Information on EC 1.3.3.12 - L-galactonolactone oxidase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.3.3.12
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RECOMMENDED NAME
GeneOntology No.
L-galactonolactone oxidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-galactono-1,4-lactone + O2 = L-ascorbate + 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
Ascorbate and aldarate metabolism
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Metabolic pathways
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SYSTEMATIC NAME
IUBMB Comments
L-galactono-1,4-lactone:oxygen 3-oxidoreductase
A flavoprotein. Acts on the 1,4-lactones of L-galactonic, D-altronic, L-fuconic, D-arabinic and D-threonic acids; not identical with EC 1.1.3.8 L-gulonolactone oxidase. (cf. EC 1.3.2.3 galactonolactone dehydrogenase).
CAS REGISTRY NUMBER
COMMENTARY hide
69403-13-0
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
D-altrono-1,4-lactone + O2
?
show the reaction diagram
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-
-
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?
D-altronolactone + O2
?
show the reaction diagram
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68% of the activity with L-galactono-1,4-lactone
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?
D-arabinono-1,4-lactone + 1,4-benzoquinone
? + 1,4-benzoquinol
show the reaction diagram
D-arabinono-1,4-lactone + O2
?
show the reaction diagram
D-arabinono-1,4-lactone + O2
? + H2O2
show the reaction diagram
D-threono-1,4-lactone + O2
?
show the reaction diagram
-
-
-
-
?
L-fucono-1,4-lactone + O2
?
show the reaction diagram
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-
-
-
?
L-galactono-1,4-lactone + 1,4-benzoquinone
L-ascorbate + 1,4-benzoquinol
show the reaction diagram
L-galactono-1,4-lactone + O2
L-ascorbate + H2O2
show the reaction diagram
L-gulonolactone + O2
?
show the reaction diagram
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32% of the activity with L-galactono-1,4-lactone
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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
L-galactono-1,4-lactone + O2
L-ascorbate + H2O2
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
FMN
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dependent on non-covalently-bound FMN
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Iron
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enzyme contains an iron-sulfur cluster
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,2'-dipyridyl disulfide
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4,4'-dipyridyl disulfide
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5,5'-dithiobis(2-nitrobenzoate)
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D-Galactono-1,4-lactone
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competitive inhibition
Hg2+
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complete inhibition at 1 mM
L-gulono-1,4-lactone
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competitive inhibition
N-ethylmaleimide
p-chloromercuribenzoate
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complete inhibition at 1 mM
p-Chloromercuriphenyl sulfonate
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Zn2+
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63% inhibition at 1 mM
additional information
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the synthesis of the enzyme is sensitive to both mitochondrial and cytoplasmic translation inhibitors
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2
D-Altrono-1,4-lactone
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0.16
D-arabino-1,4-lactone
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10.82
D-Arabinono-1,4-lactone
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in 50 mM potassium phosphate buffer, pH 8.0, with 1 mM EDTA
15
D-threono-1,4-lactone
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0.36
L-Fucono-1,4-lactone
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0.3 - 11.22
L-galactono-1,4-lactone
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.285
D-Arabinono-1,4-lactone
Trypanosoma cruzi
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in 50 mM potassium phosphate buffer, pH 8.0, with 1 mM EDTA
0.161
L-galactono-1,4-lactone
Trypanosoma cruzi
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in 50 mM potassium phosphate buffer, pH 8.0, with 1 mM EDTA
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.3
D-Galactono-1,4-lactone
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6.52
L-gulono-1,4-lactone
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1.19
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purified recombinant refolded enzyme, substrate D-arabinono-1,4-lactone. pH 8.8, temperature not specified in the publication
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.2
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assay at, substrate L-galactono-1,4-lactone
8.8
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assay at, substrate D-arabinono-1,4-lactone
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
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might be a multimeric enzyme, in which some polypeptide chains could be synthesized in the cytosol and others in the mitochondria
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Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
18000
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4 * 18000, SDS-PAGE
56000
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4 * 56000, SDS-PAGE
57000
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SDS-PAGE
70000
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gel filtration in presence of deoxycholate
74000
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non-denaturing gradient PAGE in presence of deoxycholate
290000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer or dimer
tetramer
additional information
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might be a multimeric enzyme, in which some polypeptide chains could be synthesized in the cytosol and others in the mitochondria
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
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heat-inactivated above 50°C
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
5°C, 6 months, 50% loss of activity
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Ni-NTA column chromatography
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refolded recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by anion exchange, hydroxyapatite, and hydrophobic interaction chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed as a fusion protein with glutathione S-transferase in Escherichia coli cells with the expression vector pGEX-5X-3
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expressed in Escherichia coli BL-21+ cells
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recombinant expression of the N-terminally His6-tagged enzyme in inclusion bodies in Escherichia coli strain BL21(DE3)
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A113G
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site-directed mutagenesis, in the reaction with 1,4-benzoquinone as electron acceptor, the activity of the dimeric formof A113G is about 1.5fold higher compared to the monomer, the specific activity of the A113G variant in the reaction with molecular oxygen is about 2.5-3times lower than with 1,4-benzoquinone
H447G
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the mutant is inactive and shows undetectable FMN binding
K450G
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the substitution results in an enzyme that retains an ability to bind FMN (around 50%of the wild type enzyme), but which exhibits no activity
K55G
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the mutation has only a minor inhibitory effect on both FMN binding and biochemical activity
K55H
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the mutation causes dramatically reduced FMN binding and enzyme activity (above 90%)
K55L
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the mutant is inactive and shows undetectable FMN binding
W448G
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the mutant is inactive and shows undetectable FMN binding
A113G
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site-directed mutagenesis, in the reaction with 1,4-benzoquinone as electron acceptor, the activity of the dimeric formof A113G is about 1.5fold higher compared to the monomer, the specific activity of the A113G variant in the reaction with molecular oxygen is about 2.5-3times lower than with 1,4-benzoquinone
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Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant terminally His6-tagged enzyme refolded from Escherichia coli strain BL21(DE3) inclusion bodies using a reverse micelles system. The refolded enzyme shows native-like secondary structure and is active with both L-galactono-1,4-lactone and D-arabinono-1,4-lactone
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pharmacology