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Information on EC 1.1.1.346 - 2,5-didehydrogluconate reductase (2-dehydro-L-gulonate-forming) and Organism(s) Escherichia coli and UniProt Accession Q46857

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IUBMB Comments
The enzyme is involved in ketogluconate metabolism, and catalyses the reaction in vivo in the reverse direction to that shown . It is used in the commercial microbial production of ascorbate. cf. EC 1.1.1.274, 2,5-didehydrogluconate reductase (2-dehydro-D-gluconate-forming).
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This record set is specific for:
Escherichia coli
UNIPROT: Q46857
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The taxonomic range for the selected organisms is: Escherichia coli
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Synonyms
25dkgr-a, 2,5-dkgr a, 2,5-diketo-d-gluconic acid reductase a, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2,5-diketo-D-gluconate reductase
-
2,5-diketo-D-gluconate reductase
-
-
2,5-diketo-D-gluconate-reductase
-
-
2,5-DKG reductase
-
-
YqhE reductase
-
-
PATHWAY SOURCE
PATHWAYS
-
-, -
SYSTEMATIC NAME
IUBMB Comments
2-dehydro-D-gluconate:NADP+ 2-oxidoreductase (2-dehydro-L-gulonate-forming)
The enzyme is involved in ketogluconate metabolism, and catalyses the reaction in vivo in the reverse direction to that shown [1]. It is used in the commercial microbial production of ascorbate. cf. EC 1.1.1.274, 2,5-didehydrogluconate reductase (2-dehydro-D-gluconate-forming).
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2,5-didehydro-D-gluconate + NADPH + H+
2-dehydro-L-gulonate + NADP+
show the reaction diagram
NADPH is the preferred electron donor
-
-
?
ethyl 2-acetylpent-4-enoate + NADPH + H+
ethyl (2R)-2-[(1S)-1-hydroxyethyl]pent-4-enoate + NADP+
show the reaction diagram
-
250% activity compared to ethyl 2-methylacetoacetate
-
-
?
ethyl 2-ethyl-3-oxobutanoate + NADPH + H+
ethyl (2R,3S)-2-ethyl-3-hydroxybutanoate + NADP+
show the reaction diagram
-
120% activity compared to ethyl 2-methylacetoacetate
-
-
?
ethyl 2-methylacetoacetate + NADH + H+
ethyl (2R)-methyl-(3S)-hydroxybutanoate + NAD+
show the reaction diagram
-
7% activity with NADH compared to NADPH
-
-
?
ethyl 2-methylacetoacetate + NADPH + H+
ethyl (2R)-methyl-(3S)-hydroxybutanoate + NADP+
show the reaction diagram
-
-
-
-
?
ethyl acetoacetate + NADPH + H+
ethyl (3S)-3-hydroxybutanoate + NADP+
show the reaction diagram
-
53% activity compared to ethyl 2-methylacetoacetate
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADPH
-
dependent on
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.1
ethyl 2-methylacetoacetate
-
at pH 7.0 and 30°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.6
ethyl 2-methylacetoacetate
-
at pH 7.0 and 30°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.3
-
purified enzyme, at pH 7.0 and 30°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.9
-
isoelectric focusing
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
-
the enzyme reduces 2,5-didehydro-D-gluconate, a key step in the microbial synthesis of vitamin C
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
29000
1 * 29000, SDS-PAGE
30000
gel filtration
31003
1 * 31003, calculated from amino acid sequence
31800
-
x * 31800, estimated from SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
?
-
x * 31800, estimated from SDS-PAGE
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, potassium buffer, several weeks, no loss of activity
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Ni-NTA column chromatography
ammonium sulfate precipitation, DEAE-Sepharose column chromatography, Matrex Red-A column chromatography, Sephacryl S-200 gel filtration, and hypatite C cholumn chromatography
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Yum, D.Y.; Lee, B.Y.; Pan, J.G.
Identification of the yqhE and yafB genes encoding two 2,5-diketo-D-gluconate reductases in Escherichia coli
Appl. Environ. Microbiol.
65
3341-3346
1999
Escherichia coli, Escherichia coli (Q46857)
Manually annotated by BRENDA team
Habrych, M.; Rodriguez, S.; Stewart, J.D.
Purification and identification of an Escherichia coli beta-keto ester reductase as 2,5-diketo-D-gluconate reductase YqhE
Biotechnol. Prog.
18
257-261
2002
Escherichia coli, Escherichia coli BL21-(DE3)
Manually annotated by BRENDA team