Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Pyrococcus furiosus |
Protein Variants | Comment | Organism |
---|---|---|
A18P/K40N/Q165H/D182H/E202G/R219M | mutant with increased specific activity towards 2,5-hexanedione compared to the wild type enzyme | Pyrococcus furiosus |
K209T | mutant with wild type specific activity towards 2,5-hexanedione | Pyrococcus furiosus |
L176P | mutant with the highest activity at 30°C with 2,5-hexanedione, the temperature optimum of the mutant is not changed (90°C), but the activity at lower temperature (60°C and below) is clearly increased when compared to the wild type enzyme | Pyrococcus furiosus |
N86D/R213I | mutant with increased specific activity towards 2,5-hexanedione compared to the wild type enzyme | Pyrococcus furiosus |
R11L/A180V | mutant with the highest activity at 30°C with 2,5-hexanedione, the temperature optimum of the mutant is not changed (90°C), but the activity at lower temperature (60°C and below) is clearly increased when compared to the wild type enzyme, the maximum specific activity of R11L/A180V with 2,5-hexanedione at 30°C is 10fold higher compared to the activity of the wild type enzyme | Pyrococcus furiosus |
R11L/A180V | the maximum specific activity of the mutant with 2,5-hexanedione at 30°C is 10fold higher compared to the activity of the wild type enzyme | Pyrococcus furiosus |
T153A | mutant with the highest activity at 30°C with 2,5-hexanedione, the temperature optimum of the mutant is not changed (90°C), but the activity at lower temperature (60°C and below) is clearly increased when compared to the wild type enzyme | Pyrococcus furiosus |
V66A/L176P/Y229H | mutant with increased specific activity towards 2,5-hexanedione compared to the wild type enzyme | Pyrococcus furiosus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.1 | - |
NADPH | mutant enzyme R11L/A180V, at 70°C | Pyrococcus furiosus | |
0.2 | - |
NADP+ | wild type enzyme, at 70°C | Pyrococcus furiosus | |
0.4 | - |
NADPH | wild type enzyme, at 70°C | Pyrococcus furiosus | |
0.7 | - |
NADP+ | mutant enzyme R11L/A180V, at 70°C | Pyrococcus furiosus | |
1.2 | - |
Pyruvaldehyde | wild type enzyme, at 70°C | Pyrococcus furiosus | |
4.2 | - |
Pyruvaldehyde | mutant enzyme R11L/A180V, at 70°C | Pyrococcus furiosus | |
5.4 | - |
(2S,5S)-hexanediol | wild type enzyme, at 70°C | Pyrococcus furiosus | |
30.3 | - |
2-Pentanol | wild type enzyme, at 70°C | Pyrococcus furiosus | |
51.1 | - |
2-Pentanol | mutant enzyme R11L/A180V, at 70°C | Pyrococcus furiosus | |
56.4 | - |
2,5-hexanedione | wild type enzyme, at 70°C | Pyrococcus furiosus | |
79 | - |
2,5-hexanedione | mutant enzyme R11L/A180V, at 70°C | Pyrococcus furiosus | |
327.6 | - |
(2S,5S)-hexanediol | mutant enzyme R11L/A180V, at 70°C | Pyrococcus furiosus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
26000 | - |
2 * 26000 | Pyrococcus furiosus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus furiosus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Q-Sepharose column chromatography | Pyrococcus furiosus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.06 | - |
mutant enzyme N86D/R213I, using 2-pentanol as substrate, at 30°C | Pyrococcus furiosus |
0.07 | - |
mutant enzyme N86D/R213I, using (2S,5S)-hexanediol as substrate, at 30°C | Pyrococcus furiosus |
0.07 | - |
mutant enzyme N86D/R213I, using 2-propanol as substrate, at 30°C | Pyrococcus furiosus |
0.08 | - |
mutant enzyme K209T, using 2-hexanedione as substrate, at 30°C | Pyrococcus furiosus |
0.09 | - |
mutant enzyme N86D/R213I, using 2-butanol as substrate, at 30°C | Pyrococcus furiosus |
0.09 | - |
wild type enzyme, using 2-hexanedione as substrate, at 30°C | Pyrococcus furiosus |
0.11 | - |
wild type enzyme, using 2-propanol as substrate, at 30°C | Pyrococcus furiosus |
0.12 | - |
mutant enzyme A18P/K40N/Q165H/D182H/E202G/R219M, using 2-hexadione as substrate, at 30°C | Pyrococcus furiosus |
0.12 | - |
mutant enzyme N86D/R213I, using 2-hexanedione as substrate, at 30°C | Pyrococcus furiosus |
0.14 | - |
mutant enzyme V66A/L176P/Y229H, using 2-hexanedione as substrate, at 30°C | Pyrococcus furiosus |
0.15 | - |
mutant enzyme K209T, using 2-propanol as substrate, at 30°C | Pyrococcus furiosus |
0.16 | - |
wild type enzyme, using 2-butanol as substrate, at 30°C | Pyrococcus furiosus |
0.17 | - |
mutant enzyme T153A, using 2-propanol as substrate, at 30°C | Pyrococcus furiosus |
0.18 | - |
mutant enzyme T153A, using 2-hexanedione as substrate, at 30°C | Pyrococcus furiosus |
0.19 | - |
