Protein Variants | Comment | Organism |
---|---|---|
D647A | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
D647F | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
D649A | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
D649F | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
E125A | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
E125R | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
E75A | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
E75A/R658A | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
E75F | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
E96A | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
E96A/E125A | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
E96A/K660A | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
E96F | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
K655A | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
K655R | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
K660A | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
K660F | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
R658A | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
R658A/K660A | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
R658F | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
S656F | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Serratia proteamaculans |
S656Ter | site-directed mutagenesis, mutation S656Ter gives the truncated protein PSP1-655 | Serratia proteamaculans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics of wild-type and mutant enzymes, overview | Serratia proteamaculans | |
0.0028 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant mutant E125A, pH 8.0, 25°C | Serratia proteamaculans | |
0.00473 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant wild-type enzyme, pH 8.0, 25°C | Serratia proteamaculans | |
0.00552 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant mutant D647A, pH 8.0, 25°C | Serratia proteamaculans | |
0.00861 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant mutant D649A, pH 8.0, 25°C | Serratia proteamaculans | |
0.00864 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant mutant D647A, pH 8.0, 25°C | Serratia proteamaculans | |
0.00954 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant mutant K660A, pH 8.0, 25°C | Serratia proteamaculans | |
0.0106 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant mutant D649A, pH 8.0, 25°C | Serratia proteamaculans | |
0.01086 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant mutant K660A, pH 8.0, 25°C | Serratia proteamaculans | |
0.0172 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant mutant E125A, pH 8.0, 25°C | Serratia proteamaculans | |
0.0194 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant wild-type enzyme, pH 8.0, 25°C | Serratia proteamaculans | |
0.0279 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant mutant D649A, pH 8.0, 25°C | Serratia proteamaculans | |
0.0316 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant mutant D647A, pH 8.0, 25°C | Serratia proteamaculans | |
0.0317 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant mutant K655A, pH 8.0, 25°C | Serratia proteamaculans | |
0.046 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant mutant E125A, pH 8.0, 25°C | Serratia proteamaculans | |
0.0708 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant mutant K660A, pH 8.0, 25°C | Serratia proteamaculans | |
0.0836 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant wild-type enzyme, pH 8.0, 25°C | Serratia proteamaculans | |
0.143 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant mutant K655A, pH 8.0, 25°C | Serratia proteamaculans | |
0.51 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant mutant K655A, pH 8.0, 25°C | Serratia proteamaculans |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Serratia proteamaculans | B3VI58 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide + H2O | - |
Serratia proteamaculans | Nalpha-benzoyl-L-arginyl-L-arginine + 4-nitroaniline | - |
? | |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide + H2O | - |
Serratia proteamaculans | Nalpha-benzoyl-L-lysyl-L-arginine + 4-nitroaniline | - |
? | |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide + H2O | - |
Serratia proteamaculans | Nalpha-benzoyl-DL-arginine + 4-nitroaniline | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the enzyme has a two-domain structure of the enzyme including C-terminal peptidase catalytic domain and N-terminal seven-bladed beta-propeller domain | Serratia proteamaculans |
Synonyms | Comment | Organism |
---|---|---|
OpdB | - |
Serratia proteamaculans |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Serratia proteamaculans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
4.73 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant mutant K655A, pH 8.0, 25°C | Serratia proteamaculans | |
6.27 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant mutant K660A, pH 8.0, 25°C | Serratia proteamaculans | |
