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Literature summary for 3.4.11.10 extracted from

  • Hernandez-Moreno, A.V.; Villasenor, F.; Medina-Rivero, E.; Perez, N.O.; Flores-Ortiz, L.F.; Saab-Rincon, G.; Luna-Barcenas, G.
    Kinetics and conformational stability studies of recombinant leucine aminopeptidase (2014), Int. J. Biol. Macromol., 64, 306-312 .
    View publication on PubMed
Search for more literature for 3.4.11.10

Application

Application Comment Organism
synthesis leucine aminopeptidase from Vibrio proteolyticus is a broad specificity N-terminal aminopeptidase that is widely used in pharmaceutical processes where the removal of N-terminal residues in recombinant proteins is required Vibrio proteolyticus

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of His6-tagged enzyme Vibrio proteolyticus

General Stability

General Stability Organism
urea-caused unfolding transitions of rLAP as a function of urea concentration are monitored by circular dichroism (CD) and fluorescence (FL) spectroscopy exhibiting single transitions by both techniques. Free energy change for unfolding measured by CD and FL spectroscopy are 2.8 and 3.7 kcal/mol, respectively Vibrio proteolyticus

Inhibitors

Inhibitors Comment Organism Structure
DTT complete inhibition at 20 mM Vibrio proteolyticus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information thermodynamics and Michaelis-Menten kinetics, overview Vibrio proteolyticus
0.245
-
 L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant His6-tagged enzyme, with preincubation (72 h of incubation at 37°C) Vibrio proteolyticus
0.311
-
 L-Leu-4-nitroanilide pH 8.0, 70°C, recombinant His6-tagged enzyme, with preincubation (72 h of incubation at 37°C) Vibrio proteolyticus
0.902
-
 L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant His6-tagged enzyme, without preincubation Vibrio proteolyticus

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ required, 2 Zn2+ per active site Vibrio proteolyticus

Organism

Organism UniProt Comment Textmining
Vibrio proteolyticus Q01693
-
-

Renatured (Commentary)

Renatured (Comment) Organism
activity gain of rLAP refolded after reversible denaturation by urea, at 37°C, monitoring of the secondary structure changes 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
-
 ?
additional information recombinant enzyme rLAP is selective and specific to remove N-terminal groups from amino acids Vibrio proteolyticus ?
-
 ?

Synonyms

Synonyms Comment Organism
AVP
-
 Vibrio proteolyticus
leucine aminopeptidase
-
 Vibrio proteolyticus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
70
-
-
 Vibrio proteolyticus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.817
-
 L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant His6-tagged enzyme, without preincubation Vibrio proteolyticus
15.8
-
 L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant His6-tagged enzyme, with preincubation (72 h of incubation at 37°C) Vibrio proteolyticus
48.45
-
 L-Leu-4-nitroanilide pH 8.0, 70°C, recombinant His6-tagged enzyme, with preincubation (72 h of incubation at 37°C) Vibrio proteolyticus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Vibrio proteolyticus

General Information

General Information Comment Organism
additional information temperature-dependent UV-CD spectra and fluorescent measurements Vibrio proteolyticus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.91
-
 L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant His6-tagged enzyme, without preincubation Vibrio proteolyticus
64.5
-
 L-Leu-4-nitroanilide pH 8.0, 37°C, recombinant His6-tagged enzyme, with preincubation (72 h of incubation at 37°C) Vibrio proteolyticus
155.8
-
 L-Leu-4-nitroanilide pH 8.0, 70°C, recombinant His6-tagged enzyme, with preincubation (72 h of incubation at 37°C) Vibrio proteolyticus