BRENDA - Enzyme Database

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)
EC Number
Commentary
Organism
1.5.1.11
gene ocs, DNA and amino acid sequence determination, analysis, and comparisons, genetic structure, phylogenetic analysis, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
Agrobacterium tumefaciens
1.5.1.19
gene nos, and amino acid sequence determination, analysis, and comparisons, genetic structure, phylogenetic analysis, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
Agrobacterium tumefaciens
1.5.99.B4
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
1.5.99.B4
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
EC Number
Inhibitors
Commentary
Organism
Structure
1.5.99.B4
L-arginine
substrate inhibition
Bradyrhizobium japonicum
1.5.99.B4
L-lysine
-
Bradyrhizobium japonicum
1.5.99.B4
L-ornithine
-
Bradyrhizobium japonicum
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.5.99.B4
0.0078
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
1.5.99.B4
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
1.5.99.B4
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
1.5.99.B4
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
1.5.99.B4
0.476
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
1.5.99.B4
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
1.5.99.B4
3.54
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
4.81
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
6.52
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
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
EC Number
Metals/Ions
Commentary
Organism
Structure
1.5.99.B4
Fe2+
in iron-sufur clusters, [4Fe-4S] and [2Fe-2S], overview
Bradyrhizobium japonicum
1.5.99.B4
Fe2+
in iron-sufur clusters, [4Fe-4S] and [2Fe-2S], overview
Pseudomonas putida
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.5.1.11
L-arginine + pyruvate + NADH + H+
Agrobacterium tumefaciens
-
N2-(D-1-carboxyethyl)-L-arginine + NAD+ + H2O
-
-
?
1.5.1.11
additional information
Agrobacterium tumefaciens
octopine [N2-(1-D-carboxyethyl)-L-arginine] is synthesized by a NAD(P)H-dependent soluble dehydrogenase that catalyzes the reductive condensation of pyruvate with L-arginine. Although the reaction may be reversible in vitro, the frequent use of the term synthase rather than dehydrogenase has emphasized the importance of biosynthesis, but not degradation, and distinguishes it from the mollusk octopine dehydrogenase
?
-
-
-
1.5.1.19
L-arginine + 2-oxoglutarate + NADPH + H+
Agrobacterium tumefaciens
-
N2-(D-1,3-dicarboxypropyl)-L-arginine + NADP+ + H2O
-
-
ir
1.5.1.19
L-arginine + 2-oxoglutarate + NADPH + H+
Agrobacterium tumefaciens C58 / ATCC 33970
-
N2-(D-1,3-dicarboxypropyl)-L-arginine + NADP+ + H2O
-
-
ir
1.5.99.B4
N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O
Pseudomonas putida
i.e. nopaline
L-arginine + 2-oxoglutarate + FADH2
-
-
?
1.5.99.B4
N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O
Pseudomonas putida KT 2240
i.e. nopaline
L-arginine + 2-oxoglutarate + FADH2
-
-
?
1.5.99.B4
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
Pseudomonas putida
i.e. D-octopine
L-arginine + pyruvate + FADH2
-
-
?
1.5.99.B4
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
Bradyrhizobium japonicum
i.e. D-octopine
L-arginine + pyruvate + FADH2
-
-
?
1.5.99.B4
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
Bradyrhizobium japonicum USDA110
i.e. D-octopine
L-arginine + pyruvate + FADH2
-
-
?
1.5.99.B4
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
Pseudomonas putida KT 2240
i.e. D-octopine
L-arginine + pyruvate + FADH2
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.5.1.11
Agrobacterium tumefaciens
-
-
-
1.5.1.19
Agrobacterium tumefaciens
A9CL08
-
-
1.5.1.19
Agrobacterium tumefaciens C58 / ATCC 33970
A9CL08
-
-
1.5.99.B4
Bradyrhizobium japonicum
Q89E96 AND Q89E94 AND Q89E97 AND
subunits BjOdhA (bll7191 or ooxA), BjOdhB1 (bll7193), and BjOdhB2 (bll7190 or soxB)
-
1.5.99.B4
Bradyrhizobium japonicum USDA110
Q89E96 AND Q89E94 AND Q89E97 AND
subunits BjOdhA (bll7191 or ooxA), BjOdhB1 (bll7193), and BjOdhB2 (bll7190 or soxB)
-
1.5.99.B4
Pseudomonas putida
Q88EK6 AND Q88EK5
subunits alpha and beta; isolated from soil
-
1.5.99.B4
Pseudomonas putida KT 2240
Q88EK6 AND Q88EK5
subunits alpha and beta; isolated from soil
-
Purification (Commentary)
EC Number
Commentary
Organism
1.5.1.11
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, dialysis, and gel filtration
Agrobacterium tumefaciens
1.5.1.19
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, dialysis, and gel filtration
Agrobacterium tumefaciens
1.5.99.B4
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, dialysis, and gel filtration
Bradyrhizobium japonicum
1.5.99.B4
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)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.5.1.11
L-arginine + pyruvate + NADH + H+
-
743588
Agrobacterium tumefaciens
N2-(D-1-carboxyethyl)-L-arginine + NAD+ + H2O
-
-
-
?
