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Literature summary extracted from
- 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.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.5.1.4 | expression in Escherichia coli | Saccharolobus solfataricus |
| 3.5.5.1 | expression in Escherichia coli | Saccharolobus solfataricus |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 3.5.1.4 | K96R | retains the ability to hydrolyse amides but hydrolyses nitriles more efficiently than the wild-type enzyme | Saccharolobus solfataricus |
| 3.5.5.1 | K96R | retains the ability to hydrolyse amides but hydrolyses nitriles more efficiently than the wild-type enzyme | Saccharolobus solfataricus |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.5.1.4 | phenylmethanesulfonyl fluoride | inhibits amide conversion more than 94% and completely suppresses nitrile hydrolysis | Saccharolobus solfataricus |  |
| 3.5.5.1 | phenylmethanesulfonyl fluoride | inhibits amide conversion more than 94% and completely suppresses nitrile hydrolysis | Saccharolobus solfataricus | |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.5.1.4 | 0.000093 | - |
Propionamide | 85°C, pH not specified in the publication, mutant enzyme K96R | Saccharolobus solfataricus | |
| 3.5.1.4 | 0.000099 | - |
Isobutyramide | 85°C, pH not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.1.4 | 0.000102 | - |
Propionamide | 85°C, pH not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.1.4 | 0.0002 | - |
Isobutyramide | 85°C, pH not specified in the publication, mutant enzyme K96R | Saccharolobus solfataricus | |
| 3.5.1.4 | 0.000434 | - |
cinnamamide | 85°C, pH not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.1.4 | 0.000526 | - |
cinnamamide | 85°C, pH not specified in the publication, mutant enzyme K96R | Saccharolobus solfataricus | |
| 3.5.1.4 | 0.000674 | - |
Benzamide | 85°C, pH not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.1.4 | 0.000723 | - |
Benzamide | 85°C, pH not specified in the publication, mutant enzyme K96R | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.0000000005 | - |
trichloroacetonitrile | pH 7.4, 70°C, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.000000009 | - |
phenylglycinenitrile | pH 7.4, 70°C, mutant enzyme K96R | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.0000000097 | - |
Cinnamonitrile | pH 7.4, 70°C, mutant enzyme K96R | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.000000024 | - |
phenylglycinenitrile | pH 7.4, 70°C, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.0000000495 | - |
Malononitrile | pH 7.4, 70°C, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.000000103 | - |
3-anilinoproprionitrile | pH 7.4, 70°C, mutant enzyme K96R | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.00000013 | - |
trichloroacetonitrile | pH 7.4, 70°C, mutant enzyme K96R | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.000000157 | - |
3-anilinoproprionitrile | pH 7.4, 70°C, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.000000609 | - |
Malononitrile | pH 7.4, 70°C, mutant enzyme K96R | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.000005 | - |
Benzonitrile | pH 7.4, 70°C, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.000006 | - |
Benzonitrile | pH 7.4, 70°C, mutant enzyme K96R | Saccharolobus solfataricus | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.5.1.4 | Saccharolobus solfataricus | P95896 | - |
- |
| 3.5.1.4 | Saccharolobus solfataricus P2 | P95896 | - |
- |
| 3.5.5.1 | Saccharolobus solfataricus | P95896 | - |
- |
| 3.5.5.1 | Saccharolobus solfataricus P2 | P95896 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 3.5.1.4 | - |
Saccharolobus solfataricus |
| 3.5.5.1 | - |
Saccharolobus solfataricus |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 3.5.1.4 | 1.18 | - |
85°C, pH not specified in the publication, substrate: benzamide | Saccharolobus solfataricus |
| 3.5.5.1 | 0.0006 | - |
pH 7.4, 70°C, substrate: benzonitrile | Saccharolobus solfataricus |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.5.1.4 | benzamide + H2O | - |
Saccharolobus solfataricus | benzoate + NH3 | - |
? | |
| 3.5.1.4 | benzamide + H2O | - |
