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Literature summary extracted from

  • Egorova, K.; Trauthwein, H.; Verseck, S.; Antranikian, G.
    Purification and properties of an enantioselective and thermoactive amidase from the thermophilic actinomycete Pseudonocardia thermophila (2004), Appl. Microbiol. Biotechnol., 65, 38-45.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.5.1.4 DTT 20% activation at 1 mM Pseudonocardia thermophila

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.1.4 Ba2+ 13% inhibition at 1 mM Pseudonocardia thermophila
3.5.1.4 Ca2+ 15% inhibition at 1 mM Pseudonocardia thermophila
3.5.1.4 Co2+ complete inhibition at 1 mM Pseudonocardia thermophila
3.5.1.4 Cu2+ 74% inhibition at 1 mM Pseudonocardia thermophila
3.5.1.4 Fe2+ 20% inhibition at 1 mM Pseudonocardia thermophila
3.5.1.4 Hg2+ complete inhibition at 1 mM Pseudonocardia thermophila
3.5.1.4 iodoacetamide 10% inhibition at 1 mM Pseudonocardia thermophila
3.5.1.4 iodoacetate 78% inhibition at 1 mM Pseudonocardia thermophila
3.5.1.4 Mg2+ 10% inhibition at 1 mM Pseudonocardia thermophila
3.5.1.4 additional information no inhibition by Triton X-100, SDS, EDTA, Mn2+, and Fe3+ at 1 mM Pseudonocardia thermophila
3.5.1.4 Ni2+ 56% inhibition at 1 mM Pseudonocardia thermophila
3.5.1.4 Zn2+ 7% inhibition at 1 mM Pseudonocardia thermophila

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.4 0.9
-
 2-phenylpropionamide pH 7.2, 70°C Pseudonocardia thermophila
3.5.1.4 4.9
-
 Benzamide pH 7.2, 70°C Pseudonocardia thermophila
3.5.1.4 7.4
-
 Propionamide pH 7.2, 70°C Pseudonocardia thermophila
3.5.1.4 9.2
-
 methacrylamide pH 7.2, 70°C Pseudonocardia thermophila

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.1.4 52000
-
 2 * 52000, SDS-PAGE Pseudonocardia thermophila
3.5.1.4 108000
-
 gel filtration Pseudonocardia thermophila

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.4 Pseudonocardia thermophila
-
 thermophilic actinomycete, DSMZ 43832
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.1.4 native enzyme 48fold to homogeneity by hydrophobic interaction chromatography, ultrafiltration, anion exchange chromatography, and gel filtration Pseudonocardia thermophila

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 19.4
-
 purified enzyme, substrate benzamide Pseudonocardia thermophila
3.5.1.4 23.5
-
 purified enzyme, substrate methacrylamide Pseudonocardia thermophila
3.5.1.4 25.4
-
 purified enzyme, substrate propionamide Pseudonocardia thermophila
3.5.1.4 26.8
-
 purified enzyme, substrate acrylamide Pseudonocardia thermophila

Storage Stability

EC Number Storage Stability Organism
3.5.1.4 4°C, purified enzyme, in 50 mM phosphate buffer, more than 6 months without loss of activity Pseudonocardia thermophila

