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Literature summary for 3.4.11.5 extracted from

  • Matsushita-Morita, M.; Furukawa, I.; Suzuki, S.; Yamagata, Y.; Koide, Y.; Ishida, H.; Takeuchi, M.; Kashiwagi, Y.; Kusumoto, K.I.
    Characterization of recombinant prolyl aminopeptidase from Aspergillus oryzae (2009), J. Appl. Microbiol., 109, 156-165.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
PAP is overexpressed as a His-tag fusion protein under a taka-amylase gene (amyB) promoter with a limited expressing condition in Aspergillus oryzae Aspergillus oryzae

Inhibitors

Inhibitors Comment Organism Structure
2-mercaptoethanol 1 mM ZnCl2 and 5 mM beta-mercapthoethanol, respectively, relative activity: 61% Aspergillus oryzae
bestatin 0.002 mM, relative activity: 65% Aspergillus oryzae
EDTA 1 mM, relative activity: 68% Aspergillus oryzae
leupeptin 0.04 mM, relative activity: 68% Aspergillus oryzae
N-tosyl-L-lysine chloromethyl ketone 4 mM, relative activity: 37% Aspergillus oryzae
N-tosyl-L-phenylalanyl chloromethyl ketone 0.02 mM, relative activity: 69% Aspergillus oryzae
p-chloromercuribenzenesulfonate 0.1 mM, relative activity: 0% Aspergillus oryzae
ZnCl2 1 mM ZnCl2 + and 5 mM beta-mercapthoethanol, respectively, relative activity: 61% Aspergillus oryzae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.095
-
L-Pro-4-nitroanilide pH 7.5, 30°C Aspergillus oryzae
0.31
-
Pro-Ala pH 7.5, 30°C Aspergillus oryzae

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
48000
-
SDS-PAGE analysis reveals two protein bands (51 kDa and 48 kDa) Aspergillus oryzae
51000
-
SDS-PAGE analysis reveals two protein bands (51 kDa and 48 kDa) Aspergillus oryzae
313000
-
gel filtration Aspergillus oryzae

Organism

Organism UniProt Comment Textmining
Aspergillus oryzae C6L823
-
-

Purification (Commentary)

Purification (Comment) Organism
enzyme is partially purified through two-step column chromatography using a Ni-IMAC column and a MonoQ column Aspergillus oryzae

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
PAP activity in the mycelia grown in rich medium containing glucose (repressing condition) is twice that in starch (inducing condition) Aspergillus oryzae
2.4
-
substrate: PLSRTLSVAAKK Aspergillus oryzae
4.5
-
substrate: PPGFSPFR Aspergillus oryzae
18.3
-
substrate: L-Pro-4-nitroanilide Aspergillus oryzae
20.3
-
substrate: Pro-2-naphthylamide Aspergillus oryzae
22.2
-
substrate: Pro-Leu-Gly Aspergillus oryzae
25.7
-
substrate: Pro-Ala Aspergillus oryzae
42.4
-
substrate: L-hydroxyproline-2-naphthylamide Aspergillus oryzae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-hydroxyproline-2-naphthylamide + H2O
-
Aspergillus oryzae L-hydroxyproline + 2-naphthylamine
-
?
L-Pro-4-nitroanilide + H2O
-
Aspergillus oryzae L-Pro + 4-nitroaniline
-
?
PLSRTLSVAAKK + H2O
-
Aspergillus oryzae Pro + LSRTLSVAAKK
-
?
PPGFSPFR + H2O
-
Aspergillus oryzae Pro + PGFSPFR
-
?
Pro-2-naphthylamide + H2O
-
Aspergillus oryzae Pro + 2-naphthylamide
-
?
Pro-Ala + H2O PamA releases Pro from amino terminus of a dipeptide Pro-Ala, and not from Ala-Pro Aspergillus oryzae Pro + Ala
-
?
Pro-Leu-Gly + H2O
-
Aspergillus oryzae Leu-Gly + Pro
-
?

Subunits

Subunits Comment Organism
hexamer 6 * 48000, SDS-PAGE Aspergillus oryzae

Synonyms

Synonyms Comment Organism
pamA
-
Aspergillus oryzae
PAP
-
Aspergillus oryzae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Aspergillus oryzae

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
enzyme gets inactivated above 50°C Aspergillus oryzae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at and optimal pH Aspergillus oryzae

pH Range

pH Minimum pH Maximum Comment Organism
7 11
-
Aspergillus oryzae