Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli | Thermococcus kodakarensis |
Protein Variants | Comment | Organism |
---|---|---|
D215A | Vmax is 1.8fold lower than wild-type value, kcat is 1.8 fold lower than wild-type enzyme | Thermococcus kodakarensis |
D277A | Vmax is 1.6fold higher than wild-type value, kcat is 1.6fold higher than wild-type enzyme | Thermococcus kodakarensis |
E226A | Vmax is 1.35fold higher than wild-type value, kcat is 1.3fold higher than wild-type enzyme | Thermococcus kodakarensis |
E227A | Vmax is is nearly identical to wild-type value, kcat nearly identical to wild-type enzyme | Thermococcus kodakarensis |
H191A | Vmax is 1.84fold lower than wild-type value, kcat is 1.9fold lower than wild-type enzyme | Thermococcus kodakarensis |
K150A | Vmax is 1.5fold higher than wild-type value, kcat is 1.4fold higher than wild-type enzyme | Thermococcus kodakarensis |
K209A | Vmax is 1.7fold lower than wild-type value, kcat is 1.8fold lower than wild-type enzyme | Thermococcus kodakarensis |
additional information | a truncated protein without the N-terminal 54 residues and putative transmembrane domain, exhibits high peptidase activity, and is used as the wild-type protein | Thermococcus kodakarensis |
R221A | Vmax is 7.8fold lower than wild-type value, kcat is 8fold lower than wild-type enzyme | Thermococcus kodakarensis |
R250A | Vmax is 1.5fold higher than wild-type value, kcat is 1.4fold higher than wild-type enzyme | Thermococcus kodakarensis |
S128A | Vmax is 6.9 fold higher than wild-type value, kcat is 6.2fold higher than wild-type enzyme | Thermococcus kodakarensis |
S184A | Vmax is 4fold lower than wild-type value, kcat is 4fold lower than wild-type enzyme | Thermococcus kodakarensis |
Y165A | Vmax is 2.3fold higher than wild-type value, kcat is 2.3fold higher than wild-type enzyme | Thermococcus kodakarensis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermococcus kodakarensis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
wild-type and mutant enzymes | Thermococcus kodakarensis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
Ala-Ala-Phe-4-methylcoumaryl-7-amide + H2O | - |
Thermococcus kodakarensis | Ala-Ala-Phe + 7-amino-4-methylcoumarin | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2 | - |
Ala-Ala-Phe-4-methylcoumaryl-7-amide | 60°C, pH 10.0, mutant enzyme R221A | Thermococcus kodakarensis | |
4 | - |
Ala-Ala-Phe-4-methylcoumaryl-7-amide | 60°C, pH 10.0, mutant enzyme S184A | Thermococcus kodakarensis | |
8.6 | - |
Ala-Ala-Phe-4-methylcoumaryl-7-amide | 60°C, pH 10.0, mutant enzyme H191A | Thermococcus kodakarensis | |
8.8 | - |
Ala-Ala-Phe-4-methylcoumaryl-7-amide | 60°C, pH 10.0, mutant enzyme D215A | Thermococcus kodakarensis | |
9.1 | - |
Ala-Ala-Phe-4-methylcoumaryl-7-amide | 60°C, pH 10.0, mutant enzyme K209A | Thermococcus kodakarensis | |
16 | - |
Ala-Ala-Phe-4-methylcoumaryl-7-amide | 60°C, pH 10.0, mutant enzyme E227A | Thermococcus kodakarensis | |
16 | - |
Ala-Ala-Phe-4-methylcoumaryl-7-amide | 60°C, pH 10.0, wild-type enzyme | Thermococcus kodakarensis | |
21 | - |
Ala-Ala-Phe-4-methylcoumaryl-7-amide | 60°C, pH 10.0, mutant enzyme E226A | Thermococcus kodakarensis | |
23 | - |
Ala-Ala-Phe-4-methylcoumaryl-7-amide | 60°C, pH 10.0, mutant enzyme K150A | Thermococcus kodakarensis | |
23 | - |
Ala-Ala-Phe-4-methylcoumaryl-7-amide | 60°C, pH 10.0, mutant enzyme R250A | Thermococcus kodakarensis | |
25 | - |
Ala-Ala-Phe-4-methylcoumaryl-7-amide | 60°C, pH 10.0, mutant enzyme D277A | Thermococcus kodakarensis | |
37 | - |
Ala-Ala-Phe-4-methylcoumaryl-7-amide | 60°C, pH 10.0, mutant enzyme Y165A | Thermococcus kodakarensis | |
110 | - |
Ala-Ala-Phe-4-methylcoumaryl-7-amide | 60°C, pH 10.0, mutant enzyme S128A | Thermococcus kodakarensis |