Activating Compound | Comment | Organism | Structure |
---|---|---|---|
iodoacetamide | 1 mM activates up to 25fold | Saccharolobus solfataricus |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli strain RB791 | Saccharolobus solfataricus |
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 | 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 | 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 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 | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | contains eight zinc atoms per tetramer | Saccharolobus solfataricus |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Saccharolobus solfataricus | - |
strain DSM 1617 | - |
Purification (Comment) | Organism |
---|---|
DEAE-Sepharose Fast Flow column chromatography, Matrex Gel Red A column chromatography, and Blue A column chromatography | Saccharolobus solfataricus |
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 | 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 | 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 | 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 | Comment | Organism |
---|---|---|
ADH | - |
Saccharolobus solfataricus |
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 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 Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
8.8 | 9.6 | - |
Saccharolobus solfataricus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Saccharolobus solfataricus | |
NADH | - |
Saccharolobus solfataricus |
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 |