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Literature summary extracted from

  • Allala, F.; Bouacem, K.; Boucherba, N.; Azzouz, Z.; Mechri, S.; Sahnoun, M.; Benallaoua, S.; Hacene, H.; Jaouadi, B.; Bouanane-Darenfed, A.
    Purification, biochemical, and molecular characterization of a novel extracellular thermostable and alkaline alpha-amylase from Tepidimonas fonticaldi strain HB23 (2019), Int. J. Biol. Macromol., 132, 558-574 .
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.2.1.1 chloroform activates by about 40% at 25% v/v Tepidimonas fonticaldi
3.2.1.1 Cyclohexane activates by about 15% at 25% v/v Tepidimonas fonticaldi
3.2.1.1 Dimethyl formamide activates by about 55% at 25% v/v Tepidimonas fonticaldi
3.2.1.1 DMSO activates by 10% at 25% v/v Tepidimonas fonticaldi
3.2.1.1 ethanol activates by 15% at 25% v/v Tepidimonas fonticaldi
3.2.1.1 n-hexane activates by about 13% at 25% v/v Tepidimonas fonticaldi

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.1 gene amy48, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of the extracellular fraction of enzyme TfAmy48 in Escherichia coli strain BL21(DE3)pLysS. The biochemical properties of the extracellular purified recombinant enzyme (rTfAmy48) are similar to those of the native one Tepidimonas fonticaldi

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.1 2-mercaptoethanol 59% inhibition at 10 mM Tepidimonas fonticaldi
3.2.1.1 4-bromophenacyl bromide 19% inhibition at 2 mM Tepidimonas fonticaldi
3.2.1.1 Ag+ inhibits 24% at 5 mM Tepidimonas fonticaldi
3.2.1.1 Cd2+ complete inhibition at 5 mM Tepidimonas fonticaldi
3.2.1.1 Co2+ inhibits 60% at 5 mM Tepidimonas fonticaldi
3.2.1.1 Cu2+ inhibits 75% at 5 mM Tepidimonas fonticaldi
3.2.1.1 DTT 74% inhibition at 1 mM Tepidimonas fonticaldi
3.2.1.1 Hg2+ complete inhibition at 5 mM Tepidimonas fonticaldi
3.2.1.1 Mn2+ inhibits 11% at 5 mM Tepidimonas fonticaldi
3.2.1.1 additional information enzyme TfAmy48 shows excellent compatibility with some commercial laundry detergents, overview. No or poor effects by 1 mM PMSF, 2 mM diiodopropyl fluorophosphate, 10 mM EDTA, or 1 mm EGTA Tepidimonas fonticaldi
3.2.1.1 SDS 39% inhibition at 60 mM Tepidimonas fonticaldi
3.2.1.1 Urea 51% inhibition at 6 M Tepidimonas fonticaldi
3.2.1.1 Zn2+ inhibits 79% at 5 mM Tepidimonas fonticaldi

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.1 additional information
-
 additional information classical Michaelis-Menten kinetics, Km for soluble potato starch is 1.22 mg/ml Tepidimonas fonticaldi

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.2.1.1 extracellular the enzyme harbors a predicted signal peptide Tepidimonas fonticaldi
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.1 Ba2+ activates 38% at 5 mM Tepidimonas fonticaldi
3.2.1.1 Ca2+ activates 55% at 5 mM Tepidimonas fonticaldi
3.2.1.1 Fe2+ activates 22% at 5 mM Tepidimonas fonticaldi
3.2.1.1 Mg2+ activates 13% at 5 mM Tepidimonas fonticaldi
3.2.1.1 Ni2+ activates 19% at 5 mM Tepidimonas fonticaldi

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.1 48000
-
 native PAGE and gel filtration Tepidimonas fonticaldi
3.2.1.1 48138
-
 mass spectrometry Tepidimonas fonticaldi

