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Literature summary for 1.14.20.1 extracted from
- Purification and properties of deacetoxycephalosporin C synthase from recombinant Escherichia coli and its comparison with the native enzyme purified from Streptomyces clavuligerus (1989), J. Biol. Chem., 264, 10219-10227.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| ascorbate | - |
Streptomyces clavuligerus |  |
| ascorbate | stimulates activity of native and recombinant enzyme | Streptomyces clavuligerus | |
| dithiothreitol | - |
Streptomyces clavuligerus | |
| dithiothreitol | stimulates activity of native and recombinant enzyme | Streptomyces clavuligerus | |
| additional information | no stimulation by ATP | Streptomyces clavuligerus |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
- |
Streptomyces clavuligerus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 1,10-phenanthroline | - |
Streptomyces clavuligerus | |
| 3-exomethylenecephalosporin | strong, substrate for hydroxylation reaction of enzyme, EC 1.14.11.26 | Streptomyces clavuligerus | |
| 3-exomethylenecephalosporin C | 0.28 mM, 71% inhibition of the formation of deacetoxycephalosporin C | Streptomyces clavuligerus | |
| 5,5'-dithiobis-2-nitrobenzoic acid | 0.01 mM, 30% inhibition, reactivation by dithiothreitol | Streptomyces clavuligerus | |
| ampicillin | 2.8 mM, 13% inhibition of the formation of deacetoxycephalosporin C | Streptomyces clavuligerus | |
| Co2+ | inhibition in decreasing order, Zn2+, Co2+, Ni2+ | Streptomyces clavuligerus | |
| DTNB | - |
Streptomyces clavuligerus | |
| EDTA | 0.02 mM, 35% inhibition | Streptomyces clavuligerus | |
| iodoacetic acid | 1 mM, 40% inhibition | Streptomyces clavuligerus | |
| N-ethylmaleimide | 0.01 mM, 45% inhibition | Streptomyces clavuligerus | |
| NEM | - |
Streptomyces clavuligerus | |
| Ni2+ | inhibition in decreasing order, Zn2+, Co2+, Ni2+ | Streptomyces clavuligerus | |
| o-phenanthroline | 0.02 mM, 37% inhibition | Streptomyces clavuligerus | |
| p-hydroxymercuribenzoate | 0.01 mM, 100% inhibition | Streptomyces clavuligerus | |
| penicillin G | 2.8 mM, 57% inhibition of the formation of deacetoxycephalosporin C | Streptomyces clavuligerus | |
| Penicillin V | 2.8 mM, 47% inhibition of the formation of deacetoxycephalosporin C | Streptomyces clavuligerus | |
| Zn2+ | inhibition in decreasing order, Zn2+, Co2+, Ni2+ | Streptomyces clavuligerus | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.018 | - |
2-oxoglutarate | - |
Streptomyces clavuligerus | |
| 0.018 | - |
2-oxoglutarate | recombinant enzyme | Streptomyces clavuligerus | |
| 0.022 | - |
2-oxoglutarate | native enzyme | Streptomyces clavuligerus | |
| 0.028 | - |
deacetoxycephalosporin C | - |
Streptomyces clavuligerus | |
| 0.029 | - |
Penicillin N | - |
Streptomyces clavuligerus | |
| 0.029 | - |
Penicillin N | recombinant enzyme | Streptomyces clavuligerus | |
| 0.035 | - |
Penicillin N | native enzyme | Streptomyces clavuligerus | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Fe2+ | required, Ka-value 0.008 mM | Streptomyces clavuligerus | |
| Fe3+ | can replace Fe2+ under reducing conditions | Streptomyces clavuligerus | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 35000 | - |
gel filtration | Streptomyces clavuligerus |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Streptomyces clavuligerus | - |
- |
- |
| Streptomyces clavuligerus | - |
native enzyme and recombinant enzyme expressed in Escherichia coli, enzyme catalyzes both reaction of EC 1.14.20.1 and of EC 1.14.11.26 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| native enzyme, partial, recombinant enzyme, to near homogenity | Streptomyces clavuligerus |
| recombinant enzyme expressed in Escherichia coli | Streptomyces clavuligerus |
Renatured (Commentary)
| Renatured (Comment) | Organism |
|---|---|
| enzyme inactivated by 5,5-dithiobis-2-nitrobenzoic acid, complete reactivation by dithiothreitol | Streptomyces clavuligerus |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.166 | - |
native enzyme | Streptomyces clavuligerus |
| 0.432 | - |
recombinant enzyme | Streptomyces clavuligerus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 3-exomethylenecephalosporin C + 2-oxoglutarate + O2 | 8% of the activity with cephalosporin N, recombinant enzyme | Streptomyces clavuligerus | deacetylcephalosporin C + succinate + CO2 + H2O | - |
? | |
| ampicillin + 2-oxoglutarate + O2 | no activity | Streptomyces clavuligerus | cephalexin + succinate + CO2 + H2O | - |
? | |
| additional information | no substrate: isopenicillin N, penicillin G, penicillin V, ampicillin, 6-aminopenicillanic acid | Streptomyces clavuligerus | ? | - |
? | |
| penicillin G + 2-oxoglutarate + O2 | no activity | Streptomyces clavuligerus | phenylacetyl-7-aminodeacetoxy cephalosporanic acid + succinate + CO2 + H2O | - |
? | |
| penicillin N + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | deacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
| penicillin V + 2-oxoglutarate + O2 | no activity | Streptomyces clavuligerus | ? + succinate + CO2 + H2O | - |
? | |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 36 | - |
- |
Streptomyces clavuligerus |
| 36 | - |
native and recombinant enzyme | Streptomyces clavuligerus |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | - |
stable up to | Streptomyces clavuligerus |
| 40 | - |
10 min, 50% loss of activity | Streptomyces clavuligerus |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
- |
Streptomyces clavuligerus |
| 7 | - |
in piperazineethanesulfonic acid buffer, recombinant enzyme | Streptomyces clavuligerus |
| 7.4 | - |
native enzyme | Streptomyces clavuligerus |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 6.5 | 9 | - |
Streptomyces clavuligerus |
| 6.5 | 9 | 4°C, 1 h, stable, native enzyme | Streptomyces clavuligerus |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.034 | - |
3-exomethylenecephalosporin C | - |
Streptomyces clavuligerus | |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Streptomyces clavuligerus | isoelectric focusing | - |
5.3 |
| Streptomyces clavuligerus | isoelectric focusing | - |
6.1 |
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