Cloned (Comment) | Organism |
---|---|
gene PIPA, sequence comparisons, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha | Phanerodontia chrysosporium |
Protein Variants | Comment | Organism |
---|---|---|
C267A | site-directed mutagenesis, the mutant shows reduced activity compared to wild-type, as well as reduced thermostability | Phanerodontia chrysosporium |
E198A | site-directed mutagenesis, the mutation abolishes the enzyme activity | Phanerodontia chrysosporium |
E198Q | site-directed mutagenesis, the mutation abolishes the enzyme activity | Phanerodontia chrysosporium |
E227A | site-directed mutagenesis, the mutation abolishes the enzyme activity | Phanerodontia chrysosporium |
F133A | site-directed mutagenesis, the mutant shows reduced activity compared to wild-type, as well as reduced thermostability | Phanerodontia chrysosporium |
F143A | site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type, as well as reduced thermostability | Phanerodontia chrysosporium |
F223A | site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type, as well as significantly reduced thermostability | Phanerodontia chrysosporium |
F231A | site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type, as well as highly reduced thermostability | Phanerodontia chrysosporium |
V266A | site-directed mutagenesis, almost inactive mutant | Phanerodontia chrysosporium |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.0178 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant F231A | Phanerodontia chrysosporium | |
2.0927 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant wild-type enzyme | Phanerodontia chrysosporium | |
2.1839 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant F133A | Phanerodontia chrysosporium | |
3.8809 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant C267A | Phanerodontia chrysosporium | |
6.5407 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant F223A | Phanerodontia chrysosporium | |
8.8634 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant E227A | Phanerodontia chrysosporium | |
11.8139 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant F143A | Phanerodontia chrysosporium | |
12.4863 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant E198A | Phanerodontia chrysosporium | |
13.7676 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant E198Q | Phanerodontia chrysosporium |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Phanerodontia chrysosporium | C6KI04 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) | Phanerodontia chrysosporium |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the proline iminopeptidase, PchPiPA, catalyzes specific hydrolysis of N-terminal proline from peptides | Phanerodontia chrysosporium | ? | - |
? | |
Pro-beta-naphthylamide + H2O | docking into the catalytic site of PipA | Phanerodontia chrysosporium | Pro + beta-naphthylamine | - |
? |
Synonyms | Comment | Organism |
---|---|---|
PchPiPA | - |
Phanerodontia chrysosporium |
PiPA | - |
Phanerodontia chrysosporium |
proline iminopeptidase | - |
Phanerodontia chrysosporium |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
- |
Phanerodontia chrysosporium |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
effect of heat treatment at 50°C on wild-type and mutant enzymes, overview. The most heat labile mutants are F143A, F223A, and F231A | Phanerodontia chrysosporium |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.173 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant E198A | Phanerodontia chrysosporium | |
0.192 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant E198Q | Phanerodontia chrysosporium | |
0.534 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant E227A | Phanerodontia chrysosporium | |
0.597 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant F143A | Phanerodontia chrysosporium | |
3.62 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant F231A | Phanerodontia chrysosporium | |
6.04 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant F223A | Phanerodontia chrysosporium | |
22.57 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant C267A | Phanerodontia chrysosporium | |
30.43 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant F133A | Phanerodontia chrysosporium | |
64.55 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant wild-type enzyme | Phanerodontia chrysosporium |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
- |
Phanerodontia chrysosporium |
General Information | Comment | Organism |
---|---|---|
additional information | proline iminopeptidase, PchPiPA, molecular docking of substrate Pro-beta-naphthylamide, overview. The peptide bond in Pro-beta-naphthylamide is associated with Glu198, Ser107, and Gly40, respectively, via formation of hydrogen bond. Further, some hydrophobic residues, like Phe133, Phe143, Phe223, Val266, and Cys267, are surrounding the docked substrate to form the catalytic pocket with a channel protruding out of the molecular surface. The substrate is believed to diffuse into the catalytic pocket | Phanerodontia chrysosporium |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.014 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant E198A | Phanerodontia chrysosporium | |
0.014 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant E198Q | Phanerodontia chrysosporium | |
0.051 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant F143A | Phanerodontia chrysosporium | |
0.06 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant E227A | Phanerodontia chrysosporium | |
0.92 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant F223A | Phanerodontia chrysosporium | |
1.8 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant F231A | Phanerodontia chrysosporium | |
5.82 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant C267A | Phanerodontia chrysosporium | |
13.93 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant mutant F133A | Phanerodontia chrysosporium | |
30.84 | - |
Pro-beta-naphthylamide | pH 8.0, 45°C, recombinant wild-type enzyme | Phanerodontia chrysosporium |