Cloned (Comment) | Organism |
---|---|
cloning in Escherichia coli strain DH5alpha and Saccharomyces cerevisiae strain YPH499, recombinant expression of wild-type and mutant enzymes in Bacillus megaterium strain WH320 from a shuttle expression vector carrying a ribosome-binding site and a signal sequence of extracellular metalloprotease from Bacillus megaterium downstream of xylose isomerase promoter | Bacillus licheniformis |
Protein Variants | Comment | Organism |
---|---|---|
N212G | site-directed mutagenesis, the mutant shows increased kcat compared to the wild-type enzyme | Bacillus licheniformis |
P210A | site-directed mutagenesis, the mutant shows increased kcat compared to the wild-type enzyme | Bacillus licheniformis |
P210G | site-directed mutagenesis, the mutant shows 1.5fold increased kcat compared to the wild-type enzyme | Bacillus licheniformis |
P210G/T211G | site-directed mutagenesis, the mutant shows 1.7fold increased kcat compared to the wild-type enzyme | Bacillus licheniformis |
P210G/T211G/N212G | site-directed mutagenesis, the mutant shows 1.8fold increased kcat compared to the wild-type enzyme | Bacillus licheniformis |
T211G | site-directed mutagenesis, the mutant shows 1.8fold increased kcat compared to the wild-type enzyme | Bacillus licheniformis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.227 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 8.0, 25°C, recombinant mutant N212G | Bacillus licheniformis | |
0.233 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 8.0, 25°C, recombinant wild-type enzyme | Bacillus licheniformis | |
0.278 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 8.0, 25°C, recombinant mutant P210G | Bacillus licheniformis | |
0.279 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 8.0, 25°C, recombinant wmutant T211G | Bacillus licheniformis | |
0.29 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 8.0, 25°C, recombinant mutant P210G/T211G | Bacillus licheniformis | |
0.301 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 8.0, 25°C, recombinant mutant P210G/T211G/N212G | Bacillus licheniformis | |
0.325 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 8.0, 25°C, recombinant mutant P210A | Bacillus licheniformis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | required for thermostability | Bacillus licheniformis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus licheniformis | P00780 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
N-succinyl-Ala-Ala-Pro-Leu-4-nitroanilide + H2O | - |
Bacillus licheniformis | N-succinyl-Ala-Ala-Pro-Leu + 4-nitroaniline | - |
? | |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O | - |
Bacillus licheniformis | N-succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline | - |
? | |
N-succinyl-Ala-Ala-Val-Ala-4-nitroanilide + H2O | - |
Bacillus licheniformis | N-succinyl-Ala-Ala-Val-Ala + 4-nitroaniline | - |
? |
Synonyms | Comment | Organism |
---|---|---|
subtilisin Carlsberg | - |
Bacillus licheniformis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
- |
Bacillus licheniformis |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
15 | 65 | activity range, profile overview | Bacillus licheniformis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
419 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 8.0, 25°C, recombinant wild-type enzyme | Bacillus licheniformis | |
462 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 8.0, 25°C, recombinant wmutant T211G | Bacillus licheniformis | |
475 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 8.0, 25°C, recombinant mutant N212G | Bacillus licheniformis | |
638 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 8.0, 25°C, recombinant mutant P210G | Bacillus licheniformis | |
697 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 8.0, 25°C, recombinant mutant P210G/T211G | Bacillus licheniformis | |
715 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 8.0, 25°C, recombinant mutant P210A | Bacillus licheniformis | |
759 | - |
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 8.0, 25°C, recombinant mutant P210G/T211G/N212G | Bacillus licheniformis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Bacillus licheniformis |
General Information | Comment | Organism |
---|---|---|
additional information | the catalytic triad residues consists of Ser221, Pro210 to His64 | Bacillus licheniformis |