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Literature summary extracted from

  • Turner, J.M.; Larsen, N.A.; Basran, A.; Barbas, C.F., 3rd; Bruce, N.C.; Wilson, I.A.; Lerner, R.A.
    Biochemical characterization and structural analysis of a highly proficient cocaine esterase (2002), Biochemistry, 41, 12297-12307.
    View publication on PubMed

Application

EC Number Application Comment Organism
3.1.1.84 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(Commentary)

EC Number Cloned (Comment) Organism
3.1.1.84
-
 Rhodococcus sp.

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.1.1.84 crystal structures of the S117A and Y44F mutants of cocE Rhodococcus sp.

Protein Variants

EC Number Protein Variants Comment Organism
3.1.1.84 D259N mutation results in more than 1500fold decrease in kcat Rhodococcus sp.
3.1.1.84 F261A mutant catalyzed the hydrolysis of cocaine with a 29fold lower kcat and 15fold higher KM Rhodococcus sp.
3.1.1.84 F408A mutant has 8fold increased KM and more than 100fold decrease in kcat Rhodococcus sp.
3.1.1.84 H287A mutation results in more than 1500fold decrease in kcat Rhodococcus sp.
3.1.1.84 L407A mutant has 2fold increased KM and more than 100fold decrease in kcat Rhodococcus sp.
3.1.1.84 L407A/F408A attempts to express the L407A/F408A double mutant do not result in any soluble protein Rhodococcus sp.
3.1.1.84 Q55E the mutation within the active site of cocE results in a 2fold improvement in KM, but a 14fold loss of kcat Rhodococcus sp.
3.1.1.84 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.
3.1.1.84 W151A mutant catalyzed the hydrolysis of cocaine with a 78fold lower kcat and 80fold higher KM Rhodococcus sp.
3.1.1.84 W166A mutant has a 29fold lower kcat, and a 6fold increased KM Rhodococcus sp.
3.1.1.84 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.1.84 0.00027
-
 cocaine pH 7.4, mutant enzyme Q55E Rhodococcus sp.
3.1.1.84 0.00064
-
 cocaine pH 7.4, wild-type enzyme Rhodococcus sp.
3.1.1.84 0.00075
-
 cocaine pH 7.4, mutant enzyme Q55A Rhodococcus sp.
3.1.1.84 0.0012
-
 cocaine pH 7.4, mutant enzyme L407A Rhodococcus sp.
3.1.1.84 0.0016
-
 cocaethylene pH 7.4, wild-type enzyme Rhodococcus sp.
3.1.1.84 0.0036
-
 cocaine pH 7.4, mutant enzyme W166A Rhodococcus sp.
3.1.1.84 0.0051
-
 cocaine pH 7.4, mutant enzyme F408A Rhodococcus sp.
3.1.1.84 0.0096
-
 cocaine pH 7.4, mutant enzyme F261A Rhodococcus sp.
3.1.1.84 0.046
-
 cocaine pH 7.4, mutant enzyme S117C Rhodococcus sp.
3.1.1.84 0.051
-
 cocaine pH 7.4, mutant enzyme W151A Rhodococcus sp.

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.1.1.84 cocaine + H2O Rhodococcus sp.
-
 ecgonine methyl ester + benzoate
-
 ?

Organic Solvent Stability

EC Number Organic Solvent Comment Organism
3.1.1.84 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

EC Number Organism UniProt Comment Textmining
3.1.1.84 Rhodococcus sp. Q9L9D7
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.84 cocaethylene + H2O cocaethylene is a more potent cocaine metabolite, observed in patients who concurrently abuse cocaine and alcohol Rhodococcus sp. ?
-
 ?
3.1.1.84 cocaine + H2O
-
 Rhodococcus sp. ecgonine methyl ester + benzoate
-
 ?
3.1.1.84 cocaine + H2O the bacterial cocaine esterase, cocE, hydrolyzes cocaine faster than any other reported cocaine esterase Rhodococcus sp. ecgonine methyl ester + benzoate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.1.1.84 cocE
-
 Rhodococcus sp.

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1.1.84 0.046
-
 cocaine pH 7.4, mutant enzyme S117C Rhodococcus sp.
3.1.1.84 0.057
-
 cocaine pH 7.4, mutant enzyme F408A Rhodococcus sp.
3.1.1.84 0.067
-
 cocaine pH 7.4, mutant enzyme L407A Rhodococcus sp.
3.1.1.84 0.1
-
 cocaine pH 7.4, mutant enzyme W151A Rhodococcus sp.
3.1.1.84 0.27
-
 cocaine pH 7.4, mutant enzyme F261A Rhodococcus sp.
3.1.1.84 0.27
-
 cocaine pH 7.4, mutant enzyme W166A Rhodococcus sp.
3.1.1.84 0.55
-
 cocaine pH 7.4, mutant enzyme Q55E Rhodococcus sp.
3.1.1.84 1.7
-
 cocaine pH 7.4, mutant enzyme Q55A Rhodococcus sp.
3.1.1.84 7.8
-
 cocaine pH 7.4, wild-type enzyme Rhodococcus sp.
3.1.1.84 9.4
-
 cocaethylene pH 7.4, wild-type enzyme Rhodococcus sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.1.84 9
-
-
 Rhodococcus sp.

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.1.1.84 7.8 10.5 pH 7.8: about 50% of maximal activity, pH 10.0: about 50% of maximal activity Rhodococcus sp.