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Information on EC 3.5.1.4 - amidase and Organism(s) Saccharolobus solfataricus and UniProt Accession P95896

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EC Tree
     3 Hydrolases
         3.5 Acting on carbon-nitrogen bonds, other than peptide bonds
             3.5.1 In linear amides
                3.5.1.4 amidase
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This record set is specific for:
Saccharolobus solfataricus
UNIPROT: P95896 not found.
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The taxonomic range for the selected organisms is: Saccharolobus solfataricus
The enzyme appears in selected viruses and cellular organisms
Synonyms
amidase, deaminase, acylase, acetamidase, amide hydrolase, acid transferase, n-acylethanolamine acid amidase, n-acylethanolamine-hydrolyzing acid amidase, atfaah, acylamidase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SSO2122
ordered locus name
acid transferase
-
-
-
-
acylamidase
-
-
-
-
acylase
-
-
-
-
amide hydrolase
-
-
-
-
amide transferase
-
-
-
-
amidohydrolase
-
-
-
-
deaminase
-
-
-
-
ester transferase
-
-
-
-
N-acetylaminohydrolase
-
-
-
-
signature amidase
-
-
Wide spectrum amidase
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of carboxylic acid amide
transfer of amide group
transfer of acyl group
SYSTEMATIC NAME
IUBMB Comments
acylamide amidohydrolase
-
CAS REGISTRY NUMBER
COMMENTARY hide
9012-56-0
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(S)-2-phenylpropionamide + H2O
(S)-2-phenylpropionate + NH3
show the reaction diagram
no activity with (R)-2-phenylpropionamide
-
-
?
2-aminobenzamide + H2O
2-aminobenzoic acid + NH3
show the reaction diagram
-
-
-
?
2-hydroxybenzamide + H2O
2-hydroxybenzoic acid + NH3
show the reaction diagram
-
-
-
?
2-thiophenecarboxamide + H2O
2-thiophenecarboxylate + NH3
show the reaction diagram
-
-
-
?
2-toluamide + H2O
2-toluic acid + NH3
show the reaction diagram
3-indolacetamide + H2O
3-indolacetate + NH3
show the reaction diagram
-
-
-
?
3-phenylpropionamide + H2O
3-phenylpropionate + NH3
show the reaction diagram
3-toluamide + H2O
3-toluic acid + NH3
show the reaction diagram
-
-
-
?
4-aminobenzamide + H2O
4-aminobenzoic acid + NH3
show the reaction diagram
-
-
-
?
4-hydroxybenzamide + H2O
4-hydroxybenzoate + NH3
show the reaction diagram
-
-
-
?
4-hydroxybenzamide + H2O
4-hydroxybenzoic acid + NH3
show the reaction diagram
-
-
-
?
4-methylbenzamide + H2O
4-methylbenzoic acid + NH3
show the reaction diagram
4-nitrobenzamide + H2O
4-nitrobenzoate + NH3
show the reaction diagram
-
-
-
?
4-toluamide + H2O
4-toluic acid + NH3
show the reaction diagram
acetamide + H2O
acetate + NH3
show the reaction diagram
-
-
-
?
acetamide + H2O
acetic acid + NH3
show the reaction diagram
-
-
-
?
acrylamide + H2O
acrylate + NH3
show the reaction diagram
-
-
-
?
anthranilamide + H2O
anthranilate + NH3
show the reaction diagram
-
-
-
?
benzamide + H2O
benzoate + NH3
show the reaction diagram
butyramide + H2O
butyrate + NH3
show the reaction diagram
cinnamamide + H2O
cinnamic acid + NH3
show the reaction diagram
-
-
-
?
cyclohexanamide + H2O
cyclohexanoate + NH3
show the reaction diagram
-
-
-
?
hexanamide + H2O
hexanoate + NH3
show the reaction diagram
indole-3-acetamide + H2O
indole-3-acetate + NH3
show the reaction diagram
-
-
-
?
isobutyramide + H2O
isobutyrate + NH3
show the reaction diagram
isonicotinamide + H2O
isonicotinic acid + NH3
show the reaction diagram
-
-
-
?
