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Literature summary for 3.4.11.1 extracted from

  • Chi, M.C.; Ong, P.L.; Hsu, W.H.; Chen, Y.H.; Huang, H.B.; Lin, L.L.
    Role of the invariant Asn345 and Asn435 residues in a leucine aminopeptidase from Bacillus kaustophilus as evaluated by site-directed mutagenesis (2008), Int. J. Biol. Macromol., 43, 481-487.
    View publication on PubMed
Search for more literature for 3.4.11.1

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli M15 cells Geobacillus kaustophilus

Protein Variants

Protein Variants Comment Organism
N345L the mutation results in 93% reduction in LAP activity Geobacillus kaustophilus
N345Q the mutation results in 72% reduction in LAP activity Geobacillus kaustophilus
N435E the mutation results in complete loss of the LAP activity Geobacillus kaustophilus
N435L the mutation results in complete loss of the LAP activity Geobacillus kaustophilus
N435Q the mutation results in complete loss of the LAP activity Geobacillus kaustophilus
N435R the mutation results in complete loss of the LAP activity Geobacillus kaustophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.23
-
 Leu-p-nitroanilide wild type enzyme, in 20 mM Tris-HCl buffer (pH 8.0) containing 1 mM NiCl2 Geobacillus kaustophilus
0.52
-
 Leu-p-nitroanilide mutant enzyme N345Q, in 20 mM Tris-HCl buffer (pH 8.0) containing 1 mM NiCl2 Geobacillus kaustophilus
0.97
-
 Leu-p-nitroanilide mutant enzyme N345L, in 20 mM Tris-HCl buffer (pH 8.0) containing 1 mM NiCl2 Geobacillus kaustophilus

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ LAP contains two Zn2+ ions in the active site Geobacillus kaustophilus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
54000
-
 SDS-PAGE Geobacillus kaustophilus

Organism

Organism UniProt Comment Textmining
Geobacillus kaustophilus Q7X2C8
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni2+-NTA resin column chromatography Geobacillus kaustophilus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
11.4
-
 mutant enzyme N345L, using Leu-p-nitroanilide as substrate, at pH 8.0 Geobacillus kaustophilus
28.8
-
 mutant enzyme N345Q, using Leu-p-nitroanilide as substrate, at pH 8.0 Geobacillus kaustophilus
160
-
 wild type enzyme, using Leu-p-nitroanilide as substrate, at pH 8.0 Geobacillus kaustophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Leu-4-nitroanilide + H2O
-
 Geobacillus kaustophilus Leu + 4-nitroaniline
-
 ?

Synonyms

Synonyms Comment Organism
LAP
-
 Geobacillus kaustophilus
leucine aminopeptidase
-
 Geobacillus kaustophilus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
63
-
 the melting temperature of the wild type enzyme is at 63°C Geobacillus kaustophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.17
-
 Leu-p-nitroanilide mutant enzyme N345L, in 20 mM Tris-HCl buffer (pH 8.0) containing 1 mM NiCl2 Geobacillus kaustophilus
0.63
-
 Leu-p-nitroanilide mutant enzyme N345Q, in 20 mM Tris-HCl buffer (pH 8.0) containing 1 mM NiCl2 Geobacillus kaustophilus
5.92
-
 Leu-p-nitroanilide wild type enzyme, in 20 mM Tris-HCl buffer (pH 8.0) containing 1 mM NiCl2 Geobacillus kaustophilus