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Literature summary extracted from

  • Leferink, N.G.; van den Berg, W.A.; van Berkel, W.J.
    L-galactono-gamma-lactone dehydrogenase from Arabidopsis thaliana, a flavoprotein involved in vitamin C biosynthesis (2008), FEBS J., 275, 713-726.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.3.2.3 expressed in Escherichia coli BL21(DE3) cells Arabidopsis thaliana

Protein Variants

EC Number Protein Variants Comment Organism
1.3.2.3 L56A less active than the wild type enzyme Arabidopsis thaliana
1.3.2.3 L56C less active than the wild type enzyme Arabidopsis thaliana
1.3.2.3 L56F less active than the wild type enzyme Arabidopsis thaliana
1.3.2.3 L56H less active than the wild type enzyme and releases its FAD cofactor more easily than wild type GALDH Arabidopsis thaliana
1.3.2.3 L56I the mutant displays a higher turnover rate with L-galactono-1,4-lactone than the wild type enzyme Arabidopsis thaliana

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.3.2.3 L-galactono-1,4-lactone inhibitory at concentrations above 2 mM Arabidopsis thaliana
1.3.2.3 additional information no substrate inhibition with L-gulono-1,4-lactone at concentrations up to 100 mM, D-galactono-1,4-lactone, D-gulono-1,4-lactone, L-mannono-1,4-lactone, and D-galacturonic acid do not inhibit the oxidation of L-galactono-1,4-lactone, no specific inhibition by cations or anions is observed Arabidopsis thaliana

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.3.2.3 0.026
-
 phenazine methosulfate wild type enzyme, in the presence of L-galactono-1,4-lactone, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 0.034
-
 cytochrome c wild type enzyme, in the presence of L-galactono-1,4-lactone, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 0.12
-
 L-galactono-1,4-lactone mutant enzyme L56H, in the presence of cytochrome c, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 0.17
-
 L-galactono-1,4-lactone wild type enzyme, in the presence of cytochrome c, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 0.28
-
 1,4-benzoquinone wild type enzyme, in the presence of L-galactono-1,4-lactone, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 0.32
-
 L-galactono-1,4-lactone mutant enzyme L56I, in the presence of cytochrome c, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 0.56
-
 L-galactono-1,4-lactone mutant enzyme L56F in the presence of cytochrome c, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 0.99
-
 L-galactono-1,4-lactone mutant enzyme L56C, in the presence of cytochrome c, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 1.7
-
 L-galactono-1,4-lactone mutant enzyme L56IA in the presence of cytochrome c, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 13.1
-
 L-gulono-1,4-lactone wild type enzyme, in the presence of cytochrome c, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.3.2.3 mitochondrion
-
 Arabidopsis thaliana 5739
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.3.2.3 55000
-
 mature enzyme, SDS-PAGE Arabidopsis thaliana
1.3.2.3 56000
-
 recombinant His-tagged enzyme, gel filtration Arabidopsis thaliana
1.3.2.3 58760
-
 predicted molecular mass of mature His-tagged enzyme Arabidopsis thaliana

Organism

EC Number Organism UniProt Comment Textmining
1.3.2.3 Arabidopsis thaliana
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.3.2.3 Ni-NTA agarose column chromatography and Q-Sepharose column chromatography Arabidopsis thaliana

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.3.2.3 7
-
 cell extract, at pH 8.8 Arabidopsis thaliana
1.3.2.3 76
-
 after purification, at pH 8.8 Arabidopsis thaliana

Storage Stability

EC Number Storage Stability Organism
1.3.2.3 -80°C, 50 mM sodium phosphate and 300 mM NaCl (pH 7.4), more than 12 months, 30-50% loss of activity Arabidopsis thaliana

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.2.3 L-galactono-1,4-lactone + 1,4-benzoquinone
-
 Arabidopsis thaliana L-ascorbate + reduced 1,4-benzoquinone
-
 ?
1.3.2.3 L-galactono-1,4-lactone + cytochrome c
-
 Arabidopsis thaliana L-ascorbate + reduced cytochrome c
-
 ?
1.3.2.3 L-galactono-1,4-lactone + phenazine methosulfate
-
 Arabidopsis thaliana L-ascorbate + reduced phenazine methosulfate
-
 ?
1.3.2.3 L-gulono-1,4-lactone + ferricytochrome c
-
 Arabidopsis thaliana L-ascorbate + ferrocytochrome c
-
 ?
1.3.2.3 additional information D-galactono-1,4-lactone, D-gulono-1,4-lactone, L-mannono-1,4-lactone, and D-galacturonic acid are no substrates, molecular oxygen can not serve as efficient electron acceptor for the mutant enzymes Arabidopsis thaliana ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.3.2.3 ferricytochrome c oxidoreductase
-
 Arabidopsis thaliana
1.3.2.3 GALDH
-
 Arabidopsis thaliana
1.3.2.3 L-galactono-gamma-lactone dehydrogenase catalyzes the final step in the biosynthesis of vitamin C in plants Arabidopsis thaliana

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.3.2.3 52
-
 the recombinant enzyme has a half-life of 20 min at 52°C, in the presence of excess FAD the half-life at 52°C is increased to 115 min Arabidopsis thaliana

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.3.2.3 4
-
 L-gulono-1,4-lactone in the presence of cytochrome c, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 32
-
 L-galactono-1,4-lactone mutant enzyme L56H, in the presence of cytochrome c, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 45
-
 L-galactono-1,4-lactone mutant enzyme L56IA in the presence of cytochrome c, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 64
-
 phenazine methosulfate in the presence of L-galactono-1,4-lactone, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 76
-
 L-galactono-1,4-lactone mutant enzyme L56C, in the presence of cytochrome c, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 108
-
 1,4-benzoquinone in the presence of L-galactono-1,4-lactone, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 126
-
 L-galactono-1,4-lactone mutant enzyme L56F in the presence of cytochrome c, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 134
-
 L-galactono-1,4-lactone wild type enzyme, in the presence of cytochrome c, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 151
-
 cytochrome c in the presence of L-galactono-1,4-lactone, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana
1.3.2.3 240
-
 L-galactono-1,4-lactone mutant enzyme L56I, in the presence of cytochrome c, in 50 mM sodium phosphate (pH 8.8), at 25°C Arabidopsis thaliana

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.3.2.3 8.8
-
 for activity with cytochrome c Arabidopsis thaliana

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.3.2.3 8 9.5 for activity with cytochrome c Arabidopsis thaliana

Cofactor

EC Number Cofactor Comment Organism Structure
1.3.2.3 FAD flavoenzyme with noncovalently bound FAD Arabidopsis thaliana
1.3.2.3 ferricytochrome c
-
 Arabidopsis thaliana

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.3.2.3 16.4
-
 L-galactono-1,4-lactone at 25°C Arabidopsis thaliana

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.3.2.3 Arabidopsis thaliana theoretical value for recombinant His-tagged GALDH
-
 6.8