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Literature summary for 3.5.1.4 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

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

Cloned(Commentary)

Cloned (Comment) Organism
constitutive enzyme Geobacillus subterraneus

Inhibitors

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

KM Value [mM]

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

Molecular Weight [Da]

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

Organic Solvent Stability

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

Organism

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

Purification (Commentary)

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

Source Tissue

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

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
48.66
-
purified native enzyme, pH 6.5, 70°C, substrate nicotinamide Geobacillus subterraneus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
benzamide + H2O
-
Geobacillus subterraneus benzoate + NH3
-
?
butyramide + H2O high activity Geobacillus subterraneus butyrate + NH3
-
?
cyanoacetamide + H2O low activity Geobacillus subterraneus cyanoacetate + NH3
-
?
formamide + H2O
-
Geobacillus subterraneus formate + NH3
-
?
formamide + H2O
-
Geobacillus subterraneus RL-2a formate + NH3
-
?
isobutyramide + H2O high activity Geobacillus subterraneus isobutyrate + NH3
-
?
isobutyramide + H2O high activity Geobacillus subterraneus RL-2a isobutyrate + NH3
-
?
isonicotinamide + H2O
-
Geobacillus subterraneus isonicotinate + NH3
-
?
L-leucinamide + H2O
-
Geobacillus subterraneus L-leucine + NH3
-
?
L-leucinamide + H2O
-
Geobacillus subterraneus RL-2a L-leucine + NH3
-
?
lactamide + H2O
-
Geobacillus subterraneus lactate + NH3
-
?
methacrylamide + H2O
-
Geobacillus subterraneus methacrylate + NH3
-
?
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 ?
-
?
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 ?
-
?
nicotinamide + H2O reaction of EC 3.5.1.19, nicotinamidase Geobacillus subterraneus nicotinate + NH3
-
?
prolinamide + H2O
-
Geobacillus subterraneus proline + NH3
-
?
propainamide + H2O high activity Geobacillus subterraneus ? + NH3
-
?
propionamide + H2O best substrate Geobacillus subterraneus propionate + NH3
-
?
sulfanilamide + H2O
-
Geobacillus subterraneus sulfanilate + NH3
-
?
valeramide + H2O high activity Geobacillus subterraneus valerate + NH3
-
?
valeramide + H2O high activity Geobacillus subterraneus RL-2a valerate + NH3
-
?

Subunits

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

Temperature Optimum [°C]

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

Temperature Range [°C]

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

Temperature Stability [°C]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
-
Geobacillus subterraneus

pH Range

pH Minimum pH Maximum Comment Organism
4.5 11.5 activity range Geobacillus subterraneus

General Information

General Information Comment Organism
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
additional information the enzyme is a wide-spectrum thermostable amidase Geobacillus subterraneus