Application | Comment | Organism |
---|---|---|
synthesis | LAP is an important enzyme for the industrial production of enantiomerically pure amino acids | Pseudomonas putida |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Pseudomonas putida |
Crystallization (Comment) | Organism |
---|---|
purified enzyme in complex with inhibitor bestatin at pH 5.2 and pH 9.5, hanging drop vapour diffusion, 8 mg/ml protein in 20 mM HEPESKOH, pH 8.0, 1 mM DTT, is mixed with 11% w/v PEG 8000, 0.2 M sodium formate, 0.1 M MesNaOH, pH 5.2, 1 mM NaN3 at 5°C, or 4 mg/ml protein solution is mixed with 15% w/v PEG 1500, 0.1 M propionic acid, cacodylate, bis-Tris propane cocktail buffer, pH 9.5 at 23°C, three days, X-ray diffraction structure determination and analysis at 1.5-2.75 A resolution, modelling | Pseudomonas putida |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
bestatin | binding structure and comparison, overview | Pseudomonas putida |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | at pH 9.5, the active site contains two metal ions, one identified as Mn2+ or Zn2+ at site 1, and the other as Zn2+ at site 2. Mn2+ has a significant activation effect when bound to site 1 of ppLAP | Pseudomonas putida | |
additional information | ppLAP requires the presence of divalent metal ions for its activity, in particular Zn2+ and/or Mn2+. At pH 5.2, the active site of ppLAP is highly disordered and metal ions are absent, most probably due to full protonation of one of the metal-interacting residues, Lys267 | Pseudomonas putida | |
Zn2+ | LAP is zinc-dependent. At pH 9.5, the active site contains two metal ions, one identified as Mn2+ or Zn2+ at site 1, and the other as Zn2+ at site 2 | Pseudomonas putida |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
270000 | - |
recombinant enzyme, gel filtration | Pseudomonas putida |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | O86436 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme from Escherichia coli by cation exchange chromatography and gel filtration | Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | LAP shows broad substrate specificity and high enantioselectivity, structure-function relationship, active site metal composition and metal-dependent activity, overview | Pseudomonas putida | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
LAP | - |
Pseudomonas putida |
leucine aminopeptidase | - |
Pseudomonas putida |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Pseudomonas putida |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
the enzyme is inactive at low pH | Pseudomonas putida |