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Literature summary for 3.5.1.4 extracted from

  • dAbusco, A.S.; Ammendola, S.; Scandurra, R.; Politi, L.
    Molecular and biochemical characterization of the recombinant amidase from hyperthermophilic archaeon Sulfolobus solfataricus (2001), Extremophiles, 5, 183-192.
    View publication on PubMed
Search for more literature for 3.5.1.4

Cloned(Commentary)

Cloned (Comment) Organism
overexpression in Escherichia coli as a fusion protein with an N-terminus six-histidine-residue affinity tag Saccharolobus solfataricus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.3
-
 nicotinamide pH 7.5, 70°C Saccharolobus solfataricus
0.3
-
 Benzamide pH 7.5, 70°C Saccharolobus solfataricus
0.3
-
 cyclomaltohexaose pH 7.5, 70°C Saccharolobus solfataricus
0.3
-
 2-toluamide pH 7.5, 70°C Saccharolobus solfataricus
0.3
-
 4-Aminobenzamide pH 7.5, 70°C Saccharolobus solfataricus
0.3
-
 2-Aminobenzamide pH 7.5, 70°C Saccharolobus solfataricus
0.4
-
 2-hydroxybenzamide pH 7.5, 70°C Saccharolobus solfataricus
0.8
-
 Isonicotinamide pH 7.5, 70°C Saccharolobus solfataricus
0.9
-
 3-indolacetamide pH 7.5, 70°C Saccharolobus solfataricus
1
-
 Isobutyramide pH 7.5, 70°C Saccharolobus solfataricus
1.2
-
 butyramide pH 7.5, 70°C Saccharolobus solfataricus
1.2
-
 methacrylamide pH 7.5, 70°C Saccharolobus solfataricus
1.3
-
 Propionamide pH 7.5, 70°C Saccharolobus solfataricus
1.3
-
 Pentanamide pH 7.5, 70°C Saccharolobus solfataricus
1.5
-
 4-hydroxybenzamide pH 7.5, 70°C Saccharolobus solfataricus
2.5
-
 picolinamide pH 7.5, 70°C Saccharolobus solfataricus
3
-
 Acrylamide pH 7.5, 70°C Saccharolobus solfataricus
3
-
 lactamide pH 7.5, 70°C Saccharolobus solfataricus
3.6
-
 acetamide pH 7.5, 70°C Saccharolobus solfataricus
4
-
 3-phenylpropionamide pH 7.5, 70°C Saccharolobus solfataricus
5
-
 cyclohexanamide pH 7.5, 70°C Saccharolobus solfataricus
5.8
-
 hexanamide pH 7.5, 70°C Saccharolobus solfataricus
6.7
-
 L-prolinamide pH 7.5, 70°C Saccharolobus solfataricus

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ 1 mM, about 20% activation Saccharolobus solfataricus
Fe3+ 1 mM, about 20% activation Saccharolobus solfataricus
additional information no effect: Cu2+, Mn2+, and Ca2+ Saccharolobus solfataricus
V2+ 1 mM, about 20% activation Saccharolobus solfataricus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
55000
-
 x * 55000, SDS-PAGE Saccharolobus solfataricus
55784
-
 x * 55784, calculated from sequence Saccharolobus solfataricus

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus P95896
-
-
Saccharolobus solfataricus MT4 P95896
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Saccharolobus solfataricus

