Cloned (Comment) | Organism |
---|---|
gne yqhE, DNA and amino acid sequence determination and analysis | Escherichia coli |
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 Maximum [Da] | Comment | Organism |
---|---|---|---|
31800 | - |
1 * 31800, SDS-PAGE | Escherichia coli |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
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 | 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 Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
6.3 | - |
- |
Escherichia coli |
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 | 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 | Comment | Organism |
---|---|---|
monomer | 1 * 31800, SDS-PAGE | Escherichia coli |
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 Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Escherichia coli |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | 7% of the activity with NADPH | Escherichia coli | |
NADP+ | - |
Escherichia coli | |
NADPH | - |
Escherichia coli | |
NADPH | dependent on | Escherichia coli |