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Literature summary for 1.1.1.1 extracted from

  • Raia, C.A.; Giordano, A.; Rossi, M.
    Alcohol dehydrogenase from Sulfolobus solfataricus (2001), Methods Enzymol., 331, 176-195.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
iodoacetamide 1 mM activates up to 25fold Saccharolobus solfataricus

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
N249Y shows 7fold higher specific activity compared to the wild type enzyme, is active up to 95°C and more stable than the native enzyme, has acquired an improved resistance to proteolysis by thermolysin and shows a decreased activating effect by treatment with denaturants at moderate concentration Saccharolobus solfataricus

General Stability

General Stability Organism
does not require the presence of reducing agents to mantain its stability even at high temperature, evidently due to the lack in free cysteines Saccharolobus solfataricus

Inhibitors

Inhibitors Comment Organism Structure
4-iodopyrazole competitive inhibitor Saccharolobus solfataricus
4-Methylpyrazole competitive inhibitor Saccharolobus solfataricus
EDTA loses 30% of its activity immediately on addition of EDTA Saccharolobus solfataricus
Hg2+
-
Saccharolobus solfataricus
Iodine
-
Saccharolobus solfataricus
additional information not affected by Cu2+ Saccharolobus solfataricus
o-phenanthroline loses 30% of its activity immediately on addition of o-phenanthroline Saccharolobus solfataricus
pyrazole competitive inhibitor Saccharolobus solfataricus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.03
-
NADH in 50 mM Tris-HCl (pH 7.5) containing 0.25 mM benzaldehyde, at 65°C Saccharolobus solfataricus
0.045
-
Cyclohexanol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
0.12
-
4-bromobenzylalcohol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
0.14
-
4-methoxybenzaldehyde in 50 mM Tris-HCl (pH 7.5) containing 0.2 mM NADH + H+, at 65°C Saccharolobus solfataricus
0.14
-
3-bromobenzylalcohol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
0.2
-
NAD+ in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
0.28
-
4-carboxybenzaldehyde in 50 mM Tris-HCl (pH 7.5) containing 0.2 mM NADH + H+, at 65°C Saccharolobus solfataricus
0.3
-
benzaldehyde in 50 mM Tris-HCl (pH 7.5) containing 0.2 mM NADH + H+, at 65°C Saccharolobus solfataricus
0.33
-
1-propanol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
0.6
-
2-propanol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
0.7
-
ethanol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
0.8
-
benzylalcohol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.2
-
3-methoxybenzylalcohol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.2
-
3-methoxybenzylalcohol in the presence of 5 mM benzylalcohol, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.3
-
4-methoxybenzylalcohol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ contains eight zinc atoms per tetramer Saccharolobus solfataricus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
37588
-
4 * 37588, calculated from amino acid sequence Saccharolobus solfataricus
37591
-
4 * 37591, electrospray mass spectrometry Saccharolobus solfataricus
135000
-
SDS-PAGE Saccharolobus solfataricus

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus
-
strain DSM 1617
-

Purification (Commentary)

Purification (Comment) Organism
DEAE-Sepharose Fast Flow column chromatography, Matrex Gel Red A column chromatography, and Blue A column chromatography Saccharolobus solfataricus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.64
-
wild type enzyme, crude extract, at 65°C Saccharolobus solfataricus
4.03
-
recombinant wild type enzyme, after 6fold purification, at 65°C Saccharolobus solfataricus
43
-
mutant enzyme N249Y, after 6fold purification, at 65°C Saccharolobus solfataricus

Storage Stability

Storage Stability Organism
-20°C, 1 M NaCl in 20 mM Tris-HCl (pH 8.4) in the presence of an equal volume of glycerol, 12 months, no loss of activity Saccharolobus solfataricus
4°C, 1 M NaCl in 20 mM Tris-HCl (pH 8.4) in the absence of glycerol, 4 months, 60% loss of activity Saccharolobus solfataricus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1-propanol + NAD+
-
Saccharolobus solfataricus propanal + NADH + H+
-
?
2-propanol + NAD+
-
Saccharolobus solfataricus 2-propanone + NADH + H+
-
?
3-bromobenzylalcohol + NAD+
-
Saccharolobus solfataricus 3-bromobenzaldehyde + NADH + H+
-
?
3-methoxybenzylalcohol + NAD+
-
Saccharolobus solfataricus 3-methoxybenzaldehyde + NADH + H+
-
?
4-bromobenzylalcohol + NAD+
-
Saccharolobus solfataricus 4-bromobenzaldehyde + NADH + H+
-
?
4-carboxybenzaldehyde + NADH + H+
-
Saccharolobus solfataricus 4-carboxybenzylalcohol + NAD+
-
?
4-methoxybenzylalcohol + NAD+
-
Saccharolobus solfataricus 4-methoxybenzaldehyde + NADH + H+
-
r
4-nitrobenzaldehyde + NADH + H+
-
Saccharolobus solfataricus 4-nitrobenzylalcohol + NAD+
-
?
benzylalcohol + NAD+
-
Saccharolobus solfataricus benzaldehyde + NADH + H+
-
r
cyclohexanol + NAD+
-
Saccharolobus solfataricus cyclohexanone + NADH + H+
-
?
ethanol + NAD+
-
Saccharolobus solfataricus acetaldehyde + NADH + H+
-
?

Subunits

Subunits Comment Organism
tetramer 4 * 37588, calculated from amino acid sequence Saccharolobus solfataricus
tetramer 4 * 37591, electrospray mass spectrometry Saccharolobus solfataricus
tetramer crosslinking of ADH with dimethylsuberimidate and ethylene glycol bis [succinimidylsuccinate] produces mainly dimers and tetramers, suggesting a dimer of dimers assembly Saccharolobus solfataricus

Synonyms

Synonyms Comment Organism
ADH
-
Saccharolobus solfataricus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
70
-
remains quite stable at 70°C in the absence of chelating agents Saccharolobus solfataricus
90
-
at 90°C the specific activity is about three times as high as that measured at 65°C Saccharolobus solfataricus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.25
-
2-propanol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
0.5
-
ethanol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
0.6
-
4-methoxybenzaldehyde in 50 mM Tris-HCl (pH 7.5) containing 0.2 mM NADH + H+, at 65°C Saccharolobus solfataricus
0.65
-
Cyclohexanol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
0.7
-
4-carboxybenzaldehyde in 50 mM Tris-HCl (pH 7.5) containing 0.2 mM NADH + H+, at 65°C Saccharolobus solfataricus
1.1
-
4-bromobenzylalcohol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.2
-
1-propanol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.2
-
3-bromobenzylalcohol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.2
-
3-methoxybenzylalcohol in the presence of 5 mM benzylalcohol, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.4
-
NAD+ in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.4
-
benzylalcohol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.4
-
3-methoxybenzylalcohol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.5
-
4-methoxybenzylalcohol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
3.5
-
benzaldehyde in 50 mM Tris-HCl (pH 7.5) containing 0.2 mM NADH + H+, at 65°C Saccharolobus solfataricus
4.5
-
NADH in 50 mM Tris-HCl (pH 7.5) containing 0.25 mM benzaldehyde, at 65°C Saccharolobus solfataricus
4.6
-
4-nitrobenzaldehyde in 50 mM Tris-HCl (pH 7.5) containing 0.2 mM NADH + H+, at 65°C Saccharolobus solfataricus

pH Range

pH Minimum pH Maximum Comment Organism
8.8 9.6
-
Saccharolobus solfataricus

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Saccharolobus solfataricus
NADH
-
Saccharolobus solfataricus

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0032
-
4-iodopyrazole apparent value Saccharolobus solfataricus
0.009
-
4-Methylpyrazole apparent value Saccharolobus solfataricus
0.13
-
pyrazole apparent value Saccharolobus solfataricus