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Literature summary extracted from

  • Mieda, T.; Yabuta, Y.; Rapolu, M.; Motoki, T.; Takeda, T.; Yoshimura, K.; Ishikawa, T.; Shigeoka, S.
    Feedback inhibition of spinach L-galactose dehydrogenase by L-ascorbate (2004), Plant Cell Physiol., 45, 1271-1279.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.1.1.316 dithiothreitol 1 mM, 17% stimulation Spinacia oleracea
1.1.1.316 DTT 1 mM: 17% stimulation Spinacia oleracea
1.1.1.316 glutathione 1 mM: 28% stimulation Spinacia oleracea
1.1.1.316 glutathione 1 mM, 28% stimulation Spinacia oleracea

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.316 expression in Escherichia coli Spinacia oleracea
1.1.1.316 fusion construct with glutathione S-transferase expressed in Escherichia coli BL21(DE3)pLysS Spinacia oleracea

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.316 4-chloromercuribenzoate 0.2 mM: 92% inhibition Spinacia oleracea
1.1.1.316 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
1.1.1.316 N-ethylmaleimide 5 mM: 67% inhibition; 5 mM, 67% inhibition. Inhibition is restored to 71% by incubation with 5 mM GSH Spinacia oleracea
1.1.1.316 p-chloromercuribenzoate 0.2 mM, 92% inhibition Spinacia oleracea

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.316 0.0179
-
 NAD+ pH 7.5, 37°C Spinacia oleracea
1.1.1.316 0.116
-
 L-galactose pH 7.5, 37°C Spinacia oleracea
1.1.1.316 0.1162
-
 L-galactose pH 7.5, 37°C Spinacia oleracea
1.1.1.316 1.5
-
 L-gulose pH 7.5, 37°C Spinacia oleracea

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.316 35261
-
 2 * 35261, calculated from sequence Spinacia oleracea
1.1.1.316 36000
-
 2 * 36000, SDS-PAGE Spinacia oleracea
1.1.1.316 36000
-
 2 * 36000, SDS-PAGE, gel filtration, 35261 Da calculated Spinacia oleracea
1.1.1.316 75000
-
 gel filtration Spinacia oleracea

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.316 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

EC Number Organism UniProt Comment Textmining
1.1.1.316 Spinacia oleracea Q6BDJ2
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.316
-
 Spinacia oleracea
1.1.1.316 ammonium sulfate precipitation, anion exchange chromatography, hydrophobic interaction chromatography and gel filtration, recombinant fusion protein by GST-affinity chromatography Spinacia oleracea

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.1.1.316 cotyledon
-
 Spinacia oleracea
-
1.1.1.316 leaf
-
 Spinacia oleracea
-
1.1.1.316 root
-
 Spinacia oleracea
-
1.1.1.316 stem
-
 Spinacia oleracea
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.316 0.25
-
 purified protein from spinach, pH 7.5, 37°C Spinacia oleracea
1.1.1.316 2.17
-
 pH 7.5, 37°C Spinacia oleracea
1.1.1.316 2.17
-
 purified recombinant protein, pH 7.5, 37°C Spinacia oleracea

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.316 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
1.1.1.316 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
1.1.1.316 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
1.1.1.316 L-glucose + NAD+ 11.9% of the activity with L-galactose Spinacia oleracea ?
-
 ?
1.1.1.316 L-gulose + NAD+ 71.4% of the activity with L-galactose Spinacia oleracea ?
-
 ?
1.1.1.316 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

EC Number Subunits Comment Organism
1.1.1.316 homodimer 2 * 36000, SDS-PAGE Spinacia oleracea
1.1.1.316 homodimer 2 * 36000, SDS-PAGE, gel filtration, 35261 Da calculated Spinacia oleracea
1.1.1.316 homodimer 2 * 35261, calculated from sequence Spinacia oleracea

Synonyms

EC Number Synonyms Comment Organism
1.1.1.316 GDH
-
 Spinacia oleracea
1.1.1.316 L-galactose dehydrogenase
-
 Spinacia oleracea
1.1.1.316 L-GalDH
-
 Spinacia oleracea

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.316 40
-
-
 Spinacia oleracea

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.316 9.25
-
-
 Spinacia oleracea
1.1.1.316 9.3
-
-
 Spinacia oleracea

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.316 NAD+
-
 Spinacia oleracea
1.1.1.316 NAD+ the activity with 1 mM NADP+ is 4.7% of that with 1 mM NAD+ Spinacia oleracea
1.1.1.316 NADP+ the activity with 1 mM NADP+ is 4.7% of that with 1 mM NAD+ Spinacia oleracea

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.1.1.316 0.133
-
 L-ascorbate pH 7.5, 37°C Spinacia oleracea
1.1.1.316 0.1332
-
 L-ascorbate linear-competitive inhibition, pH 7.5, 37°C Spinacia oleracea

General Information

EC Number General Information Comment Organism
1.1.1.316 metabolism key enzyme in the biosynthetic pathway of L-ascorbate in plants Spinacia oleracea