Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.1.1.2 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)

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

Protein Variants

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

Organism

Organism UniProt Comment Textmining
Pyrococcus furiosus
-
-
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

Subunits Comment Organism
homodimer 2 * 26000 Pyrococcus furiosus

Synonyms

Synonyms Comment Organism
ADH
-
Pyrococcus furiosus
AdhA
-
Pyrococcus furiosus

Temperature Optimum [°C]

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

Cofactor Comment Organism Structure
NADP+
-
Pyrococcus furiosus