Cloned (Comment) | Organism |
---|---|
expression of mutant enzmyes F10Y, F10A, F13Y, F13A, F142Y and F142A | Bacillus amyloliquefaciens |
Crystallization (Comment) | Organism |
---|---|
crystallographic analysis of enzyme-inhibitor complex with pyroglutaminal and of the mutant enzyme F142, hanging drop vapour diffusion method | Bacillus amyloliquefaciens |
Protein Variants | Comment | Organism |
---|---|---|
F10Y | the ratio of turnover number to Km-value is 4.6% of the wild-type value | Bacillus amyloliquefaciens |
F13A | the ratio of turnover number to Km-value is 0.04% of the wild-type value | Bacillus amyloliquefaciens |
F13Y | the ratio of turnover number to Km-value is 51.5% of the wild-type value | Bacillus amyloliquefaciens |
F142A | the ratio of turnover number to Km-value is 0.06% of the wild-type value | Bacillus amyloliquefaciens |
F142Y | the ratio of turnover number to Km-value is 81.3% of the wild-type value | Bacillus amyloliquefaciens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
pyroglutaminal | - |
Bacillus amyloliquefaciens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.17 | - |
L-pyroglutamyl-beta-naphthylamide | pH 7.0, 37°C, mutant enzyme F142Y | Bacillus amyloliquefaciens | |
0.2 | - |
L-pyroglutamyl-beta-naphthylamide | pH 7.0, 37°C, mutant enzyme F13Y | Bacillus amyloliquefaciens | |
0.21 | - |
L-pyroglutamyl-beta-naphthylamide | pH 7.0, 37°C, wild-type enzyme | Bacillus amyloliquefaciens | |
0.55 | - |
L-pyroglutamyl-beta-naphthylamide | pH 7.0, 37°C, mutant enzyme F142A | Bacillus amyloliquefaciens | |
0.76 | - |
L-pyroglutamyl-beta-naphthylamide | pH 7.0, 37°C, mutant enzyme F10Y | Bacillus amyloliquefaciens | |
1.46 | - |
L-pyroglutamyl-beta-naphthylamide | pH 7.0, 37°C, mutant enzyme F142Y | Bacillus amyloliquefaciens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
pyroglutamyl-peptide + H2O | Bacillus amyloliquefaciens | the enzyme is able to specifically remove the amino-terminal pyroglutamyl residue protecting proteins or peptides from aminopeptidases | pyroglutamate + peptide | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus amyloliquefaciens | - |
- |
- |
Purification (Comment) | Organism |
---|---|
purification of mutant enzmyes F10Y, F10A, F13Y, F13A, F142Y and F142A | Bacillus amyloliquefaciens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-pyroglutamyl-beta-naphthylamide + H2O | - |
Bacillus amyloliquefaciens | L-pyroglutamate + beta-naphthylamine | - |
? | |
additional information | the molecular recognition of pyroglutamic acid is achieved through two hydrogen bonds and an insertion in the hydrophobic pocket. In the pocket Phe10 is more important to the hydrophobic interaction than is Phe142 and furthermore Phe13 serves as an induced fit mechanism | Bacillus amyloliquefaciens | ? | - |
? | |
pyroglutamyl-peptide + H2O | the enzyme is able to specifically remove the amino-terminal pyroglutamyl residue protecting proteins or peptides from aminopeptidases | Bacillus amyloliquefaciens | pyroglutamate + peptide | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.5 | - |
L-pyroglutamyl-beta-naphthylamide | pH 7.0, 37°C, mutant enzyme F142A | Bacillus amyloliquefaciens | |
0.8 | - |
L-pyroglutamyl-beta-naphthylamide | pH 7.0, 37°C, mutant enzyme F142Y | Bacillus amyloliquefaciens | |
54.7 | - |
L-pyroglutamyl-beta-naphthylamide | pH 7.0, 37°C, mutant enzyme F10Y | Bacillus amyloliquefaciens | |
163 | - |
L-pyroglutamyl-beta-naphthylamide | pH 7.0, 37°C, mutant enzyme F13Y | Bacillus amyloliquefaciens | |
210 | - |
L-pyroglutamyl-beta-naphthylamide | pH 7.0, 37°C, mutant enzyme F142Y | Bacillus amyloliquefaciens | |
319 | - |
L-pyroglutamyl-beta-naphthylamide | pH 7.0, 37°C, wild-type enzyme | Bacillus amyloliquefaciens |