1.5.1.11
L-arginine + pyruvate + NADH + H+
the reduction reaction is highly preferred
743588
Agrobacterium tumefaciens
N2-(D-1-carboxyethyl)-L-arginine + NAD+ + H2O
-
-
-
?
1.5.1.11
additional information
octopine [N2-(1-D-carboxyethyl)-L-arginine] is synthesized by a NAD(P)H-dependent soluble dehydrogenase that catalyzes the reductive condensation of pyruvate with L-arginine. Although the reaction may be reversible in vitro, the frequent use of the term synthase rather than dehydrogenase has emphasized the importance of biosynthesis, but not degradation, and distinguishes it from the mollusk octopine dehydrogenase
743588
Agrobacterium tumefaciens
?
-
-
-
-
1.5.1.11
additional information
the enzyme OCS utilizes not only L-arginine, but also other amino acids, to yield the corresponding members of the octopine family. Analysis of specificity of electron acceptors using 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
Agrobacterium tumefaciens
?
-
-
-
-
1.5.1.19
L-arginine + 2-oxoglutarate + NADPH + H+
-
743588
Agrobacterium tumefaciens
N2-(D-1,3-dicarboxypropyl)-L-arginine + NADP+ + H2O
-
-
-
ir
1.5.1.19
L-arginine + 2-oxoglutarate + NADPH + H+
the reduction reaction is highly preferred
743588
Agrobacterium tumefaciens
N2-(D-1,3-dicarboxypropyl)-L-arginine + NADP+ + H2O
-
-
-
?
1.5.1.19
L-arginine + 2-oxoglutarate + NADPH + H+
-
743588
Agrobacterium tumefaciens C58 / ATCC 33970
N2-(D-1,3-dicarboxypropyl)-L-arginine + NADP+ + H2O
-
-
-
ir
1.5.1.19
L-arginine + 2-oxoglutarate + NADPH + H+
the reduction reaction is highly preferred
743588
Agrobacterium tumefaciens C58 / ATCC 33970
N2-(D-1,3-dicarboxypropyl)-L-arginine + NADP+ + H2O
-
-
-
?
1.5.1.19
additional information
analysis of specificity of electron acceptors using 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
Agrobacterium tumefaciens
?
-
-
-
-
1.5.1.19
additional information
analysis of specificity of electron acceptors using 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
Agrobacterium tumefaciens C58 / ATCC 33970
?
-
-
-
-
1.5.99.B4
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
?
-
-
-
-
1.5.99.B4
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
?
-
-
-
-
1.5.99.B4
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
?
-
-
-
-
1.5.99.B4
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
?
-
-
-
-
1.5.99.B4
N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O
i.e. nopaline
743588
Pseudomonas putida
L-arginine + 2-oxoglutarate + FADH2
-
-
-
?
1.5.99.B4
N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O
i.e. nopaline
743588
Pseudomonas putida KT 2240
L-arginine + 2-oxoglutarate + FADH2
-
-
-
?
1.5.99.B4
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
i.e. D-octopine
743588
Pseudomonas putida
L-arginine + pyruvate + FADH2
-
-
-
?
1.5.99.B4
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
i.e. D-octopine
743588
Bradyrhizobium japonicum
L-arginine + pyruvate + FADH2
-
-
-
?
1.5.99.B4
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
i.e. D-octopine
743588
Bradyrhizobium japonicum USDA110
L-arginine + pyruvate + FADH2
-
-
-
?
1.5.99.B4
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
i.e. D-octopine
743588
Pseudomonas putida KT 2240
L-arginine + pyruvate + FADH2
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.5.99.B4
heterododecamer
alpha4beta4gamma4
Bradyrhizobium japonicum
1.5.99.B4
heterododecamer
alpha4beta4gamma4, 4 * 42000, beta-subunit, + 4 * 9000, gamma-subunit, + 4 * 45000, alpha-subunit, SDS-PAGE
Pseudomonas putida
1.5.99.B4
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
1.5.99.B4
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]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.5.1.11
30
-
assay at
Agrobacterium tumefaciens
1.5.1.19
30
-
assay at
Agrobacterium tumefaciens
1.5.99.B4
30
-
assay at
Bradyrhizobium japonicum
1.5.99.B4
30
-
assay at
Pseudomonas putida
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.5.99.B4
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
1.5.99.B4
0.098
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
0.173
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
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
1.5.99.B4
0.252
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
2.3
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
1.5.99.B4
17.33
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Pseudomonas putida
1.5.99.B4
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
1.5.99.B4
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
1.5.99.B4
298.67
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Pseudomonas putida
1.5.99.B4
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
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.5.1.11
9
-
assay at
Agrobacterium tumefaciens
1.5.1.19
9
-
assay at
Agrobacterium tumefaciens
1.5.99.B4
additional information
-
pH optimum of BjOdhAB2C and BjOdhAB1C, overview
Bradyrhizobium japonicum
1.5.99.B4
9
-
assay at
Bradyrhizobium japonicum
1.5.99.B4
9
-
assay at
Pseudomonas putida
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.5.1.11
NADH
-
Agrobacterium tumefaciens
1.5.1.19
NADPH
-
Agrobacterium tumefaciens
1.5.99.B4
FAD
-
Bradyrhizobium japonicum
1.5.99.B4
FAD
-
Pseudomonas putida
1.5.99.B4
FMN
-
Bradyrhizobium japonicum
1.5.99.B4
FMN
required for activity, the gamma-subunit has no effect on the binding of FMN
Pseudomonas putida
1.5.99.B4
additional information
electron acceptor specificities of BjOdhAB2C and BjOdhAB1C, overview
Bradyrhizobium japonicum
1.5.99.B4
[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
1.5.99.B4
[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
1.5.99.B4
[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
1.5.99.B4
[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)
EC Number
Commentary
Organism
1.5.1.11
gene ocs, DNA and amino acid sequence determination, analysis, and comparisons, genetic structure, phylogenetic analysis, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
Agrobacterium tumefaciens
1.5.1.19
gene nos, and amino acid sequence determination, analysis, and comparisons, genetic structure, phylogenetic analysis, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
Agrobacterium tumefaciens
1.5.99.B4
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
1.5.99.B4
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)
EC Number
Cofactor
Commentary
Organism
Structure
1.5.1.11
NADH
-
Agrobacterium tumefaciens
1.5.1.19
NADPH
-
Agrobacterium tumefaciens
1.5.99.B4
FAD
-
Bradyrhizobium japonicum
1.5.99.B4
FAD
-
Pseudomonas putida
1.5.99.B4
FMN
-
Bradyrhizobium japonicum
1.5.99.B4
FMN
required for activity, the gamma-subunit has no effect on the binding of FMN
Pseudomonas putida
1.5.99.B4
additional information
electron acceptor specificities of BjOdhAB2C and BjOdhAB1C, overview
Bradyrhizobium japonicum
1.5.99.B4
[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
1.5.99.B4
[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
1.5.99.B4
[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
1.5.99.B4
[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)
EC Number
Inhibitors
Commentary
Organism
Structure
1.5.99.B4
L-arginine
substrate inhibition
Bradyrhizobium japonicum
1.5.99.B4
L-lysine
-
Bradyrhizobium japonicum
1.5.99.B4
L-ornithine
-
Bradyrhizobium japonicum
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.5.99.B4
0.0078
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
1.5.99.B4
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
1.5.99.B4
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
1.5.99.B4
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
1.5.99.B4
0.476
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
1.5.99.B4
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
1.5.99.B4
3.54
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
4.81
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
6.52
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
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)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.5.99.B4
Fe2+
in iron-sufur clusters, [4Fe-4S] and [2Fe-2S], overview
Bradyrhizobium japonicum
1.5.99.B4
Fe2+
in iron-sufur clusters, [4Fe-4S] and [2Fe-2S], overview
Pseudomonas putida
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.5.1.11
L-arginine + pyruvate + NADH + H+
Agrobacterium tumefaciens
-
N2-(D-1-carboxyethyl)-L-arginine + NAD+ + H2O
-
-
?
