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Literature summary for 3.4.19.1 extracted from

  • Nishimura, M.; Ikeda, K.; Sugiyama, M.
    Molecular cloning and characterization of gene encoding novel puromycin-inactivating enzyme from blasticidin S-producing Streptomyces morookaensis (2006), J. Biosci. Bioeng., 101, 63-69.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
dithiothreitol maintains catalytic activity of PMH Streptomyces morookaense

Application

Application Comment Organism
additional information amino acid sequence of PMH displays high similarity to that of the Streptomyces acyl-peptide hydrolase. PMH is an aminopeptidase carrying a putative catalytic triad, Ser511-Asp593-His625, with broad substrate specificity Streptomyces morookaense

Cloned(Commentary)

Cloned (Comment) Organism
pTrc-PMH overexpressed in Escherichia coli strain JM105 Streptomyces morookaense

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.013
-
Pro-beta-naphthylamide
-
Streptomyces morookaense
0.028
-
Phe-beta-naphthylamide
-
Streptomyces morookaense
0.067
-
Tyr-beta-naphthylamide
-
Streptomyces morookaense
0.12
-
Leu-beta-naphthylamide
-
Streptomyces morookaense
0.39
-
Ala-beta-naphthylamide
-
Streptomyces morookaense
1.91
-
puromycin
-
Streptomyces morookaense

Organism

Organism UniProt Comment Textmining
Streptomyces morookaense
-
strain JCM4673
-
Streptomyces morookaense JCM 4673
-
strain JCM4673
-

Purification (Commentary)

Purification (Comment) Organism
by gel filtration, to homogeneity Streptomyces morookaense

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.124
-
recombinant enzyme, after the induction with IPTG, at 8 h of culture in LB medium Streptomyces morookaense
0.172
-
recombinant enzyme, after the induction with IPTG, at 18 h of culture in TB medium Streptomyces morookaense
13
-
with Tyr-Leu as substrate Streptomyces morookaense
19
-
with Tyr-Phe as substrate Streptomyces morookaense
23
-
with Ala-p-nitroanilide as substrate Streptomyces morookaense
154
-
with Phe-p-nitroanilide as substrate Streptomyces morookaense
186
-
with Ala-beta-naphthylamide as substrate Streptomyces morookaense

Storage Stability

Storage Stability Organism
-30°C, 20% glycerol Streptomyces morookaense

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Ala-beta-naphthylamide + H2O
-
Streptomyces morookaense Ala + 2-naphthylamine
-
?
Ala-beta-naphthylamide + H2O
-
Streptomyces morookaense JCM 4673 Ala + 2-naphthylamine
-
?
Ala-p-nitroanilide + H2O prefered substrate for PMH Streptomyces morookaense Ala + p-nitroaniline
-
?
Ala-p-nitroanilide + H2O prefered substrate for PMH Streptomyces morookaense JCM 4673 Ala + p-nitroaniline
-
?
Leu-beta-naphthylamide + H2O
-
Streptomyces morookaense Leu + 2-naphthylamine
-
?
Leu-beta-naphthylamide + H2O
-
Streptomyces morookaense JCM 4673 Leu + 2-naphthylamine
-
?
Phe-beta-naphthylamide + H2O
-
Streptomyces morookaense Phe + 2-naphthylamine
-
?
Phe-p-nitroanilide + H2O prefered substrate for PMH Streptomyces morookaense Phe + p-nitroaniline
-
?
Pro-beta-naphthylamide + H2O
-
Streptomyces morookaense Pro + 2-naphthylamine
-
?
puromycin + H2O
-
Streptomyces morookaense ?
-
?
Tyr-beta-naphthylamide + H2O
-
Streptomyces morookaense Tyr + 2-naphthylamine
-
?
Tyr-Leu + H2O
-
Streptomyces morookaense Tyr + Leu
-
?
Tyr-Leu + H2O
-
Streptomyces morookaense JCM 4673 Tyr + Leu
-
?
Tyr-Phe + H2O
-
Streptomyces morookaense Tyr + Phe
-
?
Tyr-Phe + H2O
-
Streptomyces morookaense JCM 4673 Tyr + Phe
-
?

Synonyms

Synonyms Comment Organism
PM hydrolase
-
Streptomyces morookaense
PMH
-
Streptomyces morookaense

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.98
-
puromycin
-
Streptomyces morookaense
1.61
-
Pro-beta-naphthylamide
-
Streptomyces morookaense
1.69
-
Phe-beta-naphthylamide
-
Streptomyces morookaense
1.85
-
Tyr-beta-naphthylamide
-
Streptomyces morookaense
4.07
-
Leu-beta-naphthylamide
-
Streptomyces morookaense
8.79
-
Ala-beta-naphthylamide
-
Streptomyces morookaense