EC Number | Cloned (Comment) | Organism |
---|---|---|
3.4.24.27 | expression of wild-type and mutant enzymes in Escherichia coli strain T5KC991 | Bacillus thermoproteolyticus |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
3.4.24.27 | G117D | site-directed mutagenesis, the mutant enzyme shows reduced activity compared to the wild-type | Bacillus thermoproteolyticus |
3.4.24.27 | G117E | site-directed mutagenesis, the mutant enzyme shows increased activity compared to the wild-type enzyme, the kcat/Km value is 80% of wild-type level with N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide, but 130% with N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester | Bacillus thermoproteolyticus |
3.4.24.27 | G117K | site-directed mutagenesis, the mutant enzyme shows reduced activity compared to the wild-type, the kcat/Km value is 40% of wild-type level with N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide, but 80% with N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester | Bacillus thermoproteolyticus |
3.4.24.27 | G117R | site-directed mutagenesis, the mutant enzyme shows reduced activity compared to the wild-type, the kcat/Km value is 40% of wild-type level with N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide and N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester | Bacillus thermoproteolyticus |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.4.24.27 | additional information | - |
additional information | Michaelis-Menten kinetics, overview | Bacillus thermoproteolyticus | |
3.4.24.27 | 0.42 | - |
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester | recombinant wild-type enzyme, pH 7.5, 25°C | Bacillus thermoproteolyticus | |
3.4.24.27 | 0.44 | - |
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester | recombinant mutant G117E, pH 7.5, 25°C | Bacillus thermoproteolyticus | |
3.4.24.27 | 0.5 | - |
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester | recombinant mutant G117R, pH 7.5, 25°C | Bacillus thermoproteolyticus | |
3.4.24.27 | 0.57 | - |
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester | recombinant mutant G117K, pH 7.5, 25°C | Bacillus thermoproteolyticus |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
3.4.24.27 | Ca2+ | required | Bacillus thermoproteolyticus | |
3.4.24.27 | NaCl | induces enzyme activation, activation of mutant enzymes is reduced compared to the wild-type enzyme | Bacillus thermoproteolyticus |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.4.24.27 | Bacillus thermoproteolyticus | - |
- |
- |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
3.4.24.27 | 2000 | - |
recombinant mutant G117D, substrate casein, pH 7.5, 25°C | Bacillus thermoproteolyticus |
3.4.24.27 | 2900 | - |
recombinant mutant G117R, substrate casein, pH 7.5, 25°C | Bacillus thermoproteolyticus |
3.4.24.27 | 3200 | - |
recombinant mutant G117K, substrate casein, pH 7.5, 25°C | Bacillus thermoproteolyticus |
3.4.24.27 | 11000 | - |
recombinant wild-type enzyme, substrate casein, pH 7.5, 25°C | Bacillus thermoproteolyticus |
3.4.24.27 | 12000 | - |
recombinant mutant G117E, substrate casein, pH 7.5, 25°C | Bacillus thermoproteolyticus |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.4.24.27 | casein + H2O | - |
Bacillus thermoproteolyticus | ? | - |
? | |
3.4.24.27 | N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester + H2O | - |
Bacillus thermoproteolyticus | N-benzyloxycarbonyl-L-Asp + L-Phe-methyl ester | - |
? | |
3.4.24.27 | N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide + H2O | - |
Bacillus thermoproteolyticus | N-[3-(2-furyl)acryloyl]-glycine + L-leucine amide | - |
? |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.4.24.27 | 25 | - |
assay at | Bacillus thermoproteolyticus |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.4.24.27 | 4.8 | - |
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester | recombinant mutant G117R, pH 7.5, 25°C | Bacillus thermoproteolyticus | |
3.4.24.27 | 7.9 | - |
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester | recombinant wild-type enzyme, pH 7.5, 25°C | Bacillus thermoproteolyticus | |
3.4.24.27 | 8.4 | - |
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester | recombinant mutant G117K, pH 7.5, 25°C | Bacillus thermoproteolyticus | |
3.4.24.27 | 10.5 | - |
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester | recombinant mutant G117E, pH 7.5, 25°C | Bacillus thermoproteolyticus |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
3.4.24.27 | 7.5 | - |
assay at | Bacillus thermoproteolyticus |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.4.24.27 | 9.6 | - |
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester | recombinant mutant G117R, pH 7.5, 25°C | Bacillus thermoproteolyticus | |
3.4.24.27 | 14.7 | - |
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester | recombinant mutant G117K, pH 7.5, 25°C | Bacillus thermoproteolyticus | |
3.4.24.27 | 15 | - |
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide | recombinant mutant G117R, pH 7.5, 25°C | Bacillus thermoproteolyticus | |
3.4.24.27 | 16 | - |
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide | recombinant mutant G117K, pH 7.5, 25°C | Bacillus thermoproteolyticus | |
3.4.24.27 | 18.8 | - |
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester | recombinant wild-type enzyme, pH 7.5, 25°C | Bacillus thermoproteolyticus | |
3.4.24.27 | 23.9 | - |
N-benzyloxycarbonyl-L-Asp-L-Phe-methyl ester | recombinant mutant G117E, pH 7.5, 25°C | Bacillus thermoproteolyticus | |
3.4.24.27 | 33 | - |
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide | recombinant mutant G117E, pH 7.5, 25°C | Bacillus thermoproteolyticus | |
3.4.24.27 | 40 | - |
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide | recombinant wild-type enzyme, pH 7.5, 25°C | Bacillus thermoproteolyticus |