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Literature summary for 3.5.1.11 extracted from

  • Yuryev, R.; Kasche, V.; Ignatova, Z.; Galunsky, B.
    Improved A. faecalis penicillin amidase mutant retains the thermodynamic and pH stability of the wild type enzyme (2010), Protein J., 29, 181-187.
    View publication on PubMed

Application

Application Comment Organism
synthesis penicillin amidase from Alacaligenes faecalis is an attractive biocatalyst for hydrolysis of penicillin G for production of 6-aminopenicillanic acid, which is used in the synthesis of semi-synthetic beta-lactam antibiotics Alcaligenes faecalis

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli BL21(DE3) Alcaligenes faecalis

Protein Variants

Protein Variants Comment Organism
T206G/S213G construction of a mutant with extended C-terminus of the A-chain comprising parts of the connecting linker peptide showing almost 2fold increased activity and 3fold higher specificity compared to the wild-type enzyme, overview Alcaligenes faecalis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
penicillin G + H2O Alcaligenes faecalis
-
phenylacetic acid + 6-aminopenicillanate
-
?

Organism

Organism UniProt Comment Textmining
Alcaligenes faecalis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli BL21(DE3) to homogeneity Alcaligenes faecalis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
80
-
wild-type enzyme, pH 7.5, 25°C Alcaligenes faecalis
140
-
T206G/S213G mutant enzyme, pH 7.5, 25°C Alcaligenes faecalis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
penicillin G + H2O
-
Alcaligenes faecalis phenylacetic acid + 6-aminopenicillanate
-
?
penicillin G + H2O
-
Alcaligenes faecalis phenylacetic acid + 6-aminopenicillanate product detection by reaction with substrate 6-nitro-3-(phenylacetamido)benzoic acid ?

Synonyms

Synonyms Comment Organism
penicillin amidase belongs to the family of N-terminal nucleophile hydrolases acting on carbon nitrogen bonds in linear amides Alcaligenes faecalis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Alcaligenes faecalis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
15 95 temperature-induced unfolding studies of recombinant wild-type and mutant enzymes, kinetics, overview Alcaligenes faecalis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Alcaligenes faecalis

pH Stability

pH Stability pH Stability Maximum Comment Organism
3.9 10.7 pH-stability studies of recombinant wild-type and mutant enzymes, inactivation above pH 10.7 and below pH 3.9, overview Alcaligenes faecalis