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Literature summary extracted from

  • Machielsen, R.; Leferink, N.G.; Hendriks, A.; Brouns, S.J.; Hennemann, H.G.; Daussmann, T.; van der Oost, J.
    Laboratory evolution of Pyrococcus furiosus alcohol dehydrogenase to improve the production of (2S,5S)-hexanediol at moderate temperatures (2008), Extremophiles, 12, 587-594.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.2 expressed in Escherichia coli BL21(DE3) cells Pyrococcus furiosus

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.2 A18P/K40N/Q165H/D182H/E202G/R219M mutant with increased specific activity towards 2,5-hexanedione compared to the wild type enzyme Pyrococcus furiosus
1.1.1.2 K209T mutant with wild type specific activity towards 2,5-hexanedione Pyrococcus furiosus
1.1.1.2 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
1.1.1.2 N86D/R213I mutant with increased specific activity towards 2,5-hexanedione compared to the wild type enzyme Pyrococcus furiosus
1.1.1.2 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
1.1.1.2 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
1.1.1.2 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
1.1.1.2 V66A/L176P/Y229H mutant with increased specific activity towards 2,5-hexanedione compared to the wild type enzyme Pyrococcus furiosus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.2 0.1
-
 NADPH mutant enzyme R11L/A180V, at 70°C Pyrococcus furiosus
1.1.1.2 0.2
-
 NADP+ wild type enzyme, at 70°C Pyrococcus furiosus
1.1.1.2 0.4
-
 NADPH wild type enzyme, at 70°C Pyrococcus furiosus
1.1.1.2 0.7
-
 NADP+ mutant enzyme R11L/A180V, at 70°C Pyrococcus furiosus
1.1.1.2 1.2
-
 Pyruvaldehyde wild type enzyme, at 70°C Pyrococcus furiosus
1.1.1.2 4.2
-
 Pyruvaldehyde mutant enzyme R11L/A180V, at 70°C Pyrococcus furiosus
1.1.1.2 5.4
-
 (2S,5S)-hexanediol wild type enzyme, at 70°C Pyrococcus furiosus
1.1.1.2 30.3
-
 2-Pentanol wild type enzyme, at 70°C Pyrococcus furiosus
1.1.1.2 51.1
-
 2-Pentanol mutant enzyme R11L/A180V, at 70°C Pyrococcus furiosus
1.1.1.2 56.4
-
 2,5-hexanedione wild type enzyme, at 70°C Pyrococcus furiosus
1.1.1.2 79
-
 2,5-hexanedione mutant enzyme R11L/A180V, at 70°C Pyrococcus furiosus
1.1.1.2 327.6
-
 (2S,5S)-hexanediol mutant enzyme R11L/A180V, at 70°C Pyrococcus furiosus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.2 26000
-
 2 * 26000 Pyrococcus furiosus