Storage Stability

Storage Stability Organism
the enzyme may be stored at room temperature in the presence of 0.04% NaN3 for 1 year without any loss of activity Saccharolobus solfataricus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(S)-2-phenylpropionamide + H2O no activity with (R)-2-phenylpropionamide Saccharolobus solfataricus (S)-2-phenylpropionate + NH3
-
 ?
2-aminobenzamide + H2O
-
 Saccharolobus solfataricus 2-aminobenzoic acid + NH3
-
 ?
2-hydroxybenzamide + H2O
-
 Saccharolobus solfataricus 2-hydroxybenzoic acid + NH3
-
 ?
2-thiophenecarboxamide + H2O
-
 Saccharolobus solfataricus 2-thiophenecarboxylate + NH3
-
 ?
2-toluamide + H2O
-
 Saccharolobus solfataricus 2-toluic acid + NH3
-
 ?
3-indolacetamide + H2O
-
 Saccharolobus solfataricus 3-indolacetate + NH3
-
 ?
3-phenylpropionamide + H2O
-
 Saccharolobus solfataricus 3-phenylpropionate + NH3
-
 ?
3-toluamide + H2O
-
 Saccharolobus solfataricus 3-toluic acid + NH3
-
 ?
4-aminobenzamide + H2O
-
 Saccharolobus solfataricus 4-aminobenzoic acid + NH3
-
 ?
4-hydroxybenzamide + H2O
-
 Saccharolobus solfataricus 4-hydroxybenzoic acid + NH3
-
 ?
4-methylbenzamide + H2O best aromatic substrate Saccharolobus solfataricus 4-methylbenzoic acid + NH3
-
 ?
4-toluamide + H2O
-
 Saccharolobus solfataricus 4-toluic acid + NH3
-
 ?
acetamide + H2O
-
 Saccharolobus solfataricus acetate + NH3
-
 ?
acrylamide + H2O
-
 Saccharolobus solfataricus acrylate + NH3
-
 ?
anthranilamide + H2O
-
 Saccharolobus solfataricus anthranilate + NH3
-
 ?
benzamide + H2O
-
 Saccharolobus solfataricus benzoate + NH3
-
 ?
butyramide + H2O
-
 Saccharolobus solfataricus butyrate + NH3
-
 ?
cyclohexanamide + H2O
-
 Saccharolobus solfataricus cyclohexanoate + NH3
-
 ?
hexanamide + H2O
-
 Saccharolobus solfataricus hexanoate + NH3
-
 ?
isobutyramide + H2O best aliphatic substrate Saccharolobus solfataricus isobutyrate + NH3
-
 ?
isobutyramide + H2O best aliphatic substrate Saccharolobus solfataricus MT4 isobutyrate + NH3
-
 ?
isonicotinamide + H2O
-
 Saccharolobus solfataricus isonicotinic acid + NH3
-
 ?
L-asparagine + H2O
-
 Saccharolobus solfataricus L-aspartate + NH3
-
 ?
L-asparagine + H2O
-
 Saccharolobus solfataricus MT4 L-aspartate + NH3
-
 ?
L-glutamine + H2O
-
 Saccharolobus solfataricus L-glutamate + NH3
-
 ?
L-glutamine + H2O
-
 Saccharolobus solfataricus MT4 L-glutamate + NH3
-
 ?
L-prolinamide + H2O
-
 Saccharolobus solfataricus L-proline + NH3
-
 ?
lactamide + H2O
-
 Saccharolobus solfataricus lactate + NH3
-
 ?
methacrylamide + H2O
-
 Saccharolobus solfataricus methacrylate + NH3
-
 ?
additional information the enzyme transfers the acyl group also to hydroxylamine and hydrazine Saccharolobus solfataricus ?
-
 ?
additional information the enzyme transfers the acyl group also to hydroxylamine and hydrazine Saccharolobus solfataricus MT4 ?
-
 ?
nicotinamide + H2O
-
 Saccharolobus solfataricus nicotinic acid + NH3
-
 ?
pentanamide + H2O
-
 Saccharolobus solfataricus pentanoate + NH3
-
 ?
pentanamide + H2O
-
 Saccharolobus solfataricus MT4 pentanoate + NH3
-
 ?
picolinamide + H2O
-
 Saccharolobus solfataricus picolinate + NH3
-
 ?
propionamide + H2O
-
 Saccharolobus solfataricus propionate + NH3
-
 ?
pyrazinamide + H2O
-
 Saccharolobus solfataricus pyrazinoic acid + NH3
-
 ?
salicylamide + H2O
-
 Saccharolobus solfataricus salicylate + NH3
-
 ?

