Literature summary extracted from
Kale, A.; Pijning, T.; Sonke, T.; Dijkstra, B.W.; Thunnissen, A.M.
Crystal structure of the leucine aminopeptidase from Pseudomonas putida reveals the molecular basis for its enantioselectivity and broad substrate specificity (2010), J. Mol. Biol., 398, 703-714.
Application
EC Number |
Application |
Comment |
Organism |
---|
3.4.11.10 |
synthesis |
LAP is an important enzyme for the industrial production of enantiomerically pure amino acids |
Pseudomonas putida |
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
3.4.11.10 |
expression in Escherichia coli |
Pseudomonas putida |
Crystallization (Commentary)
EC Number |
Crystallization (Comment) |
Organism |
---|
3.4.11.10 |
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
EC Number |
Inhibitors |
Comment |
Organism |
Structure |
---|
3.4.11.10 |
bestatin |
binding structure and comparison, overview |
Pseudomonas putida |
|
Metals/Ions
EC Number |
Metals/Ions |
Comment |
Organism |
Structure |
---|
3.4.11.10 |
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 |
|
3.4.11.10 |
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 |
|
3.4.11.10 |
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]
EC Number |
Molecular Weight [Da] |
Molecular Weight Maximum [Da] |
Comment |
Organism |
---|
3.4.11.10 |
270000 |
- |
recombinant enzyme, gel filtration |
Pseudomonas putida |
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
3.4.11.10 |
Pseudomonas putida |
O86436 |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
3.4.11.10 |
recombinant enzyme from Escherichia coli by cation exchange chromatography and gel filtration |
Pseudomonas putida |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
3.4.11.10 |
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
EC Number |
Synonyms |
Comment |
Organism |
---|
3.4.11.10 |
LAP |
- |
Pseudomonas putida |
3.4.11.10 |
leucine aminopeptidase |
- |
Pseudomonas putida |
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
3.4.11.10 |
8 |
- |
assay at |
Pseudomonas putida |
pH Range
EC Number |
pH Minimum |
pH Maximum |
Comment |
Organism |
---|
3.4.11.10 |
additional information |
- |
the enzyme is inactive at low pH |
Pseudomonas putida |