Protein Variants | Comment | Organism |
---|---|---|
G75S | 96% loss of activity of mutant enzyme M184L after 30 min at 70°C, compared to 90% loss of wild-type enzyme. Residual activity after incubation with 0.2 mM H2O2 for 30 min at 25°C is 42.5% compared to 5% for the wild-type enzyme | Agrobacterium tumefaciens |
H58Y | 80% loss of activity of mutant enzyme M184L after 30 min at 70°C, compared to 90% loss of wild-type enzyme. Residual activity after incubation with 0.2 mM H2O2 for 30 min at 25°C is 20.6% compared to 5% for the wild-type enzyme | Agrobacterium tumefaciens |
M184L | 80% loss of activity of mutant enzyme M184L after 30 min at 70°C, compared to 90% loss of wild-type enzyme. Residual activity after incubation with 0.2 mM H2O2 for 30 min at 25°C is 20% compared to 5% for the wild-type enzyme | Agrobacterium tumefaciens |
M184L/T262A | 46% loss of activity of mutant enzyme M184L after 30 min at 70°C, compared to 90% loss of wild-type enzyme. Residual activity after incubation with 0.2 mM H2O2 for 30 min at 25°C is 54.8% compared to 5% for the wild-type enzyme. The ratio of turnover number to Km-value for N-carbamoyl-D-p-hydroxyphenylglycine is 58.6% of the wild-type ratio | Agrobacterium tumefaciens |
Q23L | 81% loss of activity of mutant enzyme M184L after 30 min at 70°C, compared to 90% loss of wild-type enzyme. Residual activity after incubation with 0.2 mM H2O2 for 30 min at 25°C is 63.9% compared to 5% for the wild-type enzyme | Agrobacterium tumefaciens |
Q23L/V40A/H58Y/G75S/M184L/T262A | mutant enzyme with improved oxidative and thermostability, 79.3% loss of activity of mutant enzyme M184L after 30 min at 70°C, compared to 90% loss of wild-type enzyme. The ratio of turnover number to Km-value for N-carbamoyl-D-p-hydroxyphenylglycine is 72.4% of the wild-type ratio | Agrobacterium tumefaciens |
T262A | 49% loss of activity of mutant enzyme M184L after 30 min at 70°C, compared to 90% loss of wild-type enzyme. Residual activity after incubation with 0.2 mM H2O2 for 30 min at 25°C is 40.9% compared to 5% for the wild-type enzyme | Agrobacterium tumefaciens |
V40A | 93% loss of activity of mutant enzyme M184L after 30 min at 70°C, compared to 90% loss of wild-type enzyme. Residual activity after incubation with 0.2 mM H2O2 for 30 min at 25°C is 20.6% compared to 5% for the wild-type enzyme | Agrobacterium tumefaciens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.1 | - |
N-carbamoyl-D-p-hydroxyphenylglycine | pH 7.0, 40°C, wild-type enzyme | Agrobacterium tumefaciens | |
1.1 | - |
N-carbamoyl-D-p-hydroxyphenylglycine | pH 7.0, 40°C,mutant enzyme Q23L/V40A/H58Y/G75S/M184L/T262A | Agrobacterium tumefaciens | |
2.4 | - |
N-carbamoyl-D-p-hydroxyphenylglycine | pH 7.0, 40°C, mutant enzyme M184L, T262A | Agrobacterium tumefaciens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Agrobacterium tumefaciens | - |
NRRL B11291 | - |
Agrobacterium tumefaciens NRRL B11291 | - |
NRRL B11291 | - |
Oxidation Stability | Organism |
---|---|
residual activity after incubation with 0.2 mM hydrogen peroxide for 30 min at 25°C is 5% for the wild-type enzyme, 20% for mutant enzyme M184L, 40.9% for mutant enzyme T262A, 54.8% for mutant enzyme M184L/T262A, 63.9% for mutant enzyme Q23L, 20.6% for mutant enzymes V40A and H58Y, 42.5% for mutant enzyme G75S, 79.3% for mutant enzyme Q23L/V40A/H58Y/G75S/M184L/T262A | Agrobacterium tumefaciens |
Purification (Comment) | Organism |
---|---|
- |
Agrobacterium tumefaciens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
N-carbamoyl-D-p-hydroxyphenylglycine + H2O | - |
Agrobacterium tumefaciens | D-p-hydroxyphenylglycine + NH3 + CO2 | - |
? | |
N-carbamoyl-D-p-hydroxyphenylglycine + H2O | - |
Agrobacterium tumefaciens NRRL B11291 | D-p-hydroxyphenylglycine + NH3 + CO2 | - |
? |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | - |
30 min, 90% loss of wild-type activity, 80% loss of activity of mutant enzyme M184L, 49% loss of activity of mutant enzyme T262A, 46% loss of activity of mutant enzyme M184L/T262A 81% loss of activity of mutant enzyme Q23L, 93% loss of activity of mutant enzyme V40A, 80% loss of activity of mutant enzyme H58Y, 96% loss of activity of mutant enzyme G75S, 21% loss of activity of mutant enzyme Q23L/V40A/H58Y/G75S/M184L/T262A | Agrobacterium tumefaciens |
73 | - |
30 min, mutant enzyme Q23L/V40A/H58Y/G75S/M184L/T262A, 50% loss of activity | Agrobacterium tumefaciens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
6.9 | - |
N-carbamoyl-D-p-hydroxyphenylglycine | pH 7.0, 40°C, mutant enzyme Q23L/V40A/H58Y/G75S/M184L/T262A | Agrobacterium tumefaciens | |
9.5 | - |
N-carbamoyl-D-p-hydroxyphenylglycine | pH 7.0, 40°C, wild-type enzyme | Agrobacterium tumefaciens | |
12.3 | - |
N-carbamoyl-D-p-hydroxyphenylglycine | pH 7.0, 40°C, mutant enzyme M184L, T262A | Agrobacterium tumefaciens |