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Literature summary for 3.4.11.24 extracted from

  • Arima, J.; Uesugi, Y.; Uraji, M.; Yatsushiro, S.; Tsuboi, S.; Iwabuchi, M.; Hatanaka, T.
    Modulation of Streptomyces leucine aminopeptidase by calcium: identification and functional analysis of key residues in activation and stabilization by calcium (2006), J. Biol. Chem., 281, 5885-5894.
    View publication on PubMed

Application

Application Comment Organism
analysis the enzyme is clinically important as a model for undestanding the structure and mechanism of action of other metallopeptidases Streptomyces griseus

Cloned(Commentary)

Cloned (Comment) Organism
gene Sgap, screening of 21 strains with strain NBRC12875 showing the highest activity Streptomyces griseus

Protein Variants

Protein Variants Comment Organism
D3A/D262G site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme Streptomyces griseus
E196A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Streptomyces griseus
additional information construction of chimeric enzymes from the enzymes of Streptomyces griseus and Streptomyces septatus by DNA in vivo shuffling and site-directed mutagenesis for calcium activation and binding studies, overview Streptomyces griseus

Inhibitors

Inhibitors Comment Organism Structure
L-histidine
-
Streptomyces griseus
L-serine
-
Streptomyces griseus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.54
-
L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant mutant D3A/D262G, in absence of Ca2+ Streptomyces griseus
2.61
-
L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant wild-type enzyme, in absence of Ca2+ Streptomyces griseus
3.47
-
L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant mutant E196A, in presence of Ca2+ Streptomyces griseus
3.79
-
L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant mutant E196A, in absence of Ca2+ Streptomyces griseus
3.91
-
L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant chimeric mutant, in presence of Ca2+ Streptomyces griseus
4.02
-
L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant chimeric mutant, in absence of Ca2+ Streptomyces griseus
4.02
-
L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant wild-type enzyme, in presence of Ca2+ Streptomyces griseus
4.07
-
L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant mutant D3A/D262G, in presence of Ca2+ Streptomyces griseus

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ activates and stabilizes, influences substrate specificity, Asp173 and Asp174 are key residues in Ca2+ binding and important for enzyme activity, residues Asp3, Ile4, Asp262, and Asp266 are also involved in calcium binding but are important for enzyme stabilization, overview, binding capacity of recombinant wild-type and mutant enzymes, overview Streptomyces griseus
Zn2+ the enzyme is a double-zinc aminopeptidase, metal ions are bound at the active site Streptomyces griseus

Organism

Organism UniProt Comment Textmining
Streptomyces griseus Q5WA30
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
recombinant wild-type and mutant enzymes, overview Streptomyces griseus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-Leu-4-nitroanilide + H2O
-
Streptomyces griseus L-leucine + 4-nitroaniline
-
?

Synonyms

Synonyms Comment Organism
leucine aminopeptidase
-
Streptomyces griseus
SGAP
-
Streptomyces griseus
Streptomyces griseus leucine aminopeptidase
-
Streptomyces griseus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Streptomyces griseus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.49
-
L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant mutant E196A, in presence of Ca2+ Streptomyces griseus
0.55
-
L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant mutant E196A, in absence of Ca2+ Streptomyces griseus
0.97
-
L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant wild-type enzyme, in presence of Ca2+ Streptomyces griseus
1.08
-
L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant mutant D3A/D262G, in presence of Ca2+ Streptomyces griseus
2.12
-
L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant wild-type enzyme, in absence of Ca2+ Streptomyces griseus
2.71
-
L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant mutant D3A/D262G, in absence of Ca2+ Streptomyces griseus
3 6 L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant mutant D3A/D262G, in absence of Ca2+ Streptomyces griseus
4.72
-
L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant chimeric mutant, in presence of Ca2+ Streptomyces griseus
5.2
-
L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant chimeric mutant, in absence of Ca2+ Streptomyces griseus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Streptomyces griseus