BRENDA - Enzyme Database

Role of Tryptophan 95 in substrate specificity and structural stability of Sulfolobus solfataricus alcohol dehydrogenase

Pennacchio, A.; Esposito, L.; Zagari, A.; Rossi, M.; Raia, C.A.; Extremophiles 13, 751-761 (2009)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
1.1.1.1
expressed in Escherichia coli RB791 cells
Saccharolobus solfataricus
1.1.1.B3
expressed in Escherichia coli RB791 cells
Saccharolobus solfataricus
1.1.1.B4
expressed in Escherichia coli RB791 cells
Saccharolobus solfataricus
Crystallization (Commentary)
EC Number
Crystallization (Commentary)
Organism
1.1.1.1
-
Saccharolobus solfataricus
1.1.1.B3
-
Saccharolobus solfataricus
1.1.1.B4
-
Saccharolobus solfataricus
Engineering
EC Number
Protein Variants
Commentary
Organism
1.1.1.1
W95L
the mutant displays no apparent activity with short-chain primary and secondary alcohols and poor activity with aromatic substrates and coenzyme, the substitution affects the structural stability of the archaeal ADH, decreasing its thermal stability without relevant changes in secondary structure, optimum pH is at about pH 10
Saccharolobus solfataricus
1.1.1.1
W95L/N249Y
the mutant exhibits higher activity but decreased affinity toward aliphatic alcohols, aldehydes as well as NAD+ and NADH compared to the wild type enzyme, optimum pH is at about pH 8.6
Saccharolobus solfataricus
1.1.1.B3
W95L/N249Y
the mutant exhibits higher activity but decreased affinity toward aliphatic alcohols, aldehydes as well as NAD+ and NADH compared to the wild type enzyme, optimum pH is at about pH 8.6
Saccharolobus solfataricus
1.1.1.B4
W95L/N249Y
the mutant exhibits higher activity but decreased affinity toward aliphatic alcohols, aldehydes as well as NAD+ and NADH compared to the wild type enzyme, optimum pH is at about pH 8.6
Saccharolobus solfataricus
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.1
0.008
-
NADH
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.009
-
trans-cinnamaldehyde
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.01
-
NADH
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.012
-
(S)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.027
-
1-Pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.03
-
benzaldehyde
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.03
-
Cyclohexanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.035
-
1-Hexanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.038
-
1-Heptanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.04
-
1-Hexanol
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.07
-
4-methoxybenzyl alcohol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.07
-
benzaldehyde
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.07
-
Butyraldehyde
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.07
-
(S)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.08
-
1-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.09
-
trans-cinnamaldehyde
mutant enzyme W95L,, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.12
-
4-methoxybenzyl alcohol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.12
-
benzyl alcohol
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.13
-
Butyraldehyde
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.21
-
1-Heptanol
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.22
-
(R)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.23
-
trans-cinnamaldehyde
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.24
-
1-propanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.25
-
2-ethoxyethanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.26
-
1-Pentanol
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.26
-
benzyl alcohol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.26
-
Isobutyraldehyde
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.27
-
3-Pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.27
-
Butyraldehyde
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.28
-
benzaldehyde
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.33
-
Isobutyraldehyde
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.41
-
(R)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.5
-
NAD+
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.67
-
1-Heptanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.75
-
NADH
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.76
-
Isobutyraldehyde
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.88
-
1-Hexanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.3
-
4-methoxybenzyl alcohol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.6
-
1-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.9
-
1-Pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
3.8
-
benzyl alcohol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
4
-
NAD+
