Information on EC 1.1.3.17 - choline oxidase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
1.1.3.17
-
RECOMMENDED NAME
GeneOntology No.
choline oxidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
betaine aldehyde + O2 + H2O = betaine + H2O2
show the reaction diagram
choline + 2 O2 + H2O = betaine + 2 H2O2
show the reaction diagram
choline + O2 = betaine aldehyde + H2O2
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Glycine, serine and threonine metabolism
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
choline:oxygen 1-oxidoreductase
A flavoprotein (FAD). In many bacteria, plants and animals, the osmoprotectant betaine is synthesized using different enzymes to catalyse the conversion of (1) choline into betaine aldehyde and (2) betaine aldehyde into betaine. In plants, the first reaction is catalysed by EC 1.14.15.7, choline monooxygenase, whereas in animals and many bacteria, it is catalysed by either membrane-bound choline dehydrogenase (EC 1.1.99.1) or soluble choline oxidase (EC 1.1.3.17) [6]. The enzyme involved in the second step, EC 1.2.1.8, betaine-aldehyde dehydrogenase, appears to be the same in those plants, animals and bacteria that use two separate enzymes.
CAS REGISTRY NUMBER
COMMENTARY hide
9028-67-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
gene codA
-
-
Manually annotated by BRENDA team
rat
-
-
Manually annotated by BRENDA team
102F34
-
-
Manually annotated by BRENDA team
cv. Superior, wild-type and transgenic plants expressing bacterial choline oxidase (codA) gene of Arthrobacter globiformis
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
-
replacing Asn510 with alanine or histidine negatively affects both the reductive and oxidative half-reactions catalyzed by choline oxidase. 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
physiological function
-
the reaction catalyzed by choline oxidase is of importance due to the trigger of glycine betaine accumulation in the cytoplasm of a number of plants and pathogenic bacteria in response to hyperosmotic and adverse temperature conditions to prevent dehydration and eventual cell death
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-methylaminoethanol + 2 O2 + H2O
? + 2 H2O2
show the reaction diagram
-
-
-
-
?
3,3-dimethyl-butanol + O2
3,3-dimethylbutanal + H2O2
show the reaction diagram
-
-
-
-
?
3,3-dimethylbutan-1-ol + 2 O2 + H2O2
?
show the reaction diagram
-
-
-
-
?
3,3-dimethylbutan-1-ol + O2
?
show the reaction diagram
-
10fold lower activity compared to choline
-
-
?
3-(dimethylamino)propan-1-ol + 2 O2 + H2O
? + 2 H2O2
show the reaction diagram
-
-
-
-
?
3-hydroxypropyl-trimethylamine + 2 O2 + H2O2
?
show the reaction diagram
-
-
-
-
?
3-trimethylamino-1-propanol + 2 O2 + H2O
3-trimethylaminopropanoate + 2 H2O2
show the reaction diagram
-
-
-
-
?
4-trimethylamino-1-butanol + 2 O2 + H2O
4-trimethylaminobutanoate + 2 H2O2
show the reaction diagram
-
-
-
-
?
betaine aldehyde + O2 + H2O
betaine + H2O2
show the reaction diagram
choline + 2 O2 + H2O
betaine + 2 H2O2
show the reaction diagram
choline + 2 O2 + H2O2
betaine + 2 H2O2
show the reaction diagram
choline + O2
betaine aldehyde + H2O2
show the reaction diagram
choline + O2 + H2O
choline acetate + H2O2
show the reaction diagram
-
-
-
-
?
diethanolamine + O2
?
show the reaction diagram
-
-
-
-
?
diethylamino ethanol + 2 O2 + H2O2
? + 2 H2O2
show the reaction diagram
-
-
-
-
?
diethylaminoethanol + 2 O2 + H2O
? + 2 H2O2
show the reaction diagram
-
-
-
-
?
dimethylaminoethanol + 2 O2 + H2O
? + 2 H2O2
show the reaction diagram
-
-
-
-
?
FADH2 + O2
FAD + H2O2
show the reaction diagram
-
-
-
-
?
monoethanolamine + O2
aminoacetaldehyde + H2O2
show the reaction diagram
N,N-dimethylaminoethanol + O2
(dimethylamino)acetaldehyde + H2O2
show the reaction diagram
N,N-dimethylethanolamine + O2
N,N-dimethylethanalamine + H2O2
show the reaction diagram
-
-
-
-
?