L-asparagine + H2O
L-aspartate + NH3
show the reaction diagram
-
-
-
?
L-glutamine + H2O
L-glutamate + NH3
show the reaction diagram
-
-
-
?
L-prolinamide + H2O
L-proline + NH3
show the reaction diagram
-
-
-
?
lactamide + H2O
lactate + NH3
show the reaction diagram
-
-
-
?
methacrylamide + H2O
methacrylate + NH3
show the reaction diagram
nicotinamide + H2O
nicotinic acid + NH3
show the reaction diagram
pentanamide + H2O
pentanoate + NH3
show the reaction diagram
picolinamide + H2O
picolinate + NH3
show the reaction diagram
-
-
-
?
propionamide + H2O
propionate + NH3
show the reaction diagram
propionamide + H2O
propionic acid + NH3
show the reaction diagram
-
-
-
?
pyrazinamide + H2O
pyrazinoic acid + NH3
show the reaction diagram
-
-
-
?
salicylamide + H2O
salicylate + NH3
show the reaction diagram
-
-
-
?
(R,S)-2-(3-benzoylphenyl)propanamide + H2O
(S)-2-(3-benzoylphenyl)propanoic acid + NH3
show the reaction diagram
-
enantioselective for the S-enantiomer
-
-
?
(R,S)-ketoprofen amide + H2O
(S)-ketoprofen + NH3
show the reaction diagram
-
the enzyme hydrolyzes S-ketoprofen amide to its corresponding acid in an absolutely enantioselective manner
-
-
?
(S)-2-(3-benzoylphenyl)propanamide + H2O
(S)-2-(3-benzoylphenyl)propanoic acid + NH3
show the reaction diagram
-
ketoprofen amide, R-amide is not converted
ketoprofen
-
?
(S)-ibuprofen amide + H2O
(S)-ibuprofen + NH3
show the reaction diagram
-
-
-
-
?
2-toluamide + H2O
2-toluic acid + NH3
show the reaction diagram
-
-
-
-
?
3-phenyl-propionamide + H2O
3-phenyl-propionate + NH3
show the reaction diagram
-
-
-
-
?
4-methylbenzamide + H2O
4-methylbenzoic acid + NH3
show the reaction diagram
-
-
-
-
?
acetamide + H2O
acetate + NH3
show the reaction diagram
-
-
-
-
?
benzamide + H2O
benzoate + NH3
show the reaction diagram
-
-
-
-
?
benzamide + H2O
benzoic acid + NH3
show the reaction diagram
-
-
-
-
?
butyramide + H2O
butyrate + NH3
show the reaction diagram
-
-
-
-
?
ibuprofen amide + H2O
ibuprofen + NH3
show the reaction diagram
-
-
-
-
?
indole-3-acetamide + H2O
indole-3-acetate + NH3
show the reaction diagram
-
-
-
-
?
isobutyramide + H2O
isobutyrate + NH3
show the reaction diagram
-
-
-
-
?
methacrylamide + H2O
methacrylate + NH3
show the reaction diagram
-
-
-
-
?
