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Information on EC 1.1.1.274 - 2,5-didehydrogluconate reductase (2-dehydro-D-gluconate-forming) and Organism(s) Escherichia coli and UniProt Accession P30863

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EC Tree
IUBMB Comments
The enzyme is involved in the catabolism of 2,5-didehydrogluconate. cf. EC 1.1.1.346, 2,5-didehydrogluconate reductase (2-dehydro-L-gulonate-forming).
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This record set is specific for:
Escherichia coli
UNIPROT: P30863
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Word Map
The taxonomic range for the selected organisms is: Escherichia coli
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Synonyms
2,5-dkg reductase, 2,5-dkg, 2,5-dkgr, 2,5-diketo-d-gluconate reductase, beta-keto ester reductase, yqhe reductase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2,5-diketo-D-gluconate reductase
2,5-diketo-gluconate reductase
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beta-keto ester reductase
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YqhE reductase
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-
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
2-dehydro-D-gluconate + NADP+ = 2,5-didehydro-D-gluconate + NADPH + H+
show the reaction diagram
enzyme also possesses beta-keto ester reductase activity
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
redox reaction
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-
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oxidation
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-
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reduction
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PATHWAY SOURCE
PATHWAYS
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-, -
SYSTEMATIC NAME
IUBMB Comments
2-dehydro-D-gluconate:NADP+ 2-oxidoreductase (2-dehydro-D-gluconate-forming)
The enzyme is involved in the catabolism of 2,5-didehydrogluconate. cf. EC 1.1.1.346, 2,5-didehydrogluconate reductase (2-dehydro-L-gulonate-forming).
CAS REGISTRY NUMBER
COMMENTARY hide
95725-95-4
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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
2-keto-L-gulonate + NADP+
show the reaction diagram
both isoenzymes A and B are specific for NADPH
-
?
1,2-naphthoquinone + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
?
2,5-didehydro-D-gluconate + NADPH
2-keto-L-gulonate + NADP+
show the reaction diagram
-
-
-
?
2,5-diketo-D-gluconate + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
?
2-carboxybenzaldehyde + NADPH
2-hydroxybenzoate + NADP+
show the reaction diagram
-
-
-
-
?
3-nitrobenzaldehyde + NADPH
3-nitrobenzyl alcohol + NADP+
show the reaction diagram
-
-
-
-
?
4-nitrobenzaldehyde + NADPH
4-nitrobenzyl alcohol + NADP+
show the reaction diagram
-
-
-
-
?
benzaldehyde + NADPH
benzyl alcohol + NADP+
show the reaction diagram
-
-
-
-
?
D-glucuronic acid + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
?
D-xylose + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
?
DL-glyceraldehyde + NADPH + H+
glycerol + NADP+
show the reaction diagram
-
-
-
-
?
ethyl 2-ethylacetoacetate + NADPH
ethyl (2R)-ethyl-(3S)-hydroxybutanoate + NADP+
show the reaction diagram
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stereospecific reaction
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-
?
ethyl 2-methylacetoacetate + NADPH
ethyl (2R)-methyl-(3S)-hydroxybutanoate + NADP+
show the reaction diagram
-
stereospecific reaction
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-
?
ethyl 2-methylacetoacetate + NADPH + H+
ethyl (2R)-methyl-(3S)-hydroxybutanoate + NADP+
show the reaction diagram
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7% activity with NADH
-
?
ethyl acetoacetate + NADPH
ethyl (3S)-hydroxybutanoate + NADP+
show the reaction diagram
-
stereospecific reaction
-
-
?
ethyl-(2R)-allylacetoacetate + NADPH
?
show the reaction diagram
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250% of activity with ethyl-2-methylacetoacetate
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-
?
ethyl-(2R)-ethylacetoacetate + NADPH
ethyl-(2R)-ethyl-(3S)-hydroxybutanoate + NADP+
show the reaction diagram
-
120% of activity with ethyl-2-methylacetoacetate
-
?
ethylacetoacetate + NADPH
ethyl-(3S)-hydroxybutanoate + NADP+
show the reaction diagram
-
53% of activity with ethyl-2-methylacetoacetate
-
?
methylglyoxal + NADPH
? + NADP+
show the reaction diagram
-
-
-
-
?
phenylglyoxal + NADPH
? + NADP+
show the reaction diagram
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-
-
-
?
additional information
?
