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Literature summary extracted from
- Conformational flexibility of the C terminus with implications for substrate binding and catalysis revealed in a new crystal form of deacetoxycephalosporin C synthase (2004), J. Mol. Biol., 343, 157-171.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.14.20.1 | expression of wild-type and C-terminally truncated mutant enzymes as N-terminally His-tagged proteins in Escherichia coli strain BL21(DE3) | Streptomyces clavuligerus |
Crystallization (Commentary)
| EC Number | Crystallization (Comment) | Organism |
|---|---|---|
| 1.14.20.1 | recombinant N-terminally His-tagged wild-type and C-terminally truncated mutant enzymes, free enzyme, or complexing with Fe2+, or Fe2+/ampicillin, hanging drop vapour diffusion method, 4°C, precipitant solution: 100 mM HEPES-NaOH, pH 8.0, 0.9-1.1 M ammonium sulfate, the reservoir solution is covered with oil to retard the evaporation, cryoprotection by 30% v/v ethylene glycol in precipitant solution, X-ray structure determination and analysis at 2.3 A resolution, molecular modeling | Streptomyces clavuligerus |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 1.14.20.1 | additional information | shortening of the C-terminus by more than 4 residues reduces enzyme activity and alters the crystal structure from merohedrally twinned trimeric crystals to a non-trimeric structure with a free C-terminal arm, crystals show no twinning | Streptomyces clavuligerus |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.20.1 | 0.89 | - |
penicillin G | wild-type enzyme, pH 7.5, 30°C | Streptomyces clavuligerus |  |
| 1.14.20.1 | 20.6 | - |
penicillin G | recombinant His-tagged enzyme, pH 7.5, 30°C | Streptomyces clavuligerus | |
Localization
| EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.20.1 | Fe2+ | bound to the active site, forms a reactive iron-oxygen reaction intermediate during catalysis, complex structure in native and His-tagged enzyme | Streptomyces clavuligerus | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 1.14.20.1 | additional information | - |
gel filtration | Streptomyces clavuligerus |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.20.1 | penicillin N + 2-oxoglutarate + O2 | Streptomyces clavuligerus | - |
deacetoxycephalosporin C + succinate + CO2 + H2O | - |
ir | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.14.20.1 | Streptomyces clavuligerus | P18548 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.14.20.1 | recombinant N-terminally His-tagged wild-type and C-terminally truncated mutant enzymes from Escherichia coli strain BL21(DE3) | Streptomyces clavuligerus |
Reaction
| EC Number | Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|---|
| 1.14.20.1 | penicillin N + 2-oxoglutarate + O2 = deacetoxycephalosporin C + succinate + CO2 + H2O | reaction mechanism | Streptomyces clavuligerus |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.20.1 | penicillin G + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | phenylacetyl-7-aminodeacetoxycephalosporanic acid + succinate + CO2 + H2O | - |
ir | |
| 1.14.20.1 | penicillin N + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | deacetoxycephalosporin C + succinate + CO2 + H2O | - |
ir | |
| 1.14.20.1 | penicillin N + 2-oxoglutarate + O2 | oxidative ring expansion via reactive iron-oxygen intermediate | Streptomyces clavuligerus | deacetoxycephalosporin C + succinate + CO2 + H2O | - |
ir | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.14.20.1 | monomer | His-tagged enzyme in solution | Streptomyces clavuligerus |
| 1.14.20.1 | More | active site structure in the apo-enzyme | Streptomyces clavuligerus |
| 1.14.20.1 | trimer | wild-type enzyme in crystals | Streptomyces clavuligerus |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.14.20.1 | DAOCS | - |
Streptomyces clavuligerus |
| 1.14.20.1 | deacetoxycephalosporin C synthase | - |
Streptomyces clavuligerus |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.14.20.1 | 30 | - |
assay at | Streptomyces clavuligerus |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.20.1 | 0.056 | - |
penicillin G | recombinant His-tagged enzyme, pH 7.5, 30°C | Streptomyces clavuligerus | |
| 1.14.20.1 | 0.079 | - |
penicillin G | wild-type enzyme, pH 7.5, 30°C | Streptomyces clavuligerus | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.14.20.1 | 7.5 | - |
assay at | Streptomyces clavuligerus |
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Release 2023.1 (January 2023)
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