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Literature summary for 1.14.14.9 extracted from
- A novel two-protein component flavoprotein hydroxylase. p-Hydroxyphenylacetate hydroxylase from Acinetobacter baumannii (2001), Eur. J. Biochem., 268, 5550-5561.
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| FAD | inhibitory effect at more than 0.15 mM | Acinetobacter baumannii |  |
| FMN | inhibitory effect at more than 0.15 mM | Acinetobacter baumannii | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.012 | - |
NADH | apparent, with FMN | Acinetobacter baumannii | |
| 0.014 | - |
4-hydroxyphenylacetate | apparent, with FAD | Acinetobacter baumannii | |
| 0.019 | - |
4-hydroxyphenylacetate | apparent, with FMN | Acinetobacter baumannii | |
| 0.028 | - |
NADH | apparent, with FAD | Acinetobacter baumannii | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 19000 | - |
2 * 19000 small component, 2 * 59000 large component | Escherichia coli |
| 30750 | - |
2 * 30750, small component | Pseudomonas putida |
| 32000 | - |
x * 32000, small component, SDS-PAGE | Acinetobacter baumannii |
| 38500 | - |
1 * 38500, large component | Pseudomonas putida |
| 50000 | - |
homotetramer, 4 * 50000, large component, SDS-PAGE | Acinetobacter baumannii |
| 59000 | - |
2 * 19000 small component, 2 * 59000 large component | Escherichia coli |
| 73000 | - |
component C1, gel filtration | Acinetobacter baumannii |
| 209000 | - |
component C2, gel filtration | Acinetobacter baumannii |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-hydroxyphenylacetate + NADH + O2 | Escherichia coli | - |
3,4-dihydroxyphenylacetate + NAD+ + H2O | - |
? | |
| 4-hydroxyphenylacetate + NADH + O2 | Pseudomonas putida | - |
3,4-dihydroxyphenylacetate + NAD+ + H2O | - |
? | |
| 4-hydroxyphenylacetate + NADH + O2 | Acinetobacter baumannii | biodegradation of aromatic compounds | 3,4-dihydroxyphenylacetate + NAD+ + H2O | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Acinetobacter baumannii | - |
- |
- |
| Escherichia coli | - |
- |
- |
| Pseudomonas putida | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Escherichia coli |
- |
Pseudomonas putida |
| protamine sulfate precipitation, ion-exchange, gel filtration | Acinetobacter baumannii |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| 4-hydroxyphenylacetate + FADH2 + O2 = 3,4-dihydroxyphenylacetate + FAD + H2O | different from other hydroxylases. Two-protein component enzyme: component C1 is a flavoprotein and C2 the hydroxylase component. Mechanism: 4-hydroxyphenylacetate binds to C1, then the enzyme-bound flavin is reduced by NADH, this reduced flavin is transferred to C2 and the reoxidation of the flavin occurs concurrently with the hydroxylation of the substrate | Acinetobacter baumannii |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 8.89 | - |
C2 component | Acinetobacter baumannii |
| 201 | - |
C1 component | Acinetobacter baumannii |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 3-hydroxyphenylacetate + NAD(P)H + O2 | 98% of 4-hydroxyphenylacetate activity | Acinetobacter baumannii | 3,4-hydroxyphenylacetate + NAD(P)+ + H2O | - |
? | |
| 4-hydroxyphenylacetate + NADH + O2 | - |
Escherichia coli | 3,4-dihydroxyphenylacetate + NAD+ + H2O | - |
? | |
| 4-hydroxyphenylacetate + NADH + O2 | - |
Pseudomonas putida | 3,4-dihydroxyphenylacetate + NAD+ + H2O | - |
? | |
| 4-hydroxyphenylacetate + NADH + O2 | - |
Acinetobacter baumannii | 3,4-dihydroxyphenylacetate + NAD+ + H2O | - |
? | |
| 4-hydroxyphenylacetate + NADH + O2 | biodegradation of aromatic compounds | Acinetobacter baumannii | 3,4-dihydroxyphenylacetate + NAD+ + H2O | - |
? | |
| additional information | - |
Escherichia coli | ? | - |
? | |
| additional information | - |
Pseudomonas putida | ? | - |
? | |
| additional information | a variety of aromatic compounds that contain a hydroxyl group in para-position can be hydroxylated. 4-hydroxyphenylacetate hydroxylase is an effector for C1 and substrate for C2 | Acinetobacter baumannii | ? | - |
? | |
| phenylacetic acid + ? | - |
Pseudomonas putida | ? | - |
? | |
| phenylacetic acid + ? | 97% of 4-hydroxyphenylacetate activity | Acinetobacter baumannii | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | 2 * 30750, small component | Pseudomonas putida |
| dimer | x * 32000, small component, SDS-PAGE | Acinetobacter baumannii |
| dimer | 2 * 19000 small component, 2 * 59000 large component | Escherichia coli |
| monomer | 1 * 38500, large component | Pseudomonas putida |
| tetramer | homotetramer, 4 * 50000, large component, SDS-PAGE | Acinetobacter baumannii |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
assay at | Acinetobacter baumannii |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
- |
Pseudomonas putida |
| 7.5 | - |
assay at | Acinetobacter baumannii |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| FAD | - |
Escherichia coli | |
| FAD | alternative to FMN for C1 | Acinetobacter baumannii | |
| FAD | tightly associated with small component, native cofactor | Pseudomonas putida | |
| FADH2 | - |
Escherichia coli | |
| FADH2 | - |
Acinetobacter baumannii | |
| FMN | - |
Escherichia coli | |
| FMN | native cofactor for C1 component | Acinetobacter baumannii | |
| FMNH2 | - |
Acinetobacter baumannii | |
| NADH | - |
Escherichia coli | |
| NADH | - |
Pseudomonas putida | |
| NADH | - |
Acinetobacter baumannii | |
| NADPH | - |
Escherichia coli | |
| riboflavin | - |
Escherichia coli | |
| riboflavin | component C1 | Acinetobacter baumannii | |
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Release 2023.1 (January 2023)
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