EC Number | Cloned (Comment) | Organism |
---|---|---|
1.1.1.1 | expression in Escherichia coli | Pyrococcus furiosus |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.1.1.1 | G211C | mutant enzyme exhibits almost complete reversals in cofactor specificity | Pyrococcus furiosus |
1.1.1.1 | G211InsA | with NADP+, the mutant enzyme shows a 29-fold increase in kcat as compared to the wild-type enzyme | Pyrococcus furiosus |
1.1.1.1 | G211InsC | with NAD+, the mutant enzyme exhibits 6-fold decrease in kcat as compared to wild-type enzyme. Mutant enzyme exhibits almost complete reversals in cofactor specificity | Pyrococcus furiosus |
1.1.1.1 | G211InsG | with NAD+, the mutant enzyme exhibits 2.5-fold enhancement in kcat over the wild-type enzyme. Activity with NADP+ exceeds that of the wild type enzyme | Pyrococcus furiosus |
1.1.1.1 | G211InsS | with NAD+, the mutant enzyme exhibits 2.5-fold enhancement in kcat over the wild-type enzyme. With NADP+, the mutant enzyme shows a 29-fold increase in kcat as compared to the wild-type enzyme | Pyrococcus furiosus |
1.1.1.1 | G211S | with NAD+, the mutant enzyme exhibits 1.5-fold enhancement in kcat over the wild-type. Activity with NADP+ exceeds that of the wild type enzyme | Pyrococcus furiosus |
1.1.1.2 | G211C | site-directed mutagenesis, cofactor binding and kinetic analysis compared to the wild-type enzyme | Pyrococcus furiosus |
1.1.1.2 | G211S | site-directed mutagenesis, cofactor binding and kinetic analysis compared to the wild-type enzyme | Pyrococcus furiosus |
1.1.1.2 | additional information | insertion mutants of Ala, Gly, Ser or Cys between positions 211 and 212 (termed G211InsA, G211InsG, G211InsS, and G211InsC, respectively) are created by site-directed mutagenesis of the wild-type AdhD gene in the pET-20b vector. For some mutants (G211S, G211C, G211InsA and G211InsS), with NAD+ as a cofactor, the KA or KB values are observed to be higher than the maximum concentration of substrate or cofactor utilized. So, for mutants G211S, G211C, G211InsA and G211InsS, kinetic data are also measured using assay reaction mixtures containing 1-450 mM 2,3-butanediol and 1-5500 microM NAD+. In the case of NAD+, the on-rate of cofactor (kss 1 ) is found to decrease in all of the mutants except for the G211S and insertion of Gly (G211InsG). Kinetic analysis, detailed overview | Pyrococcus furiosus |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.1 | 0.097 | - |
NAD+ | pH 8.8, 45°C, mutant enzyme G211InsC | Pyrococcus furiosus | |
1.1.1.1 | 0.1 | - |
NAD+ | pH 8.8, 45°C, mutant enzyme G211S | Pyrococcus furiosus | |
1.1.1.1 | 0.12 | - |
NAD+ | pH 8.8, 45°C, mutant enzyme G211InsA | Pyrococcus furiosus | |
1.1.1.1 | 0.23 | - |
NAD+ | pH 8.8, 45°C, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 0.29 | - |
NAD+ | pH 8.8, 45°C, mutant enzyme G211InsG | Pyrococcus furiosus | |
1.1.1.1 | 0.44 | - |
2,3-Butanediol | pH 8.8, 45°C, wild-type enzyme, cofactor: NAP+ | Pyrococcus furiosus | |
1.1.1.1 | 0.48 | - |
NAD+ | pH 8.8, 45°C, mutant enzyme G211C | Pyrococcus furiosus | |
1.1.1.1 | 0.91 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211S, cofactor: NADP+ | Pyrococcus furiosus | |
1.1.1.1 | 1.5 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211InsG, cofactor: NADP+ | Pyrococcus furiosus | |
1.1.1.1 | 1.6 | - |
NAD+ | pH 8.8, 45°C, mutant enzyme G211InsS | Pyrococcus furiosus | |
1.1.1.1 | 5.5 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211InsC, cofactor: NADP+ | Pyrococcus furiosus | |
1.1.1.1 | 7.4 | - |