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
4.5
-
Cyclohexanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
4.6
-
ethanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
6.5
-
1-propanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
8.8
-
(S)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
11.7
-
(R)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
29.8
-
(S)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
37.4
-
(R)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
50
-
NAD+
Km above 50 mM, mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
66
-
ethanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B3
0.012
-
(S)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B3
0.07
-
(S)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B3
8.8
-
(S)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B3
29.8
-
(S)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B4
0.22
-
(R)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B4
0.41
-
(R)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B4
11.7
-
(R)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B4
37.4
-
(R)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
Organism
EC Number
Organism
UniProt
Commentary
Textmining
1.1.1.1
Saccharolobus solfataricus
P39462
-
-
1.1.1.B3
Saccharolobus solfataricus
-
-
-
1.1.1.B4
Saccharolobus solfataricus
-
-
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
1.1.1.1
-
Saccharolobus solfataricus
1.1.1.B3
-
Saccharolobus solfataricus
1.1.1.B4
-
Saccharolobus solfataricus
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.1.1.1
0.5
-
purified mutant enzyme W95L, using benzyl alcohol as substrate, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
4.2
-
purified wild type enzyme, using benzyl alcohol as substrate, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
18.6
-
purified mutant enzyme W95L/N249Y, using benzyl alcohol as substrate, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
1.1.1.1
(R)-2-butanol + NAD+
-
697839
Saccharolobus solfataricus
2-butanone + NADH + H+
-
-
-
r
1.1.1.1
(R)-2-pentanol + NAD+
-
697839
Saccharolobus solfataricus
2-pentanone + NADH + H+
-
-
-
r
1.1.1.1
(S)-2-butanol + NAD+
-
697839
Saccharolobus solfataricus
2-butanone + NADH + H+
-
-
-
r
1.1.1.1
(S)-2-pentanol + NAD+
-
697839
Saccharolobus solfataricus
2-pentanone + NADH + H+
-
-
-
r
1.1.1.1
1-butanol + NAD+
-
697839
Saccharolobus solfataricus
butanal + NADH + H+
-
-
-
r
1.1.1.1
1-heptanol + NAD+
-
697839
Saccharolobus solfataricus
1-heptanal + NADH + H+
-
-
-
r
1.1.1.1
1-hexanol + NAD+
-
697839
Saccharolobus solfataricus
1-hexanal + NADH + H+
-
-
-
r
1.1.1.1
1-pentanol + NAD+
-
697839
Saccharolobus solfataricus
1-pentanal + NADH + H+
-
-
-
r
1.1.1.1
1-propanol + NAD+
-
697839
Saccharolobus solfataricus
propanal + NADH + H+
-
-
-
r
1.1.1.1
2-ethoxyethanol + NAD+
-
697839
Saccharolobus solfataricus
2-ethoxyacetaldehyde + NADH + H+
-
-
-
r
1.1.1.1
3-pentanol + NAD+
-
697839
Saccharolobus solfataricus
3-pentanone + NADH + H+
-
-
-
r
1.1.1.1
4-methoxybenzyl alcohol + NAD+
-
697839
Saccharolobus solfataricus
4-methoxybenzaldehyde + NADH + H+
-
-
-
r
1.1.1.1
benzyl alcohol + NAD+
-
697839
Saccharolobus solfataricus
benzaldehyde + NADH + H+
-
-
-
r
1.1.1.1
cyclohexanol + NAD+
-
697839
Saccharolobus solfataricus
cyclohexanone + NADH + H+
-
-
-
r
1.1.1.1
ethanol + NAD+
-
697839
Saccharolobus solfataricus
acetaldehyde + NADH + H+
-
-
-
r
1.1.1.1
isobutyraldehyde + NADH + H+
-
697839
Saccharolobus solfataricus
isobutanol + NAD+
-
-
-
r
1.1.1.1
n-butanol + NAD+
-
697839
Saccharolobus solfataricus
butyraldehyde + NADH + H+
-
-
-
r
1.1.1.1
pentan-3-ol + NAD+
-
697839
Saccharolobus solfataricus
3-pentanone + NADH + H+
-
-
-
r
1.1.1.1
trans-cinnamaldehyde + NADH + H+
-
697839
Saccharolobus solfataricus
cinnamyl alcohol + NAD+
-
-
-
r
1.1.1.B3
(S)-2-butanol + NAD+
-
697839
Saccharolobus solfataricus
2-butanone + NADH + H+
-
-
-
r
1.1.1.B3
(S)-2-pentanol + NAD+
-
697839
Saccharolobus solfataricus
2-pentanone + NADH + H+
-
-
-
r
1.1.1.B4
(R)-2-butanol + NAD+
-
697839
Saccharolobus solfataricus
2-butanone + NADH + H+
-
-
-
r
1.1.1.B4
(R)-2-pentanol + NAD+
-
697839
Saccharolobus solfataricus
2-pentanone + NADH + H+
-
-
-
r
Synonyms
EC Number
Synonyms
Commentary
Organism
1.1.1.1
ADH
-
Saccharolobus solfataricus
1.1.1.1
NAD+-dependent (S)-stereospecific alcohol dehydrogenase
-
Saccharolobus solfataricus
1.1.1.B3
ADH
-
Saccharolobus solfataricus
1.1.1.B3
NAD+-dependent (S)-stereospecific alcohol dehydrogenase
-
Saccharolobus solfataricus
1.1.1.B4
ADH
-
Saccharolobus solfataricus
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
1.1.1.1
65
88
