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Literature summary for 3.4.11.10 extracted from

  • Lin, L.L.; Chen, Y.P.; Yang, J.C.; Hua, Y.W.; Wang, W.C.; Kuo, L.Y.
    Significance of the conserved Tyr352 and Asp380 residues in the catalytic activity of Bacillus stearothermophilus aminopeptidase II as evaluated by site-directed mutagenesis (2008), Protein J., 27, 215-222.
    View publication on PubMed
Search for more literature for 3.4.11.10

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli M15 cells Geobacillus stearothermophilus

Protein Variants

Protein Variants Comment Organism
D380E the mutation causes more than 2fold decrease in kcat as compared to the wild type enzyme Geobacillus stearothermophilus
D380N the mutation causes more than 2fold decrease in kcat as compared to the wild type enzyme Geobacillus stearothermophilus
D380V the mutation causes over 5fold decrease in kcat as compared to the wild type enzyme Geobacillus stearothermophilus
Y352D the mutation results in about 3.5fold decrease in specific activity Geobacillus stearothermophilus
Y352L the mutation results in about 3.5fold decrease in specific activity Geobacillus stearothermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.95
-
 L-Leu-4-nitroanilide wild type enzyme, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C Geobacillus stearothermophilus
2.12
-
 L-Leu-4-nitroanilide mutant enzyme Y352D, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C Geobacillus stearothermophilus
2.83
-
 L-Leu-4-nitroanilide mutant enzyme Y352L, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C Geobacillus stearothermophilus
3.05
-
 L-Leu-4-nitroanilide mutant enzyme D380N, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C Geobacillus stearothermophilus
3.39
-
 L-Leu-4-nitroanilide mutant enzyme D380E, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C Geobacillus stearothermophilus
3.41
-
 L-Leu-4-nitroanilide mutant enzyme D380V, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C Geobacillus stearothermophilus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
45000
-
 SDS-PAGE Geobacillus stearothermophilus

Organism

Organism UniProt Comment Textmining
Geobacillus stearothermophilus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni2+-NTA resin chromatography Geobacillus stearothermophilus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
13.8
-
 mutant enzyme D380E, at 60°C, using L-Leu-4-nitroanilide as substrate Geobacillus stearothermophilus
15.1
-
 mutant enzyme D380V, at 60°C, using L-Leu-4-nitroanilide as substrate Geobacillus stearothermophilus
34.5
-
 mutant enzyme Y532L, at 60°C, using L-Leu-4-nitroanilide as substrate Geobacillus stearothermophilus
55.3
-
 mutant enzyme D380N, at 60°C, using L-Leu-4-nitroanilide as substrate Geobacillus stearothermophilus
104.9
-
 mutant enzyme Y532D, at 60°C, using L-Leu-4-nitroanilide as substrate Geobacillus stearothermophilus
572.5
-
 wild type enzyme, at 60°C, using L-Leu-4-nitroanilide as substrate Geobacillus stearothermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-Leu-4-nitroanilide + H2O
-
 Geobacillus stearothermophilus L-Leu + 4-nitroaniline
-
 ?

Synonyms

Synonyms Comment Organism
aminopeptidase II
-
 Geobacillus stearothermophilus
AP-II
-
 Geobacillus stearothermophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.035
-
 L-Leu-4-nitroanilide mutant enzyme D380E, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C Geobacillus stearothermophilus
0.048
-
 L-Leu-4-nitroanilide mutant enzyme D380V, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C Geobacillus stearothermophilus
0.113
-
 L-Leu-4-nitroanilide mutant enzyme Y352L, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C Geobacillus stearothermophilus
0.135
-
 L-Leu-4-nitroanilide mutant enzyme D380N, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C Geobacillus stearothermophilus
0.153
-
 L-Leu-4-nitroanilide mutant enzyme Y352D, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C Geobacillus stearothermophilus
0.303
-
 L-Leu-4-nitroanilide wild type enzyme, in 25 mM Tris-HCl buffer, pH 8.0, at 60°C Geobacillus stearothermophilus