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Literature summary extracted from

  • Mehta, P.; Bhatia, S.; Bhatia, R.; Bhalla, T.
    Purification and characterization of a novel thermo-active amidase from Geobacillus subterraneus RL-2a (2013), Extremophiles, 17, 637-648.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.5.1.4 additional information presence of mid-chain aliphatic and amino acid amides enhances the enzymatic activity Geobacillus subterraneus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.1.4 constitutive enzyme Geobacillus subterraneus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.1.4 Ag+ complete inhibition at 1 mM Geobacillus subterraneus
3.5.1.4 Ba2+ 11% inhibition at 1 mM Geobacillus subterraneus
3.5.1.4 Ca2+ 10% inhibition at 1 mM Geobacillus subterraneus
3.5.1.4 Co2+ 9% inhibition at 1 mM Geobacillus subterraneus
3.5.1.4 Cu2+ 28.5% inhibition at 1 mM Geobacillus subterraneus
3.5.1.4 DTT 21% inhibition at 1 mM Geobacillus subterraneus
3.5.1.4 EDTA 21% inhibition at 1 mM Geobacillus subterraneus
3.5.1.4 Fe3+ 87% inhibition at 1 mM Geobacillus subterraneus
3.5.1.4 Hg2+ complete inhibition at 1 mM Geobacillus subterraneus
3.5.1.4 Mg2+ 53% inhibition at 1 mM Geobacillus subterraneus
3.5.1.4 Mn2+ 12% inhibition at 1 mM Geobacillus subterraneus
3.5.1.4 Na+ 17% inhibition at 1 mM Geobacillus subterraneus
3.5.1.4 Pb2+ 8% inhibition at 1 mM Geobacillus subterraneus
3.5.1.4 PEG 8% inhibition at 1 mM Geobacillus subterraneus
3.5.1.4 PMSF 87.5% inhibition at 1 mM Geobacillus subterraneus
3.5.1.4 Zn2+ 11% inhibition at 1 mM Geobacillus subterraneus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.4 additional information
-
 additional information Michaelis–Menten kinetics Geobacillus subterraneus
3.5.1.4 6.02
-
 nicotinamide pH 6.5, 70°C Geobacillus subterraneus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.1.4 37000
-
 3 * 37000, SDS-PAGE Geobacillus subterraneus
3.5.1.4 111000
-
 gel filtration Geobacillus subterraneus

Organic Solvent Stability

EC Number Organic Solvent Comment Organism
3.5.1.4 Acetone 5-10% activity remaining at 5-10% acetone after 15 min Geobacillus subterraneus
3.5.1.4 benzene 70-80% activity remaining at 5-10% benzene after 15 min Geobacillus subterraneus
3.5.1.4 Butanol 15-25% activity remaining at 5-10% butanol after 15 min Geobacillus subterraneus
3.5.1.4 CCl4 80-90% activity remaining at 5-10% CCl4 after 15 min Geobacillus subterraneus
3.5.1.4 dimethyl sulfoxide 35-40% activity remaining at 5-10% DMSO after 15 min Geobacillus subterraneus
3.5.1.4 Ethanol 15% activity remaining at 5-10% ethanol after 15 min Geobacillus subterraneus
3.5.1.4 Ethylene glycol 60-70% activity remaining at 5-10% ethylene glycol after 15 min Geobacillus subterraneus
3.5.1.4 propanol 20-25% activity remaining at 5-10% propanol after 15 min Geobacillus subterraneus
3.5.1.4 toluene 90% activity remaining at 10% toluene after 15 min Geobacillus subterraneus

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.4 Geobacillus subterraneus
-
 isolated from Manikaran thermal spring (Himachal Pradesh, India)
-
3.5.1.4 Geobacillus subterraneus RL-2a
-
 isolated from Manikaran thermal spring (Himachal Pradesh, India)
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.1.4 native enzyme 52.04fold to homogeneity by heat treatment, ammonium sulfate fractionation, anion exchange chromatography, and gel filtration Geobacillus subterraneus

