Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Acosta, D.; Cancela, M.; Piacenza, L.; Roche, L.; Carmona, C.; Tort, J.F.
    Fasciola hepatica leucine aminopeptidase, a promising candidate for vaccination against ruminant fasciolosis (2008), Mol. Biochem. Parasitol., 158, 52-64.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.11.10 expressed in Escherichia coli TOP 10 cells Fasciola hepatica

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.11.10 1,10-phenanthroline the recombinant enzyme shows 18.8% relative activity in the presence of 1 mM 1,10-phenanthroline on hydrolysis of L-Leu-7-amido-4-methylcoumarin Fasciola hepatica
3.4.11.10 Aprotinin the recombinant enzyme shows 58.9% relative activity in the presence of 0.001 mM aprotinin on hydrolysis of L-Leu-7-amido-4-methylcoumarin Fasciola hepatica
3.4.11.10 bestatin strongly inhibited by bestatin, 20.5% relative activity in the presence of bestatin Fasciola hepatica
3.4.11.10 Ca2+ 44% relative activity at 10 mM Ca2+ Fasciola hepatica
3.4.11.10 dithiothreitol the recombinant enzyme shows 20.5% relative activity in the presence of 5 mM dithiothreitol on hydrolysis of L-Leu-7-amido-4-methylcoumarin Fasciola hepatica
3.4.11.10 E-64 the recombinant enzyme shows 43% relative activity in the presence of 0.005 mM E-64 on hydrolysis of L-Leu-7-amido-4-methylcoumarin Fasciola hepatica
3.4.11.10 L-cysteine strongly inhibited by cysteine Fasciola hepatica
3.4.11.10 leupeptin the recombinant enzyme shows 32.7% relative activity in the presence of 0.01 mM leupeptin on hydrolysis of L-Leu-7-amido-4-methylcoumarin Fasciola hepatica
3.4.11.10 N-alpha-tosyl-L-lysine chlormethyl ketone the recombinant enzyme shows 28.9% relative activity in the presence of 0.005 mM N-alpha-tosyl-L-lysine chlormethyl ketone on hydrolysis of L-Leu-7-amido-4-methylcoumarin Fasciola hepatica
3.4.11.10 pepstatin A the recombinant enzyme shows 26.1% relative activity in the presence of 1 mM pepstatin A on hydrolysis of L-Leu-7-amido-4-methylcoumarin Fasciola hepatica
3.4.11.10 phenylmethylsulfonyl fluoride the recombinant enzyme shows 24.5% relative activity in the presence of 1 mM phenylmethylsulfonyl fluoride on hydrolysis of L-Leu-7-amido-4-methylcoumarin Fasciola hepatica

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4.11.10 0.038
-
 L-Leu-7-amido-4-methylcoumarin recombinant enzyme, in 0.1 M glycine buffer, pH 8.5 containing 1 mM MnCl2, at 37°C Fasciola hepatica
3.4.11.10 0.06
-
 L-Thr-7-amido-4-methylcoumarin recombinant enzyme, in 0.1 M glycine buffer, pH 8.5 containing 1 mM MnCl2, at 37°C Fasciola hepatica
3.4.11.10 0.066
-
 L-Phe-7-amido-4-methylcoumarin recombinant enzyme, in 0.1 M glycine buffer, pH 8.5 containing 1 mM MnCl2, at 37°C Fasciola hepatica
3.4.11.10 0.077
-
 L-Cys-7-amido-4-methylcoumarin recombinant enzyme, in 0.1 M glycine buffer, pH 8.5 containing 1 mM MnCl2, at 37°C Fasciola hepatica
3.4.11.10 0.078
-
 L-Arg-7-amido-4-methylcoumarin recombinant enzyme, in 0.1 M glycine buffer, pH 8.5 containing 1 mM MnCl2, at 37°C Fasciola hepatica
3.4.11.10 0.093
-
 L-Met-7-amido-4-methylcoumarin recombinant enzyme, in 0.1 M glycine buffer, pH 8.5 containing 1 mM MnCl2, at 37°C Fasciola hepatica
3.4.11.10 0.231
-
 L-Ala-7-amido-4-methylcoumarin recombinant enzyme, in 0.1 M glycine buffer, pH 8.5 containing 1 mM MnCl2, at 37°C Fasciola hepatica

