Cloned (Comment) | Organism |
---|---|
gene PH0743, recombinant expression of StrepII-tagged enzyme in Escherichia coli strain BL21(DE3) | Pyrococcus horikoshii |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | the Fe2+-reconstituted Ni-PhADH was sensitively and rapidly inactivated by dioxygen gas. The enzyme is inactivated by the replacement of the ferrous ion in the active site with another metal ion such as a zinc ion, which has been considered an inhibitor of iron-activating group III ADHs | Pyrococcus horikoshii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Pyrococcus horikoshii | |
0.011 | - |
NADH | substrates NADH and butanal, pH 5.5, 30°C | Pyrococcus horikoshii | |
0.03 | - |
NAD+ | substrates NAD+ and 1-butanol, pH 9.0, 30°C | Pyrococcus horikoshii | |
0.09 | - |
pentanal | substrates NADH and pentanal, pH 5.5, 30°C | Pyrococcus horikoshii | |
0.25 | - |
Butanal | substrates NADH and butanal, pH 5.5, 30°C | Pyrococcus horikoshii | |
0.52 | - |
1-butanol | substrates NAD+ and 1-butanol, pH 9.0, 30°C | Pyrococcus horikoshii | |
0.52 | - |
NADPH | substrates NADPH and butanal, pH 5.5, 30°C | Pyrococcus horikoshii | |
2.77 | - |
propanal | substrates NADH and propanal, pH 5.5, 30°C | Pyrococcus horikoshii |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | moderately activating the reduction of butanal, but no activation of oxidation of 1-butanol | Pyrococcus horikoshii | |
Fe2+ | moderately activating the reduction of butanal, but only very weak activation of oxidation of 1-butanol | Pyrococcus horikoshii | |
Fe3+ | moderately activating the reduction of butanal, but no activation of oxidation of 1-butanol | Pyrococcus horikoshii | |
Mn2+ | moderately activating the reduction of butanal, but only very weak activation of oxidation of 1-butanol | Pyrococcus horikoshii | |
additional information | no activity with Ca2+, Mg2+, Zn2+, and Cu2+. The Fe2+-reconstituted Ni-PhADH is sensitively and rapidly inactivated by dioxygen gas. The activity of apoform o-PhADH is significantly lower than that of Ni2+-bound PhADH. The enzyme is inactivated by the replacement of the ferrous ion in the active site with another metal ion such as a zinc ion, which has been considered an inhibitor of iron-activating group III ADHs | Pyrococcus horikoshii | |
Ni2+ | required for reduction of butanal, activates, but only slightly in oxidation of 1-butanol | Pyrococcus horikoshii |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus horikoshii | O58517 | - |
- |
Pyrococcus horikoshii ATCC 700860 | O58517 | - |
- |
Pyrococcus horikoshii DSM 12428 | O58517 | - |
- |
Pyrococcus horikoshii JCM 9974 | O58517 | - |
- |
Pyrococcus horikoshii NBRC 100139 | O58517 | - |
- |
Pyrococcus horikoshii OT-3 | O58517 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant StrepII-tagged enzyme from Escherichia coli strain BL21(DE3) by affinity chromatography, ultrafiltration, and gel filtration | Pyrococcus horikoshii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1,2-propanediol + NAD+ | - |
Pyrococcus horikoshii | ? + NADH + H+ | - |
r | |
1,3-propanediol + NAD+ | - |
Pyrococcus horikoshii | ? + NADH + H+ | - |
r | |
1,4-butanediol + NAD+ | - |
Pyrococcus horikoshii | ? + NADH + H+ | - |
