Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli M15 cells | Geobacillus stearothermophilus |
Protein Variants | Comment | Organism |
---|---|---|
D380E | the mutation causes more than 2fold decrease in kcat as compared to the wild type enzyme | Geobacillus stearothermophilus |
D380N | the mutation causes more than 2fold decrease in kcat as compared to the wild type enzyme | Geobacillus stearothermophilus |
D380V | the mutation causes over 5fold decrease in kcat as compared to the wild type enzyme | Geobacillus stearothermophilus |
Y352D | the mutation results in about 3.5fold decrease in specific activity | Geobacillus stearothermophilus |
Y352L | the mutation results in about 3.5fold decrease in specific activity | Geobacillus stearothermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.95 | - |
L-Leu-4-nitroanilide | wild type enzyme, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C | Geobacillus stearothermophilus | |
2.12 | - |
L-Leu-4-nitroanilide | mutant enzyme Y352D, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C | Geobacillus stearothermophilus | |
2.83 | - |
L-Leu-4-nitroanilide | mutant enzyme Y352L, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C | Geobacillus stearothermophilus | |
3.05 | - |
L-Leu-4-nitroanilide | mutant enzyme D380N, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C | Geobacillus stearothermophilus | |
3.39 | - |
L-Leu-4-nitroanilide | mutant enzyme D380E, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C | Geobacillus stearothermophilus | |
3.41 | - |
L-Leu-4-nitroanilide | mutant enzyme D380V, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C | Geobacillus stearothermophilus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
45000 | - |
SDS-PAGE | Geobacillus stearothermophilus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Geobacillus stearothermophilus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Ni2+-NTA resin chromatography | Geobacillus stearothermophilus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
13.8 | - |
mutant enzyme D380E, at 60°C, using L-Leu-4-nitroanilide as substrate | Geobacillus stearothermophilus |
15.1 | - |
mutant enzyme D380V, at 60°C, using L-Leu-4-nitroanilide as substrate | Geobacillus stearothermophilus |
34.5 | - |
mutant enzyme Y532L, at 60°C, using L-Leu-4-nitroanilide as substrate | Geobacillus stearothermophilus |
55.3 | - |
mutant enzyme D380N, at 60°C, using L-Leu-4-nitroanilide as substrate | Geobacillus stearothermophilus |
104.9 | - |
mutant enzyme Y532D, at 60°C, using L-Leu-4-nitroanilide as substrate | Geobacillus stearothermophilus |
572.5 | - |
wild type enzyme, at 60°C, using L-Leu-4-nitroanilide as substrate | Geobacillus stearothermophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-Leu-4-nitroanilide + H2O | - |
Geobacillus stearothermophilus | L-Leu + 4-nitroaniline | - |
? |
Synonyms | Comment | Organism |
---|---|---|
aminopeptidase II | - |
Geobacillus stearothermophilus |
AP-II | - |
Geobacillus stearothermophilus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.035 | - |
L-Leu-4-nitroanilide | mutant enzyme D380E, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C | Geobacillus stearothermophilus | |
0.048 | - |
L-Leu-4-nitroanilide | mutant enzyme D380V, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C | Geobacillus stearothermophilus | |
0.113 | - |
L-Leu-4-nitroanilide | mutant enzyme Y352L, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C | Geobacillus stearothermophilus | |
0.135 | - |
L-Leu-4-nitroanilide | mutant enzyme D380N, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C | Geobacillus stearothermophilus | |
0.153 | - |
L-Leu-4-nitroanilide | mutant enzyme Y352D, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C | Geobacillus stearothermophilus | |
0.303 | - |
L-Leu-4-nitroanilide | wild type enzyme, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C | Geobacillus stearothermophilus |