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Literature summary extracted from
- Structural mechanism for the temperature-dependent activation of the hyperthermophilic Pf2001 esterase (2018), Structure, 26, 199-208.e3 .
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.1.1.1 | expression in Escherichia coli by using a His-tag expression vector | Pyrococcus furiosus |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 3.1.1.1 | 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 |
| 3.1.1.1 | 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]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.1.1.1 | 0.01064 | - |
4-methylumbelliferyl heptanoate | pH 7.0, 70°C, wild-type enzyme | Pyrococcus furiosus |  |
| 3.1.1.1 | 0.01502 | - |
4-methylumbelliferyl heptanoate | pH 7.0, 70°C, mutant enzyme W194A | Pyrococcus furiosus | |
| 3.1.1.1 | 0.02108 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, mutant enzyme F198A | Pyrococcus furiosus | |
| 3.1.1.1 | 0.03598 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, mutant enzyme W194A | Pyrococcus furiosus | |
| 3.1.1.1 | 0.06266 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, wild-type enzyme | Pyrococcus furiosus | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.1.1.1 | Pyrococcus furiosus | Q8TZJ1 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 3.1.1.1 | - |
Pyrococcus furiosus |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.1.1.1 | 4-methylumbelliferyl butyrate + H2O | - |
Pyrococcus furiosus | 4-methylumbelliferol + butyrate | - |
? | |
| 3.1.1.1 | 4-methylumbelliferyl heptanoate + H2O | - |
Pyrococcus furiosus | 4-methylumbelliferol + heptanoate | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 3.1.1.1 | 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 |
| 3.1.1.1 | 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
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 3.1.1.1 | Pf2001 esterase | - |
Pyrococcus furiosus |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.1.1.1 | 80 | - |
- |
Pyrococcus furiosus |
Temperature Range [°C]
| EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.1.1.1 | 50 | 80 | 50°C: about 50% of maximal activity, 80°C: temperature optimum | Pyrococcus furiosus |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.1.1.1 | 0.067 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, mutant enzyme F198A | Pyrococcus furiosus | |
| 3.1.1.1 | 0.159 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, mutant enzyme W194A | Pyrococcus furiosus | |
| 3.1.1.1 | 0.335 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, wild-type enzyme | Pyrococcus furiosus | |
| 3.1.1.1 | 0.49 | - |
4-methylumbelliferyl heptanoate | pH 7.0, 70°C, mutant enzyme W194A | Pyrococcus furiosus | |
| 3.1.1.1 | 0.883 | - |
4-methylumbelliferyl heptanoate | pH 7.0, 70°C, wild-type enzyme | Pyrococcus furiosus | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.1.1.1 | 7 | - |
assay at | Pyrococcus furiosus |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.1.1.1 | 3.2 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, mutant enzyme F198A | Pyrococcus furiosus | |
| 3.1.1.1 | 4.5 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, mutant enzyme W194A | Pyrococcus furiosus | |
| 3.1.1.1 | 5.3 | - |
4-Methylumbelliferyl butyrate | pH 7.0, 70°C, wild-type enzyme | Pyrococcus furiosus | |
| 3.1.1.1 | 6.5 | - |
4-methylumbelliferyl heptanoate | pH 7.0, 70°C, mutant enzyme F198A | Pyrococcus furiosus | |
| 3.1.1.1 | 32.6 | - |
4-methylumbelliferyl heptanoate | pH 7.0, 70°C, mutant enzyme W194A | Pyrococcus furiosus | |
| 3.1.1.1 | 82.8 | - |
4-methylumbelliferyl heptanoate | pH 7.0, 70°C, wild-type enzyme | Pyrococcus furiosus | |
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