EC Number | Cloned (Comment) | Organism |
---|---|---|
1.1.1.1 | expression of wild-type and mutant enzymes in Escherichia coli | Pyrococcus furiosus |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.1.1.1 | H255R | the H255R single mutant exhibits an increased binding affinity toward NADP+ and a concomitant reduction in affinity for NAD+. The apparent kcat for H255R is about 60% of that of the wild-type with NAD+, but it is sixfold higher than the wild-type with NADP+. Position 255 is important for recognizing NADP(H), but is not the sole determinant of cofactor specificity | Pyrococcus furiosus |
1.1.1.1 | K249G | catalytic efficiency increases 5fold for NADH, the efficiency with NADPH increases more than 30fold in comparison to the wild-type | Pyrococcus furiosus |
1.1.1.1 | K249G/H255R | the catalytic efficiency with NADH increases 4fold, the efficiency with NADPH increases more than 16fold. In the oxidation reaction, the kcat with NAD+ improves by 15fold for the double mutant over the wild type enzyme. With NADP+ the kcat is nearly two orders of magnitude larger than the wild-type. Mutant exhibits significantly improved activity and broadened cofactor specificity as compared to the wild-type enzyme | Pyrococcus furiosus |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.1 | 0.0051 | - |
NADP+ | 45°C, pH 8.8, cosubstrate: 2,3-butanediol, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 0.033 | - |
NADPH | 45°C, pH 6.1, cosubstrate: 3-hydroxy-2-butanone, mutant enzyme K249G/H255R | Pyrococcus furiosus | |
1.1.1.1 | 0.05 | - |
NADH | 45°C, pH 6.1, cosubstrate: 3-hydroxy-2-butanone, mutant enzyme K249G/H255R | Pyrococcus furiosus | |
1.1.1.1 | 0.063 | - |
NAD+ | 45°C, pH 8.8, cosubstrate: 2,3-butanediol, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 0.078 | - |
NADP+ | 45°C, pH 8.8, cosubstrate: 2,3-butanediol, mutant enzyme K249G/H255R | Pyrococcus furiosus | |
1.1.1.1 | 0.19 | - |
NADH | 45°C, pH 6.1, cosubstrate: 3-hydroxy-2-butanone, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 0.28 | - |
NADPH | 45°C, pH 6.1, cosubstrate: 3-hydroxy-2-butanone, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 0.46 | - |
NAD+ | 45°C, pH 8.8, cosubstrate: 2,3-butanediol, mutant enzyme K249G/H255R | Pyrococcus furiosus | |
1.1.1.1 | 0.9 | - |
3-hydroxy-2-butanone | 45°C, pH 6.1, cosubstrate: NADH, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 1.3 | - |
2,3-Butanediol | 45°C, pH 8.8, cosubstrate: NADP+, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 5 | - |
3-hydroxy-2-butanone | 45°C, pH 6.1, cosubstrate: NADPH, mutant enzyme K249G/H255R | Pyrococcus furiosus | |
1.1.1.1 | 6.7 | - |
3-hydroxy-2-butanone | 45°C, pH 6.1, cosubstrate: NADPH, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 13 | - |
3-hydroxy-2-butanone | 45°C, pH 6.1, cosubstrate: NADH, mutant enzyme K249G/H255R | Pyrococcus furiosus | |
1.1.1.1 | 29 | - |
2,3-Butanediol | 45°C, pH 8.8, cosubstrate: NAD+, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 200 | - |
2,3-Butanediol | 45°C, pH 8.8, cosubstrate: NADP+, mutant enzyme K249G/H255R | Pyrococcus furiosus | |
1.1.1.1 | 690 | - |
2,3-Butanediol | 45°C, pH 8.8, cosubstrate: NAD+, mutant enzyme K249G/H255R | Pyrococcus furiosus |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.1.1.1 | Pyrococcus furiosus | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.1.1.1 | - |
Pyrococcus furiosus |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.1.1 | 2,3-butanediol + NAD+ | - |
