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Literature summary for 1.14.16.1 extracted from
- Structure of phenylalanine hydroxylase from Colwellia psychrerythraea 34H, a monomeric cold active enzyme with local flexibility around the active site and high overall stability (2007), J. Biol. Chem., 282, 21973-21986.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene pah, overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Colwellia psychrerythraea 34H |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| native enzyme by hanging drop method, 0.001 ml of protein solution containing 10 mg/ml protein in 20 mM Na-HEPES, 200 mM NaCl, pH 7.0, are mixed with 0.001 ml of reservoir solution containing 1.6-1.8 M ammonium sulfate, 100 mM NaCl, and 20 mM NaHepes, pH 7.5, equilibration at 4 °C, 3 weeks, cryoprotection with 25% glycerol, X-ray diffraction structure determination and analysis | Colwellia psychrerythraea 34H |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.3 | - |
tetrahydrobiopterin | pH 7.0, 10°C, recombinant enzyme | Colwellia psychrerythraea 34H |  |
| 0.5 | - |
tetrahydrobiopterin | pH 7.0, 25°C, recombinant enzyme | Colwellia psychrerythraea 34H | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Fe2+ | 0.1 mM added as ferrous ammonium sulfate, 40-70% activation depending on assay conditions | Colwellia psychrerythraea 34H | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 29000 | - |
recombinant enzyme, gel filtration | Colwellia psychrerythraea 34H |
| 31000 | - |
1 * 31000, recombinant enzyme, SDS-PAGE | Colwellia psychrerythraea 34H |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| L-phenylalanine + tetrahydrobiopterin + O2 | Colwellia psychrerythraea 34H | - |
L-tyrosine + 4a-hydroxytetrahydrobiopterin | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Colwellia psychrerythraea 34H | - |
a psychrophilic bacterium | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged enzyme from Escherichia coli strain BL21 by metal affinity chromatography | Colwellia psychrerythraea 34H |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| additional information | optimal growth temperature is 9°C | Colwellia psychrerythraea 34H | - |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| L-phenylalanine + tetrahydrobiopterin + O2 | - |
Colwellia psychrerythraea 34H | L-tyrosine + 4a-hydroxytetrahydrobiopterin | - |
? | |
| L-phenylalanine + tetrahydrobiopterin + O2 | although the enzyme is monomeric with a single L-Phe-binding site, the substrate binds cooperatively | Colwellia psychrerythraea 34H | L-tyrosine + 4a-hydroxytetrahydrobiopterin | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 * 31000, recombinant enzyme, SDS-PAGE | Colwellia psychrerythraea 34H |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| PAH | - |
Colwellia psychrerythraea 34H |
| phenylalanine hydroxylase | - |
Colwellia psychrerythraea 34H |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
- |
Colwellia psychrerythraea 34H |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 10 | 25 | - |
Colwellia psychrerythraea 34H |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 52 | - |
Tm, inactivation | Colwellia psychrerythraea 34H |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
assay at | Colwellia psychrerythraea 34H |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| (6R)-5,6,7,8-tetrahydro-L-monapterin | 7% of the activity with tetrahydrobiopterin | Colwellia psychrerythraea 34H | |
| 6,7-dimethyltetrahydropterin | 17% of the activity with tetrahydrobiopterin | Colwellia psychrerythraea 34H | |
| tetrahydrobiopterin | - |
Colwellia psychrerythraea 34H | |
| tetrahydrofolate | 20% of the activity with tetrahydrobiopterin | Colwellia psychrerythraea 34H | |
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