Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli by using a His-tag expression vector | Pyrococcus furiosus |
Protein Variants | Comment | Organism |
---|---|---|
F198A | mutant enzyme behaves as a monodisperse peak in analytical gel-filtration experiments at either 6°C or 55°C, indicating a reduced tendency to dimerize in this mutant. Significant reduction in the relative Kcat/Km of around 60% for the C7-chain substrate in comparison with wild-type enzyme | Pyrococcus furiosus |
W194A | mutant enzyme behaves as a monodisperse peak in analytical gel-filtration experiments at either 6°C or 55°C, indicating a reduced tendency to dimerize in this mutant. Significant reduction in the relative Kcat/Km of around 90% for the C7-chain substrate in comparison with wild-type enzyme | Pyrococcus furiosus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.01064 | - |
4-methylumbelliferyl heptanoate | pH 7.0, 70°C, wild-type enzyme | Pyrococcus furiosus | |
0.01502 | - |
4-methylumbelliferyl heptanoate | pH 7.0, 70°C, mutant enzyme W194A | Pyrococcus furiosus | |
0.02108 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, mutant enzyme F198A | Pyrococcus furiosus | |
0.03598 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, mutant enzyme W194A | Pyrococcus furiosus | |
0.06266 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, wild-type enzyme | Pyrococcus furiosus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus furiosus | Q8TZJ1 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Pyrococcus furiosus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-methylumbelliferyl butyrate + H2O | - |
Pyrococcus furiosus | 4-methylumbelliferol + butyrate | - |
? | |
4-methylumbelliferyl heptanoate + H2O | - |
Pyrococcus furiosus | 4-methylumbelliferol + heptanoate | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 30000, the crystal structure of the Pf2001 esterase shows two different conformations: monomer and dimer. A temperature-dependent activation mechanism of the Pf2001 esterase is proposed via dimerization that is necessary for the substrate channel formation in the active site cleft | Pyrococcus furiosus |
monomer | 1 * 30000, the crystal structure of the Pf2001 esterase shows two different conformations: monomer and dimer. A temperature-dependent activation mechanism of the Pf2001 esterase is proposed via dimerization that is necessary for the substrate channel formation in the active site cleft | Pyrococcus furiosus |
Synonyms | Comment | Organism |
---|---|---|
Pf2001 esterase | - |
Pyrococcus furiosus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
80 | - |
- |
Pyrococcus furiosus |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | 80 | 50°C: about 50% of maximal activity, 80°C: temperature optimum | Pyrococcus furiosus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.067 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, mutant enzyme F198A | Pyrococcus furiosus | |
0.159 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, mutant enzyme W194A | Pyrococcus furiosus | |
0.335 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, wild-type enzyme | Pyrococcus furiosus | |
0.49 | - |
4-methylumbelliferyl heptanoate | pH 7.0, 70°C, mutant enzyme W194A | Pyrococcus furiosus | |
0.883 | - |
4-methylumbelliferyl heptanoate | pH 7.0, 70°C, wild-type enzyme | Pyrococcus furiosus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Pyrococcus furiosus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.2 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, mutant enzyme F198A | Pyrococcus furiosus | |
4.5 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, mutant enzyme W194A | Pyrococcus furiosus | |
5.3 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, wild-type enzyme | Pyrococcus furiosus | |
6.5 | - |
4-methylumbelliferyl heptanoate | pH 7.0, 70°C, mutant enzyme F198A | Pyrococcus furiosus | |
32.6 | - |
4-methylumbelliferyl heptanoate | pH 7.0, 70°C, mutant enzyme W194A | Pyrococcus furiosus | |
82.8 | - |
4-methylumbelliferyl heptanoate | pH 7.0, 70°C, wild-type enzyme | Pyrococcus furiosus |