N-methylethanolamine + O2
N-methylethanalamine + H2O2
show the reaction diagram
-
-
-
-
?
triethanolamine + O2
?
show the reaction diagram
tris-(2-hydroxyethyl)-methylammonium methylsulfate + 2 O2 + H2O2
? + 2 H2O2
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
betaine aldehyde + O2 + H2O
betaine + H2O2
show the reaction diagram
choline + 2 O2 + H2O
betaine + 2 H2O2
show the reaction diagram
choline + 2 O2 + H2O2
betaine + 2 H2O2
show the reaction diagram
choline + O2
betaine aldehyde + H2O2
show the reaction diagram
FADH2 + O2
FAD + H2O2
show the reaction diagram
-
-
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2-amino-ethyl)trimethylammonim chloride
-
-
2-chloro-N-(2-chloroethyl)-N-methylethanamine
-
nitrogen mustard, strong competitive inhibitor
3,3-dimethylbutyraldehyde
-
product analogue inhibition, 74-90% reduced flavin reduction dependent on pH
4-chloromercuribenzoate
-
75% inhibition at 1 mM
allyltrimethylammonium chloride
-
-
Betaine aldehyde
-
-
diethyldicarbonate
-
competitive inhibition, 10% inhibition
diethylmethylamine
-
-
Dimethylamine
-
-
dimethylethylamine
-
-
ethylparathion
-
uncompetitive versus choline, inhibition and subsequent inactivation of the enzyme, active site specific
glycine betaine
iodoacetic acid
-
10% inhibition at 1 mM
Methylethylamine
-
-
Mn2+
-
7% inhibition at 1 mM
N,N-dimethylglycine
-
-
N-methylglycine
-
-
Nicotine
-
ability of nicotine to inhibit catalytic activity of choline oxidase used for biosensorial detection of
p-chloromercuribenzoate
-
-
Paraoxon
-
uncompetitive versus choline, inhibition and subsequent inactivation of the enzyme, active site specific
PMSF
-
uncompetitive inhibition, 50% inhibition
Quinine
-
20% inhibition at 1 mM
rotenone
-
-
tetramethylamine
-
-
trimethylamine
-
-
trimethylethylammonium chloride
-
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NAD+
-
stimulation of choline oxidation
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.01
3,3-dimethyl-1-butanol
-
pH 8.0, 25C
0.39 - 8.7
Betaine aldehyde
2.3 - 2.35
betaine-aldehyde
0.25 - 213
choline
14
N,N-Dimethylaminoethanol
-
-
0.32
N,N-dimethylethanolamine
-
pH 8.0, 25C
0.52
N-Methylethanolamine
-
pH 8.0, 25C
0.055 - 703
O2
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
8
3,3-dimethyl-1-butanol
Arthrobacter globiformis
-
pH 8.0, 25C
43 - 133
Betaine aldehyde
1.71 - 11.6
betaine-aldehyde
0.02 - 86
choline
5.9
N,N-dimethylethanolamine
Arthrobacter globiformis
-
pH 8.0, 25C
0.37
N-Methylethanolamine
Arthrobacter globiformis
-
pH 8.0, 25C
0.09 - 3.4
O2
additional information
additional information
Arthrobacter globiformis
-
overall turnover of mutant variant about 60-fold decreased with choline compared to wild-type enzyme
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.031 - 2
3,3-dimethylbutan-1-ol
63084
0.00009 - 237
choline
149
0.1 - 6.36
O2
9
additional information
additional information
Arthrobacter globiformis
-
kcat/Koxygen values are independent of the pH between pH 5.0 and 10.0, average value is 1.7 mM
2
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
9
(2-amino-ethyl)trimethylammonim chloride
-
pH 8.0, 25C
15
allyltrimethylammonium chloride
-
pH 8.0, 25C
2
Betaine aldehyde
-
oxidation of choline is competitively inhibited
5
diethylmethylamine
-
pH 8.0, 25C
6
Dimethylamine
-
pH 8.0, 25C
9
dimethylethylamine
-
pH 8.0, 25C
11
ethylamine
-
pH 8.0, 25C
15
glycine betaine
-
pH 8.0, 25C
26
methylamine
-
pH 8.0, 25C
8
Methylethylamine
-
pH 8.0, 25C
57
N,N-dimethylglycine
-
pH 8.0, 25C
405
N-methylglycine
-
pH 8.0, 25C
11
tetramethylamine
-
pH 8.0, 25C
2.4
trimethylamine
-
pH 8.0, 25C
13
trimethylethylammonium chloride
-
pH 8.0, 25C
additional information
additional information
-
inhibition kinetics
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.08
-
H99N mutant, substrate choline, pH 7, 25C. Measuring the rate of oxygen consumption with a computer-interfaced Oxy-32 oxygen monitoring system
0.28
-
apparent, H99N mutant, substrate choline, pH 7, 25C. Measuring the rate of oxygen consumption with a computer-interfaced Oxy-32 oxygen monitoring system
0.3
-
V464A mutant, substrate choline (10 mM), pH 7; V464T mutant, substrate choline (10 mM), pH 7
12.5
-
lyophilized powder
12.8
-
gel filtration
32.1
-
purified enzyme, pH 10.0, 25C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 10
-
assay at
7 - 8
-
choline as substrate
7 - 9
-
assay at
8 - 11
-
H99N mutant
9.5
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 10
6 - 10
6 - 8
-
H99N mutant, lower KM compared to pH 8-11. Measuring the rate of oxygen consumption with a computer-interfaced Oxy-32 oxygen monitoring system
6
-
4C, the anionic flavosemiquinone is slowly oxidized under aerobic conditions
6.5 - 9.6
7.5 - 10
-
biosensor retains 60% of its maximum response
8 - 11
-