propionamide + H2O
propionate + NH3
show the reaction diagram
-
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
1 mM, about 20% activation
Fe3+
1 mM, about 20% activation
V2+
1 mM, about 20% activation
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
additional information
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
phenylmethanesulfonyl fluoride
inhibits amide conversion more than 94% and completely suppresses nitrile hydrolysis
phenylphosphorodiamidate
-
competitive inhibitor, the inhibitory effect is fully reversed by SSAM substrate benzamide, demonstrating the hydrophobicity of the active site
additional information
-
diethylphosphoroamidate shows no inhibitory activity
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.3
2-Aminobenzamide
pH 7.5, 70°C
0.4
2-hydroxybenzamide
pH 7.5, 70°C
0.1 - 0.3
2-toluamide
0.9
3-indolacetamide
pH 7.5, 70°C
2.5 - 4
3-phenylpropionamide
0.3
4-Aminobenzamide
pH 7.5, 70°C
0.1 - 1.5
4-hydroxybenzamide
0.2 - 0.3
4-nitrobenzamide
3.6 - 37
acetamide
3
Acrylamide
pH 7.5, 70°C
0.000674 - 0.9
Benzamide
1.2 - 14
butyramide
0.000434 - 0.000526
cinnamamide
5
cyclohexanamide
pH 7.5, 70°C
0.2 - 0.3
cyclomaltohexaose
4.5 - 5.8
hexanamide
0.8
indol-3-acetamide
0.000099 - 7
Isobutyramide
0.8
Isonicotinamide
pH 7.5, 70°C
6.7
L-prolinamide
pH 7.5, 70°C
3
lactamide
pH 7.5, 70°C
0.3 - 0.5
metacrylamide
1.2
methacrylamide
pH 7.5, 70°C
0.3 - 1.5
nicotinamide
1.3 - 2.5
Pentanamide
2.5
picolinamide
pH 7.5, 70°C
0.000093 - 10
Propionamide
0.125
2-toluamide
-
pH 5.0, 70°C, wild-type enzyme
2.325
3-phenyl-propionamide
-
pH 5.0, 70°C, wild-type enzyme
26.5
acetamide
-
pH 5.0, 70°C, wild-type enzyme
7.25
Benzamide
-
pH 5.0, 70°C, wild-type enzyme
8.55
butyramide
-
pH 5.0, 70°C, wild-type enzyme
0.25
cyclomaltohexaose
-
pH 5.0, 70°C, wild-type enzyme
1.225
indol-3-acetamide
-
pH 5.0, 70°C, wild-type enzyme
4.975
Isobutyramide
-
pH 5.0, 70°C, wild-type enzyme
0.000096
ketoprofen amide
-
-
0.55
metacrylamide
-
pH 5.0, 70°C, wild-type enzyme
12.25
Propionamide
-
pH 5.0, 70°C, wild-type enzyme
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.9
2-Aminobenzamide
pH 7.5, 70°C
1.3
2-hydroxybenzamide
pH 7.5, 70°C
0.3 - 0.8
2-toluamide
1.8
3-indolacetamide
pH 7.5, 70°C
7.1 - 18
3-phenylpropionamide
0.8
4-Aminobenzamide
pH 7.5, 70°C
1.1 - 2.1
4-hydroxybenzamide
0.8 - 2.5
4-nitrobenzamide
7.3 - 14.2
4-toluamide
-
1.6 - 30.1
acetamide
8.3
Acrylamide
pH 7.5, 70°C
6.4 - 1246
Benzamide
9.9 - 34.3
butyramide
769 - 906
cinnamamide
16.3
cyclohexanamide
pH 7.5, 70°C
5.3 - 21.6
hexanamide
1.2 - 2.2
indol-3-acetamide
21.9 - 175
Isobutyramide
1.6
Isonicotinamide
pH 7.5, 70°C
3.1
L-prolinamide
pH 7.5, 70°C
10.7
lactamide
pH 7.5, 70°C
3 - 4.8
metacrylamide
2.9
methacrylamide
pH 7.5, 70°C
1.6 - 6.4
nicotinamide
20.5 - 28.6
Pentanamide
1.8
picolinamide
pH 7.5, 70°C
3.2 - 181
Propionamide
0.425
2-toluamide
-
pH 5.0, 70°C, wild-type enzyme
12.82
3-phenyl-propionamide
-
pH 5.0, 70°C, wild-type enzyme
20.13
acetamide
-
pH 5.0, 70°C, wild-type enzyme
6.1
Benzamide
-
pH 5.0, 70°C, wild-type enzyme
19.98
butyramide
-
pH 5.0, 70°C, wild-type enzyme
11.9
cyclomaltohexaose
-
pH 5.0, 70°C, wild-type enzyme
2.05
indol-3-acetamide
-
pH 5.0, 70°C, wild-type enzyme
41.88
Isobutyramide
-
pH 5.0, 70°C, wild-type enzyme
887
ketoprofen amide
-
-
5.75
metacrylamide
-
pH 5.0, 70°C, wild-type enzyme
101.9
Propionamide
-
pH 5.0, 70°C, wild-type enzyme