Organic Solvent Stability

EC Number Organic Solvent Comment Organism
3.2.1.1 acetonitrile 75% inhibition at 25% v/v Tepidimonas fonticaldi
3.2.1.1 benzene 55% inhibition at 25% v/v Tepidimonas fonticaldi
3.2.1.1 Ethyl acetate 55% inhibition at 25% v/v Tepidimonas fonticaldi
3.2.1.1 isooctane 15% inhibition at 25% v/v Tepidimonas fonticaldi
3.2.1.1 additional information enzyme TfAmy48 shows extreme stability and tolerance towards organic solvents. After 24 h, TfAmy48 shows a stimulatory effect in the majority of hydrophilic organic solvents, including DMSO, ethanol, chloroform, and DMF with a residual activity of 110, 115, 140, and 155%, respectively. No of poor effects on enzyme activity by 25% v/v of isopropyl alcohol, isopropanol, n-decane, toluene, n-hexanol, n-butanol, and methanol Tepidimonas fonticaldi
3.2.1.1 n-heptane 25% inhibition at 25% v/v Tepidimonas fonticaldi
3.2.1.1 n-octane 10% inhibition at 25% v/v Tepidimonas fonticaldi

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.1 Tepidimonas fonticaldi A0A482EWM9 isolated from a collection obtained froma thermal spring in Hammam Righa commune (GPS coordinates: 36°22' N, 2°23' E), located in Ain Defla province (Algeria). The isolation site is located at an altitude of 525 m, in a mountainous and wooded area. It includes several sources whose water temperature varies from 26°C to 68°C
-
3.2.1.1 Tepidimonas fonticaldi HB23 A0A482EWM9 isolated from a collection obtained froma thermal spring in Hammam Righa commune (GPS coordinates: 36°22' N, 2°23' E), located in Ain Defla province (Algeria). The isolation site is located at an altitude of 525 m, in a mountainous and wooded area. It includes several sources whose water temperature varies from 26°C to 68°C
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.1 native extracellular enzyme 9.5fold by ammonium sulfate fractionation, heat treatment, gel filtration, and anion exchange chromatography Tepidimonas fonticaldi

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.2.1.1 additional information culture growth in optimized starch broth Tepidimonas fonticaldi
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.1 1275
-
 purified native extracellular enzyme, substrate soluble starch, pH 8.0, 80°C Tepidimonas fonticaldi

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.1 amylopectin + H2O 75% activity compared to soluble potato starch Tepidimonas fonticaldi ?
-
 ?
3.2.1.1 amylopectin + H2O 75% activity compared to soluble potato starch Tepidimonas fonticaldi HB23 ?
-
 ?
3.2.1.1 amylose + H2O 85% activity compared to soluble potato starch Tepidimonas fonticaldi ?
-
 ?
3.2.1.1 amylose + H2O 85% activity compared to soluble potato starch Tepidimonas fonticaldi HB23 ?
-
 ?
3.2.1.1 additional information enzyme TfAmy48 shows an endo-acting pattern. It has no activity with raw starch, alpha-cyclodextrin, and carboxymethyl cellulose Tepidimonas fonticaldi ?
-
 ?
3.2.1.1 additional information enzyme TfAmy48 shows an endo-acting pattern. It has no activity with raw starch, alpha-cyclodextrin, and carboxymethyl cellulose Tepidimonas fonticaldi HB23 ?
-
 ?
3.2.1.1 soluble maize starch + H2O 60% activity compared to soluble potato starch Tepidimonas fonticaldi ?
-
 ?
3.2.1.1 soluble maize starch + H2O 60% activity compared to soluble potato starch Tepidimonas fonticaldi HB23 ?
-
 ?
3.2.1.1 soluble potato starch + H2O best substrate Tepidimonas fonticaldi maltopentaose + maltose + maltotriose
-
 ?
3.2.1.1 soluble potato starch + H2O best substrate Tepidimonas fonticaldi HB23 maltopentaose + maltose + maltotriose
-
 ?
3.2.1.1 soluble rice starch + H2O 55% activity compared to soluble potato starch Tepidimonas fonticaldi ?
-
 ?
3.2.1.1 soluble wheat starch + H2O 79% activity compared to soluble potato starch Tepidimonas fonticaldi ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.1 monomer 1 * 48138, mass spectrometry, 1 * 48000, SDS-PAGE, 1 * 50414, sequence calculation Tepidimonas fonticaldi
3.2.1.1 More enzyme three-dimensional structure homology modelling, overview Tepidimonas fonticaldi

