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Literature summary for 3.1.1.84 extracted from

  • Narasimhan, D.; Collins, G.T.; Nance, M.R.; Nichols, J.; Edwald, E.; Chan, J.; Ko, M.C.; Woods, J.H.; Tesmer, J.J.; Sunahara, R.K.
    Subunit stabilization and pegylation of cocaine esterase improves in vivo residence time (2011), Mol. Pharmacol., 80, 1056-1065.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.1.1.84

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL-21 Gold (DE3) cells Rhodococcus sp.

Crystallization (Commentary)

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

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 [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 [Da] Molecular Weight Maximum [Da] Comment Organism
127000
-
-
 Rhodococcus sp.

Organism

Organism UniProt Comment Textmining
Rhodococcus sp.
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Talon metal chelate column chromatography and Q-Sepharose column chromatography Rhodococcus sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
cocaine + H2O
-
 Rhodococcus sp. ecgonine methyl ester + benzoate
-
 ?

Subunits

Subunits Comment Organism
homodimer x-ray crystallography Rhodococcus sp.

Synonyms

Synonyms Comment Organism
cocE
-
 Rhodococcus sp.

Temperature Stability [°C]

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 [1/s]

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 [mM/s]

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.