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Literature summary extracted from

  • Yin, J.; Deng, Z.; Zhao, G.; Huang, X.
    The N-terminal nucleophile serine of cephalosporin acylase executes the second autoproteolytic cleavage and acylpeptide hydrolysis (2011), J. Biol. Chem., 286, 24476-24486.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.1.93 expressed in Escherichia coli BL21(DE3) cells Pseudomonas sp.

Protein Variants

EC Number Protein Variants Comment Organism
3.5.1.93 E159D the mutant is still active, but significantly retarded in the second autocleavage compared to the wild type enzyme Pseudomonas sp.
3.5.1.93 G160A the mutant performs the second autocleavage more slowly than the wild type enzyme Pseudomonas sp.
3.5.1.93 G160L the mutant performs the second autocleavage more slowly than the wild type enzyme Pseudomonas sp.
3.5.1.93 S170C the mutant simultaneously loses the activities for both the second autocleavage and substrate hydrolysis Pseudomonas sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.93 0.35
-
 glutaryl-GDPP at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 0.4
-
 glutaryl-GDAADAADKG at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 0.43
-
 glutaryl-GGGGGK at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 0.5
-
 glutaryl-7-aminocephalosporanic acid at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 0.67
-
 glutaryl-GDPPDLADQG at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 0.73
-
 glutaryl-alanyl-proline at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 0.85
-
 glutaryl-glycyl-proline at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 1.68
-
 glutaryl-GGGGAA at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 2.7
-
 glutaryl-EGDPPDLADQG at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 3.1
-
 glutaryl-glycine at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 3.2
-
 succinyl-7-aminocephalosporanic acid at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 3.3
-
 gamma-Glu-Cys-Gly at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 4.5
-
 4-(glutarylamino)benzoic acid at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 6.1
-
 glutaryl-tryptophan at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 6.4
-
 glutaryl-gamma-Glu-Cys-Gly at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 22
-
 cephalosporin C at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 40
-
 glutarylaniline at 37°C, pH not specified in the publication Pseudomonas sp.

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.93 Pseudomonas sp.
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.1.93 acetone precipitation and Sepharose CM column chromatography Pseudomonas sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.93 4-(glutarylamino)benzoic acid + H2O
-
 Pseudomonas sp. glutarate + 4-aminobenzoate
-
 ?
3.5.1.93 cephalosporin C + H2O
-
 Pseudomonas sp. cephalosporamic acid + 2-amino-5-hydroxypentanoate
-
 ?
3.5.1.93 DGDPP + H2O 20% activity compared to E-GDPPDLADQG Pseudomonas sp. L-Asp + GDPP
-
 ?
3.5.1.93 EGDPP + H2O 67% activity compared to E-GDPPDLADQG Pseudomonas sp. L-Glu + GDPP
-
 ?
3.5.1.93 EGDPPDLADQG + H2O 100% activity Pseudomonas sp. L-Glu + GDPPDLADQG
-
 ?
3.5.1.93 gamma-Glu-Cys-Gly
-
 Pseudomonas sp. L-glutamate + Cys-Gly
-
 ?
3.5.1.93 GEGDPP + H2O 5% activity compared to E-GDPPDLADQG Pseudomonas sp. Gly-L-Glu + GDPP
-
 ?
3.5.1.93 glutaryl-7-aminocephalosporanic acid + H2O
-
 Pseudomonas sp. 7-aminocephalosporanate + glutarate
-
 ?
3.5.1.93 glutaryl-alanyl-proline + H2O
-
 Pseudomonas sp. glutarate + alanyl-proline
-
 ?
3.5.1.93 glutaryl-EGDPPDLADQG + H2O
-
 Pseudomonas sp. glutarate + EGDPPDLADQG
-
 ?
3.5.1.93 glutaryl-gamma-Glu-Cys-Gly + H2O
-
 Pseudomonas sp. glutarate + gamma-Glu-Cys-Gly
-
 ?
3.5.1.93 glutaryl-GDAADAADKG + H2O
-
 Pseudomonas sp. glutarate + GDAADAADKG
-
 ?
3.5.1.93 glutaryl-GDPP + H2O substrate with highest catalytic efficiency Pseudomonas sp. glutarate + GDPP
-
 ?
3.5.1.93 glutaryl-GDPPDLADQG + H2O
-
 Pseudomonas sp. glutarate + GDPPDLADQG
-
 ?
3.5.1.93 glutaryl-GGGGAA + H2O
-
 Pseudomonas sp. glutarate + GGGGAA
-
 ?
3.5.1.93 glutaryl-GGGGGK + H2O
-
 Pseudomonas sp. glutarate + GGGGGK
-
 ?
3.5.1.93 glutaryl-glycine + H2O
-
 Pseudomonas sp. glutarate + glycine
-
 ?
3.5.1.93 glutaryl-glycyl-L-proline + H2O
-
 Pseudomonas sp. glutarate + glycyl-L-proline
-
 ?
3.5.1.93 glutaryl-glycyl-proline + H2O
-
 Pseudomonas sp. glutarate + glycylproline
-
 ?
3.5.1.93 glutaryl-tryptophan + H2O
-
 Pseudomonas sp. glutarate + tryptophan
-
 ?
3.5.1.93 glutarylaniline + H2O
-
 Pseudomonas sp. glutarate + aniline
-
 ?
3.5.1.93 additional information less than 1% activity with TLGEGDPP compared to EGDPPDLADQG, no activity with adipyl-7-aminocephalosporanic acid Pseudomonas sp. ?
-
 ?
3.5.1.93 additional information the enzyme also removes N-terminal Glu or Asp from peptides. Cephalosporin acylase is an acylpeptide hydrolase rather than a previously expected acylamino acid acylase. It only exhibits exopeptidase activity for the hydrolysis of an externally added peptide, supporting the intra-molecular interaction Pseudomonas sp. ?
-
 ?
3.5.1.93 additional information the precursor cephalosporin acylase (without signal peptide) is activated by two peptide bond cleavages, which excise an internal spacer. The first cleavage occurs between Gly169 and Ser170 and the second cleavage occurs between Glu159 and Gly160 Pseudomonas sp. ?
-
 ?
3.5.1.93 succinyl-7-aminocephalosporanic acid + H2O
-
 Pseudomonas sp. 7-aminocephalosporanate + succinate
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.1.93 heterodimer
-
 Pseudomonas sp.