Pyrococcus furiosus | 3-hydroxy-2-butanone + NADH + H+ | - |
r | |
1.1.1.1 | 2,3-butanediol + NADP+ | - |
Pyrococcus furiosus | 3-hydroxy-2-butanone + NADPH + H+ | - |
? | |
1.1.1.1 | 3-hydroxy-2-butanone + NADH + H+ | - |
Pyrococcus furiosus | 2,3-butanediol + NAD+ | - |
r | |
1.1.1.1 | 3-hydroxy-2-butanone + NADPH + H+ | - |
Pyrococcus furiosus | 2,3-butanediol + NADP+ | - |
r |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.1.1.1 | AdhD | - |
Pyrococcus furiosus |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.1.1.1 | 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 | 0.03 | - |
NADP+ | 45°C, pH 8.8, cosubstrate: 2,3-butanediol, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 0.03 | - |
2,3-Butanediol | 45°C, pH 8.8, cosubstrate: NADP+, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 0.2 | - |
NADPH | 45°C, pH 6.1, cosubstrate: 3-hydroxy-2-butanone, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 0.2 | - |
3-hydroxy-2-butanone | 45°C, pH 6.1, cosubstrate: NADPH, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 0.36 | - |
NADH | 45°C, pH 6.1, cosubstrate: 3-hydroxy-2-butanone, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 0.36 | - |
3-hydroxy-2-butanone | 45°C, pH 6.1, cosubstrate: NADH, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 1 | - |
NAD+ | 45°C, pH 8.8, cosubstrate: 2,3-butanediol, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 1 | - |
2,3-Butanediol | 45°C, pH 8.8, cosubstrate: NAD+, wild-type enzyme | Pyrococcus furiosus | |
1.1.1.1 | 1.1 | - |
NADH | 45°C, pH 6.1, cosubstrate: 3-hydroxy-2-butanone, mutant enzyme K249G/H255R | Pyrococcus furiosus | |
1.1.1.1 | 1.1 | - |
3-hydroxy-2-butanone | 45°C, pH 6.1, cosubstrate: NADH, mutant enzyme K249G/H255R | Pyrococcus furiosus | |
1.1.1.1 | 1.2 | - |
NADPH | 45°C, pH 6.1, cosubstrate: 3-hydroxy-2-butanone, mutant enzyme K249G/H255R | Pyrococcus furiosus | |
1.1.1.1 | 1.2 | - |
3-hydroxy-2-butanone | 45°C, pH 6.1, cosubstrate: NADPH, mutant enzyme K249G/H255R | Pyrococcus furiosus | |
1.1.1.1 | 4.7 | - |
NADP+ | 45°C, pH 8.8, cosubstrate: 2,3-butanediol, mutant enzyme K249G/H255R | Pyrococcus furiosus | |
1.1.1.1 | 4.7 | - |
2,3-Butanediol | 45°C, pH 8.8, cosubstrate: NADP+, mutant enzyme K249G/H255R | Pyrococcus furiosus | |
1.1.1.1 | 15 | - |
NAD+ | 45°C, pH 8.8, cosubstrate: 2,3-butanediol, mutant enzyme K249G/H255R | Pyrococcus furiosus | |
1.1.1.1 | 15 | - |
2,3-Butanediol | 45°C, pH 8.8, cosubstrate: NAD+, mutant enzyme K249G/H255R | Pyrococcus furiosus |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.1.1.1 | 6.1 | - |
assay at, reduction of 3-hydroxy-2-butanone | Pyrococcus furiosus |
1.1.1.1 | 8.8 | - |
assay at, oxidation of 2,3-butanediol | Pyrococcus furiosus |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.1 | NAD+ | H255R single mutant exhibits an increased binding affinity toward NADP+ and a concomitant reduction in affinity for NAD+ | Pyrococcus furiosus | |
1.1.1.1 | NADH | - |
Pyrococcus furiosus | |
1.1.1.1 | NADP+ | H255R single mutant exhibits an increased binding affinity toward NADP+ and a concomitant reduction in affinity for NAD+ | Pyrococcus furiosus | |
1.1.1.1 | NADPH | experiments with the highly active double mutant K249G/H255R using NADPH as a cofactor demonstrate an unprecedented transient behavior where the binding mechanism appears to be dependent on cofactor concentration | Pyrococcus furiosus |