H99N mutant, no impact on KM, highest KM. Measuring the rate of oxygen consumption with a computer-interfaced Oxy-32 oxygen monitoring system
8
-
the anionic flavosemiquinone of choline oxidase is unusually insensitive to both molecular oxygen and artificial electron acceptors
additional information
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10 - 45
-
monitoring initial rates of oxygen consumption with oxygen electrode thermostated at, 0.2 mM oxygen in 50 mM sodium pyrophosphate, pH 8
18 - 30
-
similar to that of the free enzyme ranging between 25C and 35C
20 - 39
-
effect of temperature on the rate of reactivation at pH 6
additional information
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
transgenic and wild-type
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
glycine betaine production in transgenic plants
Manually annotated by BRENDA team
-
membrane-bound
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
60612
-
2 * 60614, MALDI-TOF-MS, 2 * 60612, deduced from gene sequence
60614
-
2 * 60614, MALDI-TOF-MS, 2 * 60612, deduced from gene sequence
66000
-
SDS-PAGE
71000
-
SDS-PAGE
83000
-
gel filtration
117000 - 122000
-
gel filtration
120000
128000
-
gel filtration
145000
-
sedimentation velocity method of Svedberg and Pederson
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
monomer
additional information
-
proposed catalytic base is H466
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
active site of wild-type choline oxidase, resolution of 1.86 A
-
anaylsis of biophysical properties of choline oxidase
-
crystal structure determined by synchrotron X-ray radiation, refined to a resolution of 1.86 A, data collected at 100 K
-
hanging drop vapor diffusion method, 1.2-1.8 M ammonium sulfate and 10% (v/v) DMSO in 0.1 M Bis-Tris propane (pH 8.5), flash-frozen in liquid nitrogen. Analysis of the flavin C4a-oxygen adduct in choline oxidase
-
mutant enzyme S101A, hanging drop vapor diffusion method, using 80 mM sodium cacodylate, 20% (v/v) PEG6000, 20% (v/v) glycerol, 150 mM Mg-acetate at pH 6.0
-
purified recombinnat wild-type enzyme and mutant V464A, hanging drop vapor diffusion, room temperature, from 10-15% v/v PEG 6000, 50-200 mM magnesium acetate, 200 mM trimethylamine, and 0.08 M sodium cacodylate, pH 6.0, X-ray diffraction structure determination and analysis at 2.2 A resolution
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 6
-
inactivation takes place at both 27C and 37C
695307
6 - 9
-
inactivation takes place at both 27C and 37C
695307
7 - 8.5
-
no significant inactivation at both 27C and 37C
695307
7 - 9
-
50% initial activity is lost at 40C at pH 6.0 and 9.5
389719
8 - 10.2
-
purified enzyme, 30C, 10 min, stable at
724598
additional information
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40 - 45
-
incubated for 10 min at pH 8.5 and 40C 20% initial activity is lost, 96% at 45C, only 10% initial activity is lost at 45C in presence of 10% glycerol
70 - 90
-
no degradation when incubated for 1 h
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
storage at -20C and pH 6 results in a change in the conformation, loss of catalytic activity at pH 6, reactivation of the enzyme slow at pH 6 and temperatures between 20 and 39C, facilitated by elevated pH values at 25C and pH above 6.5
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
storage at pH 6 and -20C results in a change of conformation of the enzyme, which is associated with complete loss of catalytic activity when the enzyme is assayed at pH 6.0
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native enzyme 41fold by ammonium sulfate fractionation, gel filtation, anion exchange and hydroxyapatite chromatgraphy, and again gel filtration
-
purified with the bound flavin cofactor in the fully oxidized state
-
recombinant enzyme partially from stressed and non-stressed transgenic rice plant leaves by ammonium sulfate fractionation and desalting gel filtration
-
recombinant enzyme, expression in Escherichia coli
-
recombinant protein
-
recombinant protein, purified to homogeneity, gel filtration
-
recombinant wild-type and mutants from Escherichia coli strain Rosetta(DE3)pLysS
-
recombinant wild-type enzyme in oxidized state to homogeneity
-
wild-type and mutant protein, purified enzymes have 50-70% of the enzyme-bound flavin cofactor as an air-stable anionic flavosemiquinone, converts slowly to oxidized state by extensive dialysis at pH 6 and 4C
-
wild-type and mutant variants
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Arabidopsis thaliana transformed with codA gene from Arthrobacter globiformis which encodes choline oxidase
-
Balb/c mice used for toxicity studies of choline oxidase - no significant difference from control in terms of growth, body weight, food consumption, and blood biochemical indices. Analysis of various tissues show no significant effect
-