mutant enzyme V66A/L176P/Y229H, using 2-propanol as substrate, at 30°C | Pyrococcus furiosus |
0.2 | - |
mutant enzyme A18P/K40N/Q165H/D182H/E202G/R219M, using 2-propanol as substrate, at 30°C | Pyrococcus furiosus |
0.2 | - |
mutant enzyme L176P, using 2-hexanedione as substrate, at 30°C | Pyrococcus furiosus |
0.24 | - |
mutant enzyme T153A, using 2-butanol as substrate, at 30°C | Pyrococcus furiosus |
0.33 | - |
mutant enzyme V66A/L176P/Y229H, using 2-butanol as substrate, at 30°C | Pyrococcus furiosus |
0.35 | - |
mutant enzyme K209T, using 2-butanol as substrate, at 30°C | Pyrococcus furiosus |
0.43 | - |
mutant enzyme L176P, using 2-propanol as substrate, at 30°C | Pyrococcus furiosus |
0.57 | - |
mutant enzyme L176P, using 2-butanol as substrate, at 30°C | Pyrococcus furiosus |
0.6 | - |
mutant enzyme A18P/K40N/Q165H/D182H/E202G/R219M, using 2-butanol as substrate, at 30°C | Pyrococcus furiosus |
0.65 | - |
mutant enzyme K209T, using 2-pentanol as substrate, at 30°C | Pyrococcus furiosus |
0.71 | - |
mutant enzyme A18P/K40N/Q165H/D182H/E202G/R219M, using (2S,5S)-hexanediol as substrate, at 30°C | Pyrococcus furiosus |
0.73 | - |
mutant enzyme L176P, using 2-pentanol as substrate, at 30°C | Pyrococcus furiosus |
0.73 | - |
wild type enzyme, using pyruvaldehyde as substrate, at 30°C | Pyrococcus furiosus |
0.78 | - |
mutant enzyme N86D/R213I, using pyruvaldehyde as substrate, at 30°C | Pyrococcus furiosus |
0.86 | - |
mutant enzyme T153A, using 2-pentanol as substrate, at 30°C | Pyrococcus furiosus |
0.91 | - |
wild type enzyme, using (2S,5S)-hexanediol as substrate, at 30°C | Pyrococcus furiosus |
0.95 | - |
mutant enzyme R11L/A180V, using 2-hexanedione as substrate, at 30°C | Pyrococcus furiosus |
1.07 | - |
mutant enzyme L176P, using pyruvaldehyde as substrate, at 30°C | Pyrococcus furiosus |
1.09 | - |
mutant enzyme V66A/L176P/Y229H, using (2S,5S)-hexanediol as substrate, at 30°C | Pyrococcus furiosus |
1.1 | - |
mutant enzyme K209T, using (2S,5S)-hexanediol as substrate, at 30°C | Pyrococcus furiosus |
1.11 | - |
mutant enzyme A18P/K40N/Q165H/D182H/E202G/R219M, using 2-pentanol as substrate, at 30°C | Pyrococcus furiosus |
1.11 | - |
mutant enzyme R11L/A180V, using 2-propanol as substrate, at 30°C | Pyrococcus furiosus |
1.12 | - |
mutant enzyme K209T, using pyruvaldehyde as substrate, at 30°C | Pyrococcus furiosus |
1.22 | - |
mutant enzyme V66A/L176P/Y229H, using pyruvaldehyde as substrate, at 30°C | Pyrococcus furiosus |
1.24 | - |
mutant enzyme L176P, using (2S,5S)-hexanediol as substrate, at 30°C | Pyrococcus furiosus |
1.3 | - |
mutant enzyme V66A/L176P/Y229H, using 2-pentanol as substrate, at 30°C | Pyrococcus furiosus |
1.31 | - |
mutant enzyme T153A, using (2S,5S)-hexanediol as substrate, at 30°C | Pyrococcus furiosus |
1.47 | - |
wild type enzyme, using 2-pentanol as substrate, at 30°C | Pyrococcus furiosus |
1.98 | - |
mutant enzyme R11L/A180V, using 2-butanol as substrate, at 30°C | Pyrococcus furiosus |
3 | - |
mutant enzyme T153A, using pyruvaldehyde as substrate, at 30°C | Pyrococcus furiosus |
3.23 | - |
mutant enzyme R11L/A180V, using (2S,5S)-hexanediol as substrate, at 30°C | Pyrococcus furiosus |
3.37 | - |
mutant enzyme A18P/K40N/Q165H/D182H/E202G/R219M, using pyruvaldehyde as substrate, at 30°C | Pyrococcus furiosus |
3.47 | - |
mutant enzyme R11L/A180V, using 2-pentanol as substrate, at 30°C | Pyrococcus furiosus |
7.97 | - |
mutant enzyme R11L/A180V, using pyruvaldehyde as substrate, at 30°C | Pyrococcus furiosus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,5-hexanedione + NADP+ | - |
Pyrococcus furiosus | (2S,5S)-hexanediol + NADPH + H+ | - |
r | |
2-butanone + NADP+ | - |
Pyrococcus furiosus | 2-butanol + NADPH + H+ | - |
r | |
2-pentanone + NADP+ | - |
Pyrococcus furiosus | 2-pentanol + NADPH + H+ | - |
r | |
2-propanone + NADP+ | - |
Pyrococcus furiosus | 2-propanol + NADPH + H+ | - |
r | |
pyruvaldehyde + NADP+ | - |
Pyrococcus furiosus | ? + NADPH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 26000 | Pyrococcus furiosus |
Synonyms | Comment | Organism |
---|---|---|
ADH | - |
Pyrococcus furiosus |
AdhA | - |
Pyrococcus furiosus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
preferred temperature for production of (2S,5S)-hexanediol, its activity at 30°C does not exceed 5% of its optimal activity, exhibits increased activity at 30°C on the substrate 2,5-hexanedione | Pyrococcus furiosus |
90 | 100 | the temperature optimum of the wild type enzyme is determined to be between 90 and 100°C | Pyrococcus furiosus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Pyrococcus furiosus |