BRENDA - Enzyme Database
show all sequences of 1.3.3.12

Galactonolactone oxidoreductase from Trypanosoma cruzi employs a FAD cofactor for the synthesis of vitamin C

Kudryashova, E.V.; Leferink, N.G.; Slot, I.G.; van Berkel, W.J.; Biochim. Biophys. Acta 1814, 545-552 (2011)

Data extracted from this reference:

Application
Application
Commentary
Organism
pharmacology
because humans lack the capacity to synthesize ascorbate, the trypanosomal enzymes involved in ascorbate biosynthesis are interesting targets for drug therapy
Trypanosoma cruzi
Cloned(Commentary)
Cloned (Commentary)
Organism
recombinant expression of the N-terminally His6-tagged enzyme in inclusion bodies in Escherichia coli strain BL21(DE3)
Trypanosoma cruzi
Engineering
Protein Variants
Commentary
Organism
A113G
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
Trypanosoma cruzi
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
L-galactono-1,4-lactone + O2
Trypanosoma cruzi
-
L-ascorbate + H2O2
-
-
?
L-galactono-1,4-lactone + O2
Trypanosoma cruzi X10/6
-
L-ascorbate + H2O2
-
-
?
Organism
Organism
UniProt
Commentary
Textmining
Trypanosoma cruzi
-
-
-
Trypanosoma cruzi X10/6
-
-
-
Purification (Commentary)
Purification (Commentary)
Organism
refolded recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by anion exchange, hydroxyapatite, and hydrophobic interaction chromatography
Trypanosoma cruzi
Renatured (Commentary)
Renatured (Commentary)
Organism
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
Trypanosoma cruzi
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.19
-
purified recombinant refolded enzyme, substrate D-arabinono-1,4-lactone. pH 8.8, temperature not specified in the publication
Trypanosoma cruzi
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
D-arabinono-1,4-lactone + 1,4-benzoquinone
artificial electron acceptor
724431
Trypanosoma cruzi
? + 1,4-benzoquinol
-
-
-
?
D-arabinono-1,4-lactone + 1,4-benzoquinone
artificial electron acceptor
724431
Trypanosoma cruzi X10/6
? + 1,4-benzoquinol
-
-
-
?
D-arabinono-1,4-lactone + O2
-
724431
Trypanosoma cruzi
? + H2O2
-
-
-
?
D-arabinono-1,4-lactone + O2
-
724431
Trypanosoma cruzi X10/6
? + H2O2
-
-
-
?
L-galactono-1,4-lactone + 1,4-benzoquinone
artificial electron acceptor
724431
Trypanosoma cruzi
L-ascorbate + 1,4-benzoquinol
-
-
-
?
L-galactono-1,4-lactone + 1,4-benzoquinone
artificial electron acceptor
724431
Trypanosoma cruzi X10/6
L-ascorbate + 1,4-benzoquinol
-
-
-
?
L-galactono-1,4-lactone + O2
-
724431
Trypanosoma cruzi
L-ascorbate + H2O2
-
-
-
?
L-galactono-1,4-lactone + O2
-
724431
Trypanosoma cruzi X10/6
L-ascorbate + H2O2
-
-
-
?
additional information
the refolded enzyme is active with both L-galactono-1,4-lactone and D-arabinono-1,4-lactone
724431
Trypanosoma cruzi
?
-
-
-
?
additional information
the refolded enzyme is active with both L-galactono-1,4-lactone and D-arabinono-1,4-lactone
724431
Trypanosoma cruzi X10/6
?
-
-
-
?
Subunits
Subunits
Commentary
Organism
monomer or dimer
-
Trypanosoma cruzi
Synonyms
Synonyms
Commentary
Organism
GAL
-
Trypanosoma cruzi
galactonolactone oxidoreductase
-
Trypanosoma cruzi
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.2
-
assay at, substrate L-galactono-1,4-lactone
Trypanosoma cruzi
8.8
-
assay at, substrate D-arabinono-1,4-lactone
Trypanosoma cruzi
Cofactor
Cofactor
Commentary
Organism
Structure
FAD
non-covalently bound FAD as redox-active cofactor, FAD-binding domain sequence of TcGAL and comparison to related aldonolactone oxidoreductases, overview
Trypanosoma cruzi
Application (protein specific)
Application
Commentary
Organism
pharmacology
because humans lack the capacity to synthesize ascorbate, the trypanosomal enzymes involved in ascorbate biosynthesis are interesting targets for drug therapy
Trypanosoma cruzi
Cloned(Commentary) (protein specific)
Commentary
Organism
recombinant expression of the N-terminally His6-tagged enzyme in inclusion bodies in Escherichia coli strain BL21(DE3)
Trypanosoma cruzi
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FAD
non-covalently bound FAD as redox-active cofactor, FAD-binding domain sequence of TcGAL and comparison to related aldonolactone oxidoreductases, overview
Trypanosoma cruzi
Engineering (protein specific)
Protein Variants
Commentary
Organism
A113G
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
Trypanosoma cruzi
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
L-galactono-1,4-lactone + O2
Trypanosoma cruzi
-
L-ascorbate + H2O2
-
-
?
L-galactono-1,4-lactone + O2
Trypanosoma cruzi X10/6
-
L-ascorbate + H2O2
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
refolded recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by anion exchange, hydroxyapatite, and hydrophobic interaction chromatography
Trypanosoma cruzi
Renatured (Commentary) (protein specific)
Commentary
Organism
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
Trypanosoma cruzi
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.19
-
purified recombinant refolded enzyme, substrate D-arabinono-1,4-lactone. pH 8.8, temperature not specified in the publication
Trypanosoma cruzi
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
D-arabinono-1,4-lactone + 1,4-benzoquinone
artificial electron acceptor
724431
Trypanosoma cruzi
? + 1,4-benzoquinol
-
-
-
?
D-arabinono-1,4-lactone + 1,4-benzoquinone
artificial electron acceptor
724431
Trypanosoma cruzi X10/6
? + 1,4-benzoquinol
-
-
-
?
D-arabinono-1,4-lactone + O2
-
724431
Trypanosoma cruzi
? + H2O2
-
-
-
?
D-arabinono-1,4-lactone + O2
-
724431
Trypanosoma cruzi X10/6
? + H2O2
-
-
-
?
L-galactono-1,4-lactone + 1,4-benzoquinone
artificial electron acceptor
724431
Trypanosoma cruzi
L-ascorbate + 1,4-benzoquinol
-
-
-
?
L-galactono-1,4-lactone + 1,4-benzoquinone
artificial electron acceptor
724431
Trypanosoma cruzi X10/6
L-ascorbate + 1,4-benzoquinol
-
-
-
?
L-galactono-1,4-lactone + O2
-
724431
Trypanosoma cruzi
L-ascorbate + H2O2
-
-
-
?
L-galactono-1,4-lactone + O2
-
724431
Trypanosoma cruzi X10/6
L-ascorbate + H2O2
-
-
-
?
additional information
the refolded enzyme is active with both L-galactono-1,4-lactone and D-arabinono-1,4-lactone
724431
Trypanosoma cruzi
?
-
-
-
?
additional information
the refolded enzyme is active with both L-galactono-1,4-lactone and D-arabinono-1,4-lactone
724431
Trypanosoma cruzi X10/6
?