NADP+ | pH 8.8, 45°C, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 9.6 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211C, cofactor: NADP+ | Pyrococcus furiosus | |
1.1.1.1 | 12 | - |
2,3-Butanediol | pH 8.8, 45°C, wild-type enzyme, cofactor: NAD+ | Pyrococcus furiosus | |
1.1.1.1 | 15 | - |
NADP+ | pH 8.8, 45°C, mutant enzyme G211S | Pyrococcus furiosus | |
1.1.1.1 | 17 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211InsS, cofactor: NADP+ | Pyrococcus furiosus | |
1.1.1.1 | 25 | - |
NADP+ | pH 8.8, 45°C, mutant enzyme G211C | Pyrococcus furiosus | |
1.1.1.1 | 33 | - |
NADP+ | pH 8.8, 45°C, mutant enzyme G211InsG | Pyrococcus furiosus | |
1.1.1.1 | 66 | - |
NADP+ | pH 8.8, 45°C, mutant enzyme G211InsS | Pyrococcus furiosus | |
1.1.1.1 | 76 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211InsC, cofactor: NAD+ | Pyrococcus furiosus | |
1.1.1.1 | 90 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211InsA, cofactor: NADP+ | Pyrococcus furiosus | |
1.1.1.1 | 100 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211S, cofactor: NAD+ | Pyrococcus furiosus | |
1.1.1.1 | 110 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211InsG, cofactor: NAD+ | Pyrococcus furiosus | |
1.1.1.1 | 120 | - |
NADP+ | pH 8.8, 45°C, mutant enzyme G211InsC | Pyrococcus furiosus | |
1.1.1.1 | 240 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211C, cofactor: NAD+ | Pyrococcus furiosus | |
1.1.1.1 | 260 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211InsA, cofactor: NAD+ | Pyrococcus furiosus | |
1.1.1.1 | 360 | - |
NADP+ | pH 8.8, 45°C, mutant enzyme G211InsA | Pyrococcus furiosus | |
1.1.1.1 | 1100 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211InsS, cofactor: NAD+ | Pyrococcus furiosus | |
1.1.1.2 | additional information | - |
additional information | kinetic analysis of wild-type and mutant enzymes, fit to the ordered bi-bi rate equation, steady-state kinetics, detailed overview. For some mutants (G211S, G211C, G211InsA and G211InsS), with NAD+ as a cofactor, the KA or KB values are observed to be higher than the maximum concentration of substrate or cofactor utilized. So, for mutants G211S, G211C, G211InsA and G211InsS, kinetic data is also measured using assay reaction mixtures containing 1-450 mM 2,3-butanediol and 0.001-5.50 mM NAD+ | Pyrococcus furiosus |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.1.1.1 | Pyrococcus furiosus | - |
- |
- |
1.1.1.2 | Pyrococcus furiosus | - |
- |
- |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.1.1 | 2,3-butanediol + NAD+ | - |
Pyrococcus furiosus | acetoin + NADH + H+ | - |
? | |
1.1.1.1 | 2,3-butanediol + NADP+ | - |
Pyrococcus furiosus | acetoin + NADPH + H+ | - |
? | |
1.1.1.2 | 2,3-butanediol + NADP+ | - |
Pyrococcus furiosus | acetoin + NADPH + H+ | - |
r |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.1.1.1 | AdhD | - |
Pyrococcus furiosus |
1.1.1.1 | alcohol dehydrogenase D | - |
Pyrococcus furiosus |
1.1.1.2 | AdhD | - |
Pyrococcus furiosus |
1.1.1.2 | alcohol dehydrogenase D | - |
Pyrococcus furiosus |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.1.1.1 | 45 | - |
assay at | Pyrococcus furiosus |
1.1.1.2 | 45 | - |
assay at | Pyrococcus furiosus |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.1 | 0.073 | - |
NADP+ | pH 8.8, 45°C, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 0.073 | - |
2,3-Butanediol | pH 8.8, 45°C, wild-type enzyme, cofactor: NAP+ | Pyrococcus furiosus | |
1.1.1.1 | 0.29 | - |
NADP+ | pH 8.8, 45°C, mutant enzyme G211S | Pyrococcus furiosus | |
