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Literature summary for 3.4.19.3 extracted from

  • Kabashima, T.; Li, Y.; Kanada, N.; Ito, K.; Yoshimoto, T.
    Enhancement of the thermal stability of pyroglutamyl peptidase I by introduction of an intersubunit disulfide bond (2001), Biochim. Biophys. Acta, 1547, 214-220.
    View publication on PubMed
Search for more literature for 3.4.19.3

Cloned(Commentary)

Cloned (Comment) Organism
mutant enzymes S185C and C68S/S185C expressed in Escherichia coli Bacillus amyloliquefaciens

Protein Variants

Protein Variants Comment Organism
C68S/S185C the ratio of turnover number to Km-value for the substrate L-pyroglutamyl-beta-naphthylamide is 1.3fold higher than the value of the wild-type enzyme. The disulfide bridge between the substituted C185 bonds two subunits, additional 60000 Da band detected by SDS-PAGE. Thermal stability is increased by about 30°C compared to wild-type enzyme Bacillus amyloliquefaciens
S185C the ratio of turnover number to Km-value for the substrate L-pyroglutamyl-beta-naphthylamide is 1.4fold higher than the value of the wild-type enzyme. The disulfide bridge between the substituted C185 bonds two subunits, additional 60000 Da band detected by SDS-PAGE. The mutant enzyme is more stable than the wild-type enzyme at pH 4.0 and at pH 12.0. Thermal stability is increased by about 30°C compared to wild-type enzyme Bacillus amyloliquefaciens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.18
-
 L-pyroglutamyl-beta-naphthylamide pH 7.0, 37°C, mutant enzyme C68S/S185C Bacillus amyloliquefaciens
0.18
-
 L-pyroglutamyl-beta-naphthylamide pH 7.0, 37°C, mutant enzyme S185C Bacillus amyloliquefaciens
0.25
-
 L-pyroglutamyl-beta-naphthylamide pH 7.0, 37°C, wild-type enzyme Bacillus amyloliquefaciens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
30000
-
 x * 30000, wild-type enzyme, 3 bands of 30000 Da and of 60000 Da are detected in mutant enzymes S185C and C68S/S185C, SDS-PAGE in presence and in absence of 2-mercaptoethanol Bacillus amyloliquefaciens

Organism

Organism UniProt Comment Textmining
Bacillus amyloliquefaciens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Bacillus amyloliquefaciens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-pyroglutamyl-beta-naphthylamide + H2O
-
 Bacillus amyloliquefaciens L-pyroglutamate + beta-naphthylamine
-
 ?

Subunits

Subunits Comment Organism
? x * 30000, wild-type enzyme, 3 bands of 30000 Da and of 60000 Da are detected in mutant enzymes S185C and C68S/S185C, SDS-PAGE in presence and in absence of 2-mercaptoethanol Bacillus amyloliquefaciens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30
-
 pH 7.0, 30 min, about 5% loss of activity of wild-type enzyme and of mutant enzyme S185C Bacillus amyloliquefaciens
40
-
 pH 7.0, 30 min, about 30% loss of activity of wild-type enzyme, about 5% loss of activity of mutant enzyme S185C and mutant enzyme C68S/S185C Bacillus amyloliquefaciens
50
-
 pH 7.0, 30 min, about 95% loss of activity of wild-type enzyme, about 10% loss of activity of mutant enzyme S185C and mutant enzyme C68S/S185C Bacillus amyloliquefaciens
75
-
 pH 7.0, 30 min, about 65% loss of activity of mutant enzyme S185C and about 80% loss of activity of mutant enzyme C68S/S185C Bacillus amyloliquefaciens
80
-
 pH 7.0, 30 min, complete loss of activity of mutant enzyme S185C and mutant enzyme C68S/S185C Bacillus amyloliquefaciens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
641
-
 L-pyroglutamyl-beta-naphthylamide pH 7.0, 37°C, mutant enzyme C68S/S185C Bacillus amyloliquefaciens
770
-
 L-pyroglutamyl-beta-naphthylamide pH 7.0, 37°C, mutant enzyme S185C Bacillus amyloliquefaciens
785
-
 L-pyroglutamyl-beta-naphthylamide pH 7.0, 37°C, wild-type enzyme Bacillus amyloliquefaciens