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Literature summary extracted from

  • Shi, Y.F.; Soumillion, P.; Ueda, M.
    Effects of catalytic site mutations on active expression of phage fused penicillin acylase (2010), J. Biotechnol., 145, 139-142.
    View publication on PubMed

Application

EC Number Application Comment Organism
3.5.1.11 synthesis the enzyme might be useful in the preparation of a wide range of semi-synthetic beta-lactam antibiotics from acyl side-chain precursors and beta-lactam nucleus Escherichia coli

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.1.11 expression in a phage display in Escherichia coli strain TG1 Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
3.5.1.11 additional information directed evolution of penicillin acylase using phage display technology for extending its specificity. Fusion of the penicillin acylase to fd phage coat protein III and used pIII secretion signal sequence instead of penicillin acylase, which couples gene and enzyme on phage particle and can is useful for directed evolution of penicillin acylase. Penicillin acylase is functionally displayed on phage surface. Determination by site-directed mutagenesis of the effect of Ser B1 and Asn B241 variants on post-translational maturation of phage fused penicillin acylase, overview Escherichia coli
3.5.1.11 N241G site-directed mutagensis of subunit B residue, leads to reduced activity compared to the wild-type enzyme Escherichia coli
3.5.1.11 N241S site-directed mutagensis of subunit B residue, leads to reduced activity compared to the wild-type enzyme Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.11 0.00614
-
 2-nitro-5-[(phenyl-acetyl)amino]benzoic acid pH 7.0, 25°C, mutant N241G Escherichia coli
3.5.1.11 0.0102
-
 2-nitro-5-[(phenyl-acetyl)amino]benzoic acid pH 7.0, 25°C, mutant N241S Escherichia coli
3.5.1.11 0.03
-
 2-nitro-5-[(phenyl-acetyl)amino]benzoic acid pH 7.0, 25°C Escherichia coli

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.1.11 23000
-
 1 * 23000 + 1 * 62000, alphabeta, SDS-PAGE Escherichia coli
3.5.1.11 62000
-
 1 * 23000 + 1 * 62000, alphabeta, SDS-PAGE Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.11 Escherichia coli
-
 strain Migula 1895, ATCC 11105
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.5.1.11 proteolytic modification penicillin G acylase is a 86-kDa heterodimeric protein produced by intein-mediated auto-splicing of a 92-kDa precursor Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.11 2-nitro-5-[(phenyl-acetyl)amino]benzoic acid + H2O
-
 Escherichia coli ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.1.11 heterodimer 1 * 23000 + 1 * 62000, alphabeta, SDS-PAGE Escherichia coli
3.5.1.11 More penicillin G acylase is a 86-kDa heterodimeric protein produced by intein-mediated auto-splicing of a 92-kDa precursor Escherichia coli

Synonyms

EC Number Synonyms Comment Organism
3.5.1.11 penicillin acylase
-
 Escherichia coli
3.5.1.11 penicillin G acylase
-
 Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.1.11 25
-
 assay at Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.1.11 0.204
-
 2-nitro-5-[(phenyl-acetyl)amino]benzoic acid pH 7.0, 25°C, mutant N241S Escherichia coli
3.5.1.11 0.354
-
 2-nitro-5-[(phenyl-acetyl)amino]benzoic acid pH 7.0, 25°C, mutant N241G Escherichia coli
3.5.1.11 15
-
 2-nitro-5-[(phenyl-acetyl)amino]benzoic acid pH 7.0, 25°C, wild-type enzyme Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.1.11 7
-
 assay at Escherichia coli

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.5.1.11 20
-
 2-nitro-5-[(phenyl-acetyl)amino]benzoic acid pH 7.0, 25°C, mutant N241S Escherichia coli
3.5.1.11 57.6
-
 2-nitro-5-[(phenyl-acetyl)amino]benzoic acid pH 7.0, 25°C, mutant N241G Escherichia coli
3.5.1.11 500
-
 2-nitro-5-[(phenyl-acetyl)amino]benzoic acid pH 7.0, 25°C Escherichia coli