Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Pseudomonas sp. |
Protein Variants | Comment | Organism |
---|---|---|
E159D | the mutant is still active, but significantly retarded in the second autocleavage compared to the wild type enzyme | Pseudomonas sp. |
G160A | the mutant performs the second autocleavage more slowly than the wild type enzyme | Pseudomonas sp. |
G160L | the mutant performs the second autocleavage more slowly than the wild type enzyme | Pseudomonas sp. |
S170C | the mutant simultaneously loses the activities for both the second autocleavage and substrate hydrolysis | Pseudomonas sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.35 | - |
glutaryl-GDPP | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
0.4 | - |
glutaryl-GDAADAADKG | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
0.43 | - |
glutaryl-GGGGGK | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
0.5 | - |
glutaryl-7-aminocephalosporanic acid | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
0.67 | - |
glutaryl-GDPPDLADQG | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
0.73 | - |
glutaryl-alanyl-proline | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
0.85 | - |
glutaryl-glycyl-proline | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
1.68 | - |
glutaryl-GGGGAA | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
2.7 | - |
glutaryl-EGDPPDLADQG | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
3.1 | - |
glutaryl-glycine | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
3.2 | - |
succinyl-7-aminocephalosporanic acid | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
3.3 | - |
gamma-Glu-Cys-Gly | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
4.5 | - |
4-(glutarylamino)benzoic acid | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
6.1 | - |
glutaryl-tryptophan | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
6.4 | - |
glutaryl-gamma-Glu-Cys-Gly | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
22 | - |
cephalosporin C | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
40 | - |
glutarylaniline | at 37°C, pH not specified in the publication | Pseudomonas sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. | - |
- |
- |
Purification (Comment) | Organism |
---|---|
acetone precipitation and Sepharose CM column chromatography | Pseudomonas sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-(glutarylamino)benzoic acid + H2O | - |
Pseudomonas sp. | glutarate + 4-aminobenzoate | - |
? | |
cephalosporin C + H2O | - |
Pseudomonas sp. | cephalosporamic acid + 2-amino-5-hydroxypentanoate | - |
? | |
DGDPP + H2O | 20% activity compared to E-GDPPDLADQG | Pseudomonas sp. | L-Asp + GDPP | - |
? | |
EGDPP + H2O | 67% activity compared to E-GDPPDLADQG | Pseudomonas sp. | L-Glu + GDPP | - |
? | |
EGDPPDLADQG + H2O | 100% activity | Pseudomonas sp. | L-Glu + GDPPDLADQG | - |
? | |
gamma-Glu-Cys-Gly | - |
Pseudomonas sp. | L-glutamate + Cys-Gly | - |
? | |
GEGDPP + H2O | 5% activity compared to E-GDPPDLADQG | Pseudomonas sp. | Gly-L-Glu + GDPP | - |
? | |
glutaryl-7-aminocephalosporanic acid + H2O | - |
Pseudomonas sp. | 7-aminocephalosporanate + glutarate | - |
? | |
glutaryl-alanyl-proline + H2O | - |
Pseudomonas sp. | glutarate + alanyl-proline | - |
? | |
glutaryl-EGDPPDLADQG + H2O | - |
Pseudomonas sp. | glutarate + EGDPPDLADQG | - |
? | |
glutaryl-gamma-Glu-Cys-Gly + H2O | - |
Pseudomonas sp. | glutarate + gamma-Glu-Cys-Gly | - |
? | |
glutaryl-GDAADAADKG + H2O | - |
Pseudomonas sp. | glutarate + GDAADAADKG | - |
? | |
glutaryl-GDPP + H2O | substrate with highest catalytic efficiency | Pseudomonas sp. | glutarate + GDPP | - |
? | |
glutaryl-GDPPDLADQG + H2O | - |
Pseudomonas sp. | glutarate + GDPPDLADQG | - |
? | |
glutaryl-GGGGAA + H2O | - |
Pseudomonas sp. | glutarate + GGGGAA | - |
? | |
glutaryl-GGGGGK + H2O | - |
Pseudomonas sp. | glutarate + GGGGGK | - |
? | |
glutaryl-glycine + H2O | - |
Pseudomonas sp. | glutarate + glycine | - |
? | |
glutaryl-glycyl-L-proline + H2O | - |
Pseudomonas sp. | glutarate + glycyl-L-proline | - |
? | |
glutaryl-glycyl-proline + H2O | - |
Pseudomonas sp. | glutarate + glycylproline | - |
? | |
glutaryl-tryptophan + H2O | - |
Pseudomonas sp. | glutarate + tryptophan | - |
? | |
glutarylaniline + H2O | - |
Pseudomonas sp. | glutarate + aniline | - |
? | |
additional information | less than 1% activity with TLGEGDPP compared to EGDPPDLADQG, no activity with adipyl-7-aminocephalosporanic acid | Pseudomonas sp. | ? | - |
? | |
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. | ? | - |
? | |
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. | ? | - |
? | |
succinyl-7-aminocephalosporanic acid + H2O | - |
Pseudomonas sp. | 7-aminocephalosporanate + succinate | - |
? |
Subunits | Comment | Organism |
---|---|---|
heterodimer | - |
Pseudomonas sp. |
Synonyms | Comment | Organism |
---|---|---|
cephalosporin acylase | - |
Pseudomonas sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.015 | - |
cephalosporin C | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
0.16 | - |
4-(glutarylamino)benzoic acid | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
0.33 | - |
gamma-Glu-Cys-Gly | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
0.35 | - |
glutarylaniline | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
1.28 | - |
succinyl-7-aminocephalosporanic acid | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
1.36 | - |
glutaryl-GGGGAA | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
1.65 | - |
glutaryl-glycine | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
1.73 | - |
glutaryl-GGGGGK | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
3.1 | - |
glutaryl-tryptophan | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
3.2 | - |
glutaryl-gamma-Glu-Cys-Gly | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
4.8 | - |
glutaryl-GDAADAADKG | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
5.4 | - |
glutaryl-glycyl-proline | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
6.2 | - |
glutaryl-alanyl-proline | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
7.1 | - |
glutaryl-7-aminocephalosporanic acid | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
12.8 | - |
glutaryl-EGDPPDLADQG | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
13.2 | - |
glutaryl-GDPPDLADQG | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
13.8 | - |
glutaryl-GDPP | at 37°C, pH not specified in the publication | Pseudomonas sp. |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00068 | - |
cephalosporin C | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
0.01 | - |
glutarylaniline | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
0.04 | - |
4-(glutarylamino)benzoic acid | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
0.1 | - |
gamma-Glu-Cys-Gly | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
0.4 | - |
succinyl-7-aminocephalosporanic acid | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
0.5 | - |
glutaryl-gamma-Glu-Cys-Gly | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
0.53 | - |
glutaryl-glycine | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
0.56 | - |
glutaryl-tryptophan | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
0.81 | - |
glutaryl-GGGGAA | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
4 | - |
glutaryl-GGGGGK | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
4.7 | - |
glutaryl-EGDPPDLADQG | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
6.4 | - |
glutaryl-glycyl-proline | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
8.5 | - |
glutaryl-alanyl-proline | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
12 | - |
glutaryl-GDAADAADKG | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
14.2 | - |
glutaryl-7-aminocephalosporanic acid | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
20 | - |
glutaryl-GDPPDLADQG | at 37°C, pH not specified in the publication | Pseudomonas sp. | |
40 | - |
glutaryl-GDPP | at 37°C, pH not specified in the publication | Pseudomonas sp. |