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Literature summary extracted from

  • Watanabe, S.; Sueda, R.; Fukumori, F.; Watanabe, Y.
    Characterization of flavin-containing opine dehydrogenase from bacteria (2015), PLoS ONE, 10, e0138434 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) 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 Comment 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 Comment 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 Pseudomonas putida
1.5.99.B4 0.379
-
N2-(D-1,3-dicarboxypropyl)-L-arginine 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 Comment Organism Structure
1.5.99.B4 Fe2+ in iron-sufur clusters, [4Fe-4S] and [2Fe-2S], overview Pseudomonas putida
1.5.99.B4 Fe2+ in iron-sufur clusters, [4Fe-4S] and [2Fe-2S], overview Bradyrhizobium japonicum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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 UniProt Comment 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 subunits BjOdhA (bll7191 or ooxA), BjOdhB1 (bll7193), and BjOdhB2 (bll7190 or soxB)
-
1.5.99.B4 Bradyrhizobium japonicum USDA110 Q89E96 AND Q89E94 AND Q89E97 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 Purification (Comment) 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 Pseudomonas putida
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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.11 L-arginine + pyruvate + NADH + H+
-
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 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 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)+ Agrobacterium tumefaciens ?
-
?
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+ the reduction reaction is highly preferred Agrobacterium tumefaciens N2-(D-1,3-dicarboxypropyl)-L-arginine + NADP+ + H2O
-
?
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
1.5.1.19 L-arginine + 2-oxoglutarate + NADPH + H+ the reduction reaction is highly preferred 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)+ 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)+ 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)+ 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 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 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)+ Pseudomonas putida KT 2240 ?
-
?
1.5.99.B4 N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O i.e. nopaline Pseudomonas putida L-arginine + 2-oxoglutarate + FADH2
-
?
1.5.99.B4 N2-(D-1,3-dicarboxypropyl)-L-arginine + FAD + H2O i.e. nopaline 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 Pseudomonas putida L-arginine + pyruvate + FADH2
-
?
1.5.99.B4 N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O i.e. D-octopine Bradyrhizobium japonicum L-arginine + pyruvate + FADH2
-
?
1.5.99.B4 N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O i.e. D-octopine Bradyrhizobium japonicum USDA110 L-arginine + pyruvate + FADH2
-
?
1.5.99.B4 N2-(D-1-carboxyethyl)-L-arginine + FAD + H2O i.e. D-octopine Pseudomonas putida KT 2240 L-arginine + pyruvate + FADH2
-
?

Subunits

EC Number Subunits Comment 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

Synonyms

EC Number Synonyms Comment Organism
1.5.1.11 flavin-containing opine dehydrogenase
-
Agrobacterium tumefaciens
1.5.1.11 OCS
-
Agrobacterium tumefaciens
1.5.1.11 Octopine synthase
-
Agrobacterium tumefaciens
1.5.1.11 pTi_008
-
Agrobacterium tumefaciens
1.5.1.19 Atu6015
-
Agrobacterium tumefaciens
1.5.1.19 flavin-containing opine dehydrogenase
-
Agrobacterium tumefaciens
1.5.1.19 nopaline synthase
-
Agrobacterium tumefaciens
1.5.1.19 NOS
-
Agrobacterium tumefaciens
1.5.99.B4 BjOpnDH
-
Bradyrhizobium japonicum
1.5.99.B4 bll7190
-
Bradyrhizobium japonicum
1.5.99.B4 bll7191
-
Bradyrhizobium japonicum
1.5.99.B4 bll7192
-
Bradyrhizobium japonicum
1.5.99.B4 bll7193
-
Bradyrhizobium japonicum
1.5.99.B4 flavin-containing opine dehydrogenase
-
Pseudomonas putida
1.5.99.B4 flavin-containing opine dehydrogenase
-
Bradyrhizobium japonicum
1.5.99.B4 Nox/Oox-like protein
-
Pseudomonas putida
1.5.99.B4 Nox/Oox-like protein
-
Bradyrhizobium japonicum
1.5.99.B4 OdhA
-
Pseudomonas putida
1.5.99.B4 odhB
-
Pseudomonas putida
1.5.99.B4 odhC
-
Pseudomonas putida
1.5.99.B4 ooxA
-
Bradyrhizobium japonicum
1.5.99.B4 OpnDH
-
Pseudomonas putida
1.5.99.B4 OpnDH
-
Bradyrhizobium japonicum
1.5.99.B4 SoxB
-
Bradyrhizobium japonicum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment 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 Pseudomonas putida
1.5.99.B4 30
-
assay at Bradyrhizobium japonicum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment 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 Comment 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 Pseudomonas putida
1.5.99.B4 9
-
assay at Bradyrhizobium japonicum

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.1.11 NADH
-
Agrobacterium tumefaciens
1.5.1.19 NADPH
-
Agrobacterium tumefaciens
1.5.99.B4 FAD
-
Pseudomonas putida
1.5.99.B4 FAD
-
Bradyrhizobium japonicum
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. 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 [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 [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

General Information

EC Number General Information Comment 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 Comment 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
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N2-(D-1,3-dicarboxypropyl)-L-arginine pH 9.0, 30°C, with cytochrome c, recombinant His6-tagged enzyme Pseudomonas putida