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Literature summary extracted from
- Purification and characterization of the 4-hydroxyphenylacetic acid-3-hydroxylase from Pseudomonas putida U (1997), FEMS Microbiol. Lett., 157, 47-53.
No PubMed abstract available
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.14.9 | Cibacron blue F3GA | - |
Pseudomonas putida |  |
| 1.14.14.9 | Co2+ | - |
Pseudomonas putida | |
| 1.14.14.9 | Cu2+ | - |
Pseudomonas putida | |
| 1.14.14.9 | Hg2+ | - |
Pseudomonas putida | |
| 1.14.14.9 | iodoacetate | - |
Pseudomonas putida | |
| 1.14.14.9 | additional information | SH-groups in the active site of the enzyme suggested | Pseudomonas putida | |
| 1.14.14.9 | N-ethylmaleimide | - |
Pseudomonas putida | |
| 1.14.14.9 | Zn2+ | - |
Pseudomonas putida | |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.14.9 | 0.0042 | - |
FAD | - |
Pseudomonas putida | |
| 1.14.14.9 | 0.038 | - |
4-hydroxyphenylacetate | - |
Pseudomonas putida | |
| 1.14.14.9 | 0.041 | - |
NADH | - |
Pseudomonas putida | |
| 1.14.14.9 | 0.16 | - |
phenylacetic acid | - |
Pseudomonas putida | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 1.14.14.9 | 32500 | - |
2 * 32500, alpha2 oligomeric structure, SDS-PAGE | Pseudomonas putida |
| 1.14.14.9 | 65000 | - |
gel filtration | Pseudomonas putida |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.14.9 | 4-hydroxyphenylacetate + NADH + O2 | Pseudomonas putida | degradative pathway involved in the assimilation of different aromatic compounds | 3,4-dihydroxyphenylacetate + NAD+ + H2O | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.14.14.9 | Pseudomonas putida | - |
- |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.14.14.9 | ammonium sulfate precipitation, ion-exchange, gel filtration | Pseudomonas putida |
Reaction
| EC Number | Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|---|
| 1.14.14.9 | 4-hydroxyphenylacetate + FADH2 + O2 = 3,4-dihydroxyphenylacetate + FAD + H2O | family of external flavoprotein monooxygenases. The activity is induced by the substrates | Pseudomonas putida |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 1.14.14.9 | 197.3 | - |
- |
Pseudomonas putida |
Storage Stability
| EC Number | Storage Stability | Organism |
|---|---|---|
| 1.14.14.9 | -20°C, stable for at least 5 months | Pseudomonas putida |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.14.9 | 2-phenylbutyric acid + H2 | 86% of 4-hydroxyphenylacetate activity | Pseudomonas putida | ? | - |
? | |
| 1.14.14.9 | 2-phenylpropionic acid + ? | 77% of 4-hydroxyphenylacetate activity | Pseudomonas putida | ? | - |
? | |
| 1.14.14.9 | 3,4-dihydroxyphenylacetate + FAD + H2O | - |
Pseudomonas putida | 4-hydroxyphenylacetate + FADH2 + O2 | - |
r | |
| 1.14.14.9 | 3-(4-hydroxyphenyl)propionate + ? | 83% of 4-hydroxyphenylacetate activity | Pseudomonas putida | 3-(3,4-dihydroxy)phenylpropionate + ? | - |
? | |
| 1.14.14.9 | 4-hydroxyphenylacetate + NADH + O2 | - |
Pseudomonas putida | 3,4-dihydroxyphenylacetate + NAD+ + H2O | - |
? | |
| 1.14.14.9 | 4-hydroxyphenylacetate + NADH + O2 | degradative pathway involved in the assimilation of different aromatic compounds | Pseudomonas putida | 3,4-dihydroxyphenylacetate + NAD+ + H2O | - |
? | |
| 1.14.14.9 | additional information | broad substrate range. Several other substrates show activities below 50%. The length of the acetyl moiety is very important in substrate recognition | Pseudomonas putida | ? | - |
? | |
| 1.14.14.9 | phenylacetic acid + ? | 100% of 4-hydroxyphenylacetate activity | Pseudomonas putida | ? | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.14.14.9 | dimer | 2 * 32500, alpha2 oligomeric structure, SDS-PAGE | Pseudomonas putida |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.14.14.9 | 40 | - |
- |
Pseudomonas putida |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.14.14.9 | 7.5 | - |
- |
Pseudomonas putida |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.14.9 | FAD | required for maximum activity, external flavoprotein monooxygenase | Pseudomonas putida | |
| 1.14.14.9 | NADH | unable to use NADPH | Pseudomonas putida | |
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