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Literature summary for 1.1.1.1 extracted from

  • Pennacchio, A.; Esposito, L.; Zagari, A.; Rossi, M.; Raia, C.A.
    Role of Tryptophan 95 in substrate specificity and structural stability of Sulfolobus solfataricus alcohol dehydrogenase (2009), Extremophiles, 13, 751-761.
    View publication on PubMed
Search for more literature for 1.1.1.1

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli RB791 cells Saccharolobus solfataricus

Crystallization (Commentary)

Crystallization (Comment) Organism
-
 Saccharolobus solfataricus

Protein Variants

Protein Variants Comment Organism
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
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]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.008
-
 NADH mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.009
-
 trans-cinnamaldehyde wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.01
-
 NADH wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.012
-
 (S)-2-butanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.027
-
 1-Pentanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.03
-
 benzaldehyde wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.03
-
 Cyclohexanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.035
-
 1-Hexanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.038
-
 1-Heptanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.04
-
 1-Hexanol mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.07
-
 benzaldehyde mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.07
-
 Butyraldehyde wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.07
-
 4-methoxybenzyl alcohol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.07
-
 (S)-2-pentanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.08
-
 1-butanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.09
-
 trans-cinnamaldehyde mutant enzyme W95L,, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.12
-
 benzyl alcohol mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.12
-
 4-methoxybenzyl alcohol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.13
-
 Butyraldehyde mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.21
-
 1-Heptanol mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.22
-
 (R)-2-pentanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.23
-
 trans-cinnamaldehyde mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.24
-
 1-propanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.25
-
 2-ethoxyethanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.26
-
 1-Pentanol mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.26
-
 benzyl alcohol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.26
-
 Isobutyraldehyde wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.27
-
 Butyraldehyde mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.27
-
 3-Pentanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.28
-
 benzaldehyde mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.33
-
 Isobutyraldehyde mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.41
-
 (R)-2-butanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.5
-
 NAD+ wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.67
-
 1-Heptanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.75
-
 NADH mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.76
-
 Isobutyraldehyde mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.88
-
 1-Hexanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
1.3
-
 4-methoxybenzyl alcohol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
1.6
-
 1-butanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
1.9
-
 1-Pentanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
3.8
-
 benzyl alcohol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
4
-
 NAD+ mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
4.5
-
 Cyclohexanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
4.6
-
 ethanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
6.5
-
 1-propanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
8.8
-
 (S)-2-pentanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
11.7
-
 (R)-2-pentanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
29.8
-
 (S)-2-butanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
37.4
-
 (R)-2-butanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
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
66
-
 ethanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus P39462
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Saccharolobus solfataricus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
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
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
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R)-2-butanol + NAD+
-
 Saccharolobus solfataricus 2-butanone + NADH + H+
-
 r
(R)-2-pentanol + NAD+
-
 Saccharolobus solfataricus 2-pentanone + NADH + H+
-
 r
(S)-2-butanol + NAD+
-
 Saccharolobus solfataricus 2-butanone + NADH + H+
-
 r
(S)-2-pentanol + NAD+
-
 Saccharolobus solfataricus 2-pentanone + NADH + H+
-
 r
1-butanol + NAD+
-
 Saccharolobus solfataricus butanal + NADH + H+
-
 r
1-heptanol + NAD+
-
 Saccharolobus solfataricus 1-heptanal + NADH + H+
-
 r
1-hexanol + NAD+
-
 Saccharolobus solfataricus 1-hexanal + NADH + H+
-
 r
1-pentanol + NAD+
-
 Saccharolobus solfataricus 1-pentanal + NADH + H+
-
 r
1-propanol + NAD+
-
 Saccharolobus solfataricus propanal + NADH + H+
-
 r
2-ethoxyethanol + NAD+
-
 Saccharolobus solfataricus 2-ethoxyacetaldehyde + NADH + H+
-
 r
3-pentanol + NAD+
-
 Saccharolobus solfataricus 3-pentanone + NADH + H+
-
 r
4-methoxybenzyl alcohol + NAD+
-
 Saccharolobus solfataricus 4-methoxybenzaldehyde + NADH + H+
-
 r
benzyl alcohol + NAD+
-
 Saccharolobus solfataricus benzaldehyde + NADH + H+
-
 r
cyclohexanol + NAD+
-
 Saccharolobus solfataricus cyclohexanone + NADH + H+
-
 r
ethanol + NAD+
-
 Saccharolobus solfataricus acetaldehyde + NADH + H+
-
 r
isobutyraldehyde + NADH + H+
-
 Saccharolobus solfataricus isobutanol + NAD+
-
 r
n-butanol + NAD+
-
 Saccharolobus solfataricus butyraldehyde + NADH + H+
-
 r
pentan-3-ol + NAD+
-
 Saccharolobus solfataricus 3-pentanone + NADH + H+
-
 r
trans-cinnamaldehyde + NADH + H+
-
 Saccharolobus solfataricus cinnamyl alcohol + NAD+
-
 r

Synonyms

Synonyms Comment Organism
ADH
-
 Saccharolobus solfataricus
NAD+-dependent (S)-stereospecific alcohol dehydrogenase
-
 Saccharolobus solfataricus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
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]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.21
-
 trans-cinnamaldehyde mutant enzyme W95L,, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.35
-
 benzyl alcohol mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.36
-
 benzaldehyde mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.36
-
 Butyraldehyde mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.37
-
 4-methoxybenzyl alcohol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.38
-
 NADH mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.42
-
 1-Hexanol mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.51
-
 1-Heptanol mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.52
-
 NAD+ mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.53
-
 1-Pentanol mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
0.82
-
 Isobutyraldehyde mutant enzyme W95L, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
1
-
 (R)-2-butanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
1.1
-
 (R)-2-pentanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
1.2
-
 Cyclohexanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
1.2
-
 (S)-2-butanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
1.5
-
 3-Pentanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
1.6
-
 1-Heptanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
1.6
-
 2-ethoxyethanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
1.7
-
 (R)-2-butanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
1.9
-
 (R)-2-pentanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
2
-
 1-Hexanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
2.1
-
 trans-cinnamaldehyde wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
2.5
-
 1-propanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
2.5
-
 1-Pentanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
2.6
-
 (S)-2-pentanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
2.9
-
 4-methoxybenzyl alcohol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
3.1
-
 ethanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
3.1
-
 1-butanol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
3.4
-
 NAD+ wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
3.6
-
 benzyl alcohol wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
4.1
-
 ethanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
4.8
-
 (S)-2-butanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
6.7
-
 (S)-2-pentanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
10.3
-
 NADH wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
11.3
-
 benzaldehyde wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
11.9
-
 1-Heptanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
12.4
-
 Cyclohexanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
13.1
-
 benzaldehyde mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
13.3
-
 1-Hexanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
15.1
-
 1-butanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
15.7
-
 1-propanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
16.3
-
 1-Pentanol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
16.8
-
 4-methoxybenzyl alcohol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
18.1
-
 Isobutyraldehyde mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
18.9
-
 trans-cinnamaldehyde mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
19.7
-
 Butyraldehyde mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
19.7
-
 Butyraldehyde wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
20
-
 benzyl alcohol mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
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
29
-
 Isobutyraldehyde wild type enzyme, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus
34.8
-
 NADH mutant enzyme W95L/N249Y, in 0.1 M glycine-NaOH buffer (pH 10.5), at 65°C Saccharolobus solfataricus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.8
-
 for the reduction reaction with benzaldehyde Saccharolobus solfataricus
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

Cofactor Comment Organism Structure
NAD+
-
 Saccharolobus solfataricus