Activating Compound | Comment | Organism | Structure |
---|---|---|---|
DTT | 20% activation at 1 mM | Pseudonocardia thermophila |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ba2+ | 13% inhibition at 1 mM | Pseudonocardia thermophila | |
Ca2+ | 15% inhibition at 1 mM | Pseudonocardia thermophila | |
Co2+ | complete inhibition at 1 mM | Pseudonocardia thermophila | |
Cu2+ | 74% inhibition at 1 mM | Pseudonocardia thermophila | |
Fe2+ | 20% inhibition at 1 mM | Pseudonocardia thermophila | |
Hg2+ | complete inhibition at 1 mM | Pseudonocardia thermophila | |
iodoacetamide | 10% inhibition at 1 mM | Pseudonocardia thermophila | |
iodoacetate | 78% inhibition at 1 mM | Pseudonocardia thermophila | |
Mg2+ | 10% inhibition at 1 mM | Pseudonocardia thermophila | |
additional information | no inhibition by Triton X-100, SDS, EDTA, Mn2+, and Fe3+ at 1 mM | Pseudonocardia thermophila | |
Ni2+ | 56% inhibition at 1 mM | Pseudonocardia thermophila | |
Zn2+ | 7% inhibition at 1 mM | Pseudonocardia thermophila |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.9 | - |
2-phenylpropionamide | pH 7.2, 70°C | Pseudonocardia thermophila | |
4.9 | - |
Benzamide | pH 7.2, 70°C | Pseudonocardia thermophila | |
7.4 | - |
Propionamide | pH 7.2, 70°C | Pseudonocardia thermophila | |
9.2 | - |
methacrylamide | pH 7.2, 70°C | Pseudonocardia thermophila |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
52000 | - |
2 * 52000, SDS-PAGE | Pseudonocardia thermophila |
108000 | - |
gel filtration | Pseudonocardia thermophila |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudonocardia thermophila | - |
thermophilic actinomycete, DSMZ 43832 | - |
Purification (Comment) | Organism |
---|---|
native enzyme 48fold to homogeneity by hydrophobic interaction chromatography, ultrafiltration, anion exchange chromatography, and gel filtration | Pseudonocardia thermophila |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
19.4 | - |
purified enzyme, substrate benzamide | Pseudonocardia thermophila |
23.5 | - |
purified enzyme, substrate methacrylamide | Pseudonocardia thermophila |
25.4 | - |
purified enzyme, substrate propionamide | Pseudonocardia thermophila |
26.8 | - |
purified enzyme, substrate acrylamide | Pseudonocardia thermophila |
Storage Stability | Organism |
---|---|
4°C, purified enzyme, in 50 mM phosphate buffer, more than 6 months without loss of activity | Pseudonocardia thermophila |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R,S)-2-phenylpropionamide + H2O | the enzyme is highly S-stereoselective for 2-phenylpropionamide, and the racemic amide is converted to the corresponding S-acid with an enantiomeric excess of over 95% at 50% conversion of the substrate | Pseudonocardia thermophila | (S)-2-phenylpropionic acid + NH3 | - |
? | |
2-aminobenzamide + H2O | - |
Pseudonocardia thermophila | 2-aminobenzoic acid + NH3 | - |
? | |
2-hydroxybenzamide + H2O | - |
Pseudonocardia thermophila | 2-hydroxybenzoic acid + NH3 | - |
? | |
2-tolylamide + H2O | - |
Pseudonocardia thermophila | ? + NH3 | - |
? | |
3-indolylacetamide + H2O | - |
Pseudonocardia thermophila | 3-indolylacetic acid + NH3 | - |
? | |
3-tolylamide + H2O | - |
Pseudonocardia thermophila | ? + NH3 | - |
? | |
4-aminobenzamide + H2O | - |
Pseudonocardia thermophila | 4-aminobenzoic acid + NH3 | - |
? | |
4-hydroxybenzamide + H2O | - |
Pseudonocardia thermophila | 4-hydroxybenzoic acid + NH3 | - |
? | |
4-tolylamide + H2O | - |
Pseudonocardia thermophila | ? + NH3 | - |
? | |
acetamide + H2O | - |
Pseudonocardia thermophila | acetic acid + NH3 | - |
? | |
acrylamide + H2O | - |
