agrobacterium species, PpOpnDH and BjOpnDH genes are located on the chromosome. Flavin-containing OpnDH (the beta-subunit) belongs to the D-amino acid oxidase (DAD) superfamily (pfam01266). An ancestor of this protein family may inherently possess FMN between the alpha- and beta-subunits, and OpnDHtype-1 and OpnDHtype-2 may have acquired the same substrate specificity independently, convergent evolution of OpnDH
Bradyrhizobium japonicum
evolution
opine dehydrogenase (OpnDH) belongs to a group of so-called dye-linked dehydrogenases that catalyze the oxidation of various organic acids, amino acids, and alcohols in the presence of an artificial electron acceptor, such as 2,6-dichloroindophenol, in which FAD and/or FMN is commonly contained as a prosthetic group(s). This enzyme is phylogenetically related to hydrogen cyanide synthase, EC 1.4.99.5, from bacteria, D-hydroxyproline dehydrogenase from bacteria, and L-proline dehydrogenase, EC 1.5.5.2, from archaea. In contrast to Agrobacterium species, PpOpnDH and BjOpnDH, genes are located on the chromosome. Pseudomonas putida strain KT2440 very recently acquired this ability by horizontal gene transfer (not plasmid transfer). Convergent evolution of OpnDH
Pseudomonas putida
additional information
a poor phylogenetic relationship exists between OdhB1 and OdhB2 in spite of them both functioning as octopine dehydrogenases, which provided clear evidence for the acquisition of novel functions by subunit exchange
Bradyrhizobium japonicum
additional information
the beta-subunit functions as a catalytic subunit by itself
Pseudomonas putida
physiological function
a potential physiological role in opine catabolism may be limited to (active) BjOpnDH2 because the OdhA and OdhC proteins preferentially associate with OdhB2 over OdhB1
Bradyrhizobium japonicum
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.0163
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 4-iodonitrotetrazolium violet (INT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
0.0203
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
0.0387
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
0.0489
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
0.0721
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with nitroblue tetrazolium (NBT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
45.73
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Pseudomonas putida
294.87
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
300.9
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with nitroblue tetrazolium (NBT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Pseudomonas putida
661.77
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
720.47
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with 4-iodonitrotetrazolium violet (INT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Pseudomonas putida
788.05
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Pseudomonas putida
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.0163
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 4-iodonitrotetrazolium violet (INT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
0.0203
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
0.0387
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
0.0489
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
0.0721
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with nitroblue tetrazolium (NBT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
45.73
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Pseudomonas putida
294.87
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
300.9
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with nitroblue tetrazolium (NBT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Pseudomonas putida
661.77
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
720.47
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with 4-iodonitrotetrazolium violet (INT) together with phenazine methosulfate, recombinant His6-tagged enzyme
Pseudomonas putida
788.05
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Pseudomonas putida
Other publictions for EC 1.5.99.B4
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)
742120
Watanabe
Characterization of iron-sulf ...
Bradyrhizobium japonicum, Bradyrhizobium japonicum USDA110
Biosci. Biotechnol. Biochem.
80
2371-2375
2016
-
-
1
-
2
-
-
-
-
1
-
2
-
5
-
-
1
-
-
-
2
-
4
1
1
-
1
-
1
-
-
5
-
-
-
-
-
1
5
-
2
-
-
-
-
-
-
1
-
2
-
-
-
1
-
-
2
-
4
1
1
-
1
-
1
-
-
-
-
3
3
-
2
2
743588
Watanabe
Characterization of flavin-co ...
Bradyrhizobium japonicum, Bradyrhizobium japonicum USDA110, Pseudomonas putida, Pseudomonas putida KT 2240
PLoS ONE
10
e0138434
2015
-
-
2
-
-
-
3
10
-
2
-
6
-
11
-
-
2
-
-
-
-
-
10
4
2
-
-
11
3
-
-
9
-
-
-
-
-
2
9
-
-
-
-
3
-
10
-
2
-
6
-
-
-
2
-
-
-
-
10
4
2
-
-
11
3
-
-
-
-
5
5
-
11
11