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Literature summary for 1.1.1.316 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
Search for more literature for 1.1.1.316

Activating Compound

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(Commentary)

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

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 [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 [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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Spinacia oleracea Q6BDJ2
-
-

Purification (Commentary)

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

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

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

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

Synonyms Comment Organism
GDH
-
 Spinacia oleracea
L-galactose dehydrogenase
-
 Spinacia oleracea
L-GalDH
-
 Spinacia oleracea

Temperature Optimum [°C]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9.25
-
-
 Spinacia oleracea
9.3
-
-
 Spinacia oleracea

Cofactor

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 [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

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