Application | Comment | Organism |
---|---|---|
synthesis | enzyme produces the 7-aminodeacetoxycephalosporanic acid precursor utilized in industrial applications, optimization of enzyme activity for activity with the substrate penicillin G | Streptomyces clavuligerus |
Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain ESS | Streptomyces clavuligerus |
Protein Variants | Comment | Organism |
---|---|---|
C281Y | random mutagenesis, exchange of residue surrounding the substrate, 2-6fold increased activity compared to the wild-type enzyme | Streptomyces clavuligerus |
G79E | random mutagenesis, exchange of residue surrounding the substrate, 2-6fold increased activity compared to the wild-type enzyme | Streptomyces clavuligerus |
I305L | site-directed mutagenesis, exchange of residue surrounding the substrate, 6-14fold increased activity compared to the wild-type enzyme | Streptomyces clavuligerus |
I305M | site-directed mutagenesis, exchange of residue surrounding the substrate, 6-14fold increased activity compared to the wild-type enzyme | Streptomyces clavuligerus |
N304K | site-directed mutagenesis, exchange of residue surrounding the substrate, 6-14fold increased activity compared to the wild-type enzyme | Streptomyces clavuligerus |
V275I | random mutagenesis, exchange of residue surrounding the substrate, 2-6fold increased activity compared to the wild-type enzyme | Streptomyces clavuligerus |
V275I/C281Y/I305M | site-directed mutagenesis, exchange of residue surrounding the substrate, over 32fold increased activity compared to the wild-type enzyme | Streptomyces clavuligerus |
V275I/I305M | site-directed mutagenesis, exchange of residue surrounding the substrate, 32fold increased activity compared to the wild-type enzyme | Streptomyces clavuligerus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.004 | - |
Penicillin N | pH 6.5, 30°C, recombinant mutant N304K | Streptomyces clavuligerus | |
0.006 | - |
Penicillin N | pH 6.5, 30°C, recombinant mutants C281Y and I305L | Streptomyces clavuligerus | |
0.009 | - |
Penicillin N | pH 6.5, 30°C, recombinant mutant G79E | Streptomyces clavuligerus | |
0.012 | - |
Penicillin N | pH 6.5, 30°C, recombinant mutants V275I and I305M | Streptomyces clavuligerus | |
0.013 | - |
Penicillin N | pH 6.5, 30°C, recombinant mutant V275I/I305M | Streptomyces clavuligerus | |
0.014 | - |
Penicillin N | pH 6.5, 30°C, recombinant wild-type enzyme | Streptomyces clavuligerus | |
0.22 | - |
penicillin G | pH 6.5, 30°C, recombinant mutant N304K | Streptomyces clavuligerus | |
0.25 | - |
penicillin G | pH 6.5, 30°C, recombinant mutant V275I/I305M | Streptomyces clavuligerus | |
0.66 | - |
penicillin G | pH 6.5, 30°C, recombinant mutant I305L | Streptomyces clavuligerus | |
0.68 | - |
penicillin G | pH 6.5, 30°C, recombinant mutant C281Y | Streptomyces clavuligerus | |
0.75 | - |
penicillin G | pH 6.5, 30°C, recombinant mutants G79E and I305M | Streptomyces clavuligerus | |
1.68 | - |
penicillin G | pH 6.5, 30°C, recombinant mutant V275I | Streptomyces clavuligerus | |
2.58 | - |
penicillin G | pH 6.5, 30°C, recombinant wild-type enzyme | Streptomyces clavuligerus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
penicillin G + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
phenylacetyl-7-aminodeacetoxycephalosporanic acid + succinate + CO2 + H2O | product is the precursor of 7-aminodeacetoxycephalosporanic acid, which is used in industrial applications | ir | |
penicillin N + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
deacetoxycephalosporin C + succinate + CO2 + H2O | - |
ir |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces clavuligerus | - |
gene cefE | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain ESS by ion exchange chromatography and gel filtration | Streptomyces clavuligerus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
activity of recombinant wild-type and mutant enzymes with substrate penicillin G and penicillin N | Streptomyces clavuligerus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
penicillin G + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | phenylacetyl-7-aminodeacetoxycephalosporanic acid + succinate + CO2 + H2O | - |
ir | |
penicillin G + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | phenylacetyl-7-aminodeacetoxycephalosporanic acid + succinate + CO2 + H2O | product is the precursor of 7-aminodeacetoxycephalosporanic acid, which is used in industrial applications | ir | |
penicillin N + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | deacetoxycephalosporin C + succinate + CO2 + H2O | - |
ir |
Synonyms | Comment | Organism |
---|---|---|
DAOCS | - |
Streptomyces clavuligerus |
deacetoxycephalosporin C synthase | - |
Streptomyces clavuligerus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Streptomyces clavuligerus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.032 | - |
penicillin G | pH 6.5, 30°C, recombinant mutant G79E | Streptomyces clavuligerus | |
0.045 | - |
penicillin G | pH 6.5, 30°C, recombinant wild-type enzyme | Streptomyces clavuligerus | |
0.05 | - |
penicillin G | pH 6.5, 30°C, recombinant mutant V275I | Streptomyces clavuligerus | |
0.056 | - |
penicillin G | pH 6.5, 30°C, recombinant mutant N304K | Streptomyces clavuligerus | |
0.074 | - |
penicillin G | pH 6.5, 30°C, recombinant mutant C281Y | Streptomyces clavuligerus | |
0.076 | - |
penicillin G | pH 6.5, 30°C, recombinant mutant I305L | Streptomyces clavuligerus | |
0.145 | - |
penicillin G | pH 6.5, 30°C, recombinant mutant I305M | Streptomyces clavuligerus | |
0.146 | - |
penicillin G | pH 6.5, 30°C, recombinant mutant V275I/I305M | Streptomyces clavuligerus | |
0.178 | - |
Penicillin N | pH 6.5, 30°C, recombinant mutant G79E | Streptomyces clavuligerus | |
0.252 | - |
Penicillin N | pH 6.5, 30°C, recombinant mutant V275I | Streptomyces clavuligerus | |
0.273 | - |
Penicillin N | pH 6.5, 30°C, recombinant mutant C281Y | Streptomyces clavuligerus | |
0.284 | - |
Penicillin N | pH 6.5, 30°C, recombinant mutant I305L | Streptomyces clavuligerus | |
0.307 | - |
Penicillin N | pH 6.5, 30°C, recombinant wild-type enzyme | Streptomyces clavuligerus | |
0.31 | - |
Penicillin N | pH 6.5, 30°C, recombinant mutant I305M | Streptomyces clavuligerus | |
0.316 | - |
Penicillin N | pH 6.5, 30°C, recombinant mutant V275I/I305M | Streptomyces clavuligerus | |
0.366 | - |
Penicillin N | pH 6.5, 30°C, recombinant mutant N304K | Streptomyces clavuligerus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
assay at, substrate penicillin G | Streptomyces clavuligerus |
7.5 | - |
assay at, substrate penicillin N | Streptomyces clavuligerus |