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.5.1.4 additional information constitutive enzyme Geobacillus subterraneus
-

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 48.66
-
 purified native enzyme, pH 6.5, 70°C, substrate nicotinamide Geobacillus subterraneus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.4 benzamide + H2O
-
 Geobacillus subterraneus benzoate + NH3
-
 ?
3.5.1.4 butyramide + H2O high activity Geobacillus subterraneus butyrate + NH3
-
 ?
3.5.1.4 cyanoacetamide + H2O low activity Geobacillus subterraneus cyanoacetate + NH3
-
 ?
3.5.1.4 formamide + H2O
-
 Geobacillus subterraneus formate + NH3
-
 ?
3.5.1.4 formamide + H2O
-
 Geobacillus subterraneus RL-2a formate + NH3
-
 ?
3.5.1.4 isobutyramide + H2O high activity Geobacillus subterraneus isobutyrate + NH3
-
 ?
3.5.1.4 isobutyramide + H2O high activity Geobacillus subterraneus RL-2a isobutyrate + NH3
-
 ?
3.5.1.4 isonicotinamide + H2O
-
 Geobacillus subterraneus isonicotinate + NH3
-
 ?
3.5.1.4 L-leucinamide + H2O
-
 Geobacillus subterraneus L-leucine + NH3
-
 ?
3.5.1.4 L-leucinamide + H2O
-
 Geobacillus subterraneus RL-2a L-leucine + NH3
-
 ?
3.5.1.4 lactamide + H2O
-
 Geobacillus subterraneus lactate + NH3
-
 ?
3.5.1.4 methacrylamide + H2O
-
 Geobacillus subterraneus methacrylate + NH3
-
 ?
3.5.1.4 additional information glycinamide, malonamide, acetamide, and acrylamide are poor substrates, while urea, mandelamide, N-methylacetamide, and thioacetamide are no substrates, substrate specificity, overview Geobacillus subterraneus ?
-
 ?
3.5.1.4 additional information glycinamide, malonamide, acetamide, and acrylamide are poor substrates, while urea, mandelamide, N-methylacetamide, and thioacetamide are no substrates, substrate specificity, overview Geobacillus subterraneus RL-2a ?
-
 ?
3.5.1.4 nicotinamide + H2O reaction of EC 3.5.1.19, nicotinamidase Geobacillus subterraneus nicotinate + NH3
-
 ?
3.5.1.4 prolinamide + H2O
-
 Geobacillus subterraneus proline + NH3
-
 ?
3.5.1.4 propainamide + H2O high activity Geobacillus subterraneus ? + NH3
-
 ?
3.5.1.4 propionamide + H2O best substrate Geobacillus subterraneus propionate + NH3
-
 ?
3.5.1.4 sulfanilamide + H2O
-
 Geobacillus subterraneus sulfanilate + NH3
-
 ?
3.5.1.4 valeramide + H2O high activity Geobacillus subterraneus valerate + NH3
-
 ?
3.5.1.4 valeramide + H2O high activity Geobacillus subterraneus RL-2a valerate + NH3
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.1.4 trimer 3 * 37000, SDS-PAGE Geobacillus subterraneus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.1.4 70
-
-
 Geobacillus subterraneus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.5.1.4 40 90 activity range Geobacillus subterraneus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.5.1.4 70
-
 purified native enzyme, half-life is 354 min Geobacillus subterraneus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.1.4 6.5
-
-
 Geobacillus subterraneus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.5.1.4 4.5 11.5 activity range Geobacillus subterraneus

General Information

EC Number General Information Comment Organism
3.5.1.4 evolution peptide sequences determination by MALDI-TOF analysis reveals 97% similarity with that of glutamyl-tRNA(Gln) amidotransferase subunit A (Geobacillus kaustophilus HTA426) and amidase of Geobacillus sp. C56-T3, also 68% similarity to amidase of Geobacillus sp. Y412MC61 Geobacillus subterraneus
3.5.1.4 additional information the enzyme is a wide-spectrum thermostable amidase Geobacillus subterraneus