EC Number | Cloned (Comment) | Organism |
---|---|---|
1.1.1.1 | gene adh3, sequence comparison, expression of N-terminally His-tagged enzyme in Escherichia coli strains NovaBlue Singles and M15[pREP4] | Oenococcus oeni |
1.1.1.202 | expression in Escherichia coli | Oenococcus oeni |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
1.1.1.1 | purified recombinant N-terminally His-tagged Adh3, hanging drop vapour diffusion method, mixing of 0.002 ml of 80mg/ml protein in 50 mM Tris, pH 7.0, with 0.002 ml of reservoir solution, comprising 0.1 M MES, pH 6.5, 16% w/w PEG 20000, and 0.001 ml of 50 mM CaCl2, 20°C, X-ray diffraction structure determination and analysis | Oenococcus oeni |
1.1.1.202 | to 3.2 A resolution. The electron density map around the active site shows no sign of a bound co-factor, but the active site metal could be located and identified as Ni2+ | Oenococcus oeni |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.1 | EDTA | - |
Oenococcus oeni | |
1.1.1.1 | Zn2+ | complete inhibition | Oenococcus oeni | |
1.1.1.202 | imidazole | activity is reduced down to 32% in the presence of 1 mM imidazole and addition of 250 mM results in complete inactivity | Oenococcus oeni |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.1 | 0.05 | - |
NAD+ | pH 9.0, 60°C, recombinant enzyme | Oenococcus oeni | |
1.1.1.1 | 0.2 | - |
NADH | pH 8.0, 60°C, recombinant enzyme | Oenococcus oeni | |
1.1.1.1 | 2.6 | - |
acetaldehyde | pH 8.0, 60°C, recombinant enzyme | Oenococcus oeni | |
1.1.1.1 | 10.4 | - |
ethanol | pH 9.0, 60°C, recombinant enzyme | Oenococcus oeni | |
1.1.1.1 | 11.5 | - |
1,2-propanediol | pH 8.0, 60°C, recombinant enzyme | Oenococcus oeni | |
1.1.1.1 | 26.7 | - |
1,3-Propanediol | pH 9.0, 60°C, recombinant enzyme | Oenococcus oeni | |
1.1.1.202 | 0.05 | - |
NAD+ | pH 9.0, 60°C | Oenococcus oeni | |
1.1.1.202 | 0.2 | - |
NADH | pH 8.0, 60°C | Oenococcus oeni | |
1.1.1.202 | 11.5 | - |
1,2-propanediol | pH 9.0, 60°C | Oenococcus oeni | |
1.1.1.202 | 26.7 | - |
1,3-Propanediol | pH 9.0, 60°C | Oenococcus oeni |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.1 | Mg2+ | 40% activation at 10 mM | Oenococcus oeni | |
1.1.1.1 | additional information | Adh3 exhibits tolerance to several metal ions | Oenococcus oeni | |
1.1.1.1 | Na+ | 40% activation at 1 mM | Oenococcus oeni | |
1.1.1.1 | Ni2+ | activates, the activity using 1,3-propanediol as substrate is highly dependent on the addition of NiCl2, while no change in activity is observed using ethanol in a sample with NiCl2 | Oenococcus oeni | |
1.1.1.202 | Ni2+ | located near active site, crystallization data. The activity with 1,3-propanediol is highly dependent on the presence of Ni2+ | Oenococcus oeni |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
1.1.1.1 | 74000 | - |
recombinant His-tagged enzyme, gel filtration | Oenococcus oeni |
1.1.1.202 | 42200 | - |
2 * 42200, calculated, 2 * 43000, SDS-PAGE of recombinant His-tagged protein | Oenococcus oeni |
1.1.1.202 | 43000 | - |
2 * 42200, calculated, 2 * 43000, SDS-PAGE of recombinant His-tagged protein | Oenococcus oeni |
1.1.1.202 | 74000 | - |
gel filtration | Oenococcus oeni |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.1.1 | ethanol + NAD+ | Oenococcus oeni | - |
acetaldehyde + NADH + H+ | - |
r |
EC Number | Organic Solvent | Comment | Organism |
---|---|---|---|
1.1.1.1 | additional information | Adh3 exhibits tolerance to several organic solvents | Oenococcus oeni |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.1.1.1 | Oenococcus oeni | - |
gene adh3 | - |
1.1.1.1 | Oenococcus oeni DSM 20252 | - |
gene adh3 | - |
1.1.1.202 | Oenococcus oeni | A0NIJ1 | - |
- |
1.1.1.202 | Oenococcus oeni ATCC BAA-1163 | A0NIJ1 | - |
- |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
1.1.1.1 | 0.2 | - |
substrates 1,3-propanediol and NAD+, pH 9.0, 60°C, recombinant enzyme | Oenococcus oeni |
1.1.1.1 | 0.2 | - |
substrates acetaldehyde and NADH, pH 8.0, 60°C, recombinant enzyme | Oenococcus oeni |
1.1.1.1 | 0.3 | - |
substrates 1,3-propanediol and NAD+, pH 9.0, 60°C, recombinant enzyme, with NiCl2 addition | Oenococcus oeni |
1.1.1.1 | 1.1 | - |
substrates ethanol and NAD+, pH 9.0, 60°C, recombinant enzyme | Oenococcus oeni |
1.1.1.202 | 1.1 | - |
pH 9.0, 60°C | Oenococcus oeni |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.1.1 | 1,2-propanediol + NAD+ | - |
Oenococcus oeni | ? | - |
? | |
1.1.1.1 | 1,2-propanediol + NAD+ | - |
Oenococcus oeni DSM 20252 | ? | - |
? | |
1.1.1.1 | 1,3-propanediol + NAD+ | - |
Oenococcus oeni | ? | - |
? | |
1.1.1.1 | 1,3-propanediol + NAD+ | - |
Oenococcus oeni DSM 20252 | ? | - |
? | |
1.1.1.1 | 1-butanol + NAD+ | - |
Oenococcus oeni | butanal + NADH + H+ | - |
r | |
1.1.1.1 | 1-butanol + NAD+ | - |
Oenococcus oeni DSM 20252 | butanal + NADH + H+ | - |
r | |
1.1.1.1 | 1-propanol + NAD+ | - |
Oenococcus oeni | propanaldehyde + NADH + H+ | - |
r | |
1.1.1.1 | 2-mercaptoethanol + NAD+ | - |
Oenococcus oeni | ? | - |
? | |
1.1.1.1 | acetaldehyde + NADH + H+ | - |
Oenococcus oeni | ethanol + NAD+ | - |
r | |
1.1.1.1 | acetaldehyde + NADH + H+ | - |
Oenococcus oeni DSM 20252 | ethanol + NAD+ | - |
r | |
1.1.1.1 | ethanol + NAD+ | - |
Oenococcus oeni | acetaldehyde + NADH + H+ | - |
r | |
1.1.1.1 | ethanol + NAD+ | best substrate | Oenococcus oeni | acetaldehyde + NADH + H+ | - |
r | |
1.1.1.1 | additional information | no activity with methanol, 1-butanol, glycerol or 2-propanol | Oenococcus oeni | ? | - |
? | |
1.1.1.1 | additional information | no activity with methanol, 1-butanol, glycerol or 2-propanol | Oenococcus oeni DSM 20252 | ? | - |
? | |
1.1.1.202 | 1,2-propanediol + NAD+ | - |
Oenococcus oeni | 2-hydroxypropanal + NADH | - |
? | |
1.1.1.202 | 1,2-propanediol + NAD+ | - |
Oenococcus oeni ATCC BAA-1163 | 2-hydroxypropanal + NADH | - |
? | |
1.1.1.202 | 1,3-propanediol + NAD+ | activity with 1,3-propanediol is highly dependent on the presence of Ni2+ | Oenococcus oeni | 3-hydroxypropanal + NADH | - |
? | |
1.1.1.202 | 1,3-propanediol + NAD+ | activity with 1,3-propanediol is highly dependent on the presence of Ni2+ | Oenococcus oeni ATCC BAA-1163 | 3-hydroxypropanal + NADH | - |
? | |
1.1.1.202 | additional information | enzyme additionally accepts ethanol, 1-propanol, 2-mercaptoethanol and their reduces their corresponding aldehydes. No substrates: methanol, 1-butanol, glycerol or 2-propanol | Oenococcus oeni | ? | - |
? | |
1.1.1.202 | additional information | enzyme additionally accepts ethanol, 1-propanol, 2-mercaptoethanol and their reduces their corresponding aldehydes. No substrates: methanol, 1-butanol, glycerol or 2-propanol | Oenococcus oeni ATCC BAA-1163 | ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
1.1.1.1 | homodimer | 2 * 42200-43000, recombinant His-tagged enzyme, SDS-PAGE | Oenococcus oeni |
1.1.1.1 | More | Adh3 forms a Ni2+-containing homodimer in its active form, crystal structure analysis, larger aggregates are inactive | Oenococcus oeni |
1.1.1.202 | dimer | 2 * 42200, calculated, 2 * 43000, SDