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Literature summary extracted from

  • Zaide, G.; Shallom, D.; Shulami, S.; Zolotnitsky, G.; Golan, G.; Baasov, T.; Shoham, G.; Shoham, Y.
    Biochemical characterization and identification of catalytic residues in alpha-glucuronidase from Bacillus stearothermophilus T-6 (2001), Eur. J. Biochem., 268, 3006-3016.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.131 overexpression in Escherichia coli Geobacillus stearothermophilus
3.2.1.139 overexpression in Escherichia coli using the T7 polymerase expression system Geobacillus stearothermophilus

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.131 AguADELTAC lacks the last 81 amino acids, heat labile, 0.1% of the specific activity of the wild-type enzyme at 35°C Geobacillus stearothermophilus
3.2.1.131 AguADELTAN lacks the first 126 amino acids, 1% of the specific activity of the wild-type enzyme at 55°C Geobacillus stearothermophilus
3.2.1.131 AguADELTANC lacks the first 126 and last 81 amino acids, is insoluble Geobacillus stearothermophilus
3.2.1.131 D274A no change in activity Geobacillus stearothermophilus
3.2.1.131 D364A great decrease in activity Geobacillus stearothermophilus
3.2.1.131 E158N decrease in activity Geobacillus stearothermophilus
3.2.1.131 E285N decrease in activity Geobacillus stearothermophilus
3.2.1.131 E386Q decrease in activity Geobacillus stearothermophilus
3.2.1.131 E392C great decrease in activity Geobacillus stearothermophilus
3.2.1.131 E510A decrease in activity Geobacillus stearothermophilus
3.2.1.139 D274A as active as wild-type enzyme Geobacillus stearothermophilus
3.2.1.139 D364A 0.000015% of wild-type activity Geobacillus stearothermophilus
3.2.1.139 D364A/E392C 0.0000004% of wild-type activity Geobacillus stearothermophilus
3.2.1.139 E158N 0.07% of wild-type activity Geobacillus stearothermophilus
3.2.1.139 E285N 0.0002% of wild-type activity Geobacillus stearothermophilus
3.2.1.139 E386Q 0.07% of wild-type activity Geobacillus stearothermophilus
3.2.1.139 E392C 0.00002% of wild-type activity Geobacillus stearothermophilus
3.2.1.139 E510A 20% of wild-type activity Geobacillus stearothermophilus
3.2.1.139 additional information truncated forms of the enzyme, lacking either 126 amino acids from its N-terminus or 81 amino acids from its C-terminus, exhibit low residual activity Geobacillus stearothermophilus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.131 Ag+
-
 Geobacillus stearothermophilus
3.2.1.131 Cu2+
-
 Geobacillus stearothermophilus
3.2.1.131 Hg2+
-
 Geobacillus stearothermophilus
3.2.1.131 K+
-
 Geobacillus stearothermophilus
3.2.1.131 Ni2+
-
 Geobacillus stearothermophilus
3.2.1.131 Zn2+
-
 Geobacillus stearothermophilus
3.2.1.139 Ag+ 1 mM, 90% inhibition Geobacillus stearothermophilus
3.2.1.139 Cu2+ 0.1 mM, complete inhibition Geobacillus stearothermophilus
3.2.1.139 Hg2+ 25% inhibition at 0.1 mM, complete inhibition at 1 mM Geobacillus stearothermophilus
3.2.1.139 K+ 1 mM, 15% inhibition Geobacillus stearothermophilus
3.2.1.139 Ni2+ 1 mM, 10% inhibition Geobacillus stearothermophilus
3.2.1.139 Zn2+ 1 mM, 55% inhibition Geobacillus stearothermophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.131 0.2
-
 aldotetraouronic acid 55°C, pH 5,5 Geobacillus stearothermophilus
3.2.1.139 0.2
-
 aldotetraouronic acid 55°C Geobacillus stearothermophilus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.139 Fe3+ 1 mM, 25% activation Geobacillus stearothermophilus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.131 78480
-
 alpha2, 2 * 78480, deduced from nucleotide sequence Geobacillus stearothermophilus
3.2.1.131 150000
-
 gel filtration Geobacillus stearothermophilus
3.2.1.139 78480
-
 x * 78480, calculated from sequence Geobacillus stearothermophilus

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.131 Geobacillus stearothermophilus
-
 T-6
-
3.2.1.131 Geobacillus stearothermophilus T-6
-
 T-6
-
3.2.1.139 Geobacillus stearothermophilus
-
-
-
3.2.1.139 Geobacillus stearothermophilus T-6
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.131
-
 Geobacillus stearothermophilus
3.2.1.139 recombinant Geobacillus stearothermophilus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.131 42
-
 after gel filtration Geobacillus stearothermophilus
3.2.1.139 42
-
-
 Geobacillus stearothermophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.131 aldotetraouronic acid + H2O
-
 Geobacillus stearothermophilus 4-O-methyl-D-glucuronic acid + Xylbeta(1-4)Xylbeta(1-4)Xylbeta(1-4)Xyl
-
 ?
3.2.1.131 aldotetraouronic acid + H2O
-
 Geobacillus stearothermophilus T-6 4-O-methyl-D-glucuronic acid + Xylbeta(1-4)Xylbeta(1-4)Xylbeta(1-4)Xyl
-
 ?
3.2.1.139 aldotetraouronic acid + H2O
-
 Geobacillus stearothermophilus ?
-
 ?
3.2.1.139 aldotetraouronic acid + H2O
-
 Geobacillus stearothermophilus T-6 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.131 dimer alpha2, 2 * 78480, deduced from nucleotide sequence Geobacillus stearothermophilus
3.2.1.139 ? x * 78480, calculated from sequence Geobacillus stearothermophilus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.131 65
-
-
 Geobacillus stearothermophilus
3.2.1.139 65
-
-
 Geobacillus stearothermophilus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.139 45 70 45°C: about 35% of maximal activity, 70°C: about 50% of maximal activity Geobacillus stearothermophilus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.131 70
-
 fully active after 20 min Geobacillus stearothermophilus
3.2.1.131 73.4
-
 denaturation temperature Geobacillus stearothermophilus
3.2.1.139 70
-
 20 min, no less of activity Geobacillus stearothermophilus
3.2.1.139 73.4
-
 denaturation at Geobacillus stearothermophilus
3.2.1.139 75
-
 20 min, 70% loss of activity Geobacillus stearothermophilus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.131 54.9
-
 aldotetraouronic acid 55°C, pH 5,5 Geobacillus stearothermophilus
3.2.1.139 54.9
-
 aldotetraouronic acid 55°C Geobacillus stearothermophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.131 5.5 6
-
 Geobacillus stearothermophilus
3.2.1.139 5.5 6
-
 Geobacillus stearothermophilus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.131 4.5 7.5 at pH 4.5 and 7.5 retains 40% of enzyme activity Geobacillus stearothermophilus
3.2.1.139 4.5 7.5 about 40% of maximal activity at pH 4.5 and pH 7.5 Geobacillus stearothermophilus