1.1.3.17: choline oxidase
This is an abbreviated version!
For detailed information about choline oxidase, go to the full flat file.
Word Map on EC 1.1.3.17
-
1.1.3.17
-
acetylcholine
-
electrode
-
acetylcholinesterase
-
biosensors
-
betaine
-
electrochemical
-
ache
-
amperometric
-
arthrobacter
-
globiformis
-
glycinebetaine
-
organophosphorus
-
co-immobilized
-
luminol
-
post-column
-
screen-printed
-
prussian
-
electropolymerized
-
butyrylcholine
-
4-aminoantipyrine
-
bienzymatic
-
four-electron
-
analysis
-
choline-containing
-
polypyrrole
-
alkoxide
-
3.1.1.8
-
nafion
-
enzyme-modified
-
electrodeposited
-
co-crosslinking
-
agriculture
-
synthesis
-
nutrition
-
biotechnology
- 1.1.3.17
- acetylcholine
-
electrode
- acetylcholinesterase
-
biosensors
- betaine
-
electrochemical
-
ache
-
amperometric
- arthrobacter
- globiformis
- glycinebetaine
-
organophosphorus
-
co-immobilized
- luminol
-
post-column
-
screen-printed
-
prussian
-
electropolymerized
- butyrylcholine
- 4-aminoantipyrine
-
bienzymatic
-
four-electron
- analysis
-
choline-containing
-
polypyrrole
-
alkoxide
-
3.1.1.8
-
nafion
-
enzyme-modified
-
electrodeposited
-
co-crosslinking
- agriculture
- synthesis
- nutrition
- biotechnology
Reaction
Synonyms
alkaliphilic choline oxidase, ANI01nite_22550, An_CodA, APChO-syn, CHO, choline oxidase, choline-oxygen 1-oxidoreductase, choline:oxygen 1-reductase, ChOx, ChOx protein, codA, COX
ECTree
Advanced search results
Engineering
Engineering on EC 1.1.3.17 - choline oxidase
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
E312A
generated for investigation of the negative charge on Glu312, enzyme inactive
E312D
E312Q
generated for investigation of the negative charge on Glu312, Kd value for choline about 500times larger than that of wild-type
F357A
the mutant shows about 240fold reduced catalytic efficiency compared to the wild type enzyme
H351Q
the kcat and kcat/Km values of the H351Q emutant in atmospheric oxygen are 45 and 5000fold lower than those of the wild type enzyme, respectively
H466A
H466D
-
site-directed mutagenesis, the mutation alters the flavin binding to the enzyme, while substrate choline is normally bound, binding og glycine btaine is inhibited, spectrometrical analysis, the mutant shows a different flavin-binding stoichiometry of 0.29:1, compared to 1:1 for the wild-type enzyme, stabilized at pH 6.0-10.0, overview, comparison of midpoint reduction-oxidation potentials of the enzyme-FAD form with mutant H466A and the wild-type enzyme, the mutant shows no catalytic activity
H466Q
M359R
mutant displays increased activity with hexan-1-ol, reaction of EC 1.1.3.13
M62A
the mutant shows about 3fold reduced catalytic efficiency compared to the wild type enzyme
M62A/F357A
the enzyme shows a lack of the isomerization detected in wild type choline oxidase, and a lack of saturation with an oxygen concentration as high as 1 mM, while most other kinetic parameters are similar to those of wild type choline oxidase
N510A
-
site-directed mutagenesis of a catalytic residue resulting in low incorporation of FAD into the protein, enzyme kinetics decrease of 4300fold in the kcat/Kcholine, 600fold in the kred, 660fold in the kcat, and 50fold in the kcat/Koxygen values
N510D
-
site-directed mutagenesis of a catalytic residue resulting in low incorporation of FAD into the protein, 75% of the flavin associates noncovalently, inactive mutant
N510H
-
site-directed mutagenesis of a catalytic residue resulting in low incorporation of FAD into the protein, decreases in the kcat/Kcholine, the kred, the kcat, and the kcat/Koxygen values
N510L
-
site-directed mutagenesis of a catalytic residue resulting in low incorporation of FAD into the protein
S101A
S101A/D250G/F253R/V355T/F357R/M359R
mutant displays increased activity with hexan-1-ol, reaction of EC 1.1.3.13, with a 20fold increased kcat compared to that of the wildtype enzyme. This variant enables the oxidation of 10 mM hexanol to hexanal in less than 24h with 100% conversion and catalyzes significantly improved oxidation of saturated, unsaturated, aliphatic, cyclic and benzylic alcohols
S101A/V355T/F357R
mutant displays increased activity with hexan-1-ol, reaction of EC 1.1.3.13
S101A/V355T/F357R/M359R
mutant displays increased activity with hexan-1-ol, reaction of EC 1.1.3.13
S101C
S101T