Pseudonocardia thermophila | acrylic acid + NH3 | - |
? | |
adipoamide + H2O | - |
Pseudonocardia thermophila | adipic acid + NH3 | - |
? | |
benzamide + H2O | - |
Pseudonocardia thermophila | benzoic acid + NH3 | - |
? | |
cyclohexanoamide + H2O | - |
Pseudonocardia thermophila | cyclohexanoic acid + NH3 | - |
? | |
DL-lactamide + H2O | - |
Pseudonocardia thermophila | DL-lactate + NH3 | - |
? | |
DL-methioninamide + H2O | - |
Pseudonocardia thermophila | DL-methionine + NH3 | - |
? | |
DL-tryptophanamide + H2O | - |
Pseudonocardia thermophila | DL-tryptophan + NH3 | - |
? | |
formamide + H2O | - |
Pseudonocardia thermophila | formate + NH3 | - |
? | |
hexanoamide + H2O | - |
Pseudonocardia thermophila | hexanoic acid + NH3 | - |
? | |
hydroxylamine hydrochloride + H2O | - |
Pseudonocardia thermophila | ? | - |
? | |
isobutyramide + H2O | - |
Pseudonocardia thermophila | isobutyric acid + NH3 | - |
? | |
isonicotinamide + H2O | - |
Pseudonocardia thermophila | isonicotinic acid + NH3 | - |
? | |
L-alanineamide + H2O | - |
Pseudonocardia thermophila | L-alanine + NH3 | - |
? | |
L-leucineamide + H2O | - |
Pseudonocardia thermophila | L-leucine + NH3 | - |
? | |
L-prolinamide + H2O | - |
Pseudonocardia thermophila | L-proline + NH3 | - |
? | |
L-valineamide + H2O | - |
Pseudonocardia thermophila | L-valine + NH3 | - |
? | |
malonamide + H2O | - |
Pseudonocardia thermophila | malonic acid + NH3 | - |
? | |
methacrylamide + H2O | - |
Pseudonocardia thermophila | methacrylic acid + NH3 | - |
? | |
additional information | the enzyme is enantioselective, the enzyme also shows acyl transferase and hydrolase activities, no activity with N-methylurea, fumaramide, 4-aminobenzosulfonamide, N-phenylurea, acetanilide, benzylcarbamate, and o-tolylamide, broad substrate spectrum, including aliphatic, aromatic and amino acid amides, the presence of a double bond or a methyl group near the carboxamide group of aliphatic and amino acid amides enhances the enzymatic activity, among aromatic amides with substitutions at the 2-, 3-, or 4-position, the 4-substituted amides are the preferred substrates, detailed overview | Pseudonocardia thermophila | ? | - |
? | |
nicotinamide + H2O | - |
Pseudonocardia thermophila | nicotinic acid + NH3 | - |
? | |
propionamide + H2O | - |
Pseudonocardia thermophila | propionic acid + NH3 | - |
? | |
pyrazinamide + H2O | - |
Pseudonocardia thermophila | ? + NH3 | - |
? | |
succinamide + H2O | - |
Pseudonocardia thermophila | succinic acid + NH3 | - |
? | |
trimethylacetamide + H2O | - |
Pseudonocardia thermophila | trimethylacetic acid + NH3 | - |
? | |
urea + H2O | - |
Pseudonocardia thermophila | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 52000, SDS-PAGE | Pseudonocardia thermophila |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | - |
assay at, hydrolytic activity | Pseudonocardia thermophila |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 80 | 10% of maximal activity at 30°C and 80°C | Pseudonocardia thermophila |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
purified enzyme, pH 7.2, the enzyme is heat stable, no loss of activity within the first 9 h, 20% loss of activity after 25 h | Pseudonocardia thermophila |
70 | - |
purified enzyme, half-life 1.2 h | Pseudonocardia thermophila |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.2 | - |
amidase activity | Pseudonocardia thermophila |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 9 | high activity within this range | Pseudonocardia thermophila |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Pseudonocardia thermophila | isoelectric focusing | - |
4.2 |