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Literature summary for 1.3.2.3 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
Search for more literature for 1.3.2.3

Cloned(Commentary)

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

Protein Variants

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

Inhibitors

Inhibitors Comment Organism Structure
L-galactono-1,4-lactone inhibitory at concentrations above 2 mM Arabidopsis thaliana
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]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
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
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
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
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
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
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
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
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
1.7
-
 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
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

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

Molecular Weight [Da]

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

Organism

Organism UniProt Comment Textmining
Arabidopsis thaliana
-
-
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

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

Storage Stability

Storage Stability Organism
-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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-galactono-1,4-lactone + 1,4-benzoquinone
-
 Arabidopsis thaliana L-ascorbate + reduced 1,4-benzoquinone
-
 ?
L-galactono-1,4-lactone + cytochrome c
-
 Arabidopsis thaliana L-ascorbate + reduced cytochrome c
-
 ?
L-galactono-1,4-lactone + phenazine methosulfate
-
 Arabidopsis thaliana L-ascorbate + reduced phenazine methosulfate
-
 ?
L-gulono-1,4-lactone + ferricytochrome c
-
 Arabidopsis thaliana L-ascorbate + ferrocytochrome c
-
 ?
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

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
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]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
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
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
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
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
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
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
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
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
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
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

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

pH Range

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

Cofactor

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

Ki Value [mM]

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

pI Value

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