the single and double mutant are less thermoresistant than the wild type enzyme, displaying a transition temperature of 78 and 88°C, respectively, which are 17 and 7°C lower than that of the wild-type enzyme. At 65°C the single mutant W95L is 10fold less active and the double mutant W95L/N249Y is about 6fold more active than wild type enzyme, the reaction rate catalyzed by the double mutant W95L/N249Y increases more markedly than that of the wild type enzyme up to a temperature of about 83°C and then decreases rapidly due to thermal inactivation. The reaction rate of the single mutant W95L increases more slowly up to about 80°C and then decreases rapidly. At 65°C the single mutant is 10fold less active and the double mutant is about 6fold more active than wild type enzyme.
Saccharolobus solfataricus
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.1
0.21
-
trans-cinnamaldehyde
mutant enzyme W95L,, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.35
-
benzyl alcohol
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.36
-
benzaldehyde
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.36
-
Butyraldehyde
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.37
-
4-methoxybenzyl alcohol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.38
-
NADH
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.42
-
1-Hexanol
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.51
-
1-Heptanol
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.52
-
NAD+
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.53
-
1-Pentanol
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.82
-
Isobutyraldehyde
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1
-
(R)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.1
-
(R)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.2
-
Cyclohexanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.2
-
(S)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.5
-
3-Pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.6
-
1-Heptanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.6
-
2-ethoxyethanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.7
-
(R)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.9
-
(R)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
2
-
1-Hexanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
2.1
-
trans-cinnamaldehyde
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
2.5
-
1-Pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
2.5
-
1-propanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
2.6
-
(S)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
2.9
-
4-methoxybenzyl alcohol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
3.1
-
1-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
3.1
-
ethanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
3.4
-
NAD+
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
3.6
-
benzyl alcohol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
4.1
-
ethanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
4.8
-
(S)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
6.7
-
(S)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
10.3
-
NADH
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
11.3
-
benzaldehyde
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
11.9
-
1-Heptanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
12.4
-
Cyclohexanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
13.1
-
benzaldehyde
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
13.3
-
1-Hexanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
15.1
-
1-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
15.7
-
1-propanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
16.3
-
1-Pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
16.8
-
4-methoxybenzyl alcohol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
18.1
-
Isobutyraldehyde
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
18.9
-
trans-cinnamaldehyde
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
19.7
-
Butyraldehyde
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C; wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
20
-
benzyl alcohol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
21
-
NAD+
Km above 50 mM, mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
29
-
Isobutyraldehyde
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
34.8
-
NADH
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B3
1.2
-
(S)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B3
2.6
-
(S)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B3
4.8
-
(S)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B3
6.7
-
(S)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B4
1
-
(R)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B4
1.1
-
(R)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B4
1.7
-
(R)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B4
1.9
-
(R)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.1
6.8
-
for the reduction reaction with benzaldehyde