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.3
2-Aminobenzamide
pH 7.5, 70°C
3.3
2-hydroxybenzamide
pH 7.5, 70°C
0.75 - 4
2-toluamide
2
3-indolacetamide
pH 7.5, 70°C
1.8 - 7.2
3-phenylpropionamide
0.27
4-Aminobenzamide
pH 7.5, 70°C
0.73 - 21
4-hydroxybenzamide
4 - 8.33
4-nitrobenzamide
0.44 - 1.31
acetamide
2.8
Acrylamide
pH 7.5, 70°C
7.44 - 21.3
Benzamide
2.45 - 8.25
butyramide
3.3
cyclohexanamide
pH 7.5, 70°C
24.3 - 61
cyclomaltohexaose
0.91 - 4.8
hexanamide
1.5 - 2.75
indol-3-acetamide
6.99 - 21.9
Isobutyramide
2
Isonicotinamide
pH 7.5, 70°C
0.46
L-prolinamide
pH 7.5, 70°C
3.57
lactamide
pH 7.5, 70°C
6 - 16
metacrylamide
2.4
methacrylamide
pH 7.5, 70°C
4.26 - 5.5
nicotinamide
10.7 - 19
Pentanamide
0.72
picolinamide
pH 7.5, 70°C
2.46 - 9.94
Propionamide
3.25
2-toluamide
-
pH 5.0, 70°C, wild-type enzyme
5.44
3-phenyl-propionamide
-
pH 5.0, 70°C, wild-type enzyme
0.76
acetamide
-
pH 5.0, 70°C, wild-type enzyme
10.4
Benzamide
-
pH 5.0, 70°C, wild-type enzyme
2.8
butyramide
-
pH 5.0, 70°C, wild-type enzyme
48.13
cyclomaltohexaose
-
pH 5.0, 70°C, wild-type enzyme
2.125
indol-3-acetamide
-
pH 5.0, 70°C, wild-type enzyme
7.5
Isobutyramide
-
pH 5.0, 70°C, wild-type enzyme
11.81
metacrylamide
-
pH 5.0, 70°C, wild-type enzyme
7.41
Propionamide
-
pH 5.0, 70°C, wild-type enzyme
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1.18
85°C, pH not specified in the publication, substrate: benzamide
0.19
-
>99% enantiomeric excess
16
-
purified recombinant wild-type enzyme
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5 - 8.5
pH 3.5: wild-type enzyme shows about 45% of maximal activity, mutant enzyme Y41C shows about 20% of maximal activity. pH 8.5: wild-type enzyme shows about 40% of maximal activity, mutant enzyme Y41C shows about 50% of maximal activity
4 - 9.5
pH 4: about 50% of maximal activity, pH 9.5: about 50% of maximal activity
additional information
-
wild-type and mutant enzymes, activity-pH relationships, profiles at different temperatures, overview
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
75 - 95
75°C: about 50% of maximal activity, 95°C: maximal activity, 100°C: less than 10% of activity
additional information
-
wild-type and mutant enzymes, activity-temperature relationships, profiles at different pHs, overview
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.94
protein without His-tag tail calculated from amino acid composition
6.05
protein with His-tag tail calculated from amino acid composition
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
400000
octameric enzyme form, gel filtration
55000
x * 55000, SDS-PAGE
55784
83000
dimeric enzyme form, gel filtration
110000
-
gel filtration
50000
-
2 * 50000, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
-
2 * 50000, SDS-PAGE
additional information
-
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
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
native enzyme using the hanging-drop vapour-diffudion technique, a His-tagged protein cannot be crystallized
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K96R
retains the ability to hydrolyse amides but hydrolyses nitriles more efficiently than the wild-type enzyme
Y41C
no structural modifications in the Y41C mutant, the pH-spectrum is slightly increased
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
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
