Activating Compound | Comment | Organism | Structure |
---|---|---|---|
2-mercaptoethanol | hydroxylation is not supported when NADH, NADPH, dithiothreitol, 2-mercaptoethanol and ascorbic acid are added alone to the reaction mixture, stimulation when added together with tetrahydropteridine | Pseudomonas sp. | |
ascorbic acid | hydroxylation is not supported when NADH, NADPH, dithiothreitol, 2-mercaptoethanol and ascorbic acid are added alone to the reaction mixture, stimulation when added together with tetrahydropteridine | Pseudomonas sp. | |
dithiothreitol | hydroxylation is not supported when NADH, NADPH, dithiothreitol, 2-mercaptoethanol and ascorbic acid are added alone to the reaction mixture, stimulation when added together with tetrahydropteridine | Pseudomonas sp. |
General Stability | Organism |
---|---|
freezing and thawing has no effect | Pseudomonas sp. |
quite stable in presence of benzoate | Pseudomonas sp. |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
4-Chlorobenzoate | - |
Pseudomonas sp. | |
4-Methylbenzoate | - |
Pseudomonas sp. | |
4-nitrobenzoate | 38% inhibition | Pseudomonas sp. | |
benzaldehyde | - |
Pseudomonas sp. | |
benzoate methyl ester | slight inhibition | Pseudomonas sp. | |
Cu2+ | 0.005 mM, 50% inhibition | Pseudomonas sp. | |
Hg2+ | 0.005 mM, 70% inhibition | Pseudomonas sp. | |
m-Hydroxybenzoate | 12% inhibition | Pseudomonas sp. | |
salicylate | 20% inhibition | Pseudomonas sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.045 | - |
NADH | - |
Pseudomonas sp. | |
0.045 | - |
6,7-dimethyltetrahydropterin | - |
Pseudomonas sp. | |
0.3 | - |
benzoate | - |
Pseudomonas sp. |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | preincubation for 10 min in the presence of benzoate results in maximal activity | Pseudomonas sp. |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
120000 | - |
gel filtration | Pseudomonas sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. | - |
- |
- |
Purification (Comment) | Organism |
---|---|
protamine sulfate, heat treatment, calcium phosphate gel, ammonium sulfate, DEAE-Sephadex, Sephadex G-150 | Pseudomonas sp. |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
33.92 | - |
- |
Pseudomonas sp. |
Storage Stability | Organism |
---|---|
0-4°C, enzyme concentration 0.050 mg/ml, pH 6.0-7.5, 15 days, no loss of activity | Pseudomonas sp. |
0-4°C, pH 7.2-8.0, Tris-HCl buffer, 24 h, 50% loss of activity | Pseudomonas sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-chlorobenzoate + [reduced NADPH-hemoprotein reductase] + O2 | - |
Pseudomonas sp. | ? | - |
? | |
4-methylbenzoate + [reduced NADPH-hemoprotein reductase] + O2 | - |
Pseudomonas sp. | ? | - |
? | |
benzoate + 6,7-dimethyltetrahydropterin + O2 | - |
Pseudomonas sp. | 4-hydroxybenzoate + oxidized 6,7-dimethyltetrahydropterin + H2O | - |
r | |
benzoate + NADH + H+ + O2 | - |
Pseudomonas sp. | 4-hydroxybenzoate + NAD+ + H2O | - |
r | |
benzoate + [reduced NADPH-hemoprotein reductase] + O2 | - |
Pseudomonas sp. | 4-hydroxybenzoate + [oxidized NADPH-hemoprotein reductase] + H2O | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
34 | - |
- |
Pseudomonas sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.2 | - |
- |
Pseudomonas sp. |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5.5 | 8.2 | approx. 30% of maximal activity at pH 5.5, approx. 35% of maximal activity at pH 8.2 | Pseudomonas sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
tetrahydropteridine | essential for activity, hydroxylation is not supported when NADH, NADPH, dithiothreitol, 2-mercaptoethanol and ascorbic acid are added alone to the reaction mixture, stimulation when added together with tetrahydropteridine | Pseudomonas sp. |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.75 | - |
4-nitrobenzoate | - |
Pseudomonas sp. |