Saccharolobus solfataricus
1.1.1.1
9.5
-
for the oxidation reaction, ADH activity shows a slight dependence on pH, displaying a broad peak with a maximum around 9.5
Saccharolobus solfataricus
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.1
NAD+
-
Saccharolobus solfataricus
1.1.1.B3
NAD+
-
Saccharolobus solfataricus
1.1.1.B4
NAD+
-
Saccharolobus solfataricus
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.1
expressed in Escherichia coli RB791 cells
Saccharolobus solfataricus
1.1.1.B3
expressed in Escherichia coli RB791 cells
Saccharolobus solfataricus
1.1.1.B4
expressed in Escherichia coli RB791 cells
Saccharolobus solfataricus
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.1
NAD+
-
Saccharolobus solfataricus
1.1.1.B3
NAD+
-
Saccharolobus solfataricus
1.1.1.B4
NAD+
-
Saccharolobus solfataricus
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
1.1.1.1
-
Saccharolobus solfataricus
1.1.1.B3
-
Saccharolobus solfataricus
1.1.1.B4
-
Saccharolobus solfataricus
Engineering (protein specific)
EC Number
Protein Variants
Commentary
Organism
1.1.1.1
W95L
the mutant displays no apparent activity with short-chain primary and secondary alcohols and poor activity with aromatic substrates and coenzyme, the substitution affects the structural stability of the archaeal ADH, decreasing its thermal stability without relevant changes in secondary structure, optimum pH is at about pH 10
Saccharolobus solfataricus
1.1.1.1
W95L/N249Y
the mutant exhibits higher activity but decreased affinity toward aliphatic alcohols, aldehydes as well as NAD+ and NADH compared to the wild type enzyme, optimum pH is at about pH 8.6
Saccharolobus solfataricus
1.1.1.B3
W95L/N249Y
the mutant exhibits higher activity but decreased affinity toward aliphatic alcohols, aldehydes as well as NAD+ and NADH compared to the wild type enzyme, optimum pH is at about pH 8.6
Saccharolobus solfataricus
1.1.1.B4
W95L/N249Y
the mutant exhibits higher activity but decreased affinity toward aliphatic alcohols, aldehydes as well as NAD+ and NADH compared to the wild type enzyme, optimum pH is at about pH 8.6
Saccharolobus solfataricus
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.1
0.008
-
NADH
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.009
-
trans-cinnamaldehyde
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.01
-
NADH
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.012
-
(S)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.027
-
1-Pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.03
-
benzaldehyde
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.03
-
Cyclohexanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.035
-
1-Hexanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.038
-
1-Heptanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.04
-
1-Hexanol
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.07
-
4-methoxybenzyl alcohol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.07
-
benzaldehyde
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.07
-
Butyraldehyde
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.07
-
(S)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.08
-
1-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.09
-
trans-cinnamaldehyde
mutant enzyme W95L,, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.12
-
4-methoxybenzyl alcohol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.12
-
benzyl alcohol
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.13
-
Butyraldehyde
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.21
-
1-Heptanol
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.22
-
(R)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.23
-
trans-cinnamaldehyde
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.24
-
1-propanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.25
-
2-ethoxyethanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.26
-
1-Pentanol
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.26
-
benzyl alcohol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.26
-
Isobutyraldehyde
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.27
-
3-Pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.27
-
Butyraldehyde
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.28
-
benzaldehyde
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.33
-
Isobutyraldehyde
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.41
-
(R)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.5
-
NAD+
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.67
-
1-Heptanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.75
-
NADH
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.76
-
Isobutyraldehyde
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.88
-
1-Hexanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.3
-
4-methoxybenzyl alcohol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.6
-
1-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.9
-
1-Pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
3.8
-
benzyl alcohol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
4
-
NAD+
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
4.5
-
Cyclohexanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
4.6
-
ethanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
6.5
-
1-propanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
8.8
-
(S)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
11.7
-
(R)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
29.8
-
(S)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
37.4
-
(R)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
50
-
NAD+
Km above 50 mM, mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
66
-
ethanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B3
0.012
-
(S)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B3
0.07
-
(S)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B3
8.8
-
(S)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B3
29.8
-
(S)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B4
0.22
-
(R)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B4
0.41
-
(R)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B4
11.7
-
(R)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B4
37.4
-
(R)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.1
-
Saccharolobus solfataricus
1.1.1.B3
-
Saccharolobus solfataricus
1.1.1.B4
-
Saccharolobus solfataricus
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.1.1.1
0.5
-
purified mutant enzyme W95L, using benzyl alcohol as substrate, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
4.2
-
purified wild type enzyme, using benzyl alcohol as substrate, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
18.6
-
purified mutant enzyme W95L/N249Y, using benzyl alcohol as substrate, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
1.1.1.1
(R)-2-butanol + NAD+
-
697839
Saccharolobus solfataricus
2-butanone + NADH + H+
-
-
-
r
1.1.1.1
(R)-2-pentanol + NAD+
-
697839
Saccharolobus solfataricus
2-pentanone + NADH + H+
-
-
-
r
1.1.1.1
(S)-2-butanol + NAD+
-
697839
Saccharolobus solfataricus
2-butanone + NADH + H+
-
-
-
r
1.1.1.1
(S)-2-pentanol + NAD+
-
697839
Saccharolobus solfataricus
2-pentanone + NADH + H+
-
-
-
r
1.1.1.1
1-butanol + NAD+
-
697839
Saccharolobus solfataricus
butanal + NADH + H+
-
-
-
r
1.1.1.1
1-heptanol + NAD+
-
697839
Saccharolobus solfataricus
1-heptanal + NADH + H+
-
-
-
r
1.1.1.1
1-hexanol + NAD+
-
697839
Saccharolobus solfataricus
1-hexanal + NADH + H+
-
-
-
r
1.1.1.1
1-pentanol + NAD+
-
697839
Saccharolobus solfataricus
1-pentanal + NADH + H+
-
-
-
r
1.1.1.1
1-propanol + NAD+
-
697839
Saccharolobus solfataricus
propanal + NADH + H+
-
-
-
r
1.1.1.1
2-ethoxyethanol + NAD+
-
697839
Saccharolobus solfataricus
2-ethoxyacetaldehyde + NADH + H+
-
-
-
r
1.1.1.1
3-pentanol + NAD+
-
697839
Saccharolobus solfataricus
3-pentanone + NADH + H+
-
-
-
r
1.1.1.1
4-methoxybenzyl alcohol + NAD+
-
697839
Saccharolobus solfataricus
4-methoxybenzaldehyde + NADH + H+
-
-
-
r
1.1.1.1
benzyl alcohol + NAD+
-
697839
Saccharolobus solfataricus
benzaldehyde + NADH + H+
-
-
-
r
1.1.1.1
cyclohexanol + NAD+
-
697839
Saccharolobus solfataricus
cyclohexanone + NADH + H+
-
-
-
r
1.1.1.1
ethanol + NAD+
-
697839
Saccharolobus solfataricus
acetaldehyde + NADH + H+
-
-
-
r
1.1.1.1
isobutyraldehyde + NADH + H+
-
697839
Saccharolobus solfataricus
isobutanol + NAD+
-
-
-
r
1.1.1.1
n-butanol + NAD+
-
697839
Saccharolobus solfataricus
butyraldehyde + NADH + H+
-
-
-
r
1.1.1.1
pentan-3-ol + NAD+
-
697839
Saccharolobus solfataricus
3-pentanone + NADH + H+
-
-
-
r
1.1.1.1
trans-cinnamaldehyde + NADH + H+
-
697839
Saccharolobus solfataricus
cinnamyl alcohol + NAD+
-
-
-
r
1.1.1.B3
(S)-2-butanol + NAD+
-
697839
Saccharolobus solfataricus
2-butanone + NADH + H+
-
-
-
r
1.1.1.B3
(S)-2-pentanol + NAD+
-
697839
Saccharolobus solfataricus
2-pentanone + NADH + H+
-
-
-
r
1.1.1.B4
(R)-2-butanol + NAD+
-
697839
Saccharolobus solfataricus
2-butanone + NADH + H+
-
-
-
r
1.1.1.B4
(R)-2-pentanol + NAD+
-
697839
Saccharolobus solfataricus
2-pentanone + NADH + H+
-
-
-
r
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
1.1.1.1
65
88
the single and double mutant are less thermoresistant than the wild type enzyme, displaying a transition temperature of 78 and 88°C, respectively, which are 17 and 7°C lower than that of the wild-type enzyme. At 65°C the single mutant W95L is 10fold less active and the double mutant W95L/N249Y is about 6fold more active than wild type enzyme, the reaction rate catalyzed by the double mutant W95L/N249Y increases more markedly than that of the wild type enzyme up to a temperature of about 83°C and then decreases rapidly due to thermal inactivation. The reaction rate of the single mutant W95L increases more slowly up to about 80°C and then decreases rapidly. At 65°C the single mutant is 10fold less active and the double mutant is about 6fold more active than wild type enzyme.