Subunits

Subunits Comment Organism
? x * 55000, SDS-PAGE Saccharolobus solfataricus
? x * 55784, calculated from sequence Saccharolobus solfataricus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
70
-
 assay at Saccharolobus solfataricus
95
-
-
 Saccharolobus solfataricus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
75 95 75°C: about 50% of maximal activity, 95°C: maximal activity, 100°C: less than 10% of activity Saccharolobus solfataricus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
70
-
 activity is constant for more than 72 h Saccharolobus solfataricus
80
-
 half-life: 25 h Saccharolobus solfataricus
90
-
 half-life: 0.5 h Saccharolobus solfataricus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.3
-
 2-toluamide pH 7.5, 70°C Saccharolobus solfataricus
0.8
-
 4-Aminobenzamide pH 7.5, 70°C Saccharolobus solfataricus
0.9
-
 2-Aminobenzamide pH 7.5, 70°C Saccharolobus solfataricus
1.1
-
 4-hydroxybenzamide pH 7.5, 70°C Saccharolobus solfataricus
1.3
-
 2-hydroxybenzamide pH 7.5, 70°C Saccharolobus solfataricus
1.6
-
 nicotinamide pH 7.5, 70°C Saccharolobus solfataricus
1.6
-
 acetamide pH 7.5, 70°C Saccharolobus solfataricus
1.6
-
 Isonicotinamide pH 7.5, 70°C Saccharolobus solfataricus
1.8
-
 picolinamide pH 7.5, 70°C Saccharolobus solfataricus
1.8
-
 3-indolacetamide pH 7.5, 70°C Saccharolobus solfataricus
2.9
-
 methacrylamide pH 7.5, 70°C Saccharolobus solfataricus
3.1
-
 L-prolinamide pH 7.5, 70°C Saccharolobus solfataricus
3.2
-
 Propionamide pH 7.5, 70°C Saccharolobus solfataricus
5.3
-
 hexanamide pH 7.5, 70°C Saccharolobus solfataricus
6.4
-
 Benzamide pH 7.5, 70°C Saccharolobus solfataricus
7.1
-
 3-phenylpropionamide pH 7.5, 70°C Saccharolobus solfataricus
7.3
-
 4-toluamide pH 7.5, 70°C Saccharolobus solfataricus
8.3
-
 Acrylamide pH 7.5, 70°C Saccharolobus solfataricus
9.9
-
 butyramide pH 7.5, 70°C Saccharolobus solfataricus
10.7
-
 lactamide pH 7.5, 70°C Saccharolobus solfataricus
16.3
-
 cyclohexanamide pH 7.5, 70°C Saccharolobus solfataricus
20.5
-
 Pentanamide pH 7.5, 70°C Saccharolobus solfataricus
21.9
-
 Isobutyramide pH 7.5, 70°C Saccharolobus solfataricus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
 Saccharolobus solfataricus

pH Range

pH Minimum pH Maximum Comment Organism
4 9.5 pH 4: about 50% of maximal activity, pH 9.5: about 50% of maximal activity Saccharolobus solfataricus

pI Value

Organism Comment pI Value Maximum pI Value
Saccharolobus solfataricus protein without His-tag tail calculated from amino acid composition
-
 5.94
Saccharolobus solfataricus protein with His-tag tail calculated from amino acid composition
-
 6.05

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.27
-
 4-Aminobenzamide pH 7.5, 70°C Saccharolobus solfataricus
0.3
-
 2-Aminobenzamide pH 7.5, 70°C Saccharolobus solfataricus
0.44
-
 acetamide pH 7.5, 70°C Saccharolobus solfataricus
0.46
-
 L-prolinamide pH 7.5, 70°C Saccharolobus solfataricus
0.72
-
 picolinamide pH 7.5, 70°C Saccharolobus solfataricus
0.73
-
 4-hydroxybenzamide pH 7.5, 70°C Saccharolobus solfataricus
0.75
-
 2-toluamide pH 7.5, 70°C Saccharolobus solfataricus
0.91
-
 hexanamide pH 7.5, 70°C Saccharolobus solfataricus
1.8
-
 3-phenylpropionamide pH 7.5, 70°C Saccharolobus solfataricus
2
-
 Isonicotinamide pH 7.5, 70°C Saccharolobus solfataricus
2
-
 3-indolacetamide pH 7.5, 70°C Saccharolobus solfataricus
2.4
-
 methacrylamide pH 7.5, 70°C Saccharolobus solfataricus
2.46
-
 Propionamide pH 7.5, 70°C Saccharolobus solfataricus
2.8
-
 Acrylamide pH 7.5, 70°C Saccharolobus solfataricus
3.3
-
 2-hydroxybenzamide pH 7.5, 70°C Saccharolobus solfataricus
3.3
-
 cyclohexanamide pH 7.5, 70°C Saccharolobus solfataricus
3.57
-
 lactamide pH 7.5, 70°C Saccharolobus solfataricus
5.3
-
 nicotinamide pH 7.5, 70°C Saccharolobus solfataricus
8.25
-
 butyramide pH 7.5, 70°C Saccharolobus solfataricus
15.7
-
 Pentanamide pH 7.5, 70°C Saccharolobus solfataricus
21.3
-
 Benzamide pH 7.5, 70°C Saccharolobus solfataricus
21.9
-
 Isobutyramide pH 7.5, 70°C Saccharolobus solfataricus
24.3
-
 cyclomaltohexaose pH 7.5, 70°C Saccharolobus solfataricus