1.1.1.1 | 0.29 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211S, cofactor: NADP+ | Pyrococcus furiosus | |
1.1.1.1 | 0.43 | - |
NAD+ | pH 8.8, 45°C, mutant enzyme G211InsC | Pyrococcus furiosus | |
1.1.1.1 | 0.43 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211InsC, cofactor: NAD+ | Pyrococcus furiosus | |
1.1.1.1 | 0.52 | - |
NADP+ | pH 8.8, 45°C, mutant enzyme G211InsG | Pyrococcus furiosus | |
1.1.1.1 | 0.52 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211InsG, cofactor: NADP+ | Pyrococcus furiosus | |
1.1.1.1 | 0.86 | - |
NADP+ | pH 8.8, 45°C, mutant enzyme G211InsC | Pyrococcus furiosus | |
1.1.1.1 | 0.86 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211InsC, cofactor: NADP+ | Pyrococcus furiosus | |
1.1.1.1 | 0.92 | - |
NADP+ | pH 8.8, 45°C, mutant enzyme G211C | Pyrococcus furiosus | |
1.1.1.1 | 0.92 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211C, cofactor: NADP+ | Pyrococcus furiosus | |
1.1.1.1 | 1.1 | - |
NAD+ | pH 8.8, 45°C, mutant enzyme G211C | Pyrococcus furiosus | |
1.1.1.1 | 1.1 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211C, cofactor: NAD+ | Pyrococcus furiosus | |
1.1.1.1 | 1.7 | - |
NAD+ | pH 8.8, 45°C, mutant enzyme G211InsA | Pyrococcus furiosus | |
1.1.1.1 | 1.7 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211InsA, cofactor: NAD+ | Pyrococcus furiosus | |
1.1.1.1 | 2.1 | - |
NADP+ | pH 8.8, 45°C, mutant enzyme G211InsA | Pyrococcus furiosus | |
1.1.1.1 | 2.1 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211InsA, cofactor: NADP+ | Pyrococcus furiosus | |
1.1.1.1 | 2.1 | - |
NADP+ | pH 8.8, 45°C, mutant enzyme G211InsS | Pyrococcus furiosus | |
1.1.1.1 | 2.1 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211InsS, cofactor: NADP+ | Pyrococcus furiosus | |
1.1.1.1 | 2.6 | - |
NAD+ | pH 8.8, 45°C, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 2.6 | - |
2,3-Butanediol | pH 8.8, 45°C, wild-type enzyme, cofactor: NAD+ | Pyrococcus furiosus | |
1.1.1.1 | 4 | - |
NAD+ | pH 8.8, 45°C, mutant enzyme G211S | Pyrococcus furiosus | |
1.1.1.1 | 4 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211S, cofactor: NAD+ | Pyrococcus furiosus | |
1.1.1.1 | 6.5 | - |
NAD+ | pH 8.8, 45°C, mutant enzyme G211InsG | Pyrococcus furiosus | |
1.1.1.1 | 6.5 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211InsG, cofactor: NAD+ | Pyrococcus furiosus | |
1.1.1.1 | 6.7 | - |
NAD+ | pH 8.8, 45°C, mutant enzyme G211InsS | Pyrococcus furiosus | |
1.1.1.1 | 6.7 | - |
2,3-Butanediol | pH 8.8, 45°C, mutant enzyme G211InsS, cofactor: NAD+ | Pyrococcus furiosus |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.1.1.1 | 8.8 | - |
assay at | Pyrococcus furiosus |
1.1.1.2 | 8.8 | - |
assay at | Pyrococcus furiosus |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.1 | NAD+ | - |
Pyrococcus furiosus | |
1.1.1.1 | NADP+ | - |
Pyrococcus furiosus | |
1.1.1.2 | additional information | cofactor binding energies and kinetics with wild-type and mutant enzymes, overview | Pyrococcus furiosus | |
1.1.1.2 | NADP+ | - |
Pyrococcus furiosus | |
1.1.1.2 | NADPH | - |
Pyrococcus furiosus |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.1.1.2 | additional information | structure homology model of AdhD and its mutants generated using the model of prostaglandin F synthase from Trypanosoma brucei (PDB ID 1VBJ, 40.26% identity). The homology models are aligned with the structure of 2,5-DKGR from Corynebacterium (PDB ID 1A80 with bound NADPH, and PDB ID 1M9H with bound NADH), overview | Pyrococcus furiosus |