S101V
contrary to wild-type, stopped-flow traces for the reductive half-reaction are biphasic, corresponding to the reactions of proton abstraction and hydride transfer. The rate constants for proton transfer in the S101T/C/V variants decrease logarithmically with increasing hydrophobicity of residue 101
V355T/F357R
mutant displays increased activity with hexan-1-ol, reaction of EC 1.1.3.13
V464A
V464T
A21V/G62D
-
site-directed mutagenesis, the mutant shows 1.93fold increased activity with tris-(2-hydroxyethyl)-methylammonium methylsulfate and slightly reduced activity with choline compared to the wild-type enzyme
A21V/G62D/I69V
-
site-directed mutagenesis, the mutant shows 1.68fold increased activity with tris-(2-hydroxyethyl)-methylammonium methylsulfate compared to the wild-type enzyme
A21V/G62D/I69V/S348L
-
site-directed mutagenesis, the mutant shows 3.45fold increased activity with tris-(2-hydroxyethyl)-methylammonium methylsulfate and highly reduced activity with choline compared to the wild-type enzyme
A21V/G62D/S348C
-
site-directed mutagenesis, the mutant shows 5.18fold increased activity with tris-(2-hydroxyethyl)-methylammonium methylsulfate and reduced activity with choline compared to the wild-type enzyme
A21V/G62D/S348L
-
site-directed mutagenesis, the mutant shows 3.72fold increased activity with tris-(2-hydroxyethyl)-methylammonium methylsulfate and highly reduced activity with choline compared to the wild-type enzyme
A21V/G62D/S348L/V349L
-
site-directed mutagenesis, the mutant shows 5.75fold increased activity with tris-(2-hydroxyethyl)-methylammonium methylsulfate and highly reduced activity with choline compared to the wild-type enzyme
A21V/K394R
-
site-directed mutagenesis, the mutant shows 85% activity with tris-(2-hydroxyethyl)-methylammonium methylsulfate compared to the wild-type enzyme
F351Y
-
site-directed mutagenesis in the substrate binding site, the mutant shows reduced activity with choline compared to the wild-type enzyme
G62D
-
site-directed mutagenesis in the FAD binding site, the mutant shows 2fold increased activity with tris-(2-hydroxyethyl)-methylammonium methylsulfate and reduced activity with choline compared to the wild-type enzyme
G62D/F351Y
-
site-directed mutagenesis, the mutant shows 2.14fold increased activity with tris-(2-hydroxyethyl)-methylammonium methylsulfate compared to the wild-type enzyme
G62D/R249H/F351Y
-
site-directed mutagenesis, the mutant shows 2.7fold increased activity with tris-(2-hydroxyethyl)-methylammonium methylsulfate and reduced activity with choline compared to the wild-type enzyme
I69V
-
site-directed mutagenesis in the FAD binding site, the mutant shows93% activity with tris-(2-hydroxyethyl)-methylammonium methylsulfate compared to the wild-type enzyme
P393Q/S530G
-
site-directed mutagenesis, the mutant shows 1.5fold increased activity with choline compared to the wild-type enzyme
T116I/K128M
-
site-directed mutagenesis, the mutant shows unaltered activity with choline compared to the wild-type enzyme
T116I/K128M/P393Q/S530G
-
site-directed mutagenesis, the mutant shows 2.32fold increased activity with choline compared to the wild-type enzyme
A21V
-
site-directed mutagenesis in the FAD binding site
-
G62D
-
site-directed mutagenesis in the FAD binding site, the mutant shows 2fold increased activity with tris-(2-hydroxyethyl)-methylammonium methylsulfate and reduced activity with choline compared to the wild-type enzyme
-
I69V
-
site-directed mutagenesis in the FAD binding site, the mutant shows93% activity with tris-(2-hydroxyethyl)-methylammonium methylsulfate compared to the wild-type enzyme
-
S348L
-
site-directed mutagenesis in the substrate binding site
-
V349L
-
site-directed mutagenesis in the substrate binding site
-
additional information
generated for investigation of the negative charge on Glu312, kcat values about 230times lower and and kcat/Km values about 35times lower than in the wild-type, solvent viscosity and substrate kinetic isotope effects indicates presence of internal equilibrium prior to the hydride transfer reaction
E312D