Saccharolobus solfataricus
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.1
0.21
-
trans-cinnamaldehyde
mutant enzyme W95L,, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.35
-
benzyl alcohol
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.36
-
benzaldehyde
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.36
-
Butyraldehyde
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.37
-
4-methoxybenzyl alcohol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.38
-
NADH
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.42
-
1-Hexanol
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.51
-
1-Heptanol
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.52
-
NAD+
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.53
-
1-Pentanol
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
0.82
-
Isobutyraldehyde
mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1
-
(R)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.1
-
(R)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.2
-
Cyclohexanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.2
-
(S)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.5
-
3-Pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.6
-
1-Heptanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.6
-
2-ethoxyethanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.7
-
(R)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
1.9
-
(R)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
2
-
1-Hexanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
2.1
-
trans-cinnamaldehyde
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
2.5
-
1-Pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
2.5
-
1-propanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
2.6
-
(S)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
2.9
-
4-methoxybenzyl alcohol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
3.1
-
1-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
3.1
-
ethanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
3.4
-
NAD+
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
3.6
-
benzyl alcohol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
4.1
-
ethanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
4.8
-
(S)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
6.7
-
(S)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
10.3
-
NADH
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
11.3
-
benzaldehyde
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
11.9
-
1-Heptanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
12.4
-
Cyclohexanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
13.1
-
benzaldehyde
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
13.3
-
1-Hexanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
15.1
-
1-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
15.7
-
1-propanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
16.3
-
1-Pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
16.8
-
4-methoxybenzyl alcohol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
18.1
-
Isobutyraldehyde
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
18.9
-
trans-cinnamaldehyde
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
19.7
-
Butyraldehyde
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C; wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
20
-
benzyl alcohol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
21
-
NAD+
Km above 50 mM, mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
29
-
Isobutyraldehyde
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.1
34.8
-
NADH
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B3
1.2
-
(S)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B3
2.6
-
(S)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B3
4.8
-
(S)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B3
6.7
-
(S)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B4
1
-
(R)-2-butanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B4
1.1
-
(R)-2-pentanol
wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B4
1.7
-
(R)-2-butanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
1.1.1.B4
1.9
-
(R)-2-pentanol
mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C
Saccharolobus solfataricus
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.1
6.8
-
for the reduction reaction with benzaldehyde
Saccharolobus solfataricus
1.1.1.1
9.5
-
for the oxidation reaction, ADH activity shows a slight dependence on pH, displaying a broad peak with a maximum around 9.5
Saccharolobus solfataricus