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Literature summary for 3.5.1.11 extracted from

  • Deaguero, A.; Blum, J.; Bommarius, A.
    Improving the diastereoselectivity of penicillin G acylase for ampicillin synthesis from racemic substrates (2012), Protein Eng. Des. Sel., 25, 135-144.
    View publication on PubMed
Search for more literature for 3.5.1.11

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli HB101, sequence shows one mutation resulting in a betaVal148Leu substitution in the mature protein as compared with the Swiss-Prot entry P06875 derived from Escherichia coli ATC11105 Escherichia coli

Protein Variants

Protein Variants Comment Organism
alphaF146A mutant selective for (S)-ampicillin synthesis Escherichia coli
alphaF146Q mutant selective for (S)-ampicillin synthesis Escherichia coli
alphaF146R mutant selective for (S)-ampicillin synthesis Escherichia coli
alphaF146Y mutant selective for (S)-ampicillin synthesis Escherichia coli
betaF24A mutant selective for (R)-ampicillin synthesis Escherichia coli
betaF24C mutant selective for (R)-ampicillin synthesis Escherichia coli
betaF24P mutant selective for (R)-ampicillin synthesis Escherichia coli
betaF24S mutant selective for (R)-ampicillin synthesis Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0005
-
 2-nitro-5-[(phenylacetyl)amino]-benzoic acid alphaF146Y mutant protein, pH 7.5, 25°C Escherichia coli
0.026
-
 2-nitro-5-[(phenylacetyl)amino]-benzoic acid wild type protein, pH 7.5, 25°C Escherichia coli
0.098
-
 2-nitro-5-[(phenylacetyl)amino]-benzoic acid betaF24A mutant protein, pH 7.5, 25°C Escherichia coli
0.116
-
 2-nitro-5-[(phenylacetyl)amino]-benzoic acid alphaF146A mutant protein, pH 7.5, 25°C Escherichia coli
0.272
-
 2-nitro-5-[(phenylacetyl)amino]-benzoic acid alphaF146Q mutant protein, pH 7.5, 25°C Escherichia coli
0.386
-
 2-nitro-5-[(phenylacetyl)amino]-benzoic acid alphaF146R mutant protein, pH 7.5, 25°C Escherichia coli
0.551
-
 2-nitro-5-[(phenylacetyl)amino]-benzoic acid betaF24S mutant protein, pH 7.5, 25°C Escherichia coli
0.797
-
 2-nitro-5-[(phenylacetyl)amino]-benzoic acid betaF24P mutant protein, pH 7.5, 25°C Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli P06875
-
-

Purification (Commentary)

Purification (Comment) Organism
periplasmic fraction Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.049
-
 substrate 2-nitro-5-[(phenylacetyl)amino]-benzoic acid, alphaF146Q mutant protein, pH 7.5, 25°C Escherichia coli
0.053
-
 substrate 2-nitro-5-[(phenylacetyl)amino]-benzoic acid, alphaF146R mutant protein, pH 7.5, 25°C Escherichia coli
0.073
-
 substrate 2-nitro-5-[(phenylacetyl)amino]-benzoic acid, alphaF146A mutant protein, pH 7.5, 25°C Escherichia coli
0.6
-
 substrate 2-nitro-5-[(phenylacetyl)amino]-benzoic acid, betaF24S mutant protein, pH 7.5, 25°C Escherichia coli
1
-
 substrate 2-nitro-5-[(phenylacetyl)amino]-benzoic acid, betaF24A mutant protein, pH 7.5, 25°C Escherichia coli
1.3
-
 substrate 2-nitro-5-[(phenylacetyl)amino]-benzoic acid, alphaF146Y mutant protein, pH 7.5, 25°C Escherichia coli
2.9
-
 substrate 2-nitro-5-[(phenylacetyl)amino]-benzoic acid, betaF24P mutant protein, pH 7.5, 25°C Escherichia coli
17.3
-
 substrate 2-nitro-5-[(phenylacetyl)amino]-benzoic acid, wild type protein, pH 7.5, 25°C Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-nitro-5-[(phenylacetyl)amino]-benzoic acid + H2O
-
 Escherichia coli 5-amino-2-nitrobenzoic acid + ?
-
 ?
phenylglycine methyl ester + aminopenicillanic acid
-
 Escherichia coli ampicillin + methanol
-
 ?

Synonyms

Synonyms Comment Organism
penicillin G acylase
-
 Escherichia coli