Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Rhodococcus sp. |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapor diffusion method, using 0.1 M phosphate-citrate (pH 4.2), 1.6 M sodium dihydrogen phosphate, and 0.4 M dipotassium hydrogen phosphate | Rhodococcus sp. |
Protein Variants | Comment | Organism |
---|---|---|
T172R/G173Q | the mutant has an improved in vitro half-life of about 6 h at 37°C | Rhodococcus sp. |
T172R/G173Q /G4C/S10C | the mutant shows improved catalytic efficiency against cocaine by about 20% | Rhodococcus sp. |
T172R/G173Q/L196C/I301C | the mutant has not only considerably extended the in vitro half-life at 37°C to more than 100 days, but also significantly improved catalytic efficiency against cocaine by about 150% | Rhodococcus sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0015 | - |
cocaine | mutant enzyme T172R/G173Q/L196C/I301C, at 37°C, pH not specified in the publication | Rhodococcus sp. | |
0.0021 | - |
cocaine | mutant enzyme T172R/G173Q/G4C/S10C, at 37°C, pH not specified in the publication | Rhodococcus sp. | |
0.0029 | - |
cocaine | mutant enzyme T172R/G173Q, at 37°C, pH not specified in the publication | Rhodococcus sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
cocaine + H2O | Rhodococcus sp. | - |
ecgonine methyl ester + benzoate | - |
? | |
cocaine + H2O | Rhodococcus sp. MB1 Bresler | - |
ecgonine methyl ester + benzoate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodococcus sp. | Q9L9D7 | - |
- |
Rhodococcus sp. MB1 Bresler | Q9L9D7 | - |
- |
Purification (Comment) | Organism |
---|---|
HisPur cobalt resin column chromatography | Rhodococcus sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cocaine + H2O | - |
Rhodococcus sp. | ecgonine methyl ester + benzoate | - |
? | |
cocaine + H2O | - |
Rhodococcus sp. MB1 Bresler | ecgonine methyl ester + benzoate | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | x-ray crystallography | Rhodococcus sp. |
Synonyms | Comment | Organism |
---|---|---|
cocE | - |
Rhodococcus sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
39 | - |
cocaine | mutant enzyme T172R/G173Q/G4C/S10C, at 37°C, pH not specified in the publication | Rhodococcus sp. | |
43.3 | - |
cocaine | mutant enzyme T172R/G173Q, at 37°C, pH not specified in the publication | Rhodococcus sp. | |
57.7 | - |
cocaine | mutant enzyme T172R/G173Q/L196C/I301C, at 37°C, pH not specified in the publication | Rhodococcus sp. |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
15330 | - |
cocaine | mutant enzyme T172R/G173Q, at 37°C, pH not specified in the publication | Rhodococcus sp. | |
18330 | - |
cocaine | mutant enzyme T172R/G173Q/G4C/S10C, at 37°C, pH not specified in the publication | Rhodococcus sp. | |
38330 | - |
cocaine | mutant enzyme T172R/G173Q/L196C/I301C, at 37°C, pH not specified in the publication | Rhodococcus sp. |