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Literature summary 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

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

Cloned(Commentary)

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

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.1 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

EC Number General Stability Organism
1.1.1.1 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

EC Number Inhibitors Comment Organism Structure
1.1.1.1 4-iodopyrazole competitive inhibitor Saccharolobus solfataricus
1.1.1.1 4-Methylpyrazole competitive inhibitor Saccharolobus solfataricus
1.1.1.1 EDTA loses 30% of its activity immediately on addition of EDTA Saccharolobus solfataricus
1.1.1.1 Hg2+
-
 Saccharolobus solfataricus
1.1.1.1 Iodine
-
 Saccharolobus solfataricus
1.1.1.1 additional information not affected by Cu2+ Saccharolobus solfataricus
1.1.1.1 o-phenanthroline loses 30% of its activity immediately on addition of o-phenanthroline Saccharolobus solfataricus
1.1.1.1 pyrazole competitive inhibitor Saccharolobus solfataricus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.1 0.03
-
 NADH in 50 mM Tris-HCl (pH 7.5) containing 0.25 mM benzaldehyde, at 65°C Saccharolobus solfataricus
1.1.1.1 0.045
-
 Cyclohexanol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.1.1.1 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
1.1.1.1 0.14
-
 4-methoxybenzaldehyde in 50 mM Tris-HCl (pH 7.5) containing 0.2 mM NADH + H+, at 65°C Saccharolobus solfataricus
1.1.1.1 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
1.1.1.1 0.2
-
 NAD+ in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.1.1.1 0.28
-
 4-carboxybenzaldehyde in 50 mM Tris-HCl (pH 7.5) containing 0.2 mM NADH + H+, at 65°C Saccharolobus solfataricus
1.1.1.1 0.3
-
 benzaldehyde in 50 mM Tris-HCl (pH 7.5) containing 0.2 mM NADH + H+, at 65°C Saccharolobus solfataricus
1.1.1.1 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
1.1.1.1 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
1.1.1.1 0.7
-
 ethanol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.1.1.1 0.8
-
 benzylalcohol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.1.1.1 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
1.1.1.1 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
1.1.1.1 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

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

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.1 37588
-
 4 * 37588, calculated from amino acid sequence Saccharolobus solfataricus
1.1.1.1 37591
-
 4 * 37591, electrospray mass spectrometry Saccharolobus solfataricus
1.1.1.1 135000
-
 SDS-PAGE Saccharolobus solfataricus

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.1 Saccharolobus solfataricus
-
 strain DSM 1617
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.1 0.64
-
 wild type enzyme, crude extract, at 65°C Saccharolobus solfataricus
1.1.1.1 4.03
-
 recombinant wild type enzyme, after 6fold purification, at 65°C Saccharolobus solfataricus
1.1.1.1 43
-
 mutant enzyme N249Y, after 6fold purification, at 65°C Saccharolobus solfataricus

Storage Stability

EC Number Storage Stability Organism
1.1.1.1 -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
1.1.1.1 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)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.1 1-propanol + NAD+
-
 Saccharolobus solfataricus propanal + NADH + H+
-
 ?
1.1.1.1 2-propanol + NAD+
-
 Saccharolobus solfataricus 2-propanone + NADH + H+
-
 ?
1.1.1.1 3-bromobenzylalcohol + NAD+
-
 Saccharolobus solfataricus 3-bromobenzaldehyde + NADH + H+
-
 ?
1.1.1.1 3-methoxybenzylalcohol + NAD+
-
 Saccharolobus solfataricus 3-methoxybenzaldehyde + NADH + H+
-
 ?
1.1.1.1 4-bromobenzylalcohol + NAD+
-
 Saccharolobus solfataricus 4-bromobenzaldehyde + NADH + H+
-
 ?
1.1.1.1 4-carboxybenzaldehyde + NADH + H+
-
 Saccharolobus solfataricus 4-carboxybenzylalcohol + NAD+
-
 ?
1.1.1.1 4-methoxybenzylalcohol + NAD+
-
 Saccharolobus solfataricus 4-methoxybenzaldehyde + NADH + H+
-
 r
1.1.1.1 4-nitrobenzaldehyde + NADH + H+
-
 Saccharolobus solfataricus 4-nitrobenzylalcohol + NAD+
-
 ?
1.1.1.1 benzylalcohol + NAD+
-
 Saccharolobus solfataricus benzaldehyde + NADH + H+
-
 r
1.1.1.1 cyclohexanol + NAD+
-
 Saccharolobus solfataricus cyclohexanone + NADH + H+
-
 ?
1.1.1.1 ethanol + NAD+
-
 Saccharolobus solfataricus acetaldehyde + NADH + H+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.1 tetramer 4 * 37588, calculated from amino acid sequence Saccharolobus solfataricus
1.1.1.1 tetramer 4 * 37591, electrospray mass spectrometry Saccharolobus solfataricus
1.1.1.1 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

EC Number Synonyms Comment Organism
1.1.1.1 ADH
-
 Saccharolobus solfataricus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.1 70
-
 remains quite stable at 70°C in the absence of chelating agents Saccharolobus solfataricus
1.1.1.1 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]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.1 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
1.1.1.1 0.5
-
 ethanol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.1.1.1 0.6
-
 4-methoxybenzaldehyde in 50 mM Tris-HCl (pH 7.5) containing 0.2 mM NADH + H+, at 65°C Saccharolobus solfataricus
1.1.1.1 0.65
-
 Cyclohexanol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.1.1.1 0.7
-
 4-carboxybenzaldehyde in 50 mM Tris-HCl (pH 7.5) containing 0.2 mM NADH + H+, at 65°C Saccharolobus solfataricus
1.1.1.1 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
1.1.1.1 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
1.1.1.1 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
1.1.1.1 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
1.1.1.1 1.4
-
 NAD+ in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.1.1.1 1.4
-
 benzylalcohol in the presence of 3 mM NAD+, in 0.1 M glycine-NaOH, pH 9.2, at 65°C Saccharolobus solfataricus
1.1.1.1 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
1.1.1.1 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
1.1.1.1 3.5
-
 benzaldehyde in 50 mM Tris-HCl (pH 7.5) containing 0.2 mM NADH + H+, at 65°C Saccharolobus solfataricus
1.1.1.1 4.5
-
 NADH in 50 mM Tris-HCl (pH 7.5) containing 0.25 mM benzaldehyde, at 65°C Saccharolobus solfataricus
1.1.1.1 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

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.1 8.8 9.6
-
 Saccharolobus solfataricus

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.1 NAD+
-
 Saccharolobus solfataricus
1.1.1.1 NADH
-
 Saccharolobus solfataricus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.1.1.1 0.0032
-
 4-iodopyrazole apparent value Saccharolobus solfataricus
1.1.1.1 0.009
-
 4-Methylpyrazole apparent value Saccharolobus solfataricus
1.1.1.1 0.13
-
 pyrazole apparent value Saccharolobus solfataricus