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Literature summary for 3.4.23.49 extracted from

  • Varadarajan, N.; Gam, J.; Olsen, M.J.; Georgiou, G.; Iverson, B.L.
    Engineering of protease variants exhibiting high catalytic activity and exquisite substrate selectivity (2005), Proc. Natl. Acad. Sci. USA, 102, 6855-6860.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.23.49

Application

Application Comment Organism
biotechnology engineering of enzyme variants with targeted, high substrate specificity Escherichia coli

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes Escherichia coli

Protein Variants

Protein Variants Comment Organism
D208G site-directed mutagenesis, the mutant enzyme shows increased specificity for the A-R cleavage site compared to the wild-type enzyme Escherichia coli
additional information random mutagenesis of gene ompT, screening for mutant variants with altered cleavage specificity, e.g. mutant variants 1.2.19 and 1.3.19 exhibits higher specificity for the cleavage site A-R and lower specificity for R-R than the wild-type Escherichia coli
S223R site-directed mutagenesis, the mutant enzyme shows increased specificity for the A-R cleavage site and overall reduced activity compared to the wild-type enzyme Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0073
-
 WCARVGKGRGR-NH2 22°C, recombinant mutant D208G Escherichia coli
0.009
-
 WCARVGKGRGR-NH2 22°C, recombinant mutant S223R Escherichia coli
0.009
-
 WCARVGKGRGR-NH2 22°C, recombinant mutant variant 1.2.19 Escherichia coli
0.015
-
 WCARVGKGRGR-NH2 22°C, recombinant mutant variant 1.3.19 Escherichia coli
0.016
-
 WCARVGKGRGR-NH2 22°C, recombinant wild-type enzyme Escherichia coli
0.055
-
 WEEGGRRIGRGGK-NH2 22°C, recombinant wild-type enzyme Escherichia coli
0.16
-
 WEEGGRRIGRGGK-NH2 22°C, recombinant mutant variant 1.3.19 Escherichia coli
0.24
-
 WEEGGRRIGRGGK-NH2 22°C, recombinant mutant D208G Escherichia coli
0.26
-
 WEEGGRRIGRGGK-NH2 22°C, recombinant mutant variant 1.2.19 Escherichia coli

Localization

Localization Comment Organism GeneOntology No. Textmining
outer membrane
-
 Escherichia coli 19867
-

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
WCARVGKGRGR-NH2 + H2O proteolytic cleavage of the peptide at the site A-R Escherichia coli WCA + RVGKGRGR-NH2
-
 ?
WEEGGRRIGRGGK-NH2 + H2O proteolytic cleavage of the peptide at the site R-R, no activity of mutant S223R, preferred substrate of wild-type OmpT Escherichia coli WEEGGR + RIGRGGK-NH2
-
 ?

Synonyms

Synonyms Comment Organism
ompT
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
-
 assay at room temperature Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0005
-
 WEEGGRRIGRGGK-NH2 22°C, recombinant mutant variant 1.2.19 Escherichia coli
0.031
-
 WCARVGKGRGR-NH2 22°C, recombinant wild-type enzyme Escherichia coli
0.3
-
 WEEGGRRIGRGGK-NH2 22°C, recombinant mutant D208G Escherichia coli
0.4
-
 WEEGGRRIGRGGK-NH2 22°C, recombinant mutant variant 1.3.19 Escherichia coli
1.6
-
 WCARVGKGRGR-NH2 22°C, recombinant mutant D208G Escherichia coli
1.7
-
 WCARVGKGRGR-NH2 22°C, recombinant mutant variant 1.3.19 Escherichia coli
2.2
-
 WCARVGKGRGR-NH2 22°C, recombinant mutant variant 1.2.19 Escherichia coli
2.3
-
 WCARVGKGRGR-NH2 22°C, recombinant mutant S223R Escherichia coli
8.8
-
 WEEGGRRIGRGGK-NH2 22°C, recombinant wild-type enzyme Escherichia coli