Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli LMG194 cells | Vibrio splendidus |
Protein Variants | Comment | Organism |
---|---|---|
F355W | the mutant shows slightly reduced catalytic efficiency compared to the wild type enzyme | Vibrio splendidus |
R336L | the mutant shows extreme thermo-instability and its activity drops by half at 0.1 M of urea within 1.5 h | Vibrio splendidus |
W155F | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Vibrio splendidus |
W274F | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Vibrio splendidus |
W301F | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Vibrio splendidus |
W460F | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Vibrio splendidus |
W460I | inactive | Vibrio splendidus |
W460Y | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Vibrio splendidus |
W475F | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Vibrio splendidus |
W512F | the mutant shows wild type activity | Vibrio splendidus |
General Stability | Organism |
---|---|
the enzyme is irreversibly transformed into an inactive dimer by low urea concentrations | Vibrio splendidus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.194 | - |
4-nitrophenyl phosphate | mutant enzyme W475F, at pH 9.8 and 10°C | Vibrio splendidus | |
0.194 | - |
4-nitrophenyl phosphate | wild type enzyme, at pH 9.8 and 10°C | Vibrio splendidus | |
0.212 | - |
4-nitrophenyl phosphate | mutant enzyme W512F, at pH 9.8 and 10°C | Vibrio splendidus | |
0.218 | - |
4-nitrophenyl phosphate | mutant enzyme F355W, at pH 9.8 and 10°C | Vibrio splendidus | |
0.218 | - |
4-nitrophenyl phosphate | mutant enzyme W155F, at pH 9.8 and 10°C | Vibrio splendidus | |
0.239 | - |
4-nitrophenyl phosphate | mutant enzyme W301F, at pH 9.8 and 10°C | Vibrio splendidus | |
0.252 | - |
4-nitrophenyl phosphate | mutant enzyme W460Y, at pH 9.8 and 10°C | Vibrio splendidus | |
0.354 | - |
4-nitrophenyl phosphate | mutant enzyme W460F, at pH 9.8 and 10°C | Vibrio splendidus | |
0.462 | - |
4-nitrophenyl phosphate | mutant enzyme W274F, at pH 9.8 and 10°C | Vibrio splendidus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | contains Mg2+ | Vibrio splendidus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Vibrio splendidus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Strep-Tactin affinity column chromatography, and Superdex 200 gel filtration | Vibrio splendidus |
Storage Stability | Organism |
---|---|
25°C, 25 mM MOPS buffer, pH 8.0 with 500 mM NaCl, 2 days, no loss of activity | Vibrio splendidus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl phosphate + H2O | - |
Vibrio splendidus | 4-nitrophenol + phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | - |
Vibrio splendidus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
51 | - |
melting temperature | Vibrio splendidus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
44 | - |
4-nitrophenyl phosphate | mutant enzyme W475F, at pH 9.8 and 10°C | Vibrio splendidus | |
65 | - |
4-nitrophenyl phosphate | mutant enzyme W460F, at pH 9.8 and 10°C | Vibrio splendidus | |
125 | - |
4-nitrophenyl phosphate | mutant enzyme W155F, at pH 9.8 and 10°C | Vibrio splendidus | |
137 | - |
4-nitrophenyl phosphate | mutant enzyme W301F, at pH 9.8 and 10°C | Vibrio splendidus | |
255 | - |
4-nitrophenyl phosphate | mutant enzyme F355W, at pH 9.8 and 10°C | Vibrio splendidus | |
275 | - |
4-nitrophenyl phosphate | mutant enzyme W460Y, at pH 9.8 and 10°C | Vibrio splendidus | |
300 | - |
4-nitrophenyl phosphate | mutant enzyme W274F, at pH 9.8 and 10°C | Vibrio splendidus | |
302 | - |
4-nitrophenyl phosphate | wild type enzyme, at pH 9.8 and 10°C | Vibrio splendidus | |
313 | - |
4-nitrophenyl phosphate | mutant enzyme W512F, at pH 9.8 and 10°C | Vibrio splendidus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
184 | - |
4-nitrophenyl phosphate | mutant enzyme W460F, at pH 9.8 and 10°C | Vibrio splendidus | |
230 | - |
4-nitrophenyl phosphate | mutant enzyme W475F, at pH 9.8 and 10°C | Vibrio splendidus | |
573 | - |
4-nitrophenyl phosphate | mutant enzyme W155F, at pH 9.8 and 10°C | Vibrio splendidus | |
573 | - |
4-nitrophenyl phosphate | mutant enzyme W301F, at pH 9.8 and 10°C | Vibrio splendidus | |
650 | - |
4-nitrophenyl phosphate | mutant enzyme W274F, at pH 9.8 and 10°C | Vibrio splendidus | |
869 | - |
4-nitrophenyl phosphate | mutant enzyme W460Y, at pH 9.8 and 10°C | Vibrio splendidus | |
1170 | - |
4-nitrophenyl phosphate | mutant enzyme F355W, at pH 9.8 and 10°C | Vibrio splendidus | |
1480 | - |
4-nitrophenyl phosphate | mutant enzyme W512F, at pH 9.8 and 10°C | Vibrio splendidus | |
1560 | - |
4-nitrophenyl phosphate | wild type enzyme, at pH 9.8 and 10°C | Vibrio splendidus |