EC Number | Application | Comment | Organism |
---|---|---|---|
3.5.1.93 | synthesis | the enzyme mutant may be suitable for industrial application of the mono-step process for cephalosporin C conversion to (7R)-7-aminocephalosporanate, which is the precursor for production of semisynthetic cephalosporins | Pseudomonas sp. |
EC Number | Cloned (Comment) | Organism |
---|---|---|
3.5.1.93 | phylogenetic analysis, recombinant exxpression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)pLysS | Pseudomonas sp. |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
3.5.1.93 | H57betaS/H70betaS | site-directed mutagenesis, the engineered mutant shows increased Vmax on cephalosporin C compared to the wild-type enzyme | Pseudomonas sp. |
3.5.1.93 | H57betaS/H70betaS/A215alphaY | site-directed mutagenesis, the engineered mutant shows increased Vmax on cephalosporin C compared to the wild-type enzyme | Pseudomonas sp. |
3.5.1.93 | H57betaS/H70betaS/F58betaD | random mutagenesis of mutant H57betaS/H70betaS, the mutant shows altered substrate specificity compared to the wild-type enzyme | Pseudomonas sp. |
3.5.1.93 | H57betaS/H70betaS/F58betaI | random mutagenesis of mutant H57betaS/H70betaS, the mutant shows altered substrate specificity compared to the wild-type enzyme | Pseudomonas sp. |
3.5.1.93 | H57betaS/H70betaS/F72betaR | random mutagenesis of mutant H57betaS/H70betaS, the mutant shows altered substrate specificity compared to the wild-type enzyme | Pseudomonas sp. |
3.5.1.93 | H57betaS/H70betaS/I176betaK | random mutagenesis of mutant H57betaS/H70betaS, the mutant shows altered substrate specificity compared to the wild-type enzyme | Pseudomonas sp. |
3.5.1.93 | H57betaS/H70betaS/I176betaQ | random mutagenesis of mutant H57betaS/H70betaS, the mutant shows altered substrate specificity compared to the wild-type enzyme | Pseudomonas sp. |
3.5.1.93 | H57betaS/H70betaS/I176betaT | random mutagenesis of mutant H57betaS/H70betaS, the mutant shows altered substrate specificity compared to the wild-type enzyme | Pseudomonas sp. |
3.5.1.93 | H57betaS/H70betaS/L154betaY | random mutagenesis of mutant H57betaS/H70betaS, the mutant shows altered substrate specificity compared to the wild-type enzyme | Pseudomonas sp. |
3.5.1.93 | H57betaS/H70betaS/L154betaY | site-directed mutagenesis, interest in designing a single step enzymatic conversion of cephalosporin C to (7R)-7-aminocephalosporanate catalyzed by a true cephalosporin C acylase, the engineered mutant displays highly enhanced catalytic efficiency, with a 11000fold increase in specificity constant for CephC versus glutaryl-7-amino cephalosporanic acid, on cephalosporin C over glutaryl-7-aminocephalosporanic acid compared to the wild-type enzyme under conditions resembling those used at industrial level because of its high kinetic efficiency and the absence of substrate or product inhibition effects | Pseudomonas sp. |
3.5.1.93 | H57betaS/H70betaS/L154betaY/M165alphaS | site-directed mutagenesis, the engineered mutant shows increased Vmax on cephalosporin C compared to the wild-type enzyme | Pseudomonas sp. |
3.5.1.93 | H57betaS/H70betaS/L175betaT | random mutagenesis of mutant H57betaS/H70betaS, the mutant shows altered substrate specificity compared to the wild-type enzyme | Pseudomonas sp. |
3.5.1.93 | H57betaS/H70betaS/M165alphaS | site-directed mutagenesis, the engineered mutant shows increased Vmax on cephalosporin C compared to the wild-type enzyme | Pseudomonas sp. |
3.5.1.93 | H57betaS/H70betaS/M165alphaS/I176betaS | site-directed mutagenesis, the engineered mutant shows increased Vmax on cephalosporin C compared to the wild-type enzyme | Pseudomonas sp. |
3.5.1.93 | H57betaS/H70betaS/R24betaP | random mutagenesis of mutant H57betaS/H70betaS, the mutant shows altered substrate specificity compared to the wild-type enzyme | Pseudomonas sp. |
3.5.1.93 | H57betaS/H70betaS/V25betaR | random mutagenesis of mutant H57betaS/H70betaS, the mutant shows altered substrate specificity compared to the wild-type enzyme | Pseudomonas sp. |
3.5.1.93 | additional information | mutant screening of H57betaS/H70betaS triple mutants generated by site-saturation mutagenesis, introduction of multiple mutations, overview | Pseudomonas sp. |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
3.5.1.93 | (7R)-7-(4-carboxybutanamido)cephalosporanate | substrate inhibition | Pseudomonas sp. | |
3.5.1.93 | (7R)-7-aminocephalosporanate | product inhibition | Pseudomonas sp. | |
3.5.1.93 | cephalosporin C | substrate inhibition | Pseudomonas sp. |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.5.1.93 | additional information | - |
additional information | kinetics of wild-type and mutant enzymes with cephalosporin C, overview | Pseudomonas sp. | |
3.5.1.93 | 0.4 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS/F58betaD, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 0.41 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS/M165alphaS/I176betaS, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 0.6 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS/L176betaK, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 1.5 | - |
cephalosporin C | recombinant mutant H57betaS/H70betaS/F58betaD, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 1.5 | - |
cephalosporin C | recombinant mutant H57betaS/H70betaS/L176betaC, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 1.5 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant wild-type enzyme, pH 8.0, 25°C | Pseudomonas sp. | |
3.5.1.93 | 1.8 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS/L176betaT, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 1.8 | - |
