Activating Compound | Comment | Organism | Structure |
---|---|---|---|
dithiothreitol | 4 mM, slightly increases activity | Corynebacterium glutamicum | |
EDTA | 5 mM or 20 mM, slightly increases activity | Corynebacterium glutamicum |
Protein Variants | Comment | Organism |
---|---|---|
D38A | kcat/KM for 4-hydroxybenzoate is 16.8fold higher than wild-type value | Corynebacterium glutamicum |
D38Y | kcat/KM for 4-hydroxybenzoate is 11.8fold higher than wild-type value | Corynebacterium glutamicum |
D38Y/T42R | kcat/KM for 4-hydroxybenzoate is 32fold higher than wild-type value | Corynebacterium glutamicum |
T42R | kcat/KM for 4-hydroxybenzoate is 7.2fold higher than wild-type value | Corynebacterium glutamicum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Cl- | 50 mM, significant inhibition. No inhibition at 5 mM | Corynebacterium glutamicum | |
Cu2+ | 1 mM, moderate inhibition | Corynebacterium glutamicum | |
Fe3+ | 1 mM, moderate inhibition | Corynebacterium glutamicum | |
Mn2+ | 1 mM, moderate inhibition | Corynebacterium glutamicum | |
NEM | 0.1 mM, 59% inhibition | Corynebacterium glutamicum | |
Zn2+ | 1 mM, moderate inhibition | Corynebacterium glutamicum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.008 | - |
4-hydroxybenzoate | pH 8.0, 25°C, mutant enzyme D38Y/T42R | Corynebacterium glutamicum | |
0.012 | - |
4-hydroxybenzoate | pH 8.0, 25°C, mutant enzyme D38A | Corynebacterium glutamicum | |
0.014 | - |
4-hydroxybenzoate | pH 8.0, 25°C, mutant enzyme T42R | Corynebacterium glutamicum | |
0.026 | - |
4-hydroxybenzoate | pH 8.0, 25°C, mutant enzyme D38Y | Corynebacterium glutamicum | |
0.036 | - |
4-hydroxybenzoate | pH 8.0, 25°C, wild-type enzyme | Corynebacterium glutamicum | |
0.04 | - |
NADPH | pH 8.0, 25°C, mutant enzyme D38A | Corynebacterium glutamicum | |
0.045 | - |
NADPH | pH 8.0, 25°C, mutant enzyme T42R | Corynebacterium glutamicum | |
0.047 | - |
NADPH | pH 8.0, 25°C, mutant enzyme D38Y | Corynebacterium glutamicum | |
0.084 | - |
NADPH | pH 8.0, 25°C, mutant enzyme D38Y/T42R | Corynebacterium glutamicum | |
0.195 | - |
NADPH | pH 8.0, 25°C, wild-type enzyme | Corynebacterium glutamicum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
additional information | metal ions (at 1 mM) Co2+, Ni2+, Mg2+, and Hg2+ shows little influence on hydroxylase activity | Corynebacterium glutamicum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
45000 | - |
2 * 45000, SDS-PAGE | Corynebacterium glutamicum |
80000 | - |
gel filtration, non-denaturing PAGE | Corynebacterium glutamicum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-hydroxybenzoate + NADPH + O2 | Corynebacterium glutamicum | the enzyme prefers NADPH to NADH. 4HBA hydroxylase can be induced by 4HBA, 4-cresol, and 4-hydroxybenzaldehyde | protocatechuate + NADP+ + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Corynebacterium glutamicum | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Corynebacterium glutamicum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-hydroxybenzoate + NADH + O2 | the enzyme prefers NADPH to NADH | Corynebacterium glutamicum | protocatechuate + NAD+ + H2O | - |
? | |
4-hydroxybenzoate + NADPH + O2 | the enzyme prefers NADPH to NADH. 4HBA hydroxylase can be induced by 4HBA, 4-cresol, and 4-hydroxybenzaldehyde | Corynebacterium glutamicum | protocatechuate + NADP+ + H2O | - |
? | |
4-hydroxybenzoate + NADPH + O2 | the enzyme prefers NADPH to NADH | Corynebacterium glutamicum | protocatechuate + NADP+ + H2O | - |
? | |
additional information | no hydroxylase activity is observed with 3-hydroxybenzoate, resorcinol, 3-nitrophenol, benzoate, 2,4-dihydroxybenzoate, 4-chlorobenzoate, 4-aminobenzoate, 4-aminophenol, 4-nitrophenol, and 4-dinitrobenzene | Corynebacterium glutamicum | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 45000, SDS-PAGE | Corynebacterium glutamicum |
Synonyms | Comment | Organism |
---|---|---|
4-hydroxybenzoate hydroxylase | - |
Corynebacterium glutamicum |
4HBA 3-hydroxylase | - |
Corynebacterium glutamicum |
ncgl1032 | - |
Corynebacterium glutamicum |
pobACg | - |
Corynebacterium glutamicum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.8 | - |
4-hydroxybenzoate | pH 8.0, 25°C, wild-type enzyme | Corynebacterium glutamicum | |
7.5 | - |
4-hydroxybenzoate | pH 8.0, 25°C, mutant enzyme T42R | Corynebacterium glutamicum | |
7.8 | - |
NADPH | pH 8.0, 25°C, wild-type enzyme | Corynebacterium glutamicum | |
12.4 | - |
NADPH | pH 8.0, 25°C, mutant enzyme T42R | Corynebacterium glutamicum | |
15.2 | - |
4-hydroxybenzoate | pH 8.0, 25°C, mutant enzyme D38A | Corynebacterium glutamicum | |
18.8 | - |
NADPH | pH 8.0, 25°C, mutant enzyme D38A | Corynebacterium glutamicum | |
19.6 | - |
NADPH | pH 8.0, 25°C, mutant enzyme D38Y | Corynebacterium glutamicum | |
20.4 | - |
4-hydroxybenzoate | pH 8.0, 25°C, mutant enzyme D38Y/T42R | Corynebacterium glutamicum | |
24.3 | - |
4-hydroxybenzoate | pH 8.0, 25°C, mutant enzyme D38Y | Corynebacterium glutamicum | |
46.4 | - |
NADPH | pH 8.0, 25°C, mutant enzyme D38Y/T42R | Corynebacterium glutamicum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
- |
Corynebacterium glutamicum |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
7.5 | 9 | - |
Corynebacterium glutamicum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | the enzyme prefers NADPH to NADH | Corynebacterium glutamicum | |
NADPH | the enzyme prefers NADPH to NADH | Corynebacterium glutamicum |