Cloned (Comment) | Organism |
---|---|
recombinant expression in in Escherichia coli JM109(DE3)/pET28, the enzyme performs self-activation | Pseudomonas sp. |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of 14 mutants with deletion in the alpha-C-terminal region, the majority of the fragment-deleted mutant proteins completely lose their activity due to failure of the first autocleavage, while mutant proteins D2 (227-AM-228 deletion) and D4 (212-ADLA-215 deletion) formally activate into mature enzyme with high activity. The Kcat/Kmvalues of mutant proteins D2 and D4 are 46% and 102% higher than that of wild-type control, respectively | Pseudomonas sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
51 | - |
cephalosporin C | deletion mutant D4(212-ADLA-215), pH 8.5, 37°C | Pseudomonas sp. | |
51.6 | - |
cephalosporin C | wild-type enzyme, pH 8.5, 37°C | Pseudomonas sp. | |
74.8 | - |
cephalosporin C | deletion mutant D2(227-AM-228), pH 8.5, 37°C | Pseudomonas sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. | - |
- |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
proteolytic modification | a first autoproteolytic cleavage site of the pro-enzyme is G239-S240, a second cleavage site is A232-S233, determined by tandem MS/MS analysis of the purified alpha-subunit C-terminus | Pseudomonas sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cephalosporin C + H2O | - |
Pseudomonas sp. | (7R)-7-aminocephalosporanate + 2-aminoadipate | - |
? |
Subunits | Comment | Organism |
---|---|---|
heterodimer | alphabeta | Pseudomonas sp. |
Synonyms | Comment | Organism |
---|---|---|
cephalosporin C acylase | - |
Pseudomonas sp. |
CPCAcy | - |
Pseudomonas sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Pseudomonas sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.022 | - |
cephalosporin C | wild-type enzyme and deletion mutant D4(212-ADLA-215), pH 8.5, 37°C | Pseudomonas sp. | |
0.023 | - |
cephalosporin C | deletion mutant D2(227-AM-228), pH 8.5, 37°C | Pseudomonas sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Pseudomonas sp. |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme is a member of the N-terminal nucleophile (Ntn) hydrolase superfamily, in which the precursor gene is translated into a single polypeptide chain and then folded into a self-activating pre-protein activated by intramolecular double cleavages | Pseudomonas sp. |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000315 | - |
cephalosporin C | deletion mutant D2(227-AM-228), pH 8.5, 37°C | Pseudomonas sp. | |
0.000419 | - |
cephalosporin C | wild-type enzyme, pH 8.5, 37°C | Pseudomonas sp. | |
0.000424 | - |
cephalosporin C | deletion mutant D4(212-ADLA-215), pH 8.5, 37°C | Pseudomonas sp. |