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Literature summary extracted from
- Metabolism of resorcinylic compounds by bacteria. Purification and properties of orcinol hydroxylase from Pseudomonas putida 01 (1975), J. Biol. Chem., 250, 3814-3825.
Activating Compound
| EC Number | Activating Compound | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.13.6 | m-cresol | increases activity | Pseudomonas putida |  |
Crystallization (Commentary)
| EC Number | Crystallization (Comment) | Organism |
|---|---|---|
| 1.14.13.6 | - |
Pseudomonas putida |
General Stability
| EC Number | General Stability | Organism |
|---|---|---|
| 1.14.13.6 | thiol reagents, e.g. 2-mercaptoethanol, stabilize | Pseudomonas putida |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.13.6 | 1,3-Dimethoxybenzene | 45% inhibition at 1.7 mM | Pseudomonas putida | |
| 1.14.13.6 | 2-Hydroxy-4-methoxybenzoate | 68% inhibition at 1.7 mM | Pseudomonas putida | |
| 1.14.13.6 | 2-Methylresorcinol | 27% inhibition at 1.7 mM | Pseudomonas putida | |
| 1.14.13.6 | 2-Nitroorcinol | 64% inhibition at 1.7 mM | Pseudomonas putida | |
| 1.14.13.6 | 3,4-dimethylphenol | 54% inhibition at 1.7 mM | Pseudomonas putida | |
| 1.14.13.6 | 3,5-Dihydroxybenzoate | 59% inhibition at 1.7 mM | Pseudomonas putida | |
| 1.14.13.6 | 3,5-Dimethylphenol | 68% inhibition at 1.7 mM | Pseudomonas putida | |
| 1.14.13.6 | 3-Hydroxybenzaldehyde | 23% inhibition at 1.7 mM | Pseudomonas putida | |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.13.6 | additional information | - |
additional information | - |
Pseudomonas putida | |
| 1.14.13.6 | 0.03 | - |
orcinol | cofactor: NADH | Pseudomonas putida | |
| 1.14.13.6 | 0.05 | - |
orcinol | cofactor: 3-acetylpyridine NADH | Pseudomonas putida | |
| 1.14.13.6 | 0.07 | - |
O2 | - |
Pseudomonas putida | |
| 1.14.13.6 | 0.085 | - |
orcinol | cofactor: NADPH | Pseudomonas putida | |
| 1.14.13.6 | 0.13 | - |
NADH | substrate: orcinol | Pseudomonas putida | |
| 1.14.13.6 | 0.19 | - |
resorcinol | - |
Pseudomonas putida | |
| 1.14.13.6 | 0.65 | - |
3-acetylpyridine-NADH | substrate: orcinol | Pseudomonas putida | |
| 1.14.13.6 | 2.5 | - |
NADH | substrate: resorcinol | Pseudomonas putida | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 1.14.13.6 | 63000 | - |
gel filtration | Pseudomonas putida |
| 1.14.13.6 | 65000 | - |
sedimentation equilibrium | Pseudomonas putida |
| 1.14.13.6 | 68000 | - |
1 * 68000, SDS-PAGE | Pseudomonas putida |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.14.13.6 | Pseudomonas putida | - |
01 | - |
| 1.14.13.6 | Pseudomonas putida 1 | - |
01 | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.14.13.6 | 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 (Commentary)
| EC Number | Renatured (Comment) | Organism |
|---|---|---|
| 1.14.13.6 | 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 [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 1.14.13.6 | 2.8 | - |
apoenzyme without flavin addition | Pseudomonas putida |
| 1.14.13.6 | 3.5 | - |
apoenzyme with addition of FMN | Pseudomonas putida |
| 1.14.13.6 | 14.7 | - |
apoenzyme with addition of FAD | Pseudomonas putida |
| 1.14.13.6 | 18 | - |
holoenzyme | Pseudomonas putida |
| 1.14.13.6 | 24 | - |
- |
Pseudomonas putida |
Storage Stability
| EC Number | Storage Stability | Organism |
|---|---|---|
| 1.14.13.6 | 4°C, stabilized by thiol reagents, e.g. 2-mercaptoethanol, some days | Pseudomonas putida |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.13.6 | 3-cresol + NADH + O2 | nonsubstrate effector, which increases NADH oxidase activity without being hydroxylated | Pseudomonas putida | 3-methylcatechol + NAD+ + H2O | - |
? | |
| 1.14.13.6 | 3-cresol + NADH + O2 | nonsubstrate effector, which increases NADH oxidase activity without being hydroxylated | Pseudomonas putida 1 | 3-methylcatechol + NAD+ + H2O | - |
? | |
| 1.14.13.6 | 3-ethylphenol + NADH + O2 | no substrate, mimic orcinol as effector | Pseudomonas putida | ? | - |
? | |
| 1.14.13.6 | 3-ethylphenol + NADH + O2 | no substrate, mimic orcinol as effector | Pseudomonas putida 1 | ? | - |
? | |
| 1.14.13.6 | 4-bromoresorcinol + NADH + O2 | - |
Pseudomonas putida | ? | - |
? | |
| 1.14.13.6 | 4-methylresorcinol + NADH + O2 | - |
Pseudomonas putida | ? | - |
? | |
| 1.14.13.6 | 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 | ? | - |
? | |
| 1.14.13.6 | 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 | ? | - |
? | |
| 1.14.13.6 | orcinol + 3-acetylpyridine-NADH + H+ + O2 | - |
Pseudomonas putida | 2,3,5-trihydroxytoluene + 3-acetylpyridine-NAD+ + H2O | - |
r | |
| 1.14.13.6 | orcinol + NADH + O2 | - |
Pseudomonas putida | 2,3,5-trihydroxytoluene + NAD+ + H2O | - |
? | |
| 1.14.13.6 | orcinol + NADH + O2 | - |
Pseudomonas putida 1 | 2,3,5-trihydroxytoluene + NAD+ + H2O | - |
? | |
| 1.14.13.6 | resorcinol + NADH + O2 | - |
Pseudomonas putida | hydroxyquinol + NAD+ + H2O | - |
? | |
| 1.14.13.6 | resorcinol + NADH + O2 | - |
Pseudomonas putida 1 | hydroxyquinol + NAD+ + H2O | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.14.13.6 | monomer | 1 * 68000, SDS-PAGE | Pseudomonas putida |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.13.6 | 26 | - |
orcinol | crystalline enzyme | Pseudomonas putida | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.14.13.6 | 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 |
| 1.14.13.6 | 6.8 | - |
assay at | Pseudomonas putida |
pH Stability
| EC Number | pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.14.13.6 | 7 | - |
highest stability | Pseudomonas putida |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.13.6 | 3-Acetyl pyridine nucleotide | reduced | Pseudomonas putida | |
| 1.14.13.6 | FAD | flavoprotein | Pseudomonas putida | |
| 1.14.13.6 | FAD | 1 mol of FAD per mol of protein | Pseudomonas putida | |
| 1.14.13.6 | NADH | - |
Pseudomonas putida | |
| 1.14.13.6 | NADPH | - |
Pseudomonas putida | |
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