Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli M15 cells | Geobacillus kaustophilus |
Protein Variants | Comment | Organism |
---|---|---|
N345L | the mutation results in 93% reduction in LAP activity | Geobacillus kaustophilus |
N345Q | the mutation results in 72% reduction in LAP activity | Geobacillus kaustophilus |
N435E | the mutation results in complete loss of the LAP activity | Geobacillus kaustophilus |
N435L | the mutation results in complete loss of the LAP activity | Geobacillus kaustophilus |
N435Q | the mutation results in complete loss of the LAP activity | Geobacillus kaustophilus |
N435R | the mutation results in complete loss of the LAP activity | Geobacillus kaustophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.23 | - |
Leu-p-nitroanilide | wild type enzyme, in 20 mM Tris-HCl buffer (pH 8.0) containing 1 mM NiCl2 | Geobacillus kaustophilus | |
0.52 | - |
Leu-p-nitroanilide | mutant enzyme N345Q, in 20 mM Tris-HCl buffer (pH 8.0) containing 1 mM NiCl2 | Geobacillus kaustophilus | |
0.97 | - |
Leu-p-nitroanilide | mutant enzyme N345L, in 20 mM Tris-HCl buffer (pH 8.0) containing 1 mM NiCl2 | Geobacillus kaustophilus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | LAP contains two Zn2+ ions in the active site | Geobacillus kaustophilus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
54000 | - |
SDS-PAGE | Geobacillus kaustophilus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Geobacillus kaustophilus | Q7X2C8 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni2+-NTA resin column chromatography | Geobacillus kaustophilus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
11.4 | - |
mutant enzyme N345L, using Leu-p-nitroanilide as substrate, at pH 8.0 | Geobacillus kaustophilus |
28.8 | - |
mutant enzyme N345Q, using Leu-p-nitroanilide as substrate, at pH 8.0 | Geobacillus kaustophilus |
160 | - |
wild type enzyme, using Leu-p-nitroanilide as substrate, at pH 8.0 | Geobacillus kaustophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
Leu-4-nitroanilide + H2O | - |
Geobacillus kaustophilus | Leu + 4-nitroaniline | - |
? |
Synonyms | Comment | Organism |
---|---|---|
LAP | - |
Geobacillus kaustophilus |
leucine aminopeptidase | - |
Geobacillus kaustophilus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
63 | - |
the melting temperature of the wild type enzyme is at 63°C | Geobacillus kaustophilus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.17 | - |
Leu-p-nitroanilide | mutant enzyme N345L, in 20 mM Tris-HCl buffer (pH 8.0) containing 1 mM NiCl2 | Geobacillus kaustophilus | |
0.63 | - |
Leu-p-nitroanilide | mutant enzyme N345Q, in 20 mM Tris-HCl buffer (pH 8.0) containing 1 mM NiCl2 | Geobacillus kaustophilus | |
5.92 | - |
Leu-p-nitroanilide | wild type enzyme, in 20 mM Tris-HCl buffer (pH 8.0) containing 1 mM NiCl2 | Geobacillus kaustophilus |