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
monomer or dimer
-
Trypanosoma cruzi
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.2
-
assay at, substrate L-galactono-1,4-lactone
Trypanosoma cruzi
8.8
-
assay at, substrate D-arabinono-1,4-lactone
Trypanosoma cruzi
General Information
General Information
Commentary
Organism
evolution
the terminal step in ascorbate biosynthesis is catalyzed by flavin-dependent aldonolactone oxidoreductases belonging to the vanillyl-alcohol oxidase (VAO) protein family
Trypanosoma cruzi
metabolism
the enzyme is important in vitamin C biosynthesis
Trypanosoma cruzi
General Information (protein specific)
General Information
Commentary
Organism
evolution
the terminal step in ascorbate biosynthesis is catalyzed by flavin-dependent aldonolactone oxidoreductases belonging to the vanillyl-alcohol oxidase (VAO) protein family
Trypanosoma cruzi
metabolism
the enzyme is important in vitamin C biosynthesis
Trypanosoma cruzi
Other publictions for EC 1.3.3.12
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
724431
Kudryashova
Galactonolactone oxidoreductas ...
Trypanosoma cruzi, Trypanosoma cruzi X10/6
Biochim. Biophys. Acta
1814
545-552
2011
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10
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10
1
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2
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2
2
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-
684985
Logan
The terminal step in vitamin C ...
Trypanosoma cruzi
Biochem. J.
407
419-426
2007
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1
-
6
-
4
2
-
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1
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7
-
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1
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2
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2
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2
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1
2
1
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1
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1
1
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6
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4
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2
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1
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1
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2
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2
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1
2
1
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-
-
-
-
-
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-
389748
Nishikimi
-
Identification by bacterial ex ...
Saccharomyces cerevisiae
Biochem. Mol. Biol. Int.
44
907-913
1998
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1
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1
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1
-
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-
-
-
-
-
-
-
-
-
389749
De Gara
-
The biogenesis of galactone-ga ...
Avena sativa
Phytochemistry
31
755-756
1992
-
-
-
-
-
-
1
-
1
-
-
1
-
1
-
-
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-
-
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2
1
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1
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1
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2
1
-
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-
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-
-
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-
-
-
-
389744
Bleeg
Biosynthesis of ascorbate in y ...
Saccharomyces cerevisiae
Eur. J. Biochem.
127
391-396
1982
-
-
-
-
-
-
7
6
1
1
3
1
-
3
-
-
1
-
-
-
1
1
6
1
-
-
-
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1
-
-
1
2
-
-
-
-
-
1
-
-
-
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7
2
6
1
1
3
1
-
-
-
1
-
-
1
1
6
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
389745
Noguchi
Studies on the sulfhydryl grou ...
Saccharomyces cerevisiae
J. Biochem.
90
33-38
1981
-
-
-
-
-
-
7
-
-
-
-
1
-
1
-
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1
-
-
-
-
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2
-
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1
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1
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7
-
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1
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1
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-
2
-
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-
-
-
-
-
-
-
-
-
-
-
-
-
389746
Kenney
Identification of the covalent ...
Saccharomyces cerevisiae
FEBS Lett.
97
40-42
1979
-
-
-
-
-
-
-
-
-
-
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1
-
3
-
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1
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2
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2
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2
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1
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2
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-
-
-
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-
-
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389747
Nishikimi
Occurrence in yeast of L-galac ...
Saccharomyces cerevisiae
Arch. Biochem. Biophys.
191
479-486
1978
-
-
-
-
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-
1
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1
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2
1
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5
-
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1
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1
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4
1
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1
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1
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1
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1
-
2
1
-
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1
-
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1
-
4
1
-
-
-
-
-
-
-
-
-
-
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-
-
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