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
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
additional information
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
104
Tm-value of wild-type enzyme at pH 5.0 is 104.3°C
84
Tm-value of mutant enzyme Y41C at pH 8.0 is 83.5°C
94
Tm-value of wild-type enzyme at pH 8.0 is 94.4°C
97
Tm-value of mutant enzyme Y41C at pH 5.0 is 97.3°C
additional information
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
the enzyme may be stored at room temperature in the presence of 0.04% NaN3 for 1 year without any loss of activity
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
extraction of a His-tagged enzyme from insoluble cellular fractions using sodium phosphate buffer at pH 7.8 and further purification by affinity column chromatography
-
recombinant enzyme 2.35fold from Escherichia coli strain BL21(DE3) by anion exchange chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
overexpression in Escherichia coli as a fusion protein with an N-terminus six-histidine-residue affinity tag
expression in Escherichia coli of a recombinant His-tagged protein
-
expression of gene SsAH in Escherichia coli strain BL21(DE3) as soluble, active protein, sequence comparison of wild-type and mutants enzymes, overview
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
-
the enzyme is useful for production of optically pure S-arylpropionic acids
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Nastopoulos, V.; Vallone, B.; Politi, L.; Scotto d'Abusco, A.; Scandurra, R.; Tsernoglou, D.
Crystallization and X-ray diffraction measurements of a thermophilic archaeal recombinant amidase from Sulfolobus solfataricus MT4
Acta Crystallogr. Sect. D
D57
1036-1037
2001
Saccharolobus solfataricus, Saccharolobus solfataricus MT4
Manually annotated by BRENDA team
Giordano, C.; Ammendola, S.
Characterization of mutants of Sulfolobus solfataricus signature amidase able to hydrolyse R-ketoprofen amide
Protein Pept. Lett.
15
617-623
2008
Saccharolobus solfataricus
Manually annotated by BRENDA team
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
Archaea
2
221-231
2009
Saccharolobus solfataricus
Manually annotated by BRENDA team
dAbusco, A.S.; Ammendola, S.; Scandurra, R.; Politi, L.
Molecular and biochemical characterization of the recombinant amidase from hyperthermophilic archaeon Sulfolobus solfataricus
Extremophiles
5
183-192
2001
Saccharolobus solfataricus (P95896), Saccharolobus solfataricus, Saccharolobus solfataricus MT4 (P95896)
Manually annotated by BRENDA team
D'Abusco, A.S.; Casadio, R.; Tasco, G.; Giangiacomo, L.; Giartosio, A.; Calamia, V.; Di Marco, S.; Chiaraluce, R.; Consalvi, V.; Scandurra, R.; Politi, L.
Oligomerization of Sulfolobus solfataricus signature amidase is promoted by acidic pH and high temperature
Archaea
1
411-423
2005
Saccharolobus solfataricus (P95896), Saccharolobus solfataricus, Saccharolobus solfataricus P2 (P95896)
Manually annotated by BRENDA team
Cilia, E.; Fabbri, A.; Uriani, M.; Scialdone, G.G., Ammendola, S.
The signature amidase from Sulfolobus solfataricus belongs to the CX3C subgroup of enzymes cleaving both amides and nitriles. Ser195 and Cys145 are predicted to be the active site nucleophiles
FEBS J.
272
4716-4724
2005
Saccharolobus solfataricus (P95896), Saccharolobus solfataricus, Saccharolobus solfataricus P2 (P95896)
Manually annotated by BRENDA team