Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(lambdaDE3) | Pseudomonas putida |
Crystallization (Comment) | Organism |
---|---|
hanging drop method, PEG 4000 | Pseudomonas putida |
Protein Variants | Comment | Organism |
---|---|---|
R123A | no activity | Pseudomonas putida |
R123K | retains some activity (substrate 4-hydroxy-4-methyl-2-oxoglutarate) | Pseudomonas putida |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.03 | - |
4-carboxy-4-hydroxy-2-oxoadipate | wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C | Pseudomonas putida | |
0.066 | - |
4-carboxy-4-hydroxy-2-oxoadipate | wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C | Pseudomonas putida |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Pseudomonas putida |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
26000 | - |
6 * 26000, crystal structure, gel filtration and SDS-PAGE | Pseudomonas putida |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | A5W059 | - |
- |
Purification (Comment) | Organism |
---|---|
ion exchange chromatography, hydrophobic interaction chromatography, gel filtration | Pseudomonas putida |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
37.7 | - |
wild type protein, pH 8.0, 25°C | Pseudomonas putida |
Storage Stability | Organism |
---|---|
-80°C, 20 mM sodium HEPES buffer, pH7.5, stable for 2 months | Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-carboxy-4-hydroxy-2-oxoadipate | - |
Pseudomonas putida | pyruvate + oxaloacetate | - |
r |
Subunits | Comment | Organism |
---|---|---|
homohexamer | 6 * 26000, crystal structure, gel filtration and SDS-PAGE | Pseudomonas putida |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
7.5 | - |
4-carboxy-4-hydroxy-2-oxoadipate | wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C | Pseudomonas putida | |
14.4 | - |
4-carboxy-4-hydroxy-2-oxoadipate | wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C | Pseudomonas putida |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
220 | - |
4-carboxy-4-hydroxy-2-oxoadipate | wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C | Pseudomonas putida | |
250 | - |
4-carboxy-4-hydroxy-2-oxoadipate | wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C | Pseudomonas putida |