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 (Comment) | Organism |
---|---|
- |
Streptomyces clavuligerus |
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 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 | 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 Maximum [Da] | Comment | Organism |
---|---|---|---|
35000 | - |
gel filtration | Streptomyces clavuligerus |
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 (Comment) | Organism |
---|---|
native enzyme, partial, recombinant enzyme, to near homogenity | Streptomyces clavuligerus |
recombinant enzyme expressed in Escherichia coli | Streptomyces clavuligerus |
Renatured (Comment) | Organism |
---|---|
enzyme inactivated by 5,5-dithiobis-2-nitrobenzoic acid, complete reactivation by dithiothreitol | Streptomyces clavuligerus |
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 | 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 Maximum [°C] | Comment | Organism |
---|---|---|---|
36 | - |
- |
Streptomyces clavuligerus |
36 | - |
native and recombinant enzyme | Streptomyces clavuligerus |
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 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 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 maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.034 | - |
3-exomethylenecephalosporin C | - |
Streptomyces clavuligerus |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Streptomyces clavuligerus | isoelectric focusing | - |
5.3 |
Streptomyces clavuligerus | isoelectric focusing | - |
6.1 |