Activating Compound | Comment | Organism | Structure |
---|---|---|---|
dithiothreitol | 1 mM, 17% stimulation | Spinacia oleracea | |
DTT | 1 mM: 17% stimulation | Spinacia oleracea | |
glutathione | 1 mM: 28% stimulation | Spinacia oleracea | |
glutathione | 1 mM, 28% stimulation | Spinacia oleracea |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Spinacia oleracea |
fusion construct with glutathione S-transferase expressed in Escherichia coli BL21(DE3)pLysS | Spinacia oleracea |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
4-chloromercuribenzoate | 0.2 mM: 92% inhibition | Spinacia oleracea | |
L-ascorbate | 1 mM, 41% inhibition, linear-competitive inhibition, inhibition is reversible, equilibrium-type of feedback regulation for L-ascorbate synthesis; reversible inhibition by the end-product of the biosynthetic pathway | Spinacia oleracea | |
N-ethylmaleimide | 5 mM: 67% inhibition; 5 mM, 67% inhibition. Inhibition is restored to 71% by incubation with 5 mM GSH | Spinacia oleracea | |
p-chloromercuribenzoate | 0.2 mM, 92% inhibition | Spinacia oleracea |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0179 | - |
NAD+ | pH 7.5, 37°C | Spinacia oleracea | |
0.116 | - |
L-galactose | pH 7.5, 37°C | Spinacia oleracea | |
0.1162 | - |
L-galactose | pH 7.5, 37°C | Spinacia oleracea | |
1.5 | - |
L-gulose | pH 7.5, 37°C | Spinacia oleracea |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
35261 | - |
2 * 35261, calculated from sequence | Spinacia oleracea |
36000 | - |
2 * 36000, SDS-PAGE | Spinacia oleracea |
36000 | - |
2 * 36000, SDS-PAGE, gel filtration, 35261 Da calculated | Spinacia oleracea |
75000 | - |
gel filtration | Spinacia oleracea |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-galactose + NAD+ | Spinacia oleracea | a step in the Smirnoff-Wheeler pathway, ascorbate biosynthesis in higher plants | L-galactono-1,4-lactone + NADH + H+ | - |
ir |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Spinacia oleracea | Q6BDJ2 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Spinacia oleracea |
ammonium sulfate precipitation, anion exchange chromatography, hydrophobic interaction chromatography and gel filtration, recombinant fusion protein by GST-affinity chromatography | Spinacia oleracea |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
cotyledon | - |
Spinacia oleracea | - |
leaf | - |
Spinacia oleracea | - |
root | - |
Spinacia oleracea | - |
stem | - |
Spinacia oleracea | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.25 | - |
purified protein from spinach, pH 7.5, 37°C | Spinacia oleracea |
2.17 | - |
pH 7.5, 37°C | Spinacia oleracea |
2.17 | - |
purified recombinant protein, pH 7.5, 37°C | Spinacia oleracea |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-galactose + NAD+ | activity with 1 mM NADP+ was 4.7% of that with 1 mM NAD+, no activity with L-fructose, L-mannose, L-Xylose, L-arabinose, D-galactose, D-glucose, L-fucose, D-mannose and D-arabinose | Spinacia oleracea | L-galactono-1,4-lactone + NADH + H+ | - |
ir | |
L-galactose + NAD+ | a step in the Smirnoff-Wheeler pathway, ascorbate biosynthesis in higher plants | Spinacia oleracea | L-galactono-1,4-lactone + NADH + H+ | - |
ir | |
L-galactose + NAD+ | high specificity for L-galactose. The activity with 1 mM NADP+ is 4.7% of that with 1 mM NAD+ | Spinacia oleracea | L-galactono-1,4-lactone + NADH + H+ | - |
ir | |
L-glucose + NAD+ | 11.9% of the activity with L-galactose | Spinacia oleracea | ? | - |
? | |
L-gulose + NAD+ | 71.4% of the activity with L-galactose | Spinacia oleracea | ? | - |
? | |
additional information | no activity with L-fructose, L-mannose, L-xylose, L-arabinose, D-galactose, D-glucose, L-fucose, D-mannose and D-arabinose | Spinacia oleracea | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 36000, SDS-PAGE | Spinacia oleracea |
homodimer | 2 * 36000, SDS-PAGE, gel filtration, 35261 Da calculated | Spinacia oleracea |
homodimer | 2 * 35261, calculated from sequence | Spinacia oleracea |
Synonyms | Comment | Organism |
---|---|---|
GDH | - |
Spinacia oleracea |
L-galactose dehydrogenase | - |
Spinacia oleracea |
L-GalDH | - |
Spinacia oleracea |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
- |
Spinacia oleracea |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9.25 | - |
- |
Spinacia oleracea |
9.3 | - |
- |
Spinacia oleracea |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Spinacia oleracea | |
NAD+ | the activity with 1 mM NADP+ is 4.7% of that with 1 mM NAD+ | Spinacia oleracea | |
NADP+ | the activity with 1 mM NADP+ is 4.7% of that with 1 mM NAD+ | Spinacia oleracea |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.133 | - |
L-ascorbate | pH 7.5, 37°C | Spinacia oleracea | |
0.1332 | - |
L-ascorbate | linear-competitive inhibition, pH 7.5, 37°C | Spinacia oleracea |
General Information | Comment | Organism |
---|---|---|
metabolism | key enzyme in the biosynthetic pathway of L-ascorbate in plants | Spinacia oleracea |