Information on EC 1.1.3.12 - pyridoxine 4-oxidase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.1.3.12
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RECOMMENDED NAME
GeneOntology No.
pyridoxine 4-oxidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
pyridoxine + O2 = pyridoxal + H2O2
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
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redox reaction
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reduction
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Microbial metabolism in diverse environments
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vitamin B6 degradation
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Vitamin B6 metabolism
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SYSTEMATIC NAME
IUBMB Comments
pyridoxine:oxygen 4-oxidoreductase
A flavoprotein. Can also use 2,6-dichloroindophenol as an acceptor.
CAS REGISTRY NUMBER
COMMENTARY hide
37250-82-1
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
isolate T6
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Manually annotated by BRENDA team
isolate T2
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Manually annotated by BRENDA team
MA-1
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Manually annotated by BRENDA team
MA-1
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
the enzyme belongs to the glucose methanol choline (GMC) oxidoreductase family of enzymes. Active site Pro504 in PNOX corresponds to Asn or His of the conserved His-Asn or His-His pair in other GMC oxidoreductase active sites
metabolism
first enzyme in pathway I for the degradation of pyridoxine
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
2,6-dihydroxypyridine + O2
? + H2O2
show the reaction diagram
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0.04% of the rate with pyridoxine
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?
2-demethyl-2-ethylpyridoxine + O2
2-demethyl-2-ethylpyridoxal + H2O2
show the reaction diagram
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?
4-pyridinemethanol + O2
isonicotinaldehyde + H2O2
show the reaction diagram
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0.12% of the rate with pyridoxine
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?
5-deoxypyridoxine + O2
5-deoxypyridoxal + H2O2
show the reaction diagram
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?
pyridoxine + 2,6-dichloroindophenol
pyridoxal + ?
show the reaction diagram
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?
pyridoxine + 2,6-dichloroindophenol
pyridoxol + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
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high degree of specificity for pyridoxine
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?
pyridoxine + O2
pyridoxal + H2O2
show the reaction diagram
pyridoxine + vitamin K3
pyridoxal + ?
show the reaction diagram
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?
additional information
?
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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
pyridoxine + O2
pyridoxal + H2O2
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
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no cofactor: FMN
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
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3-chloromercuribenzoate
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o-chloromercuribenzoate, m-chloromercuribenzoate, and p-chloromercuribenzoate inhibit in absence but not in presence of cyanide
alpha,alpha'-dipyridyl
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o-chloromercuribenzoate
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o-chloromercuribenzoate, m-chloromercuribenzoate, and p-chloromercuribenzoate inhibit in absence but not in presence of cyanide
p-chloromercuribenzoate
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o-chloromercuribenzoate, m-chloromercuribenzoate, and p-chloromercuribenzoate inhibit in absence but not in presence of cyanide
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0083 - 0.0097
2,6-dichloroindophenol
0.078 - 0.206
O2
0.0153 - 0.43
pyridoxine
0.05
vitamin K3
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pH 7.0, 30°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.4 - 19.9
pyridoxine
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6 - 1300
pyridoxine
1051
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0000041
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isolate T2
0.0000094
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isolate T6
0.4
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recombinant enzyme in Escherichia coli cells
19.4
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pH 8.0, 30°C
21.1
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pH 7.0, 30°C
additional information
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5 - 8
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6
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48% of maximum activity
7 - 9
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pH 7: 80% of maximal activity, pH 9: 50% of maximal acitvity
10.5
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5% of maximum activity
additional information
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
26
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assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
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30% of maximum activity
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50000
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gel filtration
54500
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gel filtration
55000
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1 * 53000, SDS-PAGE, 1 * 55000, deduced from gene sequence
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant C-terminally His6-tagged PNOX and PNOX-pyridoxamine complex, sitting drop vapour diffusion mthod and micro-seeding method, mixing of 0.002 ml of 10 mg/ml PNOX with 0.002 ml of a precipitant solution comtaining 0.1 M HEPES-NaOH, pH 7.5, 10% v/v 2-propanol, and 20% w/v PEG 4000, equilibration against 01 ml of precipitant solution, 4°C, primary crystals are crushed and suspended in 0.5 ml of a solution consisting of equal volumes of a crystallization buffer containing 50 mM Tris-HCl, pH 8.0, 10% w/v glycerol, 0.005 mM FAD, 0.1% v/v 2-mercaptoethanol, and 0.01% v/v Tween 20, and precipitant solution as seeding solution, or seeding crystals are prepared by vapor diffusion in a mixture consisting of 0.002 ml of 13 mg/ml PNOX, and 0.002 ml of the precipitant solution comprising 0.1 M HEPES-NaOH, pH 7.5, 25% v/v 2-propanol, 20% w/v PEG 4000, and 10 mM pyridoxamine, 3 weeks, X-ray diffraction structuure determination and analysis at 2.2 A and 2.1 A resolutions, respectively, molecular replacement
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, 10% glycerol, 1% 2-mercaptoethanol, 0.02 mM FAD, 0.01% Tween 20, stable
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant C-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli strain JM109
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
gene mll6785, expression of C-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain JM109
overexpression in Escherichia coli strains JM109 and BL21(DE3), co-expression with catalase, chaperonin, and pyridoxal 4-dehydrogenase for construction of a biotransformation system catalyzing the formation of 4-pyridoxolactone from pyridoxine in whole cells, overview
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H460A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
H460A/H462A
site-directed mutagenesis, inactive mutant
H462A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis