Protein Variants | Comment | Organism |
---|---|---|
H116D | slightly less heat-stable than the wild-type enzyme. Mutant enzyme shows much reduced catalytic efficiencies as compared to the wild-type enzyme. kcat/KM is 29.7fold lower than wild-type value | Vibrio sp. |
H116D/W274K | more heat-tolerant than the wild-type enzyme. Mutant enzyme shows much reduced catalytic efficiencies as compared to the wild-type enzyme. kcat/KM is 17.1fold lower than wild-type value | Vibrio sp. |
W274K | more heat-tolerant than the wild-type enzyme. Mutant enzyme shows much reduced catalytic efficiencies as compared to the wild-type enzyme. kcat/KM is 30.6fold lower than wild-type value | Vibrio sp. |
General Stability | Organism |
---|---|
loss of activity occurrs at urea concentrations below 1 M. The global structure is unfolded at urea concentrations in the range 1-3 M | Vibrio sp. |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
phosphate | - |
Escherichia coli | |
phosphate | - |
Vibrio sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.021 | - |
p-nitrophenyl phosphate | 25°C, pH 8.0, wild-type enzyme | Escherichia coli | |
0.08 | - |
p-nitrophenyl phosphate | 25°C, pH 9.8, wild-type enzyme | Vibrio sp. | |
0.19 | - |
p-nitrophenyl phosphate | 25°C, pH 9.8, mutant enzyme W274K | Vibrio sp. | |
1.25 | - |
p-nitrophenyl phosphate | 25°C, pH 9.8, mutant enzyme H116D | Vibrio sp. | |
1.44 | - |
p-nitrophenyl phosphate | 25°C, pH 9.8, mutant enzyme H116D/W274K | Vibrio sp. |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | wild-type enzyme most likely has one Mg2+ in the active site in addition to three Mg2+ that bind elsewhere. At 18°C with 10 mM Mg2+, the enzyme activity is completely stable over 160 min. With no Mg2+ in solution, the activity drops to 20% of the initial activity after 160 min | Vibrio sp. | |
Zn2+ | wild-type enzyme most likely has two Zn2+. Zn2+ release from the active site coincides with total protein structure unfolding | Vibrio sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P00634 | - |
- |
Vibrio sp. | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
p-nitrophenyl phosphate + H2O | - |
Vibrio sp. | p-nitrophenol + phosphate | - |
? | |
p-nitrophenyl phosphate + H2O | - |
Escherichia coli | p-nitrophenol + phosphate | - |
? |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
t1/2: 94 min for wild-type value | Vibrio sp. |
30 | - |
t1/2: 79 min for wild-type enzyme, 2600 min for mutant enzyme h116D | Vibrio sp. |
35 | - |
t1/2: 31 min for wild-type enzyme, 63 min for mutant enzyme H116D, 2200 min for mutant enzyme H116S/W274K | Vibrio sp. |
40 | - |
t1/2: 12 min for wild-type enzyme, 231 min for mutant enzyme W274K, 30 min for mutant enzyme H116D, 217 min for mutant enzyme H116D/W274K | Vibrio sp. |
45 | - |
t1/2: 2.3 min for wild-type enzyme, 26 min for mutant enzyme W274K, 1.1 min for mutant enzyme H116D, 96 min for mutant enzyme H116D/W274K | Vibrio sp. |
50 | - |
t1/2: 0.7 min for wild-type enzyme, 6 min for mutant enzyme W274K, 0.8 min for mutant enzyme H116D, 10 min for mutant enzyme H116D/W274K | Vibrio sp. |
55 | - |
t1/2: 1.2 min for mutant enzyme W274K, 0.2 min for mutant enzyme H116D, 5 min for mutant enzyme H116D/W274K | Vibrio sp. |
55 | - |
Tm-value for mutant enzyme H116D | Vibrio sp. |
57 | - |
Tm-value for mutant enzymeW274K is 57.2°C | Vibrio sp. |
57 | - |
Tm-value for wild-type enzyme is 56.5°C | Vibrio sp. |
59 | - |
Tm-value for mutant enzyme H116D/W274K is 58.6°C | Vibrio sp. |
60 | - |
t1/2: 0.5 min for mutant enzyme H116D/W274K | Vibrio sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
38 | - |
p-nitrophenyl phosphate | 25°C, pH 8.0, wild-type enzyme | Escherichia coli | |
220 | - |
p-nitrophenyl phosphate | 25°C, pH 9.8, mutant enzyme W274K | Vibrio sp. | |
830 | - |
p-nitrophenyl phosphate | 25°C, pH 9.8, mutant enzyme H116D | Vibrio sp. | |
930 | - |
p-nitrophenyl phosphate | 25°C, pH 9.8, mutant enzyme H116D/W274K | Vibrio sp. | |
1580 | - |
p-nitrophenyl phosphate | 25°C, pH 9.8, wild-type enzyme | Vibrio sp. |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.045 | - |
phosphate | 25°C, pH 8.0, wild-type enzyme | Escherichia coli | |
0.39 | - |
phosphate | 25°C, pH 9.8, wild-type enzyme | Vibrio sp. | |
0.77 | - |
phosphate | 25°C, pH 9.8, mutant enzyme W274K | Vibrio sp. | |
3.45 | - |
phosphate | 25°C, pH 9.8, mutant enzyme H116D/W274K | Vibrio sp. | |
5.25 | - |
phosphate | 25°C, pH 9.8, mutant enzyme H116D | Vibrio sp. |