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Literature summary extracted from

  • Hwang, G.Y.; Kuo, L.Y.; Tsai, M.R.; Yang, S.L.; Lin, L.L.
    Histidines 345 and 378 of Bacillus stearothermophilus leucine aminopeptidase II are essential for the catalytic activity of the enzyme (2005), Antonie van Leeuwenhoek, 87, 355-359.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.11.10 sequence comparisons, overexpression of His-tagged wild-type and mutant LAPII in Escherichia coli Geobacillus stearothermophilus

Protein Variants

EC Number Protein Variants Comment Organism
3.4.11.10 H191L site-directed mutagenesis, the mutant enzyme shows similar catalytic effciency compared to the wild-type enzyme, with 30% increased Km Geobacillus stearothermophilus
3.4.11.10 H227L site-directed mutagenesis, the mutant enzyme shows a 39% decreased catalytic effciency compared to the wild-type enzyme, with a 3.9fold increased Km Geobacillus stearothermophilus
3.4.11.10 H345L site-directed mutagenesis, inactive mutant enzyme Geobacillus stearothermophilus
3.4.11.10 H378L site-directed mutagenesis, inactive mutant enzyme Geobacillus stearothermophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4.11.10 0.9
-
 L-Leu-4-nitroanilide 60°C, recombinant wild-type enzyme Geobacillus stearothermophilus
3.4.11.10 1.2
-
 L-Leu-4-nitroanilide 60°C, recombinant mutant H191L Geobacillus stearothermophilus
3.4.11.10 4.4
-
 L-Leu-4-nitroanilide 60°C, recombinant mutant H227L Geobacillus stearothermophilus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.4.11.10 44500
-
 x * 44500, recombinant His-tagged wild-type and mutant LAPII, SDS-PAGE Geobacillus stearothermophilus

Organism

EC Number Organism UniProt Comment Textmining
3.4.11.10 Geobacillus stearothermophilus P24828
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.11.10 recombinant His-tagged wild-type and mutant LAPII from Escherichia coli by nickel chelate affinity chromatography Geobacillus stearothermophilus

Reaction

EC Number Reaction Comment Organism Reaction ID
3.4.11.10 Release of an N-terminal amino acid, preferentially leucine, but not glutamic or aspartic acids. residues His345 and His378 play an important role in catalysis Geobacillus stearothermophilus
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.4.11.10 60.8
-
 purified recombinant mutant H227L Geobacillus stearothermophilus
3.4.11.10 91.3
-
 purified recombinant mutant H191L Geobacillus stearothermophilus
3.4.11.10 133.4
-
 purified recombinant wild-type enzyme Geobacillus stearothermophilus

Substrates and Products (Substrate)

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

Subunits

EC Number Subunits Comment Organism
3.4.11.10 ? x * 44500, recombinant His-tagged wild-type and mutant LAPII, SDS-PAGE Geobacillus stearothermophilus

Synonyms

EC Number Synonyms Comment Organism
3.4.11.10 aminopeptidase II
-
 Geobacillus stearothermophilus
3.4.11.10 AP-II
-
 Geobacillus stearothermophilus
3.4.11.10 LAPII
-
 Geobacillus stearothermophilus
3.4.11.10 leucine aminopeptidase II
-
 Geobacillus stearothermophilus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.4.11.10 60
-
 assay at Geobacillus stearothermophilus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.4.11.10 0.29
-
 L-Leu-4-nitroanilide 60°C, recombinant wild-type enzyme Geobacillus stearothermophilus
3.4.11.10 0.46
-
 L-Leu-4-nitroanilide 60°C, recombinant mutant H191L Geobacillus stearothermophilus
3.4.11.10 0.86
-
 L-Leu-4-nitroanilide 60°C, recombinant mutant H227L Geobacillus stearothermophilus