cephalosporin C | recombinant mutant H57betaS/H70betaS/M165alphaS/I176betaS, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 2 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS/F72betaR, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 2 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS/L154betaY, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 2 | - |
cephalosporin C | recombinant mutant H57betaS/H70betaS/R24betaP, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 2.3 | - |
cephalosporin C | recombinant mutant H57betaS/H70betaS/V25betaR, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 3.1 | - |
cephalosporin C | recombinant mutant H57betaS/H70betaS/L154betaY/M165alphaS, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 3.2 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS/L176betaQ, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 3.6 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS/V25betaR, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 3.8 | - |
cephalosporin C | recombinant mutant H57betaS/H70betaS/L175betaT, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 3.8 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS/L175betaT, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 4.5 | - |
cephalosporin C | recombinant mutant H57betaS/H70betaS/L154betaY, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 5.4 | - |
cephalosporin C | recombinant mutant H57betaS/H70betaS/L176betaT, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 6.3 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS/F58betaI, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 6.6 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS/L154betaY/M165alphaS, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 6.9 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS, pH 8.0, 25°C | Pseudomonas sp. | |
3.5.1.93 | 7.5 | - |
cephalosporin C | recombinant mutant H57betaS/H70betaS/L176betaK, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 9.2 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS/R24betaP, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 9.5 | - |
cephalosporin C | recombinant mutant wild-type enzyme, pH 8.0, 25°C | Pseudomonas sp. | |
3.5.1.93 | 12.2 | - |
cephalosporin C | recombinant mutant H57betaS/H70betaS, pH 8.0, 25°C | Pseudomonas sp. | |
3.5.1.93 | 16.4 | - |
cephalosporin C | recombinant mutant H57betaS/H70betaS/F72betaR, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 21.9 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS/M165alphaS, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 27.7 | - |
cephalosporin C | recombinant mutant H57betaS/H70betaS/M165alphaS, pH 8.0, 37°C | Pseudomonas sp. | |
3.5.1.93 | 33.7 | - |
cephalosporin C | recombinant mutant H57betaS/H70betaS/F58betaI, pH 8.0, 37°C | Pseudomonas sp. |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.5.1.93 | Pseudomonas sp. | - |
- |
- |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.5.1.93 | (7R)-7-(4-carboxybutanamido)cephalosporanate + H2O | - |
Pseudomonas sp. | (7R)-7-aminocephalosporanate + glutarate | - |
? | |
3.5.1.93 | cephalosporin C + H2O | the wild-type enzyme shows weak activity with cephalosporin C, but some enzyme mutants show increased activity | Pseudomonas sp. | (7R)-7-aminocephalosporanate + 2-aminoadipate | - |
? | |
3.5.1.93 | additional information | the amino acid moiety of cephalosporin C that are potentially involved in the enzyme substrate selectivity are Met165alpha, His23beta, Arg24beta, Val25beta, Tyr32beta, Phe55beta, Phe58beta, Phe72beta, Leu154beta, Leu175beta, Ile176beta and His178beta, substrate docking in the active site, overview | Pseudomonas sp. | ? | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
3.5.1.93 | cephalosporin C acylase | - |
Pseudomonas sp. |
3.5.1.93 | glutaryl acylase | - |
Pseudomonas sp. |
3.5.1.93 | VAC acylase | - |
Pseudomonas sp. |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.5.1.93 | 25 | - |
assay at | Pseudomonas sp. |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
3.5.1.93 | 8 | - |
assay at | Pseudomonas sp. |
EC Number | Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.5.1.93 | 1 | - |
(7R)-7-aminocephalosporanate | recombinant mutant H57betaS/H70betaS/I176betaT, pH 8.0, 25°C | Pseudomonas sp. | |
3.5.1.93 | 2 | - |
(7R)-7-aminocephalosporanate | recombinant mutant H57betaS/H70betaS/R24betaP, pH 8.0, 25°C | Pseudomonas sp. | |
3.5.1.93 | 6.3 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS/F72betaR, pH 8.0, 25°C | Pseudomonas sp. | |
3.5.1.93 | 6.9 | - |
cephalosporin C | recombinant mutant H57betaS/H70betaS/L154betaY/M165alphaS, pH 8.0, 25°C | Pseudomonas sp. | |
3.5.1.93 | 21 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | wild-type enzyme, pH 8.0, 25°C | Pseudomonas sp. | |
3.5.1.93 | 21 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS/I176betaT, pH 8.0, 25°C | Pseudomonas sp. | |
3.5.1.93 | 33 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS/I176betaQ, pH 8.0, 25°C | Pseudomonas sp. | |
3.5.1.93 | 65 | - |
(7R)-7-(4-carboxybutanamido)cephalosporanate | recombinant mutant H57betaS/H70betaS/I176betaK, pH 8.0, 25°C | Pseudomonas sp. | |
3.5.1.93 | 77 | - |
cephalosporin C | recombinant mutant H57betaS/H70betaS/I176betaT, pH 8.0, 25°C | Pseudomonas sp. |
EC Number | General Information | Comment | Organism |
---|---|---|---|
3.5.1.93 | evolution | phylogenetic analysis and active site evolution concerning cephalosporin C binding, overview | Pseudomonas sp. |
3.5.1.93 | additional information | substrate binding, molecular modeling study, overview | Pseudomonas sp. |