BRENDA - Enzyme Database show
show all sequences of 3.1.1.84

Thermostable variants of cocaine esterase for long-time protection against cocaine toxicity

Gao, D.; Narasimhan, D.L.; Macdonald, J.; Brim, R.; Ko, M.C.; Landry, D.W.; Woods, J.H.; Sunahara, R.K.; Zhan, C.G.; Mol. Pharmacol. 75, 318-323 (2009)

Data extracted from this reference:

Application
Application
Commentary
Organism
medicine
enhancing cocaine metabolism by administration of cocaine esterase (CocE) is a promising treatment strategy for cocaine overdose and addiction, because CocE is the most efficient native enzyme for metabolizing the naturally occurring cocaine. A major obstacle to the clinical application of CocE is the thermoinstability of native CocE with a half-life of only a few min at physiological temperature. Computational-experimental effort yields a CocE variant with a 30fold increase in plasma half-life both in vitro and in vivo
Rhodococcus sp.
Engineering
Amino acid exchange
Commentary
Organism
G173Q
kcat and Km-value for cocaine is similar to wild-type value, half-life is increased 7fold compared to wild-type enzyme
Rhodococcus sp.
additional information
computational-experimental effort yields a CocE variant with a 30-fold increase in plasma half-life both in vitro and in vivo
Rhodococcus sp.
T172R
kcat and Km-value for cocaine is similar to wild-type value, half-life is increased 7fold compared to wild-type enzyme
Rhodococcus sp.
T172R/G173Q
kcat and Km-value for cocaine is similar to wild-type value, half-life is increased 30fold compared to wild-type enzyme
Rhodococcus sp.
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.015
-
cocaine
ph 7.4, 37°C, mutant enzyme G173Q
Rhodococcus sp.
0.021
-
cocaine
pH 7.4, 37°C, wild-type enzyme
Rhodococcus sp.
0.024
-
cocaine
ph 7.4, 37°C, mutant enzyme T172R; ph 7.4, 37°C, mutant enzyme T172R/G173Q
Rhodococcus sp.
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
cocaine + H2O
Rhodococcus sp.
CocE is the most efficient native enzyme for metabolizing the naturally occurring cocaine
ecgonine methyl ester + benzoate
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Rhodococcus sp.
Q9L9D7
-
-
Purification (Commentary)
Commentary
Organism
-
Rhodococcus sp.
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
cocaine + H2O
-
694267
Rhodococcus sp.
ecgonine methyl ester + benzoate
-
-
-
?
cocaine + H2O
CocE is the most efficient native enzyme for metabolizing the naturally occurring cocaine
694267
Rhodococcus sp.
ecgonine methyl ester + benzoate
-
-
-
?
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Rhodococcus sp.
Temperature Stability [°C]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
37
-
half-life: 11 min (wild-type enzyme), 78 min (mutant enzyme T172R), 75 min (mutant enzyme G173Q), 305 min (mutant enzyme T172R/G173Q)
Rhodococcus sp.
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2247
-
cocaine
ph 7.4, 37°C, mutant enzyme T172R/G173Q
Rhodococcus sp.
2323
-
cocaine
pH 7.4, 37°C, wild-type enzyme
Rhodococcus sp.
2384
-
cocaine
ph 7.4, 37°C, mutant enzyme G173Q
Rhodococcus sp.
2502
-
cocaine
ph 7.4, 37°C, mutant enzyme T172R
Rhodococcus sp.
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.4
-
assay at
Rhodococcus sp.
Application (protein specific)
Application
Commentary
Organism
medicine
enhancing cocaine metabolism by administration of cocaine esterase (CocE) is a promising treatment strategy for cocaine overdose and addiction, because CocE is the most efficient native enzyme for metabolizing the naturally occurring cocaine. A major obstacle to the clinical application of CocE is the thermoinstability of native CocE with a half-life of only a few min at physiological temperature. Computational-experimental effort yields a CocE variant with a 30fold increase in plasma half-life both in vitro and in vivo
Rhodococcus sp.
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
G173Q
kcat and Km-value for cocaine is similar to wild-type value, half-life is increased 7fold compared to wild-type enzyme
Rhodococcus sp.
additional information
computational-experimental effort yields a CocE variant with a 30-fold increase in plasma half-life both in vitro and in vivo
Rhodococcus sp.
T172R
kcat and Km-value for cocaine is similar to wild-type value, half-life is increased 7fold compared to wild-type enzyme
Rhodococcus sp.
T172R/G173Q
kcat and Km-value for cocaine is similar to wild-type value, half-life is increased 30fold compared to wild-type enzyme
Rhodococcus sp.
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.015
-
cocaine
ph 7.4, 37°C, mutant enzyme G173Q
Rhodococcus sp.
0.021
-
cocaine
pH 7.4, 37°C, wild-type enzyme
Rhodococcus sp.
0.024
-
cocaine
ph 7.4, 37°C, mutant enzyme T172R; ph 7.4, 37°C, mutant enzyme T172R/G173Q
Rhodococcus sp.
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
cocaine + H2O
Rhodococcus sp.
CocE is the most efficient native enzyme for metabolizing the naturally occurring cocaine
ecgonine methyl ester + benzoate
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
-
Rhodococcus sp.
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
cocaine + H2O
-
694267
Rhodococcus sp.
ecgonine methyl ester + benzoate
-
-
-
?
cocaine + H2O
CocE is the most efficient native enzyme for metabolizing the naturally occurring cocaine
694267
Rhodococcus sp.
ecgonine methyl ester + benzoate
-
-
-
?
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Rhodococcus sp.
Temperature Stability [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
37
-
half-life: 11 min (wild-type enzyme), 78 min (mutant enzyme T172R), 75 min (mutant enzyme G173Q), 305 min (mutant enzyme T172R/G173Q)
Rhodococcus sp.
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2247
-
cocaine
ph 7.4, 37°C, mutant enzyme T172R/G173Q
Rhodococcus sp.
2323
-
cocaine
pH 7.4, 37°C, wild-type enzyme
Rhodococcus sp.
2384
-
cocaine
ph 7.4, 37°C, mutant enzyme G173Q
Rhodococcus sp.
2502
-
cocaine
ph 7.4, 37°C, mutant enzyme T172R
Rhodococcus sp.
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.4
-
assay at
Rhodococcus sp.
Other publictions for EC 3.1.1.84
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
750291
Kim
Kinetic characterization of c ...
Homo sapiens
Chem. Biol. Interact.
293
107-114
2018
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1
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1
1
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3
3
728856
Fang
Rational design, preparation, ...
Rhodococcus sp., Rhodococcus sp. MB1 Bresler
ACS Chem. Biol.
9
1764-1772
2014
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1
1
3
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3
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3
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3
3
730250
Brim
The fate of bacterial cocaine ...
Rhodococcus sp.
J. Pharmacol. Exp. Ther.
340
83-95
2012
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730421
Lee