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.4.11.10 cytosol
-
 Fasciola hepatica 5829
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.4.11.10 Co2+ CoCl2 at 1mM increases the specific activity 18fold Fasciola hepatica
3.4.11.10 Mg2+ MgCl2 shows an optimal concentration at 10 mM that increases specific activity by 12fold Fasciola hepatica
3.4.11.10 Mn2+ recombinant LAP activity is progressively activated by MnCl2 (0.001-10 mM) with an optimum at 1 mM, yielding a 43fold increase Fasciola hepatica
3.4.11.10 additional information no relevant stimulatory effect with NiCl2, FeSO4 and CuSO4 Fasciola hepatica
3.4.11.10 Zn2+ ZnCl2 modestly affects the enzymatic activity Fasciola hepatica

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.4.11.10 67000
-
 recombinant enzyme, SDS-PAGE Fasciola hepatica

Organism

EC Number Organism UniProt Comment Textmining
3.4.11.10 Fasciola hepatica
-
 LAP preferentially localizes inside the epithelial cells that line the alimentary tract of the adult worm
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.11.10 native and recombinant enzymes are purified by Ni-NTA column chromatography, Superdex 200 gel filtration and bestatin-affinity chromatography Fasciola hepatica

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 0.0001
-
 recombinant enzyme, using L-Glu-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0002
-
 native enzyme, using L-Glu-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0003
-
 native enzyme, using L-Asp-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0005
-
 native enzyme, using Gly-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0006
-
 native enzyme, using L-Val-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0012
-
 recombinant enzyme, using L-Asp-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0019
-
 native enzyme, using L-Ser-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0019
-
 recombinant enzyme, using Gly-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0021
-
 recombinant enzyme, using L-Ser-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0025
-
 recombinant enzyme, using L-Val-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0032
-
 native enzyme, using L-Pro-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0054
-
 native enzyme, using L-Tyr-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0056
-
 recombinant enzyme, using L-Pro-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0062
-
 recombinant enzyme, using L-Ile-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0064
-
 recombinant enzyme, using L-Tyr-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0068
-
 recombinant enzyme, using L-Phe-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0075
-
 recombinant enzyme, using L-Ala-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0083
-
 native enzyme, using L-Ala-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0093
-
 native enzyme, using L-Ile-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0141
-
 native enzyme, using L-Phe-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0253
-
 recombinant enzyme, using L-Met-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0291
-
 recombinant enzyme, using L-Arg-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0355
-
 recombinant enzyme, using L-Thr-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0657
-
 native enzyme, using L-Met-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0694
-
 native enzyme, using L-Thr-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0752
-
 native enzyme, using L-Arg-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0831
-
 recombinant enzyme, using L-Cys-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.0953
-
 native enzyme, using L-Cys-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.1344
-
 native enzyme, using L-Leu-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica
3.4.11.10 0.2145
-
 recombinant enzyme, using L-Leu-7-amido-4-methylcoumarin as substrate, at 37°C Fasciola hepatica

Storage Stability

EC Number Storage Stability Organism
3.4.11.10 -80°C, 3 months, no appreciable loss of activity Fasciola hepatica

