Application | Comment | Organism |
---|---|---|
medicine | the high catalytic proficiency, lack of observable product inhibition, and ability to hydrolyze both cocaine and cocaethylene make cocE an attractive candidate for rapid cocaine detoxification in an emergency setting | Rhodococcus sp. |
Cloned (Comment) | Organism |
---|---|
- |
Rhodococcus sp. |
Crystallization (Comment) | Organism |
---|---|
crystal structures of the S117A and Y44F mutants of cocE | Rhodococcus sp. |
Protein Variants | Comment | Organism |
---|---|---|
D259N | mutation results in more than 1500fold decrease in kcat | Rhodococcus sp. |
F261A | mutant catalyzed the hydrolysis of cocaine with a 29fold lower kcat and 15fold higher KM | Rhodococcus sp. |
F408A | mutant has 8fold increased KM and more than 100fold decrease in kcat | Rhodococcus sp. |
H287A | mutation results in more than 1500fold decrease in kcat | Rhodococcus sp. |
L407A | mutant has 2fold increased KM and more than 100fold decrease in kcat | Rhodococcus sp. |
L407A/F408A | attempts to express the L407A/F408A double mutant do not result in any soluble protein | Rhodococcus sp. |
Q55E | the mutation within the active site of cocE results in a 2fold improvement in KM, but a 14fold loss of kcat | Rhodococcus sp. |
S117A | mutation results in more than 1500fold decrease in kcat, crystal structures of the S117A and Y44F mutants of cocE. The first urea unfolding transition in the S117A mutant is shifted from 0.5 to 1.3 M urea compared to the wild-type, while the second transition, although broader, has a similar transition point | Rhodococcus sp. |
W151A | mutant catalyzed the hydrolysis of cocaine with a 78fold lower kcat and 80fold higher KM | Rhodococcus sp. |
W166A | mutant has a 29fold lower kcat, and a 6fold increased KM | Rhodococcus sp. |
Y44F | mutation results in more than 1500fold decrease in kcat, crystal structures of the S117A and Y44F mutants of cocE. The urea unfolding curve of the Y44F mutant is very similar to the wild-type, and has almost identical transition points | Rhodococcus sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00027 | - |
cocaine | pH 7.4, mutant enzyme Q55E | Rhodococcus sp. | |
0.00064 | - |
cocaine | pH 7.4, wild-type enzyme | Rhodococcus sp. | |
0.00075 | - |
cocaine | pH 7.4, mutant enzyme Q55A | Rhodococcus sp. | |
0.0012 | - |
cocaine | pH 7.4, mutant enzyme L407A | Rhodococcus sp. | |
0.0016 | - |
cocaethylene | pH 7.4, wild-type enzyme | Rhodococcus sp. | |
0.0036 | - |
cocaine | pH 7.4, mutant enzyme W166A | Rhodococcus sp. | |
0.0051 | - |
cocaine | pH 7.4, mutant enzyme F408A | Rhodococcus sp. | |
0.0096 | - |
cocaine | pH 7.4, mutant enzyme F261A | Rhodococcus sp. | |
0.046 | - |
cocaine | pH 7.4, mutant enzyme S117C | Rhodococcus sp. | |
0.051 | - |
cocaine | pH 7.4, mutant enzyme W151A | Rhodococcus sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
cocaine + H2O | Rhodococcus sp. | - |
ecgonine methyl ester + benzoate | - |
? |
Organic Solvent | Comment | Organism |
---|---|---|
urea | urea denaturation studies of cocE by fluorescence and circular dichroism show two unfolding transitions (0.5-0.6 M and 3.2-3.7 M urea), with the first transition likely representing pertubation of the active site | Rhodococcus sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodococcus sp. | Q9L9D7 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cocaethylene + H2O | cocaethylene is a more potent cocaine metabolite, observed in patients who concurrently abuse cocaine and alcohol | Rhodococcus sp. | ? | - |
? | |
cocaine + H2O | - |
Rhodococcus sp. | ecgonine methyl ester + benzoate | - |
? | |
cocaine + H2O | the bacterial cocaine esterase, cocE, hydrolyzes cocaine faster than any other reported cocaine esterase | Rhodococcus sp. | ecgonine methyl ester + benzoate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
cocE | - |
Rhodococcus sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.046 | - |
cocaine | pH 7.4, mutant enzyme S117C | Rhodococcus sp. | |
0.057 | - |
cocaine | pH 7.4, mutant enzyme F408A | Rhodococcus sp. | |
0.067 | - |
cocaine | pH 7.4, mutant enzyme L407A | Rhodococcus sp. | |
0.1 | - |
cocaine | pH 7.4, mutant enzyme W151A | Rhodococcus sp. | |
0.27 | - |
cocaine | pH 7.4, mutant enzyme F261A | Rhodococcus sp. | |
0.27 | - |
cocaine | pH 7.4, mutant enzyme W166A | Rhodococcus sp. | |
0.55 | - |
cocaine | pH 7.4, mutant enzyme Q55E | Rhodococcus sp. | |
1.7 | - |
cocaine | pH 7.4, mutant enzyme Q55A | Rhodococcus sp. | |
7.8 | - |
cocaine | pH 7.4, wild-type enzyme | Rhodococcus sp. | |
9.4 | - |
cocaethylene | pH 7.4, wild-type enzyme | Rhodococcus sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
- |
Rhodococcus sp. |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
7.8 | 10.5 | pH 7.8: about 50% of maximal activity, pH 10.0: about 50% of maximal activity | Rhodococcus sp. |