Activating Compound | Comment | Organism | Structure |
---|---|---|---|
m-cresol | increases activity | Pseudomonas putida |
Crystallization (Comment) | Organism |
---|---|
- |
Pseudomonas putida |
General Stability | Organism |
---|---|
thiol reagents, e.g. 2-mercaptoethanol, stabilize | Pseudomonas putida |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
1,3-Dimethoxybenzene | 45% inhibition at 1.7 mM | Pseudomonas putida | |
2-Hydroxy-4-methoxybenzoate | 68% inhibition at 1.7 mM | Pseudomonas putida | |
2-Methylresorcinol | 27% inhibition at 1.7 mM | Pseudomonas putida | |
2-Nitroorcinol | 64% inhibition at 1.7 mM | Pseudomonas putida | |
3,4-dimethylphenol | 54% inhibition at 1.7 mM | Pseudomonas putida | |
3,5-Dihydroxybenzoate | 59% inhibition at 1.7 mM | Pseudomonas putida | |
3,5-Dimethylphenol | 68% inhibition at 1.7 mM | Pseudomonas putida | |
3-Hydroxybenzaldehyde | 23% inhibition at 1.7 mM | Pseudomonas putida |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | - |
Pseudomonas putida | |
0.03 | - |
orcinol | cofactor: NADH | Pseudomonas putida | |
0.05 | - |
orcinol | cofactor: 3-acetylpyridine NADH | Pseudomonas putida | |
0.07 | - |
O2 | - |
Pseudomonas putida | |
0.085 | - |
orcinol | cofactor: NADPH | Pseudomonas putida | |
0.13 | - |
NADH | substrate: orcinol | Pseudomonas putida | |
0.19 | - |
resorcinol | - |
Pseudomonas putida | |
0.65 | - |
3-acetylpyridine-NADH | substrate: orcinol | Pseudomonas putida | |
2.5 | - |
NADH | substrate: resorcinol | Pseudomonas putida |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
63000 | - |
gel filtration | Pseudomonas putida |
65000 | - |
sedimentation equilibrium | Pseudomonas putida |
68000 | - |
1 * 68000, SDS-PAGE | Pseudomonas putida |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | - |
01 | - |
Pseudomonas putida 1 | - |
01 | - |
Purification (Comment) | Organism |
---|---|
using protamine sulfate treatment, DEAE-cellulose column chromatography, ammonium sulfate treatment and column chromatography on Sephadex G-100 followed by ammonium sulfate treatment and column chromatography on hydroxylapatite | Pseudomonas putida |
Renatured (Comment) | Organism |
---|---|
most of the activity of the apoenzyme is reconstituted by the addition of FAD, FMN is a poor substitute for FAD | Pseudomonas putida |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
2.8 | - |
apoenzyme without flavin addition | Pseudomonas putida |
3.5 | - |
apoenzyme with addition of FMN | Pseudomonas putida |
14.7 | - |
apoenzyme with addition of FAD | Pseudomonas putida |
18 | - |
holoenzyme | Pseudomonas putida |
24 | - |
- |
Pseudomonas putida |
Storage Stability | Organism |
---|---|
4°C, stabilized by thiol reagents, e.g. 2-mercaptoethanol, some days | Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-cresol + NADH + O2 | nonsubstrate effector, which increases NADH oxidase activity without being hydroxylated | Pseudomonas putida | 3-methylcatechol + NAD+ + H2O | - |
? | |
3-cresol + NADH + O2 | nonsubstrate effector, which increases NADH oxidase activity without being hydroxylated | Pseudomonas putida 1 | 3-methylcatechol + NAD+ + H2O | - |
? | |
3-ethylphenol + NADH + O2 | no substrate, mimic orcinol as effector | Pseudomonas putida | ? | - |
? | |
3-ethylphenol + NADH + O2 | no substrate, mimic orcinol as effector | Pseudomonas putida 1 | ? | - |
? | |
4-bromoresorcinol + NADH + O2 | - |
Pseudomonas putida | ? | - |
? | |
4-methylresorcinol + NADH + O2 | - |
Pseudomonas putida | ? | - |
? | |
additional information | reduced orcinol hydroxylase is able to transfer reducing equivalents to a variety of electron acceptors, other than oxygen. These include free FAD, ferricyanide, cytochrome c, acetylpyridine-NAD and tetrazolium salts | Pseudomonas putida | ? | - |
? | |
additional information | reduced orcinol hydroxylase is able to transfer reducing equivalents to a variety of electron acceptors, other than oxygen. These include free FAD, ferricyanide, cytochrome c, acetylpyridine-NAD and tetrazolium salts | Pseudomonas putida 1 | ? | - |
? | |
orcinol + 3-acetylpyridine-NADH + H+ + O2 | - |
Pseudomonas putida | 2,3,5-trihydroxytoluene + 3-acetylpyridine-NAD+ + H2O | - |
r | |
orcinol + NADH + O2 | - |
Pseudomonas putida | 2,3,5-trihydroxytoluene + NAD+ + H2O | - |
? | |
orcinol + NADH + O2 | - |
Pseudomonas putida 1 | 2,3,5-trihydroxytoluene + NAD+ + H2O | - |
? | |
resorcinol + NADH + O2 | - |
Pseudomonas putida | hydroxyquinol + NAD+ + H2O | - |
? | |
resorcinol + NADH + O2 | - |
Pseudomonas putida 1 | hydroxyquinol + NAD+ + H2O | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 68000, SDS-PAGE | Pseudomonas putida |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
26 | - |
orcinol | crystalline enzyme | Pseudomonas putida |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
difficult to evaluate, because the values do not take into account the nonenzymic oxidation rate of the product of the reaction | Pseudomonas putida |
6.8 | - |
assay at | Pseudomonas putida |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
7 | - |
highest stability | Pseudomonas putida |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
3-Acetyl pyridine nucleotide | reduced | Pseudomonas putida | |
FAD | flavoprotein | Pseudomonas putida | |
FAD | 1 mol of FAD per mol of protein | Pseudomonas putida | |
NADH | - |
Pseudomonas putida | |
NADPH | - |
Pseudomonas putida |