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Literature summary for 3.5.1.93 extracted from
- Computational design of variants for cephalosporin C acylase from Pseudomonas strain N176 with improved stability and activity (2017), Appl. Microbiol. Biotechnol., 101, 621-632 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Pseudomonas sp. N176 |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| H57betaS/H70betaS | naturally occuring mutations, the two mutations increase the activity by fourfold. Structure-activity modelling using wild-type and mutant enzymes, overview. Design of N176 cephalosporin acylase at active site and structure model of predicted mutation V68betaA | Pseudomonas sp. N176 |
| L154betaF | site-directed mutagenesis, mutant M8, the mutant displays improved stability and activity compared to the wild-type | Pseudomonas sp. N176 |
| L154betaF/L180betaF | site-directed mutagenesis, mutant M10, the mutant displays improved stability and activity compared to the wild-type | Pseudomonas sp. N176 |
| L154betaF/Y167betaF/L180betaF | site-directed mutagenesis, the mutant displays improved stability and activity compared to the wild-type | Pseudomonas sp. N176 |
| L180betaF | site-directed mutagenesis, mutant M9, the mutant displays improved stability and activity compared to the wild-type | Pseudomonas sp. N176 |
| M31betaF/H57betaS/H70betaS | site-directed mutagenesis, mutant M1, the mutant displays improved stability and activity compared to the mutant M2 | Pseudomonas sp. N176 |
| M31betaF/H57betaS/V68betaA/H70betaS | site-directed mutagenesis, mutant M2, the mutant shows a decrease in stability compared to wild-type, which is largely owing to the mutation V68betaA at the active site | Pseudomonas sp. N176 |
| M31betaF/H57betaS/V68betaA/H70betaS/L154betaF/L180betaF | site-directed mutagenesis | Pseudomonas sp. N176 |
| additional information | mutation V68betaA at the active site causes a loss in protein stability. The lost stability because is recovered by introducing mutations L154betaF and L180betaF at hydrophobic core regions. Mutation V68betaA in the six-residue mutant provides more space to accommodate the bulky side chain of cephalosporin C, which can help in designing cephalosporin C acylase mutants with higher activities and the practical one-step enzymatic route to prepare 7-aminocephalosporanic acid at industrial-scale levels | Pseudomonas sp. N176 |
| Y167betaF | site-directed mutagenesis, the mutant displays improved stability and activity compared to the wild-type | Pseudomonas sp. N176 |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 9.3 | - |
cephalosporin C | recombinant mutant L154betaF/L180betaF, pH 8.0, 37°C | Pseudomonas sp. N176 |  |
| 9.73 | - |
cephalosporin C | recombinant mutant L154betaF, pH 8.0, 37°C | Pseudomonas sp. N176 | |
| 10.14 | - |
cephalosporin C | recombinant mutant L180betaF, pH 8.0, 37°C | Pseudomonas sp. N176 | |
| 12.38 | - |
cephalosporin C | recombinant mutant M31betaF/H57betaS/V68betaA/H70betaS, pH 8.0, 37°C | Pseudomonas sp. N176 | |
| 12.65 | - |
cephalosporin C | recombinant mutant M31betaF/H57betaS/H70betaS, pH 8.0, 37°C | Pseudomonas sp. N176 | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (7R)-7-(4-carboxybutanamido)cephalosporanate + H2O | Pseudomonas sp. N176 | - |
(7R)-7-aminocephalosporanate + glutarate | - |
? | |
| cephalosporin C + H2O | Pseudomonas sp. N176 | low activity | 7-aminocephalosporanic acid + 2-amino-5-hydroxypentanoate | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pseudomonas sp. N176 | A0A1D8GRD5 | - |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (7R)-7-(4-carboxybutanamido)cephalosporanate + H2O | - |
Pseudomonas sp. N176 | (7R)-7-aminocephalosporanate + glutarate | - |
? | |
| (7R)-7-(4-carboxybutanamido)cephalosporanate + H2O | GL-7-ACA | Pseudomonas sp. N176 | (7R)-7-aminocephalosporanate + glutarate | - |
? | |
| cephalosporin C + H2O | low activity | Pseudomonas sp. N176 | 7-aminocephalosporanic acid + 2-amino-5-hydroxypentanoate | - |
? | |
| cephalosporin C + H2O | CPC, low activity | Pseudomonas sp. N176 | 7-aminocephalosporanic acid + 2-amino-5-hydroxypentanoate | - |
? | |
| additional information | the D-2-aminoadipyl side chain of CPC is much larger than the glutaryl side chain of GL-7-ACA, and the activity of N176 acylase toward CPC is only 4% compared with the activity toward GL-7-ACA | Pseudomonas sp. N176 | ? | - |
- |
|
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| cephalosporin C acylase | - |
Pseudomonas sp. N176 |
| N176 cephalosporin C acylase | - |
Pseudomonas sp. N176 |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Pseudomonas sp. N176 |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
assay at | Pseudomonas sp. N176 |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | structure-activity modelling using wild-type and mutant enzymes, active site structures, overview | Pseudomonas sp. N176 |
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