Activating Compound | Comment | Organism | Structure |
---|---|---|---|
2,5-Dihydroxybenzoate | i.e. gentisate, strongly stimulates the NADH oxidase activity of the enzyme | Rhodococcus jostii | |
3-hydroxybenzoate | strongly stimulates activity | Rhodococcus jostii | |
additional information | enzyme activity is not stimulated by salicylate | Rhodococcus jostii |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli Top10 cells | Rhodococcus jostii |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Cl- | - |
Rhodococcus jostii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.046 | - |
3-hydroxybenzoate | in 50 mM Tris-SO4 (pH 8.0), at 25°C | Rhodococcus jostii | |
0.048 | - |
NADH | in 50 mM Tris-SO4 (pH 8.0), at 25°C | Rhodococcus jostii | |
0.051 | - |
2,3-Dihydroxybenzoate | in 50 mM Tris-SO4 (pH 8.0), at 25°C | Rhodococcus jostii | |
0.14 | - |
2,5-Dihydroxybenzoate | in 50 mM Tris-SO4 (pH 8.0), at 25°C | Rhodococcus jostii | |
0.25 | - |
3,5-Dihydroxybenzoate | in 50 mM Tris-SO4 (pH 8.0), at 25°C | Rhodococcus jostii | |
0.39 | - |
NADPH | in 50 mM Tris-SO4 (pH 8.0), at 25°C | Rhodococcus jostii |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
47000 | - |
2 * 47000, SDS-PAGE | Rhodococcus jostii |
108000 | - |
gel filtration | Rhodococcus jostii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-hydroxybenzoate + NADH + H+ + O2 | Rhodococcus jostii | the enzyme clearly prefers NADH as external electron donor over NADPH | 2,5-dihydroxybenzoate + NAD+ + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodococcus jostii | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA agarose column chromatography and Q-Sepharose column chromatography | Rhodococcus jostii |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
6 | - |
crude extract, in 50 mM Tris-SO4 (pH 8.0), at 25°C | Rhodococcus jostii |
24 | - |
after purification, in 50 mM Tris-SO4 (pH 8.0), at 25°C | Rhodococcus jostii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,3,4-trihydroxybenzoate + NADH + H+ + O2 | - |
Rhodococcus jostii | 2,3,4,6-tetrahydroxybenzoate + NAD+ + H2O | - |
? | |
2,3-dihydroxybenzoate + NADH + H+ + O2 | - |
Rhodococcus jostii | 2,3,6-trihydroxybenzoate + NAD+ + H2O | - |
? | |
3,4-dihydroxybenzoate + NADH + H+ + O2 | very poor substrate | Rhodococcus jostii | ? | - |
? | |
3,5-dihydroxybenzoate + NADH + H+ + O2 | - |
Rhodococcus jostii | 2,3,5,6-tetrahydroxybenzoate + NAD+ + H2O | - |
? | |
3-hydroxy-4-aminobenzoate + NADH + H+ + O2 | - |
Rhodococcus jostii | 4-amino-2,5-dihydroxybenzoate + NAD+ + H2O | - |
? | |
3-hydroxy-4-methoxybenzoate + NADH + H+ + O2 | - |
Rhodococcus jostii | 4-methoxy-2,5-dihydroxybenzoate + NAD+ + H2O | - |
? | |
3-hydroxy-4-methylbenzoate + NADH + H+ + O2 | - |
Rhodococcus jostii | 4-methyl-2,5-dihydroxybenzoate + NAD+ + H2O | - |
? | |
3-hydroxybenzoate + NADH + H+ + O2 | the enzyme clearly prefers NADH as external electron donor over NADPH | Rhodococcus jostii | 2,5-dihydroxybenzoate + NAD+ + H2O | - |
? | |
3-hydroxybenzoate + NADPH + H+ + O2 | the enzyme clearly prefers NADH as external electron donor over NADPH | Rhodococcus jostii | 2,5-dihydroxybenzoate + NADP+ + H2O | - |
? | |
additional information | no activity with 2,5-dihydroxybenzoate | Rhodococcus jostii | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 47000, SDS-PAGE | Rhodococcus jostii |
Synonyms | Comment | Organism |
---|---|---|
3-hydroxybenzoate 6-hydroxylase | - |
Rhodococcus jostii |
3HB6H | - |
Rhodococcus jostii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
7 | - |
2,5-Dihydroxybenzoate | in 50 mM Tris-SO4 (pH 8.0), at 25°C | Rhodococcus jostii | |
10 | - |
2,3-Dihydroxybenzoate | in 50 mM Tris-SO4 (pH 8.0), at 25°C | Rhodococcus jostii | |
13 | - |
3,5-Dihydroxybenzoate | in 50 mM Tris-SO4 (pH 8.0), at 25°C | Rhodococcus jostii | |
35 | - |
3-hydroxybenzoate | in 50 mM Tris-SO4 (pH 8.0), at 25°C | Rhodococcus jostii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
around pH 8.0 | Rhodococcus jostii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | the enzyme contains a non-covalently bound FAD cofactor | Rhodococcus jostii | |
NADH | - |
Rhodococcus jostii |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
50 | - |
2,5-Dihydroxybenzoate | in 50 mM Tris-SO4 (pH 8.0), at 25°C | Rhodococcus jostii | |
50 | - |
3,5-Dihydroxybenzoate | in 50 mM Tris-SO4 (pH 8.0), at 25°C | Rhodococcus jostii | |
200 | - |
2,3-Dihydroxybenzoate | in 50 mM Tris-SO4 (pH 8.0), at 25°C | Rhodococcus jostii | |
800 | - |
3-hydroxybenzoate | in 50 mM Tris-SO4 (pH 8.0), at 25°C | Rhodococcus jostii |