r | |
1-butanol + NAD+ | - |
Pyrococcus horikoshii | butanal + NADH + H+ | - |
r | |
1-butanol + NAD+ | - |
Pyrococcus horikoshii DSM 12428 | butanal + NADH + H+ | - |
r | |
1-butanol + NAD+ | - |
Pyrococcus horikoshii NBRC 100139 | butanal + NADH + H+ | - |
r | |
1-butanol + NAD+ | - |
Pyrococcus horikoshii JCM 9974 | butanal + NADH + H+ | - |
r | |
1-butanol + NAD+ | - |
Pyrococcus horikoshii ATCC 700860 | butanal + NADH + H+ | - |
r | |
1-butanol + NAD+ | - |
Pyrococcus horikoshii OT-3 | butanal + NADH + H+ | - |
r | |
1-butanol + NADP+ | - |
Pyrococcus horikoshii | butanal + NADPH + H+ | - |
r | |
1-heptanol + NAD+ | - |
Pyrococcus horikoshii | heptanal + NADH + H+ | - |
r | |
1-heptanol + NAD+ | - |
Pyrococcus horikoshii DSM 12428 | heptanal + NADH + H+ | - |
r | |
1-heptanol + NAD+ | - |
Pyrococcus horikoshii NBRC 100139 | heptanal + NADH + H+ | - |
r | |
1-heptanol + NAD+ | - |
Pyrococcus horikoshii JCM 9974 | heptanal + NADH + H+ | - |
r | |
1-heptanol + NAD+ | - |
Pyrococcus horikoshii ATCC 700860 | heptanal + NADH + H+ | - |
r | |
1-heptanol + NAD+ | - |
Pyrococcus horikoshii OT-3 | heptanal + NADH + H+ | - |
r | |
1-hexanol + NAD+ | - |
Pyrococcus horikoshii | hexanal + NADH + H+ | - |
r | |
1-hexanol + NAD+ | - |
Pyrococcus horikoshii DSM 12428 | hexanal + NADH + H+ | - |
r | |
1-hexanol + NAD+ | - |
Pyrococcus horikoshii NBRC 100139 | hexanal + NADH + H+ | - |
r | |
1-hexanol + NAD+ | - |
Pyrococcus horikoshii JCM 9974 | hexanal + NADH + H+ | - |
r | |
1-hexanol + NAD+ | - |
Pyrococcus horikoshii ATCC 700860 | hexanal + NADH + H+ | - |
r | |
1-hexanol + NAD+ | - |
Pyrococcus horikoshii OT-3 | hexanal + NADH + H+ | - |
r | |
1-octanol + NAD+ | - |
Pyrococcus horikoshii | octanal + NADH + H+ | - |
r | |
1-pentanol + NAD+ | - |
Pyrococcus horikoshii | pentanal + NADH + H+ | - |
r | |
1-propanol + NAD+ | - |
Pyrococcus horikoshii | propanal + NADH + H+ | - |
r | |
benzylalcohol + NAD+ | - |
Pyrococcus horikoshii | benzaldehyde + NADH + H+ | - |
r | |
cyclohexanol + NAD+ | - |
Pyrococcus horikoshii | cyclohexanone + NADH + H+ | - |
r | |
ethanol + NAD+ | - |
Pyrococcus horikoshii | acetaldehyde + NADH + H+ | - |
r | |
methanol + NAD+ | - |
Pyrococcus horikoshii | formaldehyde + NADH + H+ | - |
r | |
methanol + NAD+ | - |
Pyrococcus horikoshii DSM 12428 | formaldehyde + NADH + H+ | - |
r | |
methanol + NAD+ | - |
Pyrococcus horikoshii NBRC 100139 | formaldehyde + NADH + H+ | - |
r | |
methanol + NAD+ | - |
Pyrococcus horikoshii JCM 9974 | formaldehyde + NADH + H+ | - |
r | |
methanol + NAD+ | - |
Pyrococcus horikoshii ATCC 700860 | formaldehyde + NADH + H+ | - |
r | |
methanol + NAD+ | - |
Pyrococcus horikoshii OT-3 | formaldehyde + NADH + H+ | - |
r | |
additional information | substrate specificity of Ni-PhADH for alcohol oxidation reactions and aldehyde reduction reactions, overview. No activity with 2-butanol, tert-butanol, and 2,3-butanediol | Pyrococcus horikoshii | ? | - |
- |
|
additional information | substrate specificity of Ni-PhADH for alcohol oxidation reactions and aldehyde reduction reactions, overview. No activity with 2-butanol, tert-butanol, and 2,3-butanediol | Pyrococcus horikoshii DSM 12428 | ? | - |
- |
|