Synonyms

EC Number Synonyms Comment Organism
3.5.1.93 cephalosporin acylase
-
 Pseudomonas sp.

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.1.93 0.015
-
 cephalosporin C at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 0.16
-
 4-(glutarylamino)benzoic acid at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 0.33
-
 gamma-Glu-Cys-Gly at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 0.35
-
 glutarylaniline at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 1.28
-
 succinyl-7-aminocephalosporanic acid at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 1.36
-
 glutaryl-GGGGAA at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 1.65
-
 glutaryl-glycine at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 1.73
-
 glutaryl-GGGGGK at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 3.1
-
 glutaryl-tryptophan at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 3.2
-
 glutaryl-gamma-Glu-Cys-Gly at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 4.8
-
 glutaryl-GDAADAADKG at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 5.4
-
 glutaryl-glycyl-proline at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 6.2
-
 glutaryl-alanyl-proline at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 7.1
-
 glutaryl-7-aminocephalosporanic acid at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 12.8
-
 glutaryl-EGDPPDLADQG at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 13.2
-
 glutaryl-GDPPDLADQG at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 13.8
-
 glutaryl-GDPP at 37°C, pH not specified in the publication Pseudomonas sp.

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.5.1.93 0.00068
-
 cephalosporin C at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 0.01
-
 glutarylaniline at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 0.04
-
 4-(glutarylamino)benzoic acid at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 0.1
-
 gamma-Glu-Cys-Gly at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 0.4
-
 succinyl-7-aminocephalosporanic acid at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 0.5
-
 glutaryl-gamma-Glu-Cys-Gly at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 0.53
-
 glutaryl-glycine at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 0.56
-
 glutaryl-tryptophan at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 0.81
-
 glutaryl-GGGGAA at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 4
-
 glutaryl-GGGGGK at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 4.7
-
 glutaryl-EGDPPDLADQG at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 6.4
-
 glutaryl-glycyl-proline at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 8.5
-
 glutaryl-alanyl-proline at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 12
-
 glutaryl-GDAADAADKG at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 14.2
-
 glutaryl-7-aminocephalosporanic acid at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 20
-
 glutaryl-GDPPDLADQG at 37°C, pH not specified in the publication Pseudomonas sp.
3.5.1.93 40
-
 glutaryl-GDPP at 37°C, pH not specified in the publication Pseudomonas sp.