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Literature summary for 3.5.1.4 extracted from

  • Politi, L.; Chiancone, E.; Giangiacomo, L.; Cervoni, L.; Scotto dAbusco, A.; Scorsino, S.; Scandurra, R.
    pH-, temperature- and ion-dependent oligomerization of Sulfolobus solfataricus recombinant amidase: a study with site-specific mutants (2009), Archaea, 2, 221-231.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.5.1.4

Protein Variants

Protein Variants Comment Organism
D487N site-specific mutagenesis, the D487N mutation is located in the center of the protein, far distant from the 33-48 segment. The mutant amidase, like the wild-type enzyme, responds to changes in pH and temperature with a conformational change affecting the dimer-octamer equilibrium Saccharolobus solfataricus
L34P site-specific mutagenesis, the mutant amidase, in contrast to the wild-type enzyme, is unaffected by pH and temperature remaining always in the dimeric state Saccharolobus solfataricus
T319I site-specific mutagenesis, the T319I mutation is located in a strand on the surface of the protein, which is far from, and opposite to, the N-terminal segment. The mutant amidase, like the wild-type enzyme, responds to changes in pH and temperature with a conformational change affecting the dimer-octamer equilibrium Saccharolobus solfataricus
Y41C site-specific mutagenesis, the mutant amidase, in contrast to the wild-type enzyme, is unaffected by pH and temperature remaining always in the dimeric state Saccharolobus solfataricus

Inhibitors

Inhibitors Comment Organism Structure
additional information diethylphosphoroamidate shows no inhibitory activity Saccharolobus solfataricus
phenylphosphorodiamidate competitive inhibitor, the inhibitory effect is fully reversed by SSAM substrate benzamide, demonstrating the hydrophobicity of the active site Saccharolobus solfataricus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.125
-
 2-toluamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
0.25
-
 cyclomaltohexaose pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
0.55
-
 metacrylamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
1.225
-
 indol-3-acetamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
2.325
-
 3-phenyl-propionamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
4.975
-
 Isobutyramide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
7.25
-
 Benzamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
8.55
-
 butyramide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
12.25
-
 Propionamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
26.5
-
 acetamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus

Metals/Ions

Metals/Ions Comment Organism Structure
NaCl influences the oligomerization status of the enzyme. 60% octameric and 40% dimeric enzyme in absence of NaCl, 5% octameric and 95% dimeric enzyme at and above 150 mM NaCl, profile overview Saccharolobus solfataricus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
50000
-
 2 * 50000, SDS-PAGE Saccharolobus solfataricus
110000
-
 gel filtration Saccharolobus solfataricus

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-toluamide + H2O
-
 Saccharolobus solfataricus 2-toluic acid + NH3
-
 ?
3-phenyl-propionamide + H2O
-
 Saccharolobus solfataricus 3-phenyl-propionate + NH3
-
 ?
4-methylbenzamide + H2O
-
 Saccharolobus solfataricus 4-methylbenzoic acid + NH3
-
 ?
acetamide + H2O
-
 Saccharolobus solfataricus acetate + NH3
-
 ?
benzamide + H2O
-
 Saccharolobus solfataricus benzoate + NH3
-
 ?
butyramide + H2O
-
 Saccharolobus solfataricus butyrate + NH3
-
 ?
indole-3-acetamide + H2O
-
 Saccharolobus solfataricus indole-3-acetate + NH3
-
 ?
isobutyramide + H2O
-
 Saccharolobus solfataricus isobutyrate + NH3
-
 ?
methacrylamide + H2O
-
 Saccharolobus solfataricus methacrylate + NH3
-
 ?
propionamide + H2O
-
 Saccharolobus solfataricus propionate + NH3
-
 ?

Subunits

Subunits Comment Organism
dimer 2 * 50000, SDS-PAGE Saccharolobus solfataricus
More at pHs above pH 7.0 only dimers are present, but with decreasing pH dimers associate into octamers with complete oligomerization occurring at pH 3.0. Oligomerization also shows reversible temperature-dependence with octamer formation increasing with temperature from 36°C to between 70°C and 80°C. Increasing salt concentrations favor dissociation of the octamers, oligomerization profiles under different conditions, mechanism, dimer structure, modelling, overview Saccharolobus solfataricus

Synonyms

Synonyms Comment Organism
SSAM
-
 Saccharolobus solfataricus

Temperature Optimum [°C]

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

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
additional information
-
 wild-type and mutant enzymes, activity-temperature relationships, profiles at different pHs, overview Saccharolobus solfataricus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.425
-
 2-toluamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
2.05
-
 indol-3-acetamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
5.75
-
 metacrylamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
6.1
-
 Benzamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
11.9
-
 cyclomaltohexaose pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
12.82
-
 3-phenyl-propionamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
19.98
-
 butyramide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
20.13
-
 acetamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
41.88
-
 Isobutyramide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
101.9
-
 Propionamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5
-
 assay at Saccharolobus solfataricus

pH Range

pH Minimum pH Maximum Comment Organism
additional information
-
 wild-type and mutant enzymes, activity-pH relationships, profiles at different temperatures, overview Saccharolobus solfataricus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.76
-
 acetamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
2.125
-
 indol-3-acetamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
2.8
-
 butyramide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
3.25
-
 2-toluamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
5.44
-
 3-phenyl-propionamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
7.41
-
 Propionamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
7.5
-
 Isobutyramide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
10.4
-
 Benzamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
11.81
-
 metacrylamide pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus
48.13
-
 cyclomaltohexaose pH 5.0, 70°C, wild-type enzyme Saccharolobus solfataricus