choline oxidase from Arthrobacter pascens facilitates adaptation to osmotic stress in Escherichia coli
-
cloning of Sinorhizobium meliloti bet genes, Tn5 mutant LtS23-1020 is completed by plasmid pCHO34
-
codA gene introduced into Synecochoccus sp. PCC 7942 and Arabidopsis thaliana leads to accumulation of glycinebetaine and enhanced tolerance to salt and cold stress, rice Oryza sativa L. genetically engineered, Agrobacterium-mediated transformation, ability to synthesize glycinebetaine is established by introducing the codA gene for choline oxidase from Arthrobacter globiformis
-
expressed in Escherichia coli from plasmid pET/codA1
-
expressed in Escherichia coli, plant expression vector pGAH/codA,codA gene and nos terminator fragments excised from pGAH/codA, ligated to stress-inducible SWPA2 promoter, fused into pCAMBIA3300 binary vector, resulting pScodA transformed into Solanum tuberosum by using Agrobacterium tumefaciens strain AGL0
-
expressed in Escherichia coli, plasmid pET/codA1
-
expressed in Escherichia coli, strain Rosetta(DE3)pLysS, pET/codAmg plasmid, wild-type and Glu312 variants generated by site-directed mutagenesis
-
expressed in Escherichia coli, wild-type and H351A variant, plasmid pET/codAmg1-H351A
-
expression of wild-type and mutant enzymes in Escherichia coli
-
expression of wild-type and mutant enzymes in Escherichia coli strain Rosetta(DE3)pLysS
-
expression of wild-type and mutants in Escherichia coli strain Rosetta(DE3)pLysS
-
gene codA, expression in Euccalyptus globulus via Agrobaccterium tumefaciens strain EHA105 transfection method using vectors pBI121 and pGW23codA
-
gene codA, expression of Strep-tagged wild-type and mutant enzymes in Escherichia coli strain BL21-Gold
-
heterologous expression in Escherichia coli DH5 alpha and BL21(DE3), respectively. Systematic exchange of individual segments of the Arthrobacter pascens gene with segments from a choline oxidase-encoding gene from Arthrobacter globiformis yielding a functional chimeric enzyme
-
mutant enzyme S101A is expressed in Escherichia coli Rosetta(DE3)pLysS cells
-
overexpression of the enzyme, fused to a chloroplast-transit peptide from Pisum sativum, in transgenic Oryza sativa plants using the Agrobacterium tumefaciens transfection system
-
recombinant expression in Escherichia coli
-
recombinant expression in Escherichia coli strain Rosetta (DE3)pLysS
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E312A
-
generated for investigation of the negative charge on Glu312, enzyme inactive
E312Q
-
generated for investigation of the negative charge on Glu312, Kd value for choline about 500times larger than that of wild-type
H351A
-
generated by site-directed mutagenesis
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
H99N
-
mutant, analysis of kinetic parameters
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
-
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
A21V
-
site-directed mutagenesis in the FAD binding site
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
S348L
-
site-directed mutagenesis in the substrate binding site
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
V349L
-
site-directed mutagenesis in the substrate binding site
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
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additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
conformational change reverting the enzyme to the native form, rate of approaching steady state independent of concentrations of choline and enzyme, increased to a limiting value with increasing pH
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fully oxidized form of the enzyme is obtained via slow oxidation of the flavosemiquinone by extensive dialysis at pH 6 and 4C
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incubation of the inactive enzyme at pH values of pH 6.5 and above, and 25C results in a fast and partial reactivation of the enzyme, which occurrs with slow onset of steady state during enzymatic turnover, overview
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Systematic exchange of individual segments of the Arthrobacter pascens gene with segments from a choline oxidase-encoding gene from Arthrobacter globiformis yielding a functional chimeric enzyme when produced in Escherichia coli
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
analysis
biotechnology
nutrition
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introducing of the codA gene into a cereal crop allows the biosynthesis of glycinebetaine
additional information
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enzyme is of both biotechnological and medical interest, since glycine betaine can be accumulated in the cytoplasm of cells to prevent dehydration and plasmolysis in adverse hyperosmotic environments in pathogenic bacteria