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Literature summary for 3.4.11.20 extracted from

  • Gonzalez-Bacerio, J.; Osuna, J.; Ponce, A.; Fando, R.; Figarella, K.; Mendez, Y.; Charli, J.L.; Chavez, M.D.
    High-level expression in Escherichia coli, purification and kinetic characterization of Plasmodium falciparum M1-aminopeptidase (2014), Protein Expr. Purif., 104C, 103-114.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Plasmodium falciparum

Protein Variants

Protein Variants Comment Organism
additional information synthesis and cloning of an optimized gene, with a codon adaptation index and an average G/C content higher than the native gene. Optimal expression is achieved by induction with 1 mM IPTG at 37┬░C for 18 h. This allows obtaining 100 mg of recombinant enzyme per liter of culture medium. The protein shows only limited signs of proteolytic degradation, and purification in a single step by immobilized metal ion affinity chromatography increases purity 27fold Plasmodium falciparum

Inhibitors

Inhibitors Comment Organism Structure
1,10-phenanthroline 1 mM, 80% inhibition Plasmodium falciparum
Zn2+ 0.1 mM, 80% inhibition Plasmodium falciparum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.248
-
L-Lys 4-nitroanilide pH 7.2, 37┬░C Plasmodium falciparum
0.27
-
L-Arg 4-nitroanilide pH 7.2, 37┬░C Plasmodium falciparum
0.304
-
L-Ala 4-nitroanilide pH 7.2, 37┬░C Plasmodium falciparum

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
106130
-
x * 106130, calculated for recombinant protein including His-tag Plasmodium falciparum

Organism

Organism UniProt Comment Textmining
Plasmodium falciparum Q8IEK1
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-Ala 4-nitroanilide + H2O
-
Plasmodium falciparum L-Ala + 4-nitroaniline
-
?
L-Arg 4-nitroanilide + H2O
-
Plasmodium falciparum L-Arg + 4-nitroaniline
-
?
L-Leu 4-nitroanilide + H2O
-
Plasmodium falciparum L-Leu + 4-nitroaniline
-
?
L-Lys 4-nitroanilide + H2O
-
Plasmodium falciparum L-Lys + 4-nitroaniline
-
?
additional information no substrates: Ile-4-nitroanilide, Val-4-nitroanilide, Gly-4-nitroanilide, Pro-4-nitroanilide Plasmodium falciparum ?
-
?

Subunits

Subunits Comment Organism
? x * 106130, calculated for recombinant protein including His-tag Plasmodium falciparum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.15
-
L-Ala 4-nitroanilide pH 7.2, 37┬░C Plasmodium falciparum
1.55
-
L-Lys 4-nitroanilide pH 7.2, 37┬░C Plasmodium falciparum
1.58
-
L-Arg 4-nitroanilide pH 7.2, 37┬░C Plasmodium falciparum
1.65
-
L-Leu 4-nitroanilide pH 7.2, 37┬░C Plasmodium falciparum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2 7.4
-
Plasmodium falciparum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.24
-
L-Ala 4-nitroanilide pH 7.2, 37┬░C Plasmodium falciparum
5.42
-
L-Leu 4-nitroanilide pH 7.2, 37┬░C Plasmodium falciparum
5.84
-
L-Arg 4-nitroanilide pH 7.2, 37┬░C Plasmodium falciparum
6.24
-
L-Lys 4-nitroanilide pH 7.2, 37┬░C Plasmodium falciparum