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Literature summary extracted from

  • Wu, G.P.; Cao, M.J.; Chen, Y.; Liu, B.X.; Su, W.J.
    Leucine aminopeptidase from red sea bream (Pagrus major) skeletal muscle: purification, characterization, cellular location, and tissue distribution (2008), J. Agric. Food Chem., 56, 9653-9660.
    View publication on PubMed

General Stability

EC Number General Stability Organism
3.4.11.1 the enzyme completely loses its activity upon freezing and thawing Pagrus major

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.11.1 1,10-phenanthroline 31.1% residual activity at 0.2 mM Pagrus major
3.4.11.1 Ba2+ slight inhibitory effect at 1 mM Pagrus major
3.4.11.1 bestatin 22.3% residual activity at 0.01 mM Pagrus major
3.4.11.1 Ca2+ slight inhibitory effect at 1 mM Pagrus major
3.4.11.1 Cd2+ at concentration of 1 mM, the enzyme is strongly inactivated by Cd2+ (38.2% residual activity) Pagrus major
3.4.11.1 Cu2+ at concentration of 1 mM, the enzyme is strongly inactivated by Cu2+ (2.3% residual activity) Pagrus major
3.4.11.1 E-64 40.4% residual activity at 0.015 mM Pagrus major
3.4.11.1 EDTA strong inhibitor, the enzyme loses 83% of its activity when it is dialyzed extensively against 1 mM EDTA in 25 mM sodium phosphate buffer (pH 7.0), whereas the loss can be restored by Zn2+ to 97% at 0.05 mM and to 81% at 0.1 mM of its original activity, the recovery of enzymatic activity by Mn2+ reaches 94% at 0.5 mM and 79% at 1 mM, on the other hand, the enzymatic activity recovery by Ca2+ and Mg2+ is lower than that by Zn2+ and Mn2+, the former reaches 46% and the latter, 26%, at 1 mM Pagrus major
3.4.11.1 EGTA 25.4% residual activity at 5 mM Pagrus major
3.4.11.1 Fe2+ partially inactivated by Fe2+ (53.9% residual activity at 1 mM) Pagrus major
3.4.11.1 Mn2+ slight inhibitory effect at 1 mM Pagrus major
3.4.11.1 pepstatin 83.4% residual activity at 0.15 mM Pagrus major
3.4.11.1 phenylmethanesulfonyl fluoride 94.94% residual activity at 1 mM Pagrus major
3.4.11.1 Zn2+ at concentration of 1 mM, the enzyme is strongly inactivated by Zn2+ (35.1% residual activity) Pagrus major

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4.11.1 0.00155
-
 L-leucyl-7-amido-4-methylcoumarin in 25 mM sodium phosphate buffer (pH 7.0) containing 1 mM dithiothreitol and 0.02% NaN3, at 37°C Pagrus major

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.4.11.1 Ba2+ shows slight activation at 0.1 mM Pagrus major
3.4.11.1 Ca2+ shows slight activation at 0.1 mM Pagrus major
3.4.11.1 Mg2+ slightly activates the enzymatic activity at concentrations from 0.1 to 1 mM Pagrus major
3.4.11.1 Mn2+ shows slight activation at 0.1 mM Pagrus major
3.4.11.1 Zn2+ Zn2+ seem to be its metal cofactor Pagrus major

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.4.11.1 96000
-
 approximately 96000 Da, SDS-PAGE and gel filtration Pagrus major

Organism

EC Number Organism UniProt Comment Textmining
3.4.11.1 Pagrus major
-
 red sea bream
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.11.1 ammonium sulfate fractionation, DEAE-Sephacel column chromatography, Sephacryl S-200 gel filtration, hydroxyapatite column chromatography, and phenyl-Sepharose column chromatography Pagrus major

