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Literature summary for 1.1.1.274 extracted from

  • 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 (2002), Biotechnol. Prog., 18, 257-261.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gne yqhE, DNA and amino acid sequence determination and analysis Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information
-
Escherichia coli
3.1
-
ethyl-2-methylacetoacetate
-
Escherichia coli
3.1
-
ethyl 2-methylacetoacetate pH 7.0, 30°C, with a cofactor level 20fold higher than the Km value Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
31800
-
1 * 31800, SDS-PAGE Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ethyl 2-ethylacetoacetate + NADPH Escherichia coli stereospecific reaction ethyl (2R)-ethyl-(3S)-hydroxybutanoate + NADP+
-
?
ethyl 2-methylacetoacetate + NADPH Escherichia coli stereospecific reaction ethyl (2R)-methyl-(3S)-hydroxybutanoate + NADP+
-
?
ethyl acetoacetate + NADPH Escherichia coli stereospecific reaction ethyl (3S)-hydroxybutanoate + NADP+
-
?
additional information Escherichia coli substrate specificity, no activity with ethyl propionyl acetate and ethyl butyryl acetate ?
-
?
additional information Escherichia coli BL21-(DE3) substrate specificity, no activity with ethyl propionyl acetate and ethyl butyryl acetate ?
-
?

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate, DEAE-Sepharose, Matrix Red-A, Sephacryl S-200, hypatite C Escherichia coli
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 Escherichia coli

Reaction

Reaction Comment Organism Reaction ID
2-dehydro-D-gluconate + NADP+ = 2,5-didehydro-D-gluconate + NADPH + H+ enzyme also possesses beta-keto ester reductase activity Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.3
-
-
Escherichia coli

Storage Stability

Storage Stability Organism
4°C, purified enzyme, 400 mM potassium phoshate buffer, pH 7.0, several weeks without appreciable loss of activity Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,5-didehydro-D-gluconate + NADPH
-
Escherichia coli 2-keto-L-gulonate + NADP+
-
?
2,5-didehydro-D-gluconate + NADPH
-
Escherichia coli BL21-(DE3) 2-keto-L-gulonate + NADP+
-
?
ethyl 2-ethylacetoacetate + NADPH stereospecific reaction Escherichia coli ethyl (2R)-ethyl-(3S)-hydroxybutanoate + NADP+
-
?
ethyl 2-methylacetoacetate + NADPH stereospecific reaction Escherichia coli ethyl (2R)-methyl-(3S)-hydroxybutanoate + NADP+
-
?
ethyl 2-methylacetoacetate + NADPH + H+ 7% activity with NADH Escherichia coli ethyl (2R)-methyl-(3S)-hydroxybutanoate + NADP+
-
?
ethyl 2-methylacetoacetate + NADPH + H+ 7% activity with NADH Escherichia coli BL21-(DE3) ethyl (2R)-methyl-(3S)-hydroxybutanoate + NADP+
-
?
ethyl acetoacetate + NADPH stereospecific reaction Escherichia coli ethyl (3S)-hydroxybutanoate + NADP+
-
?
ethyl-(2R)-allylacetoacetate + NADPH 250% of activity with ethyl-2-methylacetoacetate Escherichia coli ?
-
?
ethyl-(2R)-ethylacetoacetate + NADPH 120% of activity with ethyl-2-methylacetoacetate Escherichia coli ethyl-(2R)-ethyl-(3S)-hydroxybutanoate + NADP+
-
?
ethyl-(2R)-ethylacetoacetate + NADPH 120% of activity with ethyl-2-methylacetoacetate Escherichia coli BL21-(DE3) ethyl-(2R)-ethyl-(3S)-hydroxybutanoate + NADP+
-
?
ethylacetoacetate + NADPH 53% of activity with ethyl-2-methylacetoacetate Escherichia coli ethyl-(3S)-hydroxybutanoate + NADP+
-
?
ethylacetoacetate + NADPH 53% of activity with ethyl-2-methylacetoacetate Escherichia coli BL21-(DE3) ethyl-(3S)-hydroxybutanoate + NADP+
-
?
additional information substrate specificity, no activity with ethyl propionyl acetate and ethyl butyryl acetate Escherichia coli ?
-
?
additional information substrate specificity, no activity with ethyl propionyl acetate and ethyl butyryl acetate Escherichia coli BL21-(DE3) ?
-
?

Subunits

Subunits Comment Organism
monomer 1 * 31800, SDS-PAGE Escherichia coli

Synonyms

Synonyms Comment Organism
2,5-diketo-D-gluconate reductase
-
Escherichia coli
2,5-DKG
-
Escherichia coli
beta-keto ester reductase
-
Escherichia coli
YqhE
-
Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.6
-
ethyl-2-methylacetoacetate
-
Escherichia coli
2.6
-
ethyl 2-methylacetoacetate pH 7.0, 30°C Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
NADH 7% of the activity with NADPH Escherichia coli
NADP+
-
Escherichia coli
NADPH
-
Escherichia coli
NADPH dependent on Escherichia coli