additional information | substrate specificity of Ni-PhADH for alcohol oxidation reactions and aldehyde reduction reactions, overview. No activity with 2-butanol, tert-butanol, and 2,3-butanediol | Pyrococcus horikoshii NBRC 100139 | ? | - |
- |
|
additional information | substrate specificity of Ni-PhADH for alcohol oxidation reactions and aldehyde reduction reactions, overview. No activity with 2-butanol, tert-butanol, and 2,3-butanediol | Pyrococcus horikoshii JCM 9974 | ? | - |
- |
|
additional information | substrate specificity of Ni-PhADH for alcohol oxidation reactions and aldehyde reduction reactions, overview. No activity with 2-butanol, tert-butanol, and 2,3-butanediol | Pyrococcus horikoshii ATCC 700860 | ? | - |
- |
|
additional information | substrate specificity of Ni-PhADH for alcohol oxidation reactions and aldehyde reduction reactions, overview. No activity with 2-butanol, tert-butanol, and 2,3-butanediol | Pyrococcus horikoshii OT-3 | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
PH0743 | - |
Pyrococcus horikoshii |
PhADH | - |
Pyrococcus horikoshii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Pyrococcus horikoshii |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | 95 | the enzyme activity is slightly decreased by 1 h incubation at 70°C (2.6%), 80°C (4.7%), 90°C (7.9%), and 95°C (18.8%), 50% loss of activity after approximately 6 h at 95°C | Pyrococcus horikoshii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.48 | - |
1-butanol | substrates NAD+ and 1-butanol, pH 9.0, 30°C | Pyrococcus horikoshii | |
0.5 | - |
NAD+ | substrates NAD+ and 1-butanol, pH 9.0, 30°C | Pyrococcus horikoshii | |
0.82 | - |
Butanal | substrates NADH and butanal, pH 5.5, 30°C | Pyrococcus horikoshii | |
6.66 | - |
propanal | substrates NADH and propanal, pH 5.5, 30°C | Pyrococcus horikoshii | |
6.96 | - |
pentanal | substrates NADH and pentanal, pH 5.5, 30°C | Pyrococcus horikoshii | |
7.4 | - |
NADH | substrates NADH and butanal, pH 5.5, 30°C | Pyrococcus horikoshii | |
7.87 | - |
NADPH | substrates NADPH and butanal, pH 5.5, 30°C | Pyrococcus horikoshii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.5 | - |
aldehyde reduction assay at | Pyrococcus horikoshii |
9 | - |
alcohol oxidation assay at | Pyrococcus horikoshii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Pyrococcus horikoshii | |
NADH | - |
Pyrococcus horikoshii |
General Information | Comment | Organism |
---|---|---|
evolution | NAD(P)-dependent group III alcohol dehydrogenases (ADHs), well known as iron-activated enzymes | Pyrococcus horikoshii |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.92 | - |
1-butanol | substrates NAD+ and 1-butanol, pH 9.0, 30°C | Pyrococcus horikoshii | |
2.4 | - |
propanal | substrates NADH and propanal, pH 5.5, 30°C | Pyrococcus horikoshii | |
3.82 | - |
Butanal | substrates NADH and butanal, pH 5.5, 30°C | Pyrococcus horikoshii | |
15.14 | - |
NADPH | substrates NADPH and butanal, pH 5.5, 30°C | Pyrococcus horikoshii | |
16.67 | - |
NAD+ | substrates NAD+ and 1-butanol, pH 9.0, 30°C | Pyrococcus horikoshii | |
77.33 | - |
pentanal | substrates NADH and pentanal, pH 5.5, 30°C | Pyrococcus horikoshii | |
672.7 | - |
NADH | substrates NADH and butanal, pH 5.5, 30°C | Pyrococcus horikoshii |