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Literature summary extracted from

  • Fang, L.; Chow, K.M.; Hou, S.; Xue, L.; Chen, X.; Rodgers, D.W.; Zheng, F.; Zhan, C.G.
    Rational design, preparation, and characterization of a therapeutic enzyme mutant with improved stability and function for cocaine detoxification (2014), ACS Chem. Biol., 9, 1764-1772.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.1.84 expressed in Escherichia coli BL21(DE3) cells Rhodococcus sp.

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.1.1.84 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

EC Number Protein Variants Comment Organism
3.1.1.84 T172R/G173Q the mutant has an improved in vitro half-life of about 6 h at 37°C Rhodococcus sp.
3.1.1.84 T172R/G173Q /G4C/S10C the mutant shows improved catalytic efficiency against cocaine by about 20% Rhodococcus sp.
3.1.1.84 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.1.84 0.0015
-
 cocaine mutant enzyme T172R/G173Q/L196C/I301C, at 37°C, pH not specified in the publication Rhodococcus sp.
3.1.1.84 0.0021
-
 cocaine mutant enzyme T172R/G173Q/G4C/S10C, at 37°C, pH not specified in the publication Rhodococcus sp.
3.1.1.84 0.0029
-
 cocaine mutant enzyme T172R/G173Q, at 37°C, pH not specified in the publication 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
-
 ?
3.1.1.84 cocaine + H2O Rhodococcus sp. MB1 Bresler
-
 ecgonine methyl ester + benzoate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.1.1.84 Rhodococcus sp. Q9L9D7
-
-
3.1.1.84 Rhodococcus sp. MB1 Bresler Q9L9D7
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.1.84 HisPur cobalt resin column chromatography Rhodococcus sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.84 cocaine + H2O
-
 Rhodococcus sp. ecgonine methyl ester + benzoate
-
 ?
3.1.1.84 cocaine + H2O
-
 Rhodococcus sp. MB1 Bresler ecgonine methyl ester + benzoate
-
 ?

Subunits

EC Number Subunits Comment Organism
3.1.1.84 homodimer x-ray crystallography Rhodococcus sp.

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 39
-
 cocaine mutant enzyme T172R/G173Q/G4C/S10C, at 37°C, pH not specified in the publication Rhodococcus sp.
3.1.1.84 43.3
-
 cocaine mutant enzyme T172R/G173Q, at 37°C, pH not specified in the publication Rhodococcus sp.
3.1.1.84 57.7
-
 cocaine mutant enzyme T172R/G173Q/L196C/I301C, at 37°C, pH not specified in the publication Rhodococcus sp.

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.1.1.84 15330
-
 cocaine mutant enzyme T172R/G173Q, at 37°C, pH not specified in the publication Rhodococcus sp.
3.1.1.84 18330
-
 cocaine mutant enzyme T172R/G173Q/G4C/S10C, at 37°C, pH not specified in the publication Rhodococcus sp.
3.1.1.84 38330
-
 cocaine mutant enzyme T172R/G173Q/L196C/I301C, at 37°C, pH not specified in the publication Rhodococcus sp.