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Literature summary 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

Cloned(Commentary)

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

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.4.11.10 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

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4.11.10 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]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.4.11.10 32000
-
 SDS-PAGE Vibrio proteolyticus

Organism

EC Number Organism UniProt Comment Textmining
3.4.11.10 Vibrio proteolyticus Q01693 formerly Aeromonas proteolytica
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.11.10 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]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.4.11.10 0.003
-
 supernatant clarification, in 10 mM Tricine buffer, pH 8.0, at 25°C Vibrio proteolyticus
3.4.11.10 120
-
 purified enzyme, in 10 mM Tricine buffer, pH 8.0, at 25°C Vibrio proteolyticus

Substrates and Products (Substrate)

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

Synonyms

EC Number Synonyms Comment Organism
3.4.11.10 Aminopeptidase
-
 Vibrio proteolyticus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.4.11.10 0.017
-
 L-Leu-4-nitroanilide apparent value, mutant enzyme D99M, in 10 mM Tricine buffer, pH 8.0, at 25°C Vibrio proteolyticus