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.4 (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
-
 ?
3.5.1.4 2-aminobenzamide + H2O
-
 Pseudonocardia thermophila 2-aminobenzoic acid + NH3
-
 ?
3.5.1.4 2-hydroxybenzamide + H2O
-
 Pseudonocardia thermophila 2-hydroxybenzoic acid + NH3
-
 ?
3.5.1.4 2-tolylamide + H2O
-
 Pseudonocardia thermophila ? + NH3
-
 ?
3.5.1.4 3-indolylacetamide + H2O
-
 Pseudonocardia thermophila 3-indolylacetic acid + NH3
-
 ?
3.5.1.4 3-tolylamide + H2O
-
 Pseudonocardia thermophila ? + NH3
-
 ?
3.5.1.4 4-aminobenzamide + H2O
-
 Pseudonocardia thermophila 4-aminobenzoic acid + NH3
-
 ?
3.5.1.4 4-hydroxybenzamide + H2O
-
 Pseudonocardia thermophila 4-hydroxybenzoic acid + NH3
-
 ?
3.5.1.4 4-tolylamide + H2O
-
 Pseudonocardia thermophila ? + NH3
-
 ?
3.5.1.4 acetamide + H2O
-
 Pseudonocardia thermophila acetic acid + NH3
-
 ?
3.5.1.4 acrylamide + H2O
-
 Pseudonocardia thermophila acrylic acid + NH3
-
 ?
3.5.1.4 adipoamide + H2O
-
 Pseudonocardia thermophila adipic acid + NH3
-
 ?
3.5.1.4 benzamide + H2O
-
 Pseudonocardia thermophila benzoic acid + NH3
-
 ?
3.5.1.4 cyclohexanoamide + H2O
-
 Pseudonocardia thermophila cyclohexanoic acid + NH3
-
 ?
3.5.1.4 DL-lactamide + H2O
-
 Pseudonocardia thermophila DL-lactate + NH3
-
 ?
3.5.1.4 DL-methioninamide + H2O
-
 Pseudonocardia thermophila DL-methionine + NH3
-
 ?
3.5.1.4 DL-tryptophanamide + H2O
-
 Pseudonocardia thermophila DL-tryptophan + NH3
-
 ?
3.5.1.4 formamide + H2O
-
 Pseudonocardia thermophila formate + NH3
-
 ?
3.5.1.4 hexanoamide + H2O
-
 Pseudonocardia thermophila hexanoic acid + NH3
-
 ?
3.5.1.4 hydroxylamine hydrochloride + H2O
-
 Pseudonocardia thermophila ?
-
 ?
3.5.1.4 isobutyramide + H2O
-
 Pseudonocardia thermophila isobutyric acid + NH3
-
 ?
3.5.1.4 isonicotinamide + H2O
-
 Pseudonocardia thermophila isonicotinic acid + NH3
-
 ?
3.5.1.4 L-alanineamide + H2O
-
 Pseudonocardia thermophila L-alanine + NH3
-
 ?
3.5.1.4 L-leucineamide + H2O
-
 Pseudonocardia thermophila L-leucine + NH3
-
 ?
3.5.1.4 L-prolinamide + H2O
-
 Pseudonocardia thermophila L-proline + NH3
-
 ?
3.5.1.4 L-valineamide + H2O
-
 Pseudonocardia thermophila L-valine + NH3
-
 ?
3.5.1.4 malonamide + H2O
-
 Pseudonocardia thermophila malonic acid + NH3
-
 ?
3.5.1.4 methacrylamide + H2O
-
 Pseudonocardia thermophila methacrylic acid + NH3
-
 ?
3.5.1.4 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 ?
-
 ?
3.5.1.4 nicotinamide + H2O
-
 Pseudonocardia thermophila nicotinic acid + NH3
-
 ?
3.5.1.4 propionamide + H2O
-
 Pseudonocardia thermophila propionic acid + NH3
-
 ?
3.5.1.4 pyrazinamide + H2O
-
 Pseudonocardia thermophila ? + NH3
-
 ?
3.5.1.4 succinamide + H2O
-
 Pseudonocardia thermophila succinic acid + NH3
-
 ?
3.5.1.4 trimethylacetamide + H2O
-
 Pseudonocardia thermophila trimethylacetic acid + NH3
-
 ?
3.5.1.4 urea + H2O
-
 Pseudonocardia thermophila ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.1.4 dimer 2 * 52000, SDS-PAGE Pseudonocardia thermophila

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.1.4 70
-
 assay at, hydrolytic activity Pseudonocardia thermophila

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.5.1.4 30 80 10% of maximal activity at 30°C and 80°C Pseudonocardia thermophila

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.5.1.4 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
3.5.1.4 70
-
 purified enzyme, half-life 1.2 h Pseudonocardia thermophila

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.1.4 7.2
-
 amidase activity Pseudonocardia thermophila

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.5.1.4 5 9 high activity within this range Pseudonocardia thermophila

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.5.1.4 Pseudonocardia thermophila isoelectric focusing
-
 4.2