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Literature summary for 3.1.1.1 extracted from

  • Varejao, N.; De-Andrade, R.; Almeida, R.; Anobom, C.; Foguel, D.; Reverter, D.
    Structural mechanism for the temperature-dependent activation of the hyperthermophilic Pf2001 esterase (2018), Structure, 26, 199-208.e3 .
    View publication on PubMed
Search for more literature for 3.1.1.1

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli by using a His-tag expression vector Pyrococcus furiosus

Protein Variants

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 [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

Organism UniProt Comment Textmining
Pyrococcus furiosus Q8TZJ1
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Pyrococcus furiosus

Substrates and Products (Substrate)

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

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

Synonyms Comment Organism
Pf2001 esterase
-
 Pyrococcus furiosus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
80
-
-
 Pyrococcus furiosus

Temperature Range [°C]

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 [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 assay at Pyrococcus furiosus

kcat/KM [mM/s]

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