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Literature summary for 3.1.1.84 extracted from

  • Lee, T.Y.; Park, Y.S.; Garcia, G.A.; Sunahara, R.K.; Woods, J.H.; Yang, V.C.
    Cell permeable cocaine esterases constructed by chemical conjugation and genetic recombination (2012), Mol. Pharm., 9, 1361-1373.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21 Star (DE3) cells Rhodococcus sp.

Protein Variants

Protein Variants Comment Organism
LMWP-S-S-T172R/G173Q the mutant shows reduced catalytic efficiency compared to the wild type enzyme Rhodococcus sp.
LMWP-T172R/G173Q the mutant shows reduced catalytic efficiency compared to the wild type enzyme Rhodococcus sp.
T172R/G173Q -LMWP the mutant shows reduced catalytic efficiency compared to the wild type enzyme Rhodococcus sp.
T172R/G173Q -YGRKKRRQRRR the mutant shows reduced catalytic efficiency compared to the wild type enzyme Rhodococcus sp.
YGRKKRRQRRR-T172R/G173Q the mutant still maintains 52% of its cocaine-hydrolyzing efficiency even after incubation at 37°C for 24 h Rhodococcus sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0288
-
cocaine wild type enzyme, at pH 7.4 and 37°C Rhodococcus sp.
0.0305
-
cocaine mutant enzyme LMWP-S-S-T172R/G173Q, at pH 7.4 and 37°C Rhodococcus sp.
0.0306
-
cocaine mutant enzyme T172R/G173Q -YGRKKRRQRRR, at pH 7.4 and 37°C Rhodococcus sp.
0.0396
-
cocaine mutant enzyme YGRKKRRQRRR-T172R/G173Q, at pH 7.4 and 37°C Rhodococcus sp.
0.0415
-
cocaine mutant enzyme T172R/G173Q -LMWP, at pH 7.4 and 37°C Rhodococcus sp.
0.049
-
cocaine mutant enzyme LMWP-T172R/G173Q, at pH 7.4 and 37°C Rhodococcus sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
70000
-
x * 70000, SDS-PAGE Rhodococcus sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
cocaine + H2O Rhodococcus sp.
-
ecgonine methyl ester + benzoate
-
?
cocaine + H2O Rhodococcus sp. MB1
-
ecgonine methyl ester + benzoate
-
?

Organism

Organism UniProt Comment Textmining
Rhodococcus sp. Q9L9D7
-
-
Rhodococcus sp. MB1 Q9L9D7
-
-

Purification (Commentary)

Purification (Comment) Organism
Talon metal affinity 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
-
?
cocaine + H2O
-
Rhodococcus sp. MB1 ecgonine methyl ester + benzoate
-
?

Subunits

Subunits Comment Organism
? x * 70000, SDS-PAGE Rhodococcus sp.

Synonyms

Synonyms Comment Organism
cocE
-
Rhodococcus sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
37
-
the in vitro half-life at 37°C is about 13.7 min Rhodococcus sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
31.45
-
cocaine mutant enzyme T172R/G173Q -YGRKKRRQRRR, at pH 7.4 and 37°C Rhodococcus sp.
32.62
-
cocaine mutant enzyme YGRKKRRQRRR-T172R/G173Q, at pH 7.4 and 37°C Rhodococcus sp.
34.42
-
cocaine mutant enzyme LMWP-T172R/G173Q, at pH 7.4 and 37°C Rhodococcus sp.
35.17
-
cocaine mutant enzyme T172R/G173Q -LMWP, at pH 7.4 and 37°C Rhodococcus sp.
39.17
-
cocaine mutant enzyme LMWP-S-S-T172R/G173Q, at pH 7.4 and 37°C Rhodococcus sp.
44.85
-
cocaine wild type enzyme, at pH 7.4 and 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
702
-
cocaine mutant enzyme LMWP-T172R/G173Q, at pH 7.4 and 37°C Rhodococcus sp.
823
-
cocaine mutant enzyme YGRKKRRQRRR-T172R/G173Q, at pH 7.4 and 37°C Rhodococcus sp.
847
-
cocaine mutant enzyme T172R/G173Q -LMWP, at pH 7.4 and 37°C Rhodococcus sp.
1028
-
cocaine mutant enzyme T172R/G173Q -YGRKKRRQRRR, at pH 7.4 and 37°C Rhodococcus sp.
1283
-
cocaine mutant enzyme LMWP-S-S-T172R/G173Q, at pH 7.4 and 37°C Rhodococcus sp.
1560
-
cocaine wild type enzyme, at pH 7.4 and 37°C Rhodococcus sp.