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.4.11.1 brain
-
 Pagrus major
-
3.4.11.1 gill
-
 Pagrus major
-
3.4.11.1 gut
-
 Pagrus major
-
3.4.11.1 kidney
-
 Pagrus major
-
3.4.11.1 liver
-
 Pagrus major
-
3.4.11.1 pyloric cecum
-
 Pagrus major
-
3.4.11.1 skeletal muscle
-
 Pagrus major
-
3.4.11.1 stomach
-
 Pagrus major
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.4.11.1 0.8
-
 crude cell extract, at 37°C Pagrus major
3.4.11.1 3977
-
 after 4850.5fold purification, at 37°C Pagrus major

Storage Stability

EC Number Storage Stability Organism
3.4.11.1 -80°C, 10% sucrose, 6 months, more than 90% of the enzyme activity remain Pagrus major

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.11.1 L-alanyl-7-amido-4-methylcoumarin + H2O 20.5% activity compared to L-leucyl-7-amido-4-methylcoumarin Pagrus major L-alanine + 7-amino-4-methylcoumarin
-
 ?
3.4.11.1 L-arginyl-7-amido-4-methylcoumarin + H2O 70% activity compared to L-leucyl-7-amido-4-methylcoumarin Pagrus major L-arginine + 7-amino-4-methylcoumarin
-
 ?
3.4.11.1 L-leucyl-7-amido-4-methylcoumarin + H2O preferred substrate Pagrus major L-leucine + 7-amino-4-methylcoumarin
-
 ?
3.4.11.1 L-tyrosyl-7-amido-4-methylcoumarin + H2O 7.4% activity compared to L-leucyl-7-amido-4-methylcoumarin Pagrus major L-tyrosine + 7-amino-4-methylcoumarin
-
 ?
3.4.11.1 additional information no activity towards L-prolyl-7-amido-4-methylcoumarin, L-valyl-7-amido-4-methylcoumarin, L-glycyl-7-amido-4-methylcoumarin, tert-butyloxycarbonyl-Phe-Ser-Arg-7-amido-4-methylcoumarin, tert-butyloxycarbonyl-Leu-Arg-Arg-7-amido-4-methylcoumarin, and tert-butyloxycarbonyl-Leu-Lys-Arg-7-amido-4-methylcoumarin Pagrus major ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.4.11.1 monomer 1 * 96000, gel filtration Pagrus major

Synonyms

EC Number Synonyms Comment Organism
3.4.11.1 LAP
-
 Pagrus major
3.4.11.1 leucine aminopeptidase
-
 Pagrus major

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.4.11.1 45
-
-
 Pagrus major

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.4.11.1 10 50 activity decreases above 50°C, the enzyme retains 10% of its optimal activity at 10°C Pagrus major

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.4.11.1 30
-
 LAP activity decreases slightly after incubation below 30°C Pagrus major
3.4.11.1 40
-
 LAP activity decreases rapidly above 40°C Pagrus major

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.4.11.1 17.3
-
 L-arginyl-7-amido-4-methylcoumarin in 25 mM sodium phosphate buffer (pH 7.0) containing 1 mM dithiothreitol and 0.02% NaN3, at 37°C Pagrus major
3.4.11.1 26.4
-
 L-leucyl-7-amido-4-methylcoumarin in 25 mM sodium phosphate buffer (pH 7.0) containing 1 mM dithiothreitol and 0.02% NaN3, at 37°C Pagrus major

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.4.11.1 7.5
-
-
 Pagrus major

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.4.11.1 5.5 8.5 the enzymatic activity is negligible below pH 5.5 and above 8.5 Pagrus major

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.4.11.1 6 7.5 moderate activity loss after incubation below pH 6.0, but rapid loss after incubation above pH 7.5 for 48 h Pagrus major

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
3.4.11.1 0.00144
-
 in 25 mM sodium phosphate buffer (pH 7.0) containing 1 mM dithiothreitol and 0.02% NaN3, at 37°C Pagrus major bestatin