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Literature summary for 3.4.11.5 extracted from

  • Ito, K.; Inoue, T.; Kabashima, T.; Kanada, N.; Huang, H.S.; Ma, X.; Azmi, N.; Azab, E.; Yoshimoto, T.
    Substrate recognition mechanism of prolyl aminopeptidase from Serratia marcescens (2000), J. Biochem., 128, 673-678.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutants in Escherichia coli Serratia marcescens

Protein Variants

Protein Variants Comment Organism
C271A site-directed mutagenesis, sensitive to inhibition by 4-chloromercuribenzoic acid Serratia marcescens
C74A site-directed mutagenesis, sensitive to inhibition by 4-chloromercuribenzoic acid Serratia marcescens
C74A/C271A site-directed mutagenesis, not sensitive to inhibition by 4-chloromercuribenzoic acid Serratia marcescens
E204Q site-directed mutagenesis, 4% of the wild-type catalytic efficiency Serratia marcescens
F139A site-directed mutagenesis, 80fold decreased catalytic activity compared to the wild-type enzyme Serratia marcescens
R136A site-directed mutagenesis, shows decreased activity compared to the wild-type enzyme, but retains arylamidase activity Serratia marcescens
Y149A site-directed mutagenesis, unaltered catalytic efficiency compared to the wild-type enzyme Serratia marcescens

Inhibitors

Inhibitors Comment Organism Structure
4-chloromercuribenzoic acid
-
Serratia marcescens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.07
-
proline beta-naphthylamide pH 8.0, 37°C, mutant Y149A Serratia marcescens
0.42
-
proline beta-naphthylamide pH 8.0, 37°C, wild-type enzyme Serratia marcescens
0.46
-
proline beta-naphthylamide pH 8.0, 37°C, mutant R136A Serratia marcescens
0.82
-
proline beta-naphthylamide pH 8.0, 37°C, mutant E204Q Serratia marcescens
1.38
-
proline beta-naphthylamide pH 8.0, 37°C, mutant F139A Serratia marcescens
2.07
-
L-Pro-L-Ala pH 8.0, 37°C, wild-type enzyme Serratia marcescens

Organism

Organism UniProt Comment Textmining
Serratia marcescens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli Serratia marcescens

Reaction

Reaction Comment Organism Reaction ID
release of N-terminal proline from a peptide substrate recognition mechanism, Phe139 and Glu204 recognize the pyrrolidine ring and the amino group of proline at the S1 site, Arg136 recognizes the P'1 site Serratia marcescens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Serratia marcescens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-Pro-L-Ala + H2O no activity of mutant R136A Serratia marcescens L-Pro + L-Ala
-
?
additional information substrate specificity and binding mechanism Serratia marcescens ?
-
?
proline beta-naphthylamide + H2O
-
Serratia marcescens proline + beta-naphthylamine
-
?

Subunits

Subunits Comment Organism
More enzyme structure, active site structure Serratia marcescens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Serratia marcescens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.04 1.97 proline beta-naphthylamide pH 8.0, 37°C, mutant R136A Serratia marcescens
0.052 2.1 proline beta-naphthylamide pH 8.0, 37°C, wild-type enzyme Serratia marcescens
0.08
-
proline beta-naphthylamide pH 8.0, 37°C, mutant E204Q Serratia marcescens
0.41
-
proline beta-naphthylamide pH 8.0, 37°C, mutant F139A Serratia marcescens
1.4
-
proline beta-naphthylamide pH 8.0, 37°C, mutant Y149A Serratia marcescens
8.87
-
proline beta-naphthylamide pH 8.0, 37°C, mutant R136A Serratia marcescens
9.81
-
proline beta-naphthylamide pH 8.0, 37°C, wild-type enzyme Serratia marcescens
16.8
-
L-Pro-L-Ala pH 8.0, 37°C, wild-type enzyme Serratia marcescens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Serratia marcescens