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Literature summary extracted from

  • Sugimoto, C.; Takeda, K.; Kariya, Y.; Matsumura, H.; Yohda, M.; Ohno, H.; Nakamura, N.
    A method of expression for an oxygen-tolerant group III alcohol dehydrogenase from Pyrococcus horikoshii OT3 (2017), J. Biol. Inorg. Chem., 22, 527-534 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.71 gene PH0743, recombinant expression of StrepII-tagged enzyme in Escherichia coli strain BL21(DE3) Pyrococcus horikoshii

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.71 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.71 additional information
-
 additional information Michaelis-Menten kinetics Pyrococcus horikoshii
1.1.1.71 0.011
-
 NADH substrates NADH and butanal, pH 5.5, 30°C Pyrococcus horikoshii
1.1.1.71 0.03
-
 NAD+ substrates NAD+ and 1-butanol, pH 9.0, 30°C Pyrococcus horikoshii
1.1.1.71 0.09
-
 pentanal substrates NADH and pentanal, pH 5.5, 30°C Pyrococcus horikoshii
1.1.1.71 0.25
-
 Butanal substrates NADH and butanal, pH 5.5, 30°C Pyrococcus horikoshii
1.1.1.71 0.52
-
 1-butanol substrates NAD+ and 1-butanol, pH 9.0, 30°C Pyrococcus horikoshii
1.1.1.71 0.52
-
 NADPH substrates NADPH and butanal, pH 5.5, 30°C Pyrococcus horikoshii
1.1.1.71 2.77
-
 propanal substrates NADH and propanal, pH 5.5, 30°C Pyrococcus horikoshii

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.71 Co2+ moderately activating the reduction of butanal, but no activation of oxidation of 1-butanol Pyrococcus horikoshii
1.1.1.71 Fe2+ moderately activating the reduction of butanal, but only very weak activation of oxidation of 1-butanol Pyrococcus horikoshii
1.1.1.71 Fe3+ moderately activating the reduction of butanal, but no activation of oxidation of 1-butanol Pyrococcus horikoshii
1.1.1.71 Mn2+ moderately activating the reduction of butanal, but only very weak activation of oxidation of 1-butanol Pyrococcus horikoshii
1.1.1.71 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
1.1.1.71 Ni2+ required for reduction of butanal, activates, but only slightly in oxidation of 1-butanol Pyrococcus horikoshii

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.71 Pyrococcus horikoshii O58517
-
-
1.1.1.71 Pyrococcus horikoshii ATCC 700860 O58517
-
-
1.1.1.71 Pyrococcus horikoshii DSM 12428 O58517
-
-
1.1.1.71 Pyrococcus horikoshii JCM 9974 O58517
-
-
1.1.1.71 Pyrococcus horikoshii NBRC 100139 O58517
-
-
1.1.1.71 Pyrococcus horikoshii OT-3 O58517
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.71 recombinant StrepII-tagged enzyme from Escherichia coli strain BL21(DE3) by affinity chromatography, ultrafiltration, and gel filtration Pyrococcus horikoshii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.71 1,2-propanediol + NAD+
-
 Pyrococcus horikoshii ? + NADH + H+
-
 r
1.1.1.71 1,3-propanediol + NAD+
-
 Pyrococcus horikoshii ? + NADH + H+
-
 r
1.1.1.71 1,4-butanediol + NAD+
-
 Pyrococcus horikoshii ? + NADH + H+
-
 r
1.1.1.71 1-butanol + NAD+
-
 Pyrococcus horikoshii butanal + NADH + H+
-
 r
1.1.1.71 1-butanol + NAD+
-
 Pyrococcus horikoshii DSM 12428 butanal + NADH + H+
-
 r
1.1.1.71 1-butanol + NAD+
-
 Pyrococcus horikoshii NBRC 100139 butanal + NADH + H+
-
 r
1.1.1.71 1-butanol + NAD+
-
 Pyrococcus horikoshii JCM 9974 butanal + NADH + H+
-
 r
1.1.1.71 1-butanol + NAD+
-
 Pyrococcus horikoshii ATCC 700860 butanal + NADH + H+
-
 r
1.1.1.71 1-butanol + NAD+
-
 Pyrococcus horikoshii OT-3 butanal + NADH + H+
-
 r
1.1.1.71 1-butanol + NADP+
-
 Pyrococcus horikoshii butanal + NADPH + H+
-
 r
1.1.1.71 1-heptanol + NAD+
-
 Pyrococcus horikoshii heptanal + NADH + H+
-
 r
1.1.1.71 1-heptanol + NAD+
-
 Pyrococcus horikoshii DSM 12428 heptanal + NADH + H+
-
 r
1.1.1.71 1-heptanol + NAD+
-
 Pyrococcus horikoshii NBRC 100139 heptanal + NADH + H+
-
 r
1.1.1.71 1-heptanol + NAD+
-
 Pyrococcus horikoshii JCM 9974 heptanal + NADH + H+
-
 r
1.1.1.71 1-heptanol + NAD+
-
 Pyrococcus horikoshii ATCC 700860 heptanal + NADH + H+
-
 r
1.1.1.71 1-heptanol + NAD+
-
 Pyrococcus horikoshii OT-3 heptanal + NADH + H+
-
 r
1.1.1.71 1-hexanol + NAD+
-
 Pyrococcus horikoshii hexanal + NADH + H+
-
 r
1.1.1.71 1-hexanol + NAD+
-
 Pyrococcus horikoshii DSM 12428 hexanal + NADH + H+
-
 r
1.1.1.71 1-hexanol + NAD+
-
 Pyrococcus horikoshii NBRC 100139 hexanal + NADH + H+
-
 r
1.1.1.71 1-hexanol + NAD+
-
 Pyrococcus horikoshii JCM 9974 hexanal + NADH + H+
-
 r
1.1.1.71 1-hexanol + NAD+
-
 Pyrococcus horikoshii ATCC 700860 hexanal + NADH + H+
-
 r
1.1.1.71 1-hexanol + NAD+
-
 Pyrococcus horikoshii OT-3 hexanal + NADH + H+
-
 r
1.1.1.71 1-octanol + NAD+
-
 Pyrococcus horikoshii octanal + NADH + H+
-
 r
1.1.1.71 1-pentanol + NAD+
-
 Pyrococcus horikoshii pentanal + NADH + H+
-
 r
1.1.1.71 1-propanol + NAD+
-
 Pyrococcus horikoshii propanal + NADH + H+
-
 r
1.1.1.71 benzylalcohol + NAD+
-
 Pyrococcus horikoshii benzaldehyde + NADH + H+
-
 r
1.1.1.71 cyclohexanol + NAD+
-
 Pyrococcus horikoshii cyclohexanone + NADH + H+
-
 r
1.1.1.71 ethanol + NAD+
-
 Pyrococcus horikoshii acetaldehyde + NADH + H+
-
 r
1.1.1.71 methanol + NAD+
-
 Pyrococcus horikoshii formaldehyde + NADH + H+
-
 r
1.1.1.71 methanol + NAD+
-
 Pyrococcus horikoshii DSM 12428 formaldehyde + NADH + H+
-
 r
1.1.1.71 methanol + NAD+
-
 Pyrococcus horikoshii NBRC 100139 formaldehyde + NADH + H+
-
 r
1.1.1.71 methanol + NAD+
-
 Pyrococcus horikoshii JCM 9974 formaldehyde + NADH + H+
-
 r
1.1.1.71 methanol + NAD+
-
 Pyrococcus horikoshii ATCC 700860 formaldehyde + NADH + H+
-
 r
1.1.1.71 methanol + NAD+
-
 Pyrococcus horikoshii OT-3 formaldehyde + NADH + H+
-
 r
1.1.1.71 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 ?
-
-
1.1.1.71 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 ?
-
-
1.1.1.71 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 ?
-
-
1.1.1.71 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 ?
-
-
1.1.1.71 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 ?
-
-
1.1.1.71 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

EC Number Synonyms Comment Organism
1.1.1.71 PH0743
-
 Pyrococcus horikoshii
1.1.1.71 PhADH
-
 Pyrococcus horikoshii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.71 30
-
 assay at Pyrococcus horikoshii

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.71 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 [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.71 0.48
-
 1-butanol substrates NAD+ and 1-butanol, pH 9.0, 30°C Pyrococcus horikoshii
1.1.1.71 0.5
-
 NAD+ substrates NAD+ and 1-butanol, pH 9.0, 30°C Pyrococcus horikoshii
1.1.1.71 0.82
-
 Butanal substrates NADH and butanal, pH 5.5, 30°C Pyrococcus horikoshii
1.1.1.71 6.66
-
 propanal substrates NADH and propanal, pH 5.5, 30°C Pyrococcus horikoshii
1.1.1.71 6.96
-
 pentanal substrates NADH and pentanal, pH 5.5, 30°C Pyrococcus horikoshii
1.1.1.71 7.4
-
 NADH substrates NADH and butanal, pH 5.5, 30°C Pyrococcus horikoshii
1.1.1.71 7.87
-
 NADPH substrates NADPH and butanal, pH 5.5, 30°C Pyrococcus horikoshii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.71 5.5
-
 aldehyde reduction assay at Pyrococcus horikoshii
1.1.1.71 9
-
 alcohol oxidation assay at Pyrococcus horikoshii

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.71 NAD+
-
 Pyrococcus horikoshii
1.1.1.71 NADH
-
 Pyrococcus horikoshii

General Information

EC Number General Information Comment Organism
1.1.1.71 evolution NAD(P)-dependent group III alcohol dehydrogenases (ADHs), well known as iron-activated enzymes Pyrococcus horikoshii

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.71 0.92
-
 1-butanol substrates NAD+ and 1-butanol, pH 9.0, 30°C Pyrococcus horikoshii
1.1.1.71 2.4
-
 propanal substrates NADH and propanal, pH 5.5, 30°C Pyrococcus horikoshii
1.1.1.71 3.82
-
 Butanal substrates NADH and butanal, pH 5.5, 30°C Pyrococcus horikoshii
1.1.1.71 15.14
-
 NADPH substrates NADPH and butanal, pH 5.5, 30°C Pyrococcus horikoshii
1.1.1.71 16.67
-
 NAD+ substrates NAD+ and 1-butanol, pH 9.0, 30°C Pyrococcus horikoshii
1.1.1.71 77.33
-
 pentanal substrates NADH and pentanal, pH 5.5, 30°C Pyrococcus horikoshii
1.1.1.71 672.7
-
 NADH substrates NADH and butanal, pH 5.5, 30°C Pyrococcus horikoshii