Information on EC 1.2.3.3 - pyruvate oxidase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.2.3.3
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RECOMMENDED NAME
GeneOntology No.
pyruvate oxidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
pyruvate + phosphate + O2 = acetyl phosphate + CO2 + H2O2
show the reaction diagram
a flavoprotein, FAD; A flavoprotein, FAD, requiring thiamine diphosphate
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
decarboxylation
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oxidation
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-
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oxidative decarboxylation
redox reaction
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-
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reduction
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Pyruvate metabolism
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Metabolic pathways
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SYSTEMATIC NAME
IUBMB Comments
pyruvate:oxygen 2-oxidoreductase (phosphorylating)
A flavoprotein (FAD) requiring thiamine diphosphate. Two reducing equivalents are transferred from the resonant carbanion/enamine forms of 2-hydroxyethyl-thiamine-diphosphate to the adjacent flavin cofactor, yielding 2-acetyl-thiamine diphosphate (AcThDP) and reduced flavin. FADH2 is reoxidized by O2 to yield H2O2 and FAD and AcThDP is cleaved phosphorolytically to acetyl phosphate and thiamine diphosphate [2].
CAS REGISTRY NUMBER
COMMENTARY hide
9001-96-1
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Aerococcus sp.
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Manually annotated by BRENDA team
strain ATCC10400
UniProt
Manually annotated by BRENDA team
strain JM101
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Manually annotated by BRENDA team
strain MG1655
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Manually annotated by BRENDA team
strain PB11
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Manually annotated by BRENDA team
strain PB12
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Manually annotated by BRENDA team
strain Lp80
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
Pediococcus sp.
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Manually annotated by BRENDA team
strain UAMS-1
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Manually annotated by BRENDA team
strain UAMS-1
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Manually annotated by BRENDA team
strain GTC13809
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2 pyruvate + phosphate + 2 O2 + H2O
acetate + acetyl phosphate + 2 CO2 + 2 H2O2
show the reaction diagram
acetaldehyde + phosphate + O2 + H2O
? + H2O2
show the reaction diagram
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15% of the activity compared to pyruvate as substrate
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?
alpha-ketobutyrate + phosphate + O2 + H2O
? + H2O2
show the reaction diagram
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very low activity
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?
methylglyoxal + phosphate + O2 + H2O
? + H2O2
show the reaction diagram
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20% of the activity compared to pyruvate as substrate
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?
pyruvate + arsenate + O2 + H2O
acetyl arsenate + CO2 + H2O2
show the reaction diagram
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can replace phosphate, 25% higher activity compared to phosphate as cosubstrate
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?
pyruvate + H2O
acetate + CO2
show the reaction diagram
pyruvate + phosphate + 2,6-dichlorophenolindophenol + H2O
acetyl phosphate + CO2 + reduced 2,6-dichlorophenolindophenol + H2O2
show the reaction diagram
pyruvate + phosphate + ferricyanide + H2O
acetyl phosphate + ferrocyanide + H2O2
show the reaction diagram
pyruvate + phosphate + O2
acetyl phosphate + CO2 + H2O2
show the reaction diagram
pyruvate + phosphate + O2 + H2O
acetyl phosphate + CO2 + H2O2
show the reaction diagram
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
2 pyruvate + phosphate + 2 O2 + H2O
acetate + acetyl phosphate + 2 CO2 + 2 H2O2
show the reaction diagram
pyruvate + H2O
acetate + CO2
show the reaction diagram
pyruvate + phosphate + O2
acetyl phosphate + CO2 + H2O2
show the reaction diagram
pyruvate + phosphate + O2 + H2O
acetyl phosphate + CO2 + H2O2
show the reaction diagram
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
thiamine diphosphate
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
EDTA
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11% inhibition at 10 mM
KCN
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25% inhibition at 10 mM
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
thiamine
Aerococcus sp.
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0.0001 mM thiamine leads to 18.2% increase in activity; activates the enzyme. Can be used as an activator on the biosensor responses
additional information
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.02
2,6-dichlorophenolindophenol
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artificial electron acceptor
1.25
arsenate
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electron acceptor: oxygen
2.3 - 4.5
phosphate
0.4 - 1.6
pyruvate
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
9.6 - 26
pyruvate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.5
ADP
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pH 6.0, 25°C, mutant V265A
3
AMP
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pH 6.0, 25°C, mutant V265A
1.5
ATP
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pH 6.0, 25°C, mutant V265A
6
NAD+
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pH 6.0, 25°C, mutant V265A
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5
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50% of the activity at pH 6.0
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
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a temperature of 25°C gives the highest sensitivity, the enzyme loses activity above 25°C
35
Aerococcus sp.
