BRENDA - Enzyme Database show
show all sequences of 3.1.1.84

Design, preparation, and characterization of high-activity mutants of human butyrylcholinesterase specific for detoxification of cocaine

Xue, L.; Ko, M.; Tong, M.; Yang, W.; Hou, S.; Fang, L.; Liu, J.; Zheng, F.; Woods, J.; Tai, H.; Zhan, C.; Mol. Pharmacol. 79, 290-297 (2011)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
wild type and mutant enzymes are expressed in HEK-293T/17 cells
Homo sapiens
Engineering
Amino acid exchange
Commentary
Organism
A199S/F227A/S287G/A328W/E441D
the mutant shows 1730fold improved (-)-cocaine-hydrolyzing activity compared to the wild type enzyme
Homo sapiens
A199S/F227A/S287G/A328W/Y332G/E441D
the mutant shows 1390fold improved (-)-cocaine-hydrolyzing activity compared to the wild type enzyme
Homo sapiens
A199S/S287G/A328W/Y332G
the mutant shows 1080fold improved (-)-cocaine-hydrolyzing activity compared to the wild type enzyme
Homo sapiens
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0011
-
(-)-cocaine
mutant enzyme A199S/F227A/S287G/A328W/E441D, pH not specified in the publication, at 25°C
Homo sapiens
0.0031
-
(-)-cocaine
mutant enzyme A199S/S287G/A328W/Y332G, pH not specified in the publication, at 25°C
Homo sapiens
0.0035
-
(-)-cocaine
mutant enzyme A199S/F227A/S287G/A328W/Y332G/E441D, pH not specified in the publication, at 25°C
Homo sapiens
0.0045
-
(-)-cocaine
wild type enzyme, pH not specified in the publication, at 25°C
Homo sapiens
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Homo sapiens
-
-
-
Purification (Commentary)
Commentary
Organism
QFF resin anion-exchange chromatography and Hypatite C column chromatography
Homo sapiens
Source Tissue
Source Tissue
Commentary
Organism
Textmining
blood plasma
-
Homo sapiens
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(-)-cocaine + H2O
-
716278
Homo sapiens
benzoic acid + methyl (1R,2R,3S,5S)-3-hydroxy-8-methyl-8-azabicyclo[3.2.1]-octane-2-carboxylate
i.e. ecgonine methyl ester
-
-
?
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.002
-
(-)-cocaine
wild type enzyme, pH not specified in the publication, at 25°C
Homo sapiens
28.83
-
(-)-cocaine
mutant enzyme A199S/F227A/S287G/A328W/E441D, pH not specified in the publication, at 25°C
Homo sapiens
51
-
(-)-cocaine
mutant enzyme A199S/S287G/A328W/Y332G, pH not specified in the publication, at 25°C
Homo sapiens
73.83
-
(-)-cocaine
mutant enzyme A199S/F227A/S287G/A328W/Y332G/E441D, pH not specified in the publication, at 25°C
Homo sapiens
Cloned(Commentary) (protein specific)
Commentary
Organism
wild type and mutant enzymes are expressed in HEK-293T/17 cells
Homo sapiens
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
A199S/F227A/S287G/A328W/E441D
the mutant shows 1730fold improved (-)-cocaine-hydrolyzing activity compared to the wild type enzyme
Homo sapiens
A199S/F227A/S287G/A328W/Y332G/E441D
the mutant shows 1390fold improved (-)-cocaine-hydrolyzing activity compared to the wild type enzyme
Homo sapiens
A199S/S287G/A328W/Y332G
the mutant shows 1080fold improved (-)-cocaine-hydrolyzing activity compared to the wild type enzyme
Homo sapiens
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0011
-
(-)-cocaine
mutant enzyme A199S/F227A/S287G/A328W/E441D, pH not specified in the publication, at 25°C
Homo sapiens
0.0031
-
(-)-cocaine
mutant enzyme A199S/S287G/A328W/Y332G, pH not specified in the publication, at 25°C
Homo sapiens
0.0035
-
(-)-cocaine
mutant enzyme A199S/F227A/S287G/A328W/Y332G/E441D, pH not specified in the publication, at 25°C
Homo sapiens
0.0045
-
(-)-cocaine
wild type enzyme, pH not specified in the publication, at 25°C
Homo sapiens
Purification (Commentary) (protein specific)
Commentary
Organism
QFF resin anion-exchange chromatography and Hypatite C column chromatography
Homo sapiens
Source Tissue (protein specific)
Source Tissue
Commentary
Organism
Textmining
blood plasma
-
Homo sapiens
-
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(-)-cocaine + H2O
-
716278
Homo sapiens
benzoic acid + methyl (1R,2R,3S,5S)-3-hydroxy-8-methyl-8-azabicyclo[3.2.1]-octane-2-carboxylate
i.e. ecgonine methyl ester
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.002
-
(-)-cocaine
wild type enzyme, pH not specified in the publication, at 25°C
Homo sapiens
28.83
-
(-)-cocaine
mutant enzyme A199S/F227A/S287G/A328W/E441D, pH not specified in the publication, at 25°C
Homo sapiens
51
-
(-)-cocaine
mutant enzyme A199S/S287G/A328W/Y332G, pH not specified in the publication, at 25°C
Homo sapiens
73.83
-
(-)-cocaine
mutant enzyme A199S/F227A/S287G/A328W/Y332G/E441D, pH not specified in the publication, at 25°C
Homo sapiens
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
15.17
-
(-)-cocaine
wild type enzyme, pH not specified in the publication, at 25°C
Homo sapiens
16500
-
(-)-cocaine
mutant enzyme A199S/S287G/A328W/Y332G, pH not specified in the publication, at 25°C
Homo sapiens
21670
-
(-)-cocaine
mutant enzyme A199S/F227A/S287G/A328W/Y332G/E441D, pH not specified in the publication, at 25°C
Homo sapiens
26670
-
(-)-cocaine
mutant enzyme A199S/F227A/S287G/A328W/E441D, pH not specified in the publication, at 25°C
Homo sapiens
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
15.17
-
(-)-cocaine
wild type enzyme, pH not specified in the publication, at 25°C
Homo sapiens
16500
-
(-)-cocaine
mutant enzyme A199S/S287G/A328W/Y332G, pH not specified in the publication, at 25°C
Homo sapiens
21670
-
(-)-cocaine
mutant enzyme A199S/F227A/S287G/A328W/Y332G/E441D, pH not specified in the publication, at 25°C
Homo sapiens
26670
-
(-)-cocaine
mutant enzyme A199S/F227A/S287G/A328W/E441D, pH not specified in the publication, at 25°C
Homo sapiens
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
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3
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3
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5
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1
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3
1
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1
1
-
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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2
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3
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1
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2
1
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3
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1
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1
3
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3
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2
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1
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2
1
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3
-
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-
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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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1
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-
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1
1
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4
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1
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1
1
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-
-
-
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-
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-
-
-
-
-
730421
Lee
Cell permeable cocaine esteras ...
Rhodococcus sp., Rhodococcus sp. MB1
Mol. Pharm.
9
1361-1373
2012
-
-
1
-
5
-
-
6
-
-
1
2
-
6
-
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1
-
-
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2
1
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1
6
-
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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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1
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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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3
-
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4
-
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1
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1
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1
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4
-
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-
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-
-
-
4
4
716281
Narasimhan
Subunit stabilization and pegy ...
Rhodococcus sp.
Mol. Pharmacol.
80
1056-1065
2011
-
-
1
1
3
-
-
4
-
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1
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3
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1
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1
1
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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
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1
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1
1
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1
4
-
-
-
-
-
-
-
-
4
4
714226
Zheng
Design of high-activity mutant ...
Homo sapiens
Biochemistry
49
9113-9119
2010
-
-
1
1
10
-
-
-
-
-
-
-
-
1
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1
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1
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1
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
-
-
-
-
1
-
-
2
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1
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1
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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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2
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1
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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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1
5
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1
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2
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1
5
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5
5
692028
Ko
Effects of cocaine esterase fo ...
Rhodococcus sp.
Drug Alcohol Depend.
101
202-209
2009
-
1
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694267
Gao
Thermostable variants of cocai ...
Rhodococcus sp.
Mol. Pharmacol.
75
318-323
2009
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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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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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1
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4
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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
1
1
11
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10
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9
1
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11
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9
1
1
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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
-
-
1
-
-
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-
1
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2
2
-
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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1
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2
2
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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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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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1
4
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2
1
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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
1
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1
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1
1
4
-
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2
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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