6.5 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant mutant D647A, pH 8.0, 25°C | Serratia proteamaculans | |
11.31 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant mutant K655A, pH 8.0, 25°C | Serratia proteamaculans | |
14.35 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant wild-type enzyme, pH 8.0, 25°C | Serratia proteamaculans | |
16.8 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant mutant D649A, pH 8.0, 25°C | Serratia proteamaculans | |
24.05 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant mutant K660A, pH 8.0, 25°C | Serratia proteamaculans | |
25.23 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant mutant D647A, pH 8.0, 25°C | Serratia proteamaculans | |
27.35 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant mutant K655A, pH 8.0, 25°C | Serratia proteamaculans | |
29.97 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant mutant K660A, pH 8.0, 25°C | Serratia proteamaculans | |
30.84 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant mutant D647A, pH 8.0, 25°C | Serratia proteamaculans | |
32.36 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant mutant D649A, pH 8.0, 25°C | Serratia proteamaculans | |
33.975 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant wild-type enzyme, pH 8.0, 25°C | Serratia proteamaculans | |
45.37 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant mutant E125A, pH 8.0, 25°C | Serratia proteamaculans | |
51.22 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant mutant D649A, pH 8.0, 25°C | Serratia proteamaculans | |
52.34 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant wild-type enzyme, pH 8.0, 25°C | Serratia proteamaculans | |
63.71 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant mutant E125A, pH 8.0, 25°C | Serratia proteamaculans | |
85.55 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant mutant E125A, pH 8.0, 25°C | Serratia proteamaculans |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Serratia proteamaculans |
General Information | Comment | Organism |
---|---|---|
evolution | oligopeptidase B is a trypsin-like peptidase belonging to the family of serine prolyl oligopeptidases with a two-domain structure of the enzyme including C-terminal peptidase catalytic domain and N-terminal seven-bladed beta-propeller domain | Serratia proteamaculans |
malfunction | the removal of negatively charged Asp647 and Asp649 leads therefore to better binding of lysine at P2 substrate position and decreased binding of arginine at the same position | Serratia proteamaculans |
additional information | comparative structure of the enzyme using the structures of both closed form of OpdB from Leishmania major (PDB ID 2XE4) and open form of OpdB from Trypanosoma brucei (PDB ID 4BP8) as templates, overview | Serratia proteamaculans |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
9.28 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant mutant K655A, pH 8.0, 25°C | Serratia proteamaculans | |
88.56 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant mutant K660A, pH 8.0, 25°C | Serratia proteamaculans | |
171.65 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant wild-type enzyme, pH 8.0, 25°C | Serratia proteamaculans | |
191.3 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant mutant K655A, pH 8.0, 25°C | Serratia proteamaculans | |
205.7 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant mutant D647A, pH 8.0, 25°C | Serratia proteamaculans | |
356.8 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant mutant K655A, pH 8.0, 25°C | Serratia proteamaculans | |
602.15 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant mutant D649A, pH 8.0, 25°C | Serratia proteamaculans | |
1385 | - |
Nalpha-benzoyl-DL-argininyl-4-nitroanilide | recombinant mutant E125A, pH 8.0, 25°C | Serratia proteamaculans | |
2521 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant mutant K660A, pH 8.0, 25°C | Serratia proteamaculans | |
2697.9 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant wild-type enzyme, pH 8.0, 25°C | Serratia proteamaculans | |
2759.7 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant mutant K660A, pH 8.0, 25°C | Serratia proteamaculans | |
3052.8 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant mutant D649A, pH 8.0, 25°C | Serratia proteamaculans | |
3569.4 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant mutant D647A, pH 8.0, 25°C | Serratia proteamaculans | |
4588.8 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant mutant D647A, pH 8.0, 25°C | Serratia proteamaculans | |
4973.8 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant mutant E125A, pH 8.0, 25°C | Serratia proteamaculans | |
5948.9 | - |
benzyloxycarbonyl-L-lysyl-L-arginyl-4-nitroanilide | recombinant mutant D649A, pH 8.0, 25°C | Serratia proteamaculans | |
7182.9 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant wild-type enzyme, pH 8.0, 25°C | Serratia proteamaculans | |
16203.6 | - |
benzyloxycarbonyl-L-argininyl-L-arginyl-4-nitroanilide | recombinant mutant E125A, pH 8.0, 25°C | Serratia proteamaculans |