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
PDB
SCOP
CATH
ORGANISM
UNIPROT
Lactobacillus plantarum (strain ATCC BAA-793 / NCIMB 8826 / WCFS1)
Lactobacillus plantarum (strain ATCC BAA-793 / NCIMB 8826 / WCFS1)
Lactobacillus plantarum (strain ATCC BAA-793 / NCIMB 8826 / WCFS1)
Lactobacillus plantarum (strain ATCC BAA-793 / NCIMB 8826 / WCFS1)
Lactobacillus plantarum (strain ATCC BAA-793 / NCIMB 8826 / WCFS1)
Lactobacillus plantarum (strain ATCC BAA-793 / NCIMB 8826 / WCFS1)
Lactobacillus plantarum (strain ATCC BAA-793 / NCIMB 8826 / WCFS1)
Lactobacillus plantarum (strain ATCC BAA-793 / NCIMB 8826 / WCFS1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
63700
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SDS-PAGE and deduced from sequence
262000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homotetramer
tetramer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by the hanging-drop vapour-diffusion method. POX complexed with FAD, with FAD and thiamine diphosphate, and with FAD and the 2-acetyl-thiamine diphosphate intermediate at 1.6, 1.8 and 1.9 A resolution, respectively. Each subunit of the homotetrameric POX enzyme consists of three domains. FAD is bound within one subunit in the elongated conformation and with the flavin moiety being planar in the oxidized form, while thiamine diphosphate is bound in a conserved V-conformation at the subunit-subunit interface. The structures reveal flexible regions in the active-site tunnel which may undergo conformational changes to allow the entrance of the substrates and the exit of the reaction products. Lys478, the side chain of which may be bent or extended depending on the stage of catalysis
the full-length form and a proteolytically activated C-terminal truncation variant which lacks the last 23 amino acids, by the hanging-drop vapour-diffusion method using either protamine sulfate (fulllength POX) or 2-methyl-2,4-pentanediol (DELTA23 POX) as precipitants. To 2.9 A resolution. Fulllength POX crystallizes in the tetragonal space group P43212 with two monomers per asymmetric unit, the crystals of DELTA23 POX belong to the orthorhombic space group P212121 and contain 12 monomers per asymmetric unit
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in complex with the thiamin diphosphate carbanion-enamine and two-electron reduced FAD, hanging drop vapor diffusion method
C6VNC6
triple mutant at 2.1 A resolution
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wild-type at 2.5 A resolution, triple mutant at 2.1 A resolution, hanging drop vapour diffusion technique
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 9
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10% activity remaining at pH 4.5 and 9.0
671504
5.6 - 5.8
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highest stability at 4°C and highest remaining activity of 67%
390437
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
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unstable above
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
substrate pyruvate can significantly stabilize the enzyme against pressure
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, pH 5.7, several weeks
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by anion-exchange chromatography, more than 95% pure
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by gel filtration
ion exchange column chromatography, immobilization on aminoalkylated controlled pore glass by crosslinking with glutaraldehyde
Pediococcus sp.
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to homogeneity, ultracentrifugation and chromatography techniques
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli DH1
expressed in the chloroplast of Chlamydomonas reinhardtii
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expression in Escherichia coli
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expression in Escherichia coli BL21
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expression in Escherichia coli DH10B
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into the expression vector pSML104 to generate a N-terminus His-tagged fusion expression plasmid pSMLPyODp and expressed in Escherichia coli DH5alpha
overexpressed in Escherichia coli strain ZK126 carrying the plasmid pYYC102 which encodes the poxB gene
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the expression of spxB is decreased in the acetate kinase and SpxR (a regulator of spxB) mutants
the expression of spxB is significantly increased in the thiamine pyrophosphokinase mutant
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
P178S/S188N/A458V
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stabilizes quaternary rather than tertiary structure
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
synthesis
additional information
Show AA Sequence (554 entries)
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