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substrate specificity, no activity with ethyl propionyl acetate and ethyl butyryl acetate
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-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ethyl 2-ethylacetoacetate + NADPH
ethyl (2R)-ethyl-(3S)-hydroxybutanoate + NADP+
show the reaction diagram
-
stereospecific reaction
-
-
?
ethyl 2-methylacetoacetate + NADPH
ethyl (2R)-methyl-(3S)-hydroxybutanoate + NADP+
show the reaction diagram
-
stereospecific reaction
-
-
?
ethyl acetoacetate + NADPH
ethyl (3S)-hydroxybutanoate + NADP+
show the reaction diagram
-
stereospecific reaction
-
-
?
additional information
?
-
-
substrate specificity, no activity with ethyl propionyl acetate and ethyl butyryl acetate
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADH
-
7% of the activity with NADPH
NADPH
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.1
ethyl 2-methylacetoacetate
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pH 7.0, 30°C, with a cofactor level 20fold higher than the Km value
3.1
ethyl-2-methylacetoacetate
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2.05 - 2.46
methylglyoxal
additional information
additional information
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.6
ethyl 2-methylacetoacetate
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pH 7.0, 30°C
2.6
ethyl-2-methylacetoacetate
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27.62 - 81
methylglyoxal
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0153
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YafB, 10 mM D-glucuronic acid as substrate
0.0247
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YqhE, 10 mM D-glucuronic acid as substrate
0.0401
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YafB, 1 mM 2,5-diketo-D-gluconate as substrate
0.0424
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YafB, 0.1 mM 1,2-naphthoquinone as substrate
0.0519
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YqhE, 1 mM 2,5-diketo-D-gluconate as substrate
0.0949
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YqhE, 10 mM D-xylose as substrate
0.127
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YqhE, 0.1 mM 1,2-naphthoquinone as substrate
1.36
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YafB, 1 mM DL-glyceraldehyde as substrate
10.3
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YqhE, 1 mM 4-nitrobenzaldehyde as substrate
14.9
-
YafB, 1 mM methylglyoxal as substrate
2.21
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YqhE, 1 mM DL-glyceraldehyde as substrate
2.23
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YafB, 1 mM 3-nitrobenzaldehyde as substrate
2.5
-
YqhE, 1 mM 3-nitrobenzaldehyde as substrate
2.79
-
YafB, 1 mM benzaldehyde as substrate
21
-
YqhE, 1 mM methylglyoxal as substrate
3.88
-
YqhE, 1 mM benzaldehyde as substrate
39.1
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YafB, 1 mM phenylglyoxal as substrate
6.53
-
YafB, 1 mM 4-nitrobenzaldehyde as substrate
64.4
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YqhE, 1 mM phenylglyoxal as substrate
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5
isoenzyme A
7
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
assay at
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
Uniprot
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
29000
1 * 29000, isoenzymes A and B, SDS-PAGE
30000
isoenzymes A and B, gel filtration
31800
-
1 * 31800, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
1 * 29000, isoenzymes A and B, SDS-PAGE
monomer
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1 * 31800, SDS-PAGE
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, purified enzyme, 400 mM potassium phoshate buffer, pH 7.0, several weeks without appreciable loss of activity
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant isoenzymes A and B, affinity chitin-binding tag
ammonium sulfate, DEAE-Sepharose, Matrix Red-A, Sephacryl S-200, hypatite C
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DEAE-Sepharose column chromatography and Superdex 200 gel filtration
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native enzyme from strain BL21(DE3), 730fold to homogeneity by ammonium sulfate fractionation, DEAE ion exchange and Red-A affinity chromatography, ultrafiltration, hypatite C-resin chromatography, and a second ultrafiltration step
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
overexpression of isoenzymes A and B
gne yqhE, DNA and amino acid sequence determination and analysis
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
enzyme may be useful for the synthesis of the ascorbate precursor 2-keto-L-gulonate
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 (P30863)
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
Manually annotated by BRENDA team
Ko, J.; Kim, I.; Yoo, S.; Min, B.; Kim, K.; Park, C.
Conversion of methylglyoxal to acetol by Escherichia coli aldo-keto reductases
J. Bacteriol.
187
5782-5789
2005
Escherichia coli
Manually annotated by BRENDA team