Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21-Gold (DE3) cells | Pseudomonas putida |
Protein Variants | Comment | Organism |
---|---|---|
H244A | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Pseudomonas putida |
H351A | inactive | Pseudomonas putida |
Q242A | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Pseudomonas putida |
R290A | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Pseudomonas putida |
R386A | inactive | Pseudomonas putida |
S389A | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Pseudomonas putida |
T259A | inactive | Pseudomonas putida |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
L-asparagine | - |
Pseudomonas putida |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.26 | - |
L-aspartate | wild type enzyme, at pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
14.25 | - |
L-aspartate | mutant enzyme S389A, at pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
26.11 | - |
L-aspartate | mutant enzyme R290A, at pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
63.27 | - |
L-aspartate | mutant enzyme Q242A, at pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
75.44 | - |
L-aspartate | mutant enzyme H244A, at pH 7.5, temperature not specified in the publication | Pseudomonas putida |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
59900 | - |
x * 59900, calculated from amino acid sequence | Pseudomonas putida |
60000 | - |
x * 60000, SDS-PAGE | Pseudomonas putida |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-asparagine + H2O + O2 | Pseudomonas putida | low activity | 4-amino-2,4-dioxobutanoate + NH3 + H2O2 | - |
? | |
L-aspartate + H2O + O2 | Pseudomonas putida | best substrate | oxaloacetate + NH3 + H2O2 | - |
? | |
L-glutamate + H2O + O2 | Pseudomonas putida | low activity | 2-oxopentanedioate + NH3 + H2O2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | - |
- |
- |
Purification (Comment) | Organism |
---|---|
HiTrap nickel chelating column chromatography and Superdex 75 gel filtration | Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-asparagine + H2O + O2 | low activity | Pseudomonas putida | 4-amino-2,4-dioxobutanoate + NH3 + H2O2 | - |
? | |
L-aspartate + H2O + O2 | best substrate | Pseudomonas putida | oxaloacetate + NH3 + H2O2 | - |
? | |
L-glutamate + H2O + O2 | low activity | Pseudomonas putida | 2-oxopentanedioate + NH3 + H2O2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 60000, SDS-PAGE | Pseudomonas putida |
? | x * 59900, calculated from amino acid sequence | Pseudomonas putida |
Synonyms | Comment | Organism |
---|---|---|
LASPO | - |
Pseudomonas putida |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | 50 | the enzyme is stable for 60 min at up to 40°C, but rapid losses in activity are observed at 50°C after 5 min | Pseudomonas putida |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.23 | - |
L-aspartate | mutant enzyme R290A, at pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
2.28 | - |
L-aspartate | mutant enzyme Q242A, at pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
7.83 | - |
L-aspartate | mutant enzyme S389A, at pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
10.6 | - |
L-aspartate | wild type enzyme, at pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
23.69 | - |
L-aspartate | mutant enzyme H244A, at pH 7.5, temperature not specified in the publication | Pseudomonas putida |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
- |
Pseudomonas putida |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 8 | about 40% activity at pH 6.0, about 50% activity at pH 8.0 | Pseudomonas putida |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Pseudomonas putida |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00884 | - |
L-aspartate | mutant enzyme R290A, at pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
0.036 | - |
L-aspartate | mutant enzyme Q242A, at pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
0.314 | - |
L-aspartate | mutant enzyme H244A, at pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
0.549 | - |
L-aspartate | mutant enzyme S389A, at pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
4.69 | - |
L-aspartate | wild type enzyme, at pH 7.5, temperature not specified in the publication | Pseudomonas putida |