Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL-21 Gold (DE3) cells | Rhodococcus sp. |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapor diffusion method, using 20% (w/v) PEG 3350, 100 mM 2-(N-morpholino)-ethane sulfonic acid, pH 6.0, and 1 M NaCl | Rhodococcus sp. |
Protein Variants | Comment | Organism |
---|---|---|
G173Q/L169K | the mutant has a half-life of 370 min and 2.9 days at 37°C | Rhodococcus sp. |
G4C/S10C | the mutant shows about 4fold reduced catalytic efficiency compared to the wild type enzyme. The mutant retains almost all activity after 7 days of 37°C treatment | Rhodococcus sp. |
T172R/G173Q | the mutant shows about 4fold reduced catalytic efficiency compared to the wild type enzyme. The mutant remains more than 90% active for longer than 40 days at 37°C, representing a more than 4700fold improvement over wild type. PEGylated mutant enzyme retains full in vitro enzymatic activity | Rhodococcus sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0057 | - |
cocaine | wild type enzyme, pH not specified in the publication, 37°C | Rhodococcus sp. | |
0.019 | - |
cocaine | pegylated mutant enzyme T172R/G173Q, pH not specified in the publication, 37°C | Rhodococcus sp. | |
0.021 | - |
cocaine | mutant enzyme G4C/S10C, pH not specified in the publication, 37°C | Rhodococcus sp. | |
0.026 | - |
cocaine | mutant enzyme T172R/G173Q, pH not specified in the publication, 37°C | Rhodococcus sp. |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
127000 | - |
- |
Rhodococcus sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodococcus sp. | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Talon metal chelate column chromatography and Q-Sepharose column chromatography | Rhodococcus sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cocaine + H2O | - |
Rhodococcus sp. | ecgonine methyl ester + benzoate | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | x-ray crystallography | Rhodococcus sp. |
Synonyms | Comment | Organism |
---|---|---|
cocE | - |
Rhodococcus sp. |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
the wild type enzyme has a half-life of 12.2 min at 37°C | Rhodococcus sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
40.1 | - |
cocaine | pegylated mutant enzyme T172R/G173Q, pH not specified in the publication, 37°C | Rhodococcus sp. | |
49.9 | - |
cocaine | mutant enzyme G4C/S10C, pH not specified in the publication, 37°C | Rhodococcus sp. | |
51.4 | - |
cocaine | wild type enzyme, pH not specified in the publication, 37°C | Rhodococcus sp. | |
56.6 | - |
cocaine | mutant enzyme T172R/G173Q, pH not specified in the publication, 37°C | Rhodococcus sp. |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2110 | - |
cocaine | pegylated mutant enzyme T172R/G173Q, pH not specified in the publication, 37°C | Rhodococcus sp. | |
2120 | - |
cocaine | mutant enzyme T172R/G173Q, pH not specified in the publication, 37°C | Rhodococcus sp. | |
2380 | - |
cocaine | mutant enzyme G4C/S10C, pH not specified in the publication, 37°C | Rhodococcus sp. | |
8990 | - |
cocaine | wild type enzyme, pH not specified in the publication, 37°C | Rhodococcus sp. |