Cloned (Comment) | Organism |
---|---|
recombinant expression of mutant enzyme sin Bacillus subtilis strain DB104, subcloning in Escherichia coli strain DH5alpha | Bacillus licheniformis |
Protein Variants | Comment | Organism |
---|---|---|
G165X | site-directed mutagenesis, catalytic constants of subtilisin Carlsberg for perhydrolysis of methylpropionate, methylbutyrate and methylpentanoate are increased up to 3.5fold, 5.4fold and 5.5fold, respectively, while proteolysis is decreased up to 100fold for N-succinyl-Ala-Ala-Pro-Phe-pnitroanilide substrate | Bacillus licheniformis |
additional information | site saturation mutagenesis of the subtilisin Carlsberg variant T58A/L216W at positionsThr33, Val67, Ser124, Gly126, Ser155, Gly165, Ala168, Trp216, Asn217, Gly218, Met221 | Bacillus licheniformis |
T58A/L216W | site-directed mutagenesis, the mutant catalyzes in addition to its proteolytic activity the generation of peroxycarboxylic acids from corresponding esters in the presence of hydrogen peroxide | Bacillus licheniformis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetic analysis of the perhydrolytic wild-type and mutant enzyme activities, overview | Bacillus licheniformis | |
0.15 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | recombinant mutant T58A/L216W, pH 7.5, 30°C | Bacillus licheniformis | |
0.2 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | recombinant mutant T58A/G165Y/L216W, pH 7.5, 30°C | Bacillus licheniformis | |
0.42 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | recombinant mutant T58A/G165L/L216W, pH 7.5, 30°C | Bacillus licheniformis | |
0.54 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | recombinant mutant T58A/G165I/L216W, pH 7.5, 30°C | Bacillus licheniformis | |
0.59 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | recombinant wild-type enzyme, pH 7.5, 30°C | Bacillus licheniformis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus licheniformis | P00780 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | besides ester hydrolysis the enzyme also performs ester perhydrolysis reacting an ester and a hydroperpxide, residue Gl165 in the S1 binding pocket of the enzyme is involved. Perhydrolysis reaction analysis and kinetcis, overview | Bacillus licheniformis | ? | - |
? | |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O | - |
Bacillus licheniformis | N-succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline | - |
? |
Synonyms | Comment | Organism |
---|---|---|
subtilisin Carlsberg | - |
Bacillus licheniformis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Bacillus licheniformis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.7 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | recombinant mutant T58A/G165I/L216W, pH 7.5, 30°C | Bacillus licheniformis | |
15.1 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | recombinant mutant T58A/G165L/L216W, pH 7.5, 30°C | Bacillus licheniformis | |
22.8 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | recombinant mutant T58A/G165Y/L216W, pH 7.5, 30°C | Bacillus licheniformis | |
33.1 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | recombinant mutant T58A/L216W, pH 7.5, 30°C | Bacillus licheniformis | |
172.6 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | recombinant wild-type enzyme, pH 7.5, 30°C | Bacillus licheniformis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Bacillus licheniformis |
General Information | Comment | Organism |
---|---|---|
additional information | molecular modeling and docking analysis using crystal structure PDB ID 1yu6 | Bacillus licheniformis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.1 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | recombinant mutant T58A/G165I/L216W, pH 7.5, 30°C | Bacillus licheniformis | |
36 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | recombinant mutant T58A/G165L/L216W, pH 7.5, 30°C | Bacillus licheniformis | |
114 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | recombinant mutant T58A/G165Y/L216W, pH 7.5, 30°C | Bacillus licheniformis | |
220.7 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | recombinant mutant T58A/L216W, pH 7.5, 30°C | Bacillus licheniformis | |
292.5 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | recombinant wild-type enzyme, pH 7.5, 30°C | Bacillus licheniformis |