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Literature summary for 1.6.5.4 extracted from

  • Sano, S.; Tao, S.; Endo, Y.; Inaba, T.; Hossain, M.A.; Miyake, C.; Matsuo, M.; Aoki, H.; Asada, K.; Saito, K.
    Purification and cDNA cloning of chloroplastic monodehydroascorbate reductase from spinach (2005), Biosci. Biotechnol. Biochem., 69, 762-772.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Spinacia oleracea

Inhibitors

Inhibitors Comment Organism Structure
dicoumarol
-
Spinacia oleracea
iodoacetate 5 mM, 58% inhibition Spinacia oleracea
additional information not inhibited by 5 mM N-ethylmaleimide Spinacia oleracea
p-chloromercuribenzoate 0.1 mM, 90% inhibition Spinacia oleracea

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0063
-
NADH at 0.003 mM monodehydroascorbate Spinacia oleracea

Localization

Localization Comment Organism GeneOntology No. Textmining
chloroplast
-
Spinacia oleracea 9507
-
cytosol
-
Spinacia oleracea 5829
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
50000
-
gel filtration, chloroplastic enzyme Spinacia oleracea
53000
-
SDS-PAGE, cytosolic enzyme Spinacia oleracea

Organism

Organism UniProt Comment Textmining
Spinacia oleracea
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Butyl-Toyopearl column chromatography, HiTrap Q column chromatography and Blue Sepharose 6FF column chromatography Spinacia oleracea

Source Tissue

Source Tissue Comment Organism Textmining
leaf
-
Spinacia oleracea
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.056
-
cell extract, wild type enzyme Spinacia oleracea
13.1
-
cell extract, recombinant enzyme Spinacia oleracea
24
-
after 429fold purification, wild type enzyme Spinacia oleracea
100
-
after 7.6fold purification, recombinant enzyme Spinacia oleracea

Storage Stability

Storage Stability Organism
-80°C, buffer containing 10 mM potassium phosphate at pH 6.5 10 mM 2-mercaptoethanol and 0.1 mM EDTA, several months, no loss of activity Spinacia oleracea

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2 ferricyanide + NADPH
-
Spinacia oleracea 2 ferrocyanide + NADP+ + H+
-
?
NADH + 2,6-dichlorophenol indophenol
-
Spinacia oleracea NAD+ + ?
-
?
NADH + monodehydroascorbate
-
Spinacia oleracea NAD+ + ascorbate
-
?
NADH + monodehydroisoascorbate
-
Spinacia oleracea NAD+ + ?
-
?
NADH + p-benzoquinone
-
Spinacia oleracea NAD+ + ?
-
?

Synonyms

Synonyms Comment Organism
MDA reductase
-
Spinacia oleracea
monodehydroascorbate radical reductase
-
Spinacia oleracea

pH Range

pH Minimum pH Maximum Comment Organism
6.5 9
-
Spinacia oleracea

Cofactor

Cofactor Comment Organism Structure
FAD
-
Spinacia oleracea
FMN
-
Spinacia oleracea
NADH
-
Spinacia oleracea

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.095
-
dicoumarol
-
Spinacia oleracea