site-directed mutagenesis, the mutant enzyme shows a 260fold decrease in the rate constant for the hydride transfer reaction and did not transfer the hydride ion in a full quantum mechanical tunneling fashion. The overall turnover of the Glu312Asp enzyme at saturating concentrations of 3-hydroxypropyl-trimethylamine and oxygen is predominantly controlled by the hydride transfer reaction that results in the reduction of the enzyme-bound flavin. The kred value with 3-hydroxypropyl-trimethylamine is 20times higher than the value determined with choline as substrate for the Glu312Asp enzyme. The reductive half-reaction can be effectively, but not completely,rescued upon introducing on the substrate the methylenethat is missing from the side chain of residue 312
-
decrease in kcat and kcat/Km-value for choline, but not for oxygen, partial rescue of activity in presence of imidazolium
H466A
-
site-directed mutagenesis, comparison of midpoint reduction-oxidation potentials of the enzyme-FAD form with mutant H466D and the wild-type enzyme
H466A
-
site-directed mutagenesis, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme, the enzyme activity in the mutant strain can partially be rescued by addition of exogenous imidazolium, but not by imidazole, overview
H466Q
the unusual fluorescence behavior of the enzyme is lost in the mutant
mutant enzyme with increased efficiencies in the oxidative half-reactions and decreased efficiencies in the reductive half-reactions accompanied by a significant decrease in the overall rate of turnover with choline. Ser101 is important, but not essential, for catalysis
S101A
mutant displays increased activity with hexan-1-ol, reaction of EC 1.1.3.13
S101A
similar to wild-type mutant displays in 20 mM Tris-HCl flavin maxima in the near-UV and visible regions typical of oxidized flavins
S101A
stopped-flow traces for the reductive half-reaction are monophasic like wild-type choline oxidase
S101A
the unusual fluorescence behavior of the enzyme persists in the mutant
contrary to wild-type, stopped-flow traces for the reductive half-reaction are biphasic, corresponding to the reactions of proton abstraction and hydride transfer. The rate constants for proton transfer in the S101T/C/V variants decrease logarithmically with increasing hydrophobicity of residue 101
S101C
in presence of protonated Tris in the active site, the reversible formation of a C4a-S-cysteinyl8alpha-N3-histidyl flavin is observed
contrary to wild-type, stopped-flow traces for the reductive half-reaction are biphasic, corresponding to the reactions of proton abstraction and hydride transfer. The rate constants for proton transfer in the S101T/C/V variants decrease logarithmically with increasing hydrophobicity of residue 101
S101T
similar to wild-type mutant displays in 20 mM Tris-HCl flavin maxima in the near-UV and visible regions typical of oxidized flavins
V464A
site-directed mutagenesis, mutation of the residue near the flavin C(4a) atom and the hydrophobic cavity, replacement of Val464 with alanine or threonine does not affect the reductive half-reaction, but it reduces the oxidative half-reaction by about 50fold, and the 3D structure of the Val464Ala enzyme is essentially identical to that of the wild-type enzyme
V464T
site-directed mutagenesis, mutation of the residue near the flavin C(4a) atom and the hydrophobic cavity, replacement of Val464 with alanine or threonine does not affect the reductive half-reaction, but it reduces the oxidative half-reaction by about 50fold
-
enzyme immobilization by 6-O-ethoxytrimethylammoniumchitosan chloride, and construction of amperometric choline biosensors prepared by layer-by-layer deposition of choline oxidase on the Prussian blue-modified platinum electrode, method optimization
additional information
-
engineering of Eucalyptus plants via expression of bacterial choline oxidase to develop environmental-stress resistant Eucalyptus globulus for expanding the plantation area of this species. Glycine betaine accumulation in transgenic plants
additional information
-
substitution of Asn510 with alanine, but not with histidine, results in a change from stepwise to concerted mechanisms for the cleavages of the OH and CH bonds of choline catalyzed by the enzyme
additional information
-
evaluation of the stress-inducible production of Arthrobacter pascens choline oxidase in transgenic rice as a strategy for producing the stress-protectant glycine betaine, stress induction by 150 mM NaCl, biomass production in wild-type and transgenic plants