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Literature summary for 3.5.1.4 extracted from

  • Suzuki, Y.; Ohta, H.
    Identification of a thermostable and enantioselective amidase from the thermoacidophilic archaeon Sulfolobus tokodaii strain 7 (2006), Protein Expr. Purif., 45, 368-373.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene ST0478, DNA and amino acid sequence determination and analyis, expression in Escherichia coli strain BL21(DE3) as insoluble aggregated His6-tagged protein Sulfurisphaera tokodaii

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
47000
-
x * 47000, recombinant His6-tagged enzyme, SDS-PAGE Sulfurisphaera tokodaii

Organism

Organism UniProt Comment Textmining
Sulfurisphaera tokodaii
-
gene ST0478
-
Sulfurisphaera tokodaii 7
-
gene ST0478
-

Purification (Commentary)

Purification (Comment) Organism
recombinant insoluble aggregated His6-tagged protein from Escherichia coli by denaturation, refolding during nickel affinity chromatography, gel filtration, and heat treatment, 2.9fold to homogeneity Sulfurisphaera tokodaii

Renatured (Commentary)

Renatured (Comment) Organism
denaturation and refolding during nickel affinity chromatography of the recombinant insoluble aggregated His6-tagged protein from Escherichia coli Sulfurisphaera tokodaii

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
14.8
-
purified recombinant His6-tagged enzyme Sulfurisphaera tokodaii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R,S)-2-phenylpropionamide + H2O enantioselective reaction with preference for the S-enantiomer Sulfurisphaera tokodaii (S)-2-phenylpropionic acid + (R)-2-phenylpropionamide + NH3
-
?
(R,S)-2-phenylpropionamide + H2O enantioselective reaction with preference for the S-enantiomer Sulfurisphaera tokodaii 7 (S)-2-phenylpropionic acid + (R)-2-phenylpropionamide + NH3
-
?
benzamide + H2O
-
Sulfurisphaera tokodaii benzoic acid + NH3
-
?
benzamide + H2O
-
Sulfurisphaera tokodaii 7 benzoic acid + NH3
-
?

Subunits

Subunits Comment Organism
? x * 47000, recombinant His6-tagged enzyme, SDS-PAGE Sulfurisphaera tokodaii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
75
-
about, recombinant enzyme Sulfurisphaera tokodaii

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
37 80 25% of maximal activity at 37°C, maximal activity at 75°C, about 90% of maximal activity at 80°C Sulfurisphaera tokodaii

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
the purified recombinant enzyme is highly thermostable Sulfurisphaera tokodaii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7 8 recombinant enzyme Sulfurisphaera tokodaii

pH Range

pH Minimum pH Maximum Comment Organism
3 9 15% of maximal activity at pH 3.0, 65% at pH 9.0 Sulfurisphaera tokodaii