Synonyms

EC Number Synonyms Comment Organism
3.2.1.1 TfAmy48
-
 Tepidimonas fonticaldi

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.1 70 75 without Ca2+ Tepidimonas fonticaldi
3.2.1.1 80
-
 in presence of 5 mM Ca2+ Tepidimonas fonticaldi

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.1 70
-
 purified enzyme, 5 mM Ca2+, completely stable for 14 h, loss of 50% activity after 19 h, inactivation after 23 h Tepidimonas fonticaldi
3.2.1.1 70
-
 purified enzyme, without Ca2+, completely stable for 10 h, loss of 50% activity after 16 h, inactivation after 19 h Tepidimonas fonticaldi
3.2.1.1 80
-
 purified enzyme, 5 mM Ca2+, completely stable for 8 h, loss of 50% activity after 13 h, inactivation after 16 h Tepidimonas fonticaldi
3.2.1.1 80
-
 purified enzyme, without Ca2+, completely stable for 6 h, loss of 50% activity after 10 h, inactivation after 13 h Tepidimonas fonticaldi
3.2.1.1 90
-
 purified enzyme, 5 mM Ca2+, completely stable for 5 h, loss of 50% activity after 8 h, inactivation after 10 h Tepidimonas fonticaldi
3.2.1.1 90
-
 purified enzyme, without Ca2+, completely stable for 3 h, loss of 50% activity after 6 h, inactivation after 8 h Tepidimonas fonticaldi
3.2.1.1 100
-
 purified enzyme, 5 mM Ca2+, completely stable for 1 h, loss of 50% activity after 3 h, inactivation after 4 h Tepidimonas fonticaldi
3.2.1.1 100
-
 purified enzyme, without Ca2+, loss of 50% activity after 2 h, inactivation after 3h Tepidimonas fonticaldi

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.1 5.9
-
 soluble potato starch pH 8.0, 80°C Tepidimonas fonticaldi

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.1 8
-
-
 Tepidimonas fonticaldi

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.1 4 12 over 50% of maximal activity within this range, profile overview Tepidimonas fonticaldi

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.2.1.1 6
-
 purified enzyme, completely stable for 16 h, loss of 50% activity after 21 h, inactivation after 24 h Tepidimonas fonticaldi
3.2.1.1 7
-
 purified enzyme, completely stable for 12.5 h, loss of 50% activity after 18 h, inactivation after 21 h Tepidimonas fonticaldi
3.2.1.1 8
-
 purified enzyme, completely stable for 9 h, loss of 50% activity after 15 h, inactivation after 18 h Tepidimonas fonticaldi
3.2.1.1 9
-
 purified enzyme, completely stable for 8 h, loss of 50% activity after 11 h, inactivation after 13 h Tepidimonas fonticaldi
3.2.1.1 10
-
 purified enzyme, completely stable for 6 h, loss of 50% activity after 9 h, inactivation after 11 h Tepidimonas fonticaldi
3.2.1.1 11
-
 purified enzyme, completely stable for 3 h, loss of 50% activity after 6 h, inactivation after 8 h Tepidimonas fonticaldi

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.2.1.1 Tepidimonas fonticaldi sequence calculation
-
 6.04

General Information

EC Number General Information Comment Organism
3.2.1.1 additional information enzyme three-dimensional structure homology modelling, overview Tepidimonas fonticaldi