Literature summary for 3.5.5.1 extracted from

  • 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 (2005), FEBS J., 272, 4716-4724.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Saccharolobus solfataricus

Engineering

Protein Variants Comment Organism
K96R retains the ability to hydrolyse amides but hydrolyses nitriles more efficiently than the wild-type enzyme Saccharolobus solfataricus

Inhibitors

Inhibitors Comment Organism Structure
phenylmethanesulfonyl fluoride inhibits amide conversion more than 94% and completely suppresses nitrile hydrolysis Saccharolobus solfataricus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.000006
-
Benzonitrile pH 7.4, 70°C, mutant enzyme K96R Saccharolobus solfataricus
0.00000013
-
trichloroacetonitrile pH 7.4, 70°C, mutant enzyme K96R Saccharolobus solfataricus
0.000005
-
Benzonitrile pH 7.4, 70°C, wild-type enzyme Saccharolobus solfataricus
0.0000000495
-
Malononitrile pH 7.4, 70°C, wild-type enzyme Saccharolobus solfataricus
0.0000000005
-
trichloroacetonitrile pH 7.4, 70°C, wild-type enzyme Saccharolobus solfataricus
0.000000024
-
phenylglycinenitrile pH 7.4, 70°C, wild-type enzyme Saccharolobus solfataricus
0.000000157
-
3-anilinoproprionitrile pH 7.4, 70°C, wild-type enzyme Saccharolobus solfataricus
0.000000609
-
Malononitrile pH 7.4, 70°C, mutant enzyme K96R Saccharolobus solfataricus
0.000000009
-
phenylglycinenitrile pH 7.4, 70°C, mutant enzyme K96R Saccharolobus solfataricus
0.0000000097
-
Cinnamonitrile pH 7.4, 70°C, mutant enzyme K96R Saccharolobus solfataricus
0.000000103
-
3-anilinoproprionitrile pH 7.4, 70°C, mutant enzyme K96R Saccharolobus solfataricus

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus P95896
-
-
Saccharolobus solfataricus P2 P95896
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Saccharolobus solfataricus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0006
-
pH 7.4, 70°C, substrate: benzonitrile Saccharolobus solfataricus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
benzonitrile + 2 H2O
-
Saccharolobus solfataricus benzoate + NH3
-
?
benzonitrile + 2 H2O
-
Saccharolobus solfataricus P2 benzoate + NH3
-
?
3-anilinoproprionitrile + 2 H2O
-
Saccharolobus solfataricus 3-anilinopropionate + NH3
-
?
cinnamonitrile + 2 H2O cinnamonitrile is a substrate for mutant enzyme K96R, no activity with wild-type enzyme Saccharolobus solfataricus cinnamic acid + NH3
-
?
malononitrile + 2 H2O
-
Saccharolobus solfataricus malonate + NH3
-
?
phenylglycinenitrile + 2 H2O
-
Saccharolobus solfataricus phenylglycine + NH3
-
?
trichloroacetonitrile + 2 H2O
-
Saccharolobus solfataricus trichloroacetate + NH3
-
?

Synonyms

Synonyms Comment Organism
SsAH
-
Saccharolobus solfataricus
SSO2122 ordered locus name Saccharolobus solfataricus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
70
-
enzyme remains active for about 6 days Saccharolobus solfataricus
95
-
half-life: 5 h Saccharolobus solfataricus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.016
-
Cinnamonitrile pH 7.4, 70°C, mutant enzyme K96R Saccharolobus solfataricus
0.016
-
phenylglycinenitrile pH 7.4, 70°C, mutant enzyme K96R Saccharolobus solfataricus
0.04
-
phenylglycinenitrile pH 7.4, 70°C, wild-type enzyme Saccharolobus solfataricus
0.01
-
trichloroacetonitrile pH 7.4, 70°C, wild-type enzyme Saccharolobus solfataricus
1.05
-
Malononitrile pH 7.4, 70°C, mutant enzyme K96R Saccharolobus solfataricus
0.28
-
3-anilinoproprionitrile pH 7.4, 70°C, wild-type enzyme Saccharolobus solfataricus
0.14
-
Malononitrile pH 7.4, 70°C, wild-type enzyme Saccharolobus solfataricus
0.18
-
3-anilinoproprionitrile pH 7.4, 70°C, mutant enzyme K96R Saccharolobus solfataricus
0.09
-
Benzonitrile pH 7.4, 70°C, wild-type enzyme Saccharolobus solfataricus
0.225
-
trichloroacetonitrile pH 7.4, 70°C, mutant enzyme K96R Saccharolobus solfataricus