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.11.10 Gly-7-amido-4-methylcoumarin + H2O very weak activity Fasciola hepatica Gly + 7-amino-4-methylcoumarin
-
 ?
3.4.11.10 L-Ala-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica L-Ala + 7-amino-4-methylcoumarin
-
 ?
3.4.11.10 L-Arg-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica L-Arg + 7-amino-4-methylcoumarin
-
 ?
3.4.11.10 L-Asp-7-amido-4-methylcoumarin + H2O very weak activity Fasciola hepatica L-Asp + 7-amino-4-methylcoumarin
-
 ?
3.4.11.10 L-Cys-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica L-Cys + 7-amino-4-methylcoumarin
-
 ?
3.4.11.10 L-Glu-7-amido-4-methylcoumarin + H2O very weak activity Fasciola hepatica L-Glu + 7-amino-4-methylcoumarin
-
 ?
3.4.11.10 L-Ile-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica L-Ile + 7-amino-4-methylcoumarin
-
 ?
3.4.11.10 L-Leu-7-amido-4-methylcoumarin + H2O preferred substrate Fasciola hepatica L-Leu + 7-amino-4-methylcoumarin
-
 ?
3.4.11.10 L-Met-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica L-Met + 7-amino-4-methylcoumarin
-
 ?
3.4.11.10 L-Phe-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica L-Phe + 7-amino-4-methylcoumarin
-
 ?
3.4.11.10 L-Pro-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica L-Pro + 7-amino-4-methylcoumarin
-
 ?
3.4.11.10 L-Ser-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica L-Ser + 7-amino-4-methylcoumarin
-
 ?
3.4.11.10 L-Thr-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica L-Thr + 7-amino-4-methylcoumarin
-
 ?
3.4.11.10 L-Tyr-7-amido-4-methylcoumarin + H2O
-
 Fasciola hepatica L-Tyr + 7-amino-4-methylcoumarin
-
 ?
3.4.11.10 L-Val-7-amido-4-methylcoumarin + H2O very weak activity Fasciola hepatica L-Val + 7-amino-4-methylcoumarin
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.4.11.10 LAP
-
 Fasciola hepatica
3.4.11.10 leucine aminopeptidase
-
 Fasciola hepatica
3.4.11.10 Leucyl aminopeptidase
-
 Fasciola hepatica

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.4.11.10 37
-
 the recombinant enzyme is active at a wide range of temperatures, with two peaks, at 37 and 50°C Fasciola hepatica
3.4.11.10 50
-
 the recombinant enzyme is active at a wide range of temperatures, with two peaks, at 37 and 50°C Fasciola hepatica

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.4.11.10 50 60 preheating of recombinant LAP at 50°C produces the maximum specific activity while at higher temperatures (above 60°C) the enzyme results inactivated Fasciola hepatica

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.4.11.10 16200
-
 L-Thr-7-amido-4-methylcoumarin recombinant enzyme, in 0.1 M glycine buffer, pH 8.5 containing 1mM MnCl2, at 37°C Fasciola hepatica
3.4.11.10 33800
-
 L-Phe-7-amido-4-methylcoumarin recombinant enzyme, in 0.1 M glycine buffer, pH 8.5 containing 1mM MnCl2, at 37°C Fasciola hepatica
3.4.11.10 59900
-
 L-Arg-7-amido-4-methylcoumarin recombinant enzyme, in 0.1 M glycine buffer, pH 8.5 containing 1mM MnCl2, at 37°C Fasciola hepatica
3.4.11.10 192000
-
 L-Ala-7-amido-4-methylcoumarin recombinant enzyme, in 0.1 M glycine buffer, pH 8.5 containing 1mM MnCl2, at 37°C Fasciola hepatica
3.4.11.10 246000
-
 L-Leu-7-amido-4-methylcoumarin recombinant enzyme, in 0.1 M glycine buffer, pH 8.5 containing 1mM MnCl2, at 37°C Fasciola hepatica
3.4.11.10 283200
-
 L-Met-7-amido-4-methylcoumarin recombinant enzyme, in 0.1 M glycine buffer, pH 8.5 containing 1mM MnCl2, at 37°C Fasciola hepatica
3.4.11.10 288100
-
 L-Cys-7-amido-4-methylcoumarin recombinant enzyme, in 0.1 M glycine buffer, pH 8.5 containing 1mM MnCl2, at 37°C Fasciola hepatica

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.4.11.10 8 8.5
-
 Fasciola hepatica

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.4.11.10 7 9 optimal activity is observed at pH 8-8.5, exhibiting a rapid decline under mildly acidic (below pH 7) and basic (above pH 9) conditions Fasciola hepatica

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.4.11.10 0.000001
-
 bestatin recombinant enzyme, at 37°C Fasciola hepatica
3.4.11.10 0.000025
-
 L-cysteine recombinant enzyme, at 37°C Fasciola hepatica