1.5.1.11
additional information
Agrobacterium tumefaciens
octopine [N2-(1-D-carboxyethyl)-L-arginine] is synthesized by a NAD(P)H-dependent soluble dehydrogenase that catalyzes the reductive condensation of pyruvate with L-arginine. Although the reaction may be reversible in vitro, the frequent use of the term synthase rather than dehydrogenase has emphasized the importance of biosynthesis, but not degradation, and distinguishes it from the mollusk octopine dehydrogenase
?
-
-
-
1.5.1.19
L-arginine + 2-oxoglutarate + NADPH + H+
Agrobacterium tumefaciens
-
N2-(D-1,3-dicarboxypropyl)-L-arginine + NADP+ + H2O
-
-
ir
1.5.1.19
L-arginine + 2-oxoglutarate + NADPH + H+
Agrobacterium tumefaciens C58 / ATCC 33970
-
N2-(D-1,3-dicarboxypropyl)-L-arginine + NADP+ + H2O
-
-
ir
1.5.99.B4
N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O
Pseudomonas putida
i.e. nopaline
L-arginine + 2-oxoglutarate + FADH2
-
-
?
1.5.99.B4
N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O
Pseudomonas putida KT 2240
i.e. nopaline
L-arginine + 2-oxoglutarate + FADH2
-
-
?
1.5.99.B4
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
Pseudomonas putida
i.e. D-octopine
L-arginine + pyruvate + FADH2
-
-
?
1.5.99.B4
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
Bradyrhizobium japonicum
i.e. D-octopine
L-arginine + pyruvate + FADH2
-
-
?
1.5.99.B4
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
Bradyrhizobium japonicum USDA110
i.e. D-octopine
L-arginine + pyruvate + FADH2
-
-
?
1.5.99.B4
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
Pseudomonas putida KT 2240
i.e. D-octopine
L-arginine + pyruvate + FADH2
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.5.1.11
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, dialysis, and gel filtration
Agrobacterium tumefaciens
1.5.1.19
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, dialysis, and gel filtration
Agrobacterium tumefaciens
1.5.99.B4
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, dialysis, and gel filtration
Bradyrhizobium japonicum
1.5.99.B4
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)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.5.1.11
L-arginine + pyruvate + NADH + H+
-
743588
Agrobacterium tumefaciens
N2-(D-1-carboxyethyl)-L-arginine + NAD+ + H2O
-
-
-
?
1.5.1.11
L-arginine + pyruvate + NADH + H+
the reduction reaction is highly preferred
743588
Agrobacterium tumefaciens
N2-(D-1-carboxyethyl)-L-arginine + NAD+ + H2O
-
-
-
?
1.5.1.11
additional information
octopine [N2-(1-D-carboxyethyl)-L-arginine] is synthesized by a NAD(P)H-dependent soluble dehydrogenase that catalyzes the reductive condensation of pyruvate with L-arginine. Although the reaction may be reversible in vitro, the frequent use of the term synthase rather than dehydrogenase has emphasized the importance of biosynthesis, but not degradation, and distinguishes it from the mollusk octopine dehydrogenase
743588
Agrobacterium tumefaciens
?
-
-
-
-
1.5.1.11
additional information
the enzyme OCS utilizes not only L-arginine, but also other amino acids, to yield the corresponding members of the octopine family. Analysis of specificity of electron acceptors using 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
Agrobacterium tumefaciens
?
-
-
-
-
1.5.1.19
L-arginine + 2-oxoglutarate + NADPH + H+
-
743588
Agrobacterium tumefaciens
N2-(D-1,3-dicarboxypropyl)-L-arginine + NADP+ + H2O
-
-
-
ir
1.5.1.19
L-arginine + 2-oxoglutarate + NADPH + H+
the reduction reaction is highly preferred
743588
Agrobacterium tumefaciens
N2-(D-1,3-dicarboxypropyl)-L-arginine + NADP+ + H2O
-
-
-
?
1.5.1.19
L-arginine + 2-oxoglutarate + NADPH + H+
-
743588
Agrobacterium tumefaciens C58 / ATCC 33970
N2-(D-1,3-dicarboxypropyl)-L-arginine + NADP+ + H2O
-
-
-
ir
1.5.1.19
L-arginine + 2-oxoglutarate + NADPH + H+
the reduction reaction is highly preferred
743588
Agrobacterium tumefaciens C58 / ATCC 33970
N2-(D-1,3-dicarboxypropyl)-L-arginine + NADP+ + H2O
-
-
-
?
1.5.1.19
additional information
analysis of specificity of electron acceptors using 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
Agrobacterium tumefaciens
?
-
-
-
-
1.5.1.19
additional information
analysis of specificity of electron acceptors using 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
Agrobacterium tumefaciens C58 / ATCC 33970
?
-
-
-
-
1.5.99.B4
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
?
-
-
-
-
1.5.99.B4
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
?
-
-
-
-
1.5.99.B4
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
?
-
-
-
-
1.5.99.B4
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
?
-
-
-
-
1.5.99.B4
N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O
i.e. nopaline
743588
Pseudomonas putida
L-arginine + 2-oxoglutarate + FADH2
-
-
-
?
1.5.99.B4
N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O
i.e. nopaline
743588
Pseudomonas putida KT 2240
L-arginine + 2-oxoglutarate + FADH2
-
-
-
?
1.5.99.B4
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
i.e. D-octopine
743588
Pseudomonas putida
L-arginine + pyruvate + FADH2
-
-
-
?
1.5.99.B4
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
i.e. D-octopine
743588
Bradyrhizobium japonicum
L-arginine + pyruvate + FADH2
-
-
-
?
1.5.99.B4
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
i.e. D-octopine
743588
Bradyrhizobium japonicum USDA110
L-arginine + pyruvate + FADH2
-
-
-
?
1.5.99.B4
N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O
i.e. D-octopine
743588
Pseudomonas putida KT 2240
L-arginine + pyruvate + FADH2
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.5.99.B4
heterododecamer
alpha4beta4gamma4
Bradyrhizobium japonicum
1.5.99.B4
heterododecamer
alpha4beta4gamma4, 4 * 42000, beta-subunit, + 4 * 9000, gamma-subunit, + 4 * 45000, alpha-subunit, SDS-PAGE
Pseudomonas putida
1.5.99.B4
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
1.5.99.B4
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)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.5.1.11
30
-
assay at
Agrobacterium tumefaciens
1.5.1.19
30
-
assay at
Agrobacterium tumefaciens
1.5.99.B4
30
-
assay at
Bradyrhizobium japonicum
1.5.99.B4
30
-
assay at
Pseudomonas putida
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.5.99.B4
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
1.5.99.B4
0.098
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
0.173
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
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
1.5.99.B4
0.252
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
2.3
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
1.5.99.B4
17.33
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Pseudomonas putida
1.5.99.B4
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
1.5.99.B4
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
1.5.99.B4
298.67
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Pseudomonas putida
1.5.99.B4
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)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.5.1.11
9
-
assay at
Agrobacterium tumefaciens
1.5.1.19
9
-
assay at
Agrobacterium tumefaciens
1.5.99.B4
additional information
-
pH optimum of BjOdhAB2C and BjOdhAB1C, overview
Bradyrhizobium japonicum
1.5.99.B4
9
-
assay at
Bradyrhizobium japonicum
1.5.99.B4
9
-
assay at
Pseudomonas putida
General Information
EC Number
General Information
Commentary
Organism
1.5.99.B4
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
1.5.99.B4
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
1.5.99.B4
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
1.5.99.B4
additional information
the beta-subunit functions as a catalytic subunit by itself
Pseudomonas putida
1.5.99.B4
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)
EC Number
General Information
Commentary
Organism
1.5.99.B4
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
1.5.99.B4
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
1.5.99.B4
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
1.5.99.B4
additional information
the beta-subunit functions as a catalytic subunit by itself
Pseudomonas putida
1.5.99.B4
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]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.5.99.B4
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
1.5.99.B4
0.0203
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
0.0387
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
0.0489
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
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
1.5.99.B4
45.73
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Pseudomonas putida
1.5.99.B4
294.87
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
1.5.99.B4
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
1.5.99.B4
661.77
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
1.5.99.B4
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
1.5.99.B4
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)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.5.99.B4
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
1.5.99.B4
0.0203
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
0.0387
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
0.0489
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Bradyrhizobium japonicum
1.5.99.B4
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
1.5.99.B4
45.73
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with ferricyanide, recombinant His6-tagged enzyme
Pseudomonas putida
1.5.99.B4
294.87
-
N2-(D-1-carboxyethyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
1.5.99.B4
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
1.5.99.B4
661.77
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with 2,6-dichloroindophenol, recombinant His6-tagged enzyme
Pseudomonas putida
1.5.99.B4
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
1.5.99.B4
788.05
-
N2-(D-1,3-dicarboxypropyl)-L-arginine
pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme
Pseudomonas putida