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.2 Pyrococcus furiosus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.2 Q-Sepharose column chromatography Pyrococcus furiosus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.2 0.06
-
 mutant enzyme N86D/R213I, using 2-pentanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.07
-
 mutant enzyme N86D/R213I, using (2S,5S)-hexanediol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.07
-
 mutant enzyme N86D/R213I, using 2-propanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.08
-
 mutant enzyme K209T, using 2-hexanedione as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.09
-
 mutant enzyme N86D/R213I, using 2-butanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.09
-
 wild type enzyme, using 2-hexanedione as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.11
-
 wild type enzyme, using 2-propanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.12
-
 mutant enzyme A18P/K40N/Q165H/D182H/E202G/R219M, using 2-hexadione as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.12
-
 mutant enzyme N86D/R213I, using 2-hexanedione as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.14
-
 mutant enzyme V66A/L176P/Y229H, using 2-hexanedione as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.15
-
 mutant enzyme K209T, using 2-propanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.16
-
 wild type enzyme, using 2-butanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.17
-
 mutant enzyme T153A, using 2-propanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.18
-
 mutant enzyme T153A, using 2-hexanedione as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.19
-
 mutant enzyme V66A/L176P/Y229H, using 2-propanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.2
-
 mutant enzyme A18P/K40N/Q165H/D182H/E202G/R219M, using 2-propanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.2
-
 mutant enzyme L176P, using 2-hexanedione as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.24
-
 mutant enzyme T153A, using 2-butanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.33
-
 mutant enzyme V66A/L176P/Y229H, using 2-butanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.35
-
 mutant enzyme K209T, using 2-butanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.43
-
 mutant enzyme L176P, using 2-propanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.57
-
 mutant enzyme L176P, using 2-butanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.6
-
 mutant enzyme A18P/K40N/Q165H/D182H/E202G/R219M, using 2-butanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.65
-
 mutant enzyme K209T, using 2-pentanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.71
-
 mutant enzyme A18P/K40N/Q165H/D182H/E202G/R219M, using (2S,5S)-hexanediol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.73
-
 mutant enzyme L176P, using 2-pentanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.73
-
 wild type enzyme, using pyruvaldehyde as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.78
-
 mutant enzyme N86D/R213I, using pyruvaldehyde as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.86
-
 mutant enzyme T153A, using 2-pentanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.91
-
 wild type enzyme, using (2S,5S)-hexanediol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 0.95
-
 mutant enzyme R11L/A180V, using 2-hexanedione as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 1.07
-
 mutant enzyme L176P, using pyruvaldehyde as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 1.09
-
 mutant enzyme V66A/L176P/Y229H, using (2S,5S)-hexanediol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 1.1
-
 mutant enzyme K209T, using (2S,5S)-hexanediol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 1.11
-
 mutant enzyme A18P/K40N/Q165H/D182H/E202G/R219M, using 2-pentanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 1.11
-
 mutant enzyme R11L/A180V, using 2-propanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 1.12
-
 mutant enzyme K209T, using pyruvaldehyde as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 1.22
-
 mutant enzyme V66A/L176P/Y229H, using pyruvaldehyde as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 1.24
-
 mutant enzyme L176P, using (2S,5S)-hexanediol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 1.3
-
 mutant enzyme V66A/L176P/Y229H, using 2-pentanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 1.31
-
 mutant enzyme T153A, using (2S,5S)-hexanediol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 1.47
-
 wild type enzyme, using 2-pentanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 1.98
-
 mutant enzyme R11L/A180V, using 2-butanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 3
-
 mutant enzyme T153A, using pyruvaldehyde as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 3.23
-
 mutant enzyme R11L/A180V, using (2S,5S)-hexanediol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 3.37
-
 mutant enzyme A18P/K40N/Q165H/D182H/E202G/R219M, using pyruvaldehyde as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 3.47
-
 mutant enzyme R11L/A180V, using 2-pentanol as substrate, at 30°C Pyrococcus furiosus
1.1.1.2 7.97
-
 mutant enzyme R11L/A180V, using pyruvaldehyde as substrate, at 30°C Pyrococcus furiosus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.2 2,5-hexanedione + NADP+
-
 Pyrococcus furiosus (2S,5S)-hexanediol + NADPH + H+
-
 r
1.1.1.2 2-butanone + NADP+
-
 Pyrococcus furiosus 2-butanol + NADPH + H+
-
 r
1.1.1.2 2-pentanone + NADP+
-
 Pyrococcus furiosus 2-pentanol + NADPH + H+
-
 r
1.1.1.2 2-propanone + NADP+
-
 Pyrococcus furiosus 2-propanol + NADPH + H+
-
 r
1.1.1.2 pyruvaldehyde + NADP+
-
 Pyrococcus furiosus ? + NADPH + H+
-
 r

Subunits

EC Number Subunits Comment Organism
1.1.1.2 homodimer 2 * 26000 Pyrococcus furiosus

Synonyms

EC Number Synonyms Comment Organism
1.1.1.2 ADH
-
 Pyrococcus furiosus
1.1.1.2 AdhA
-
 Pyrococcus furiosus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.2 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
1.1.1.2 90 100 the temperature optimum of the wild type enzyme is determined to be between 90 and 100°C Pyrococcus furiosus

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.2 NADP+
-
 Pyrococcus furiosus