Application | Comment | Organism |
---|---|---|
medicine | enzyme can activate prodrug 7-ethyl-10-[4-(1-piperidino)-1-piperidino] carbonyloxycamptothecin to give 7-ethyl-10-hydroxycampothecin, i.e. SN-38, a topoisomerase inhibitor used in cancer therapy | Geobacillus stearothermophilus |
Crystallization (Comment) | Organism |
---|---|
at 2.0 and 1.58 A resolution. Enzyme folds into a catalytic domain, an alpha/beta domain and a regulatory domain. Residue C408 adjacent to the active site is triply oxidized and may have a regulatory role | Geobacillus stearothermophilus |
Protein Variants | Comment | Organism |
---|---|---|
C408A | kinetics similar to wild-type | Geobacillus stearothermophilus |
C408G | 4fold decrease in kcat value | Geobacillus stearothermophilus |
C408P | kinetics similar to wild-type | Geobacillus stearothermophilus |
C408S | 4fold increase in Km value | Geobacillus stearothermophilus |
C408V | kinetics similar to wild-type | Geobacillus stearothermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0186 | - |
7-ethyl-10-[4-(1-piperidino)-1-piperidino] carbonyloxycamptothecin | wild-type, pH 8.5 | Geobacillus stearothermophilus | |
0.0206 | - |
7-ethyl-10-[4-(1-piperidino)-1-piperidino] carbonyloxycamptothecin | mutant C408P, pH 8.5 | Geobacillus stearothermophilus | |
0.0223 | - |
7-ethyl-10-[4-(1-piperidino)-1-piperidino] carbonyloxycamptothecin | mutant C408V, pH 8.5 | Geobacillus stearothermophilus | |
0.0224 | - |
7-ethyl-10-[4-(1-piperidino)-1-piperidino] carbonyloxycamptothecin | mutant C408G, pH 8.5 | Geobacillus stearothermophilus | |
0.0231 | - |
4-nitrophenyl butanoate | mutant C408G, pH 8.5 | Geobacillus stearothermophilus | |
0.0242 | - |
4-nitrophenyl butanoate | wild-type, pH 8.5 | Geobacillus stearothermophilus | |
0.026 | - |
4-nitrophenyl butanoate | mutant C408P, pH 8.5 | Geobacillus stearothermophilus | |
0.0283 | - |
4-nitrophenyl butanoate | mutant C408V, pH 8.5 | Geobacillus stearothermophilus | |
0.0328 | - |
4-nitrophenyl butanoate | mutant C408A, pH 8.5 | Geobacillus stearothermophilus | |
0.0453 | - |
7-ethyl-10-[4-(1-piperidino)-1-piperidino] carbonyloxycamptothecin | mutant C408A, pH 8.5 | Geobacillus stearothermophilus | |
0.0971 | - |
4-nitrophenyl butanoate | mutant C408S, pH 8.5 | Geobacillus stearothermophilus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Geobacillus stearothermophilus | - |
isoform Est55 | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl butanoate + H2O | - |
Geobacillus stearothermophilus | 4-nitrophenol + butanoate | - |
? | |
7-ethyl-10-[4-(1-piperidino)-1-piperidino] carbonyloxycamptothecin + H2O | i.e. irinotecan, CPT-11, prodrug | Geobacillus stearothermophilus | 7-ethyl-10-hydroxycampothecin + ? | i.e. SN-38, topoisomerase inhibitor used in cancer therapy | ? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000022 | - |
4-nitrophenyl butanoate | mutant C408G, pH 8.5 | Geobacillus stearothermophilus | |
0.000065 | - |
4-nitrophenyl butanoate | mutant C408P, pH 8.5 | Geobacillus stearothermophilus | |
0.000079 | - |
4-nitrophenyl butanoate | wild-type, pH 8.5 | Geobacillus stearothermophilus | |
0.000107 | - |
4-nitrophenyl butanoate | mutant C408V, pH 8.5 | Geobacillus stearothermophilus | |
0.000127 | - |
4-nitrophenyl butanoate | mutant C408S, pH 8.5 | Geobacillus stearothermophilus | |
0.000137 | - |
4-nitrophenyl butanoate | mutant C408A, pH 8.5 | Geobacillus stearothermophilus |