Saccharolobus solfataricus P2 | benzoate + NH3 | - |
? | |
| 3.5.1.4 | cinnamamide + H2O | - |
Saccharolobus solfataricus | cinnamic acid + NH3 | - |
? | |
| 3.5.1.4 | cinnamamide + H2O | - |
Saccharolobus solfataricus P2 | cinnamic acid + NH3 | - |
? | |
| 3.5.1.4 | isobutyramide + H2O | - |
Saccharolobus solfataricus | isobutyrate + NH3 | - |
? | |
| 3.5.1.4 | isobutyramide + H2O | - |
Saccharolobus solfataricus P2 | isobutyrate + NH3 | - |
? | |
| 3.5.1.4 | propionamide + H2O | - |
Saccharolobus solfataricus | propionate + NH3 | - |
? | |
| 3.5.1.4 | propionamide + H2O | - |
Saccharolobus solfataricus P2 | propionate + NH3 | - |
? | |
| 3.5.5.1 | 3-anilinoproprionitrile + 2 H2O | - |
Saccharolobus solfataricus | 3-anilinopropionate + NH3 | - |
? | |
| 3.5.5.1 | benzonitrile + 2 H2O | - |
Saccharolobus solfataricus | benzoate + NH3 | - |
? | |
| 3.5.5.1 | benzonitrile + 2 H2O | - |
Saccharolobus solfataricus P2 | benzoate + NH3 | - |
? | |
| 3.5.5.1 | cinnamonitrile + 2 H2O | cinnamonitrile is a substrate for mutant enzyme K96R, no activity with wild-type enzyme | Saccharolobus solfataricus | cinnamic acid + NH3 | - |
? | |
| 3.5.5.1 | malononitrile + 2 H2O | - |
Saccharolobus solfataricus | malonate + NH3 | - |
? | |
| 3.5.5.1 | phenylglycinenitrile + 2 H2O | - |
Saccharolobus solfataricus | phenylglycine + NH3 | - |
? | |
| 3.5.5.1 | trichloroacetonitrile + 2 H2O | - |
Saccharolobus solfataricus | trichloroacetate + NH3 | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 3.5.1.4 | SsAH | - |
Saccharolobus solfataricus |
| 3.5.1.4 | SSO2122 | ordered locus name | Saccharolobus solfataricus |
| 3.5.5.1 | SsAH | - |
Saccharolobus solfataricus |
| 3.5.5.1 | SSO2122 | ordered locus name | Saccharolobus solfataricus |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.5.1.4 | 70 | - |
enzyme remains active for about 6 days | Saccharolobus solfataricus |
| 3.5.1.4 | 95 | - |
half-life: 5 h | Saccharolobus solfataricus |
| 3.5.5.1 | 70 | - |
enzyme remains active for about 6 days | Saccharolobus solfataricus |
| 3.5.5.1 | 95 | - |
half-life: 5 h | Saccharolobus solfataricus |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.5.1.4 | 160 | - |
Propionamide | 85°C, pH not specified in the publication, mutant enzyme K96R | Saccharolobus solfataricus | |
| 3.5.1.4 | 160 | - |
Isobutyramide | 85°C, pH not specified in the publication, mutant enzyme K96R | Saccharolobus solfataricus | |
| 3.5.1.4 | 175 | - |
Isobutyramide | 85°C, pH not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.1.4 | 181 | - |
Propionamide | 85°C, pH not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.1.4 | 769 | - |
cinnamamide | 85°C, pH not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.1.4 | 906 | - |
cinnamamide | 85°C, pH not specified in the publication, mutant enzyme K96R | Saccharolobus solfataricus | |
| 3.5.1.4 | 1192 | - |
Benzamide | 85°C, pH not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.1.4 | 1246 | - |
Benzamide | 85°C, pH not specified in the publication, mutant enzyme K96R | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.01 | - |
trichloroacetonitrile | pH 7.4, 70°C, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.016 | - |
Cinnamonitrile | pH 7.4, 70°C, mutant enzyme K96R | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.016 | - |
phenylglycinenitrile | pH 7.4, 70°C, mutant enzyme K96R | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.04 | - |
phenylglycinenitrile | pH 7.4, 70°C, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.09 | - |
Benzonitrile | pH 7.4, 70°C, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.14 | - |
Malononitrile | pH 7.4, 70°C, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.18 | - |
3-anilinoproprionitrile | pH 7.4, 70°C, mutant enzyme K96R | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.225 | - |
trichloroacetonitrile | pH 7.4, 70°C, mutant enzyme K96R | Saccharolobus solfataricus | |
| 3.5.5.1 | 0.28 | - |
3-anilinoproprionitrile | pH 7.4, 70°C, wild-type enzyme | Saccharolobus solfataricus | |
| 3.5.5.1 | 1.05 | - |
Malononitrile | pH 7.4, 70°C, mutant enzyme K96R | Saccharolobus solfataricus | |
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