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Literature summary for 3.4.11.10 extracted from

  • Hartley, M.; Bennett, B.
    Heterologous expression and purification of Vibrio proteolyticus (Aeromonas proteolytica) aminopeptidase: a rapid protocol (2009), Protein Expr. Purif., 66, 91-101.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.11.10

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21 Star DE3 cells Vibrio proteolyticus

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop vapour diffusion method, aminopeptidase in 10 mM Tris buffer, pH 8.0, containing 10 mM KSCN and 400 mM NaCl is equilibrated with 100 mM Tris buffer, pH 8.0, containing 100 mM KSCN and 4.5 M NaCl, at 19°C Vibrio proteolyticus

Protein Variants

Protein Variants Comment Organism
D99A mutant shows very low activity Vibrio proteolyticus
D99H mutant shows very low activity Vibrio proteolyticus
D99M mutant shows reduced activity Vibrio proteolyticus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0317
-
 L-Leu-4-nitroanilide apparent value, mutant enzyme D99M, in 10 mM Tricine buffer, pH 8.0, at 25°C Vibrio proteolyticus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
32000
-
 SDS-PAGE Vibrio proteolyticus

Organism

Organism UniProt Comment Textmining
Vibrio proteolyticus Q01693 formerly Aeromonas proteolytica
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate fractionation, PK agarose bead chromatography, TALON Co2+-affinity resin column chromatography, and TALON Superflow metal affinity resin column chromatography Vibrio proteolyticus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.003
-
 supernatant clarification, in 10 mM Tricine buffer, pH 8.0, at 25°C Vibrio proteolyticus
120
-
 purified enzyme, in 10 mM Tricine buffer, pH 8.0, at 25°C Vibrio proteolyticus

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
Aminopeptidase
-
 Vibrio proteolyticus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.017
-
 L-Leu-4-nitroanilide apparent value, mutant enzyme D99M, in 10 mM Tricine buffer, pH 8.0, at 25°C Vibrio proteolyticus