Cell permeable cocaine esteras ...
Rhodococcus sp., Rhodococcus sp. MB1
Mol. Pharm.
9
1361-1373
2012
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1
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5
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6
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1
2
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6
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1
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1
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1
6
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2
1
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1
6
-
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6
6
716278
Xue
Design, preparation, and chara ...
Homo sapiens
Mol. Pharmacol.
79
290-297
2011
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1
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4
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4
4
716281
Narasimhan
Subunit stabilization and pegy ...
Rhodococcus sp.
Mol. Pharmacol.
80
1056-1065
2011
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1
1
3
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4
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1
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3
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4
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1
1
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1
4
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4
4
714226
Zheng
Design of high-activity mutant ...
Homo sapiens
Biochemistry
49
9113-9119
2010
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1
1
10
-
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1
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1
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10
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1
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9
9
714702
Yang
Characterization of a high-act ...
Homo sapiens
Chem. Biol. Interact.
187
148-152
2010
-
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1
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2
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2
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2
2
716276
Brim
A thermally stable form of bac ...
Rhodococcus sp., Rhodococcus sp. MB1
Mol. Pharmacol.
77
593-600
2010
-
1
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1
1
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4
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7
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1
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4
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2
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1
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716824
Narasimhan
Structural analysis of thermos ...
Rhodococcus sp., Rhodococcus sp. MB1
Protein Eng.
23
537-547
2010
-
1
1
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5
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4
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6
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1
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5
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5
5
692028
Ko
Effects of cocaine esterase fo ...
Rhodococcus sp.
Drug Alcohol Depend.
101
202-209
2009
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1
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694267
Gao
Thermostable variants of cocai ...
Rhodococcus sp.
Mol. Pharmacol.
75
318-323
2009
-
1
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4
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3
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1
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1
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1
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699760
Collins
Zhan. C.G.; Woods. J.H.: Cocai ...
Rhodococcus sp.
J. Pharmacol. Exp. Ther.
331
445-455
2009
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1
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698483
Liu
Fundamental reaction mechanism ...
Rhodococcus sp.
J. Am. Chem. Soc.
131
11964-11975
2003
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650108
Turner
Biochemical characterization a ...
Rhodococcus sp.
Biochemistry
41
12297-12307
2002
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1
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11
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10
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11
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653322
Larsen
Crystal structure of a bacteri ...
Rhodococcus sp.
Nat. Struct. Biol.
9
17-21
2002
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1
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1
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695729
Bresler
Gene cloning and nucleotide se ...
Rhodococcus sp., Rhodococcus sp. MB1
Appl. Environ. Microbiol.
66
904-908
2000
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1
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1
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2
2
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11
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1
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1
1
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6
1
1
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1
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1
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2
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6
1
1
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1
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171019
Pindel
Purification and cloning of a ...
Homo sapiens
J. Biol. Chem.
272
14769-14775
1997
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1
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4
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1
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1
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1
1
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1
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1
4
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1
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1
1
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1
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5
1
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1
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-
698540
Britt
Identification of a cocaine es ...
Stenotrophomonas maltophilia, Stenotrophomonas maltophilia MB11L
J. Bacteriol.
174
2087-2094
1992
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2
3
-
-
3
2
-
4
-
-
1
-
-
-
1
-
10
2
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
3
-
-
3
2
-
-
-
1
-
-
1
-
10
2
1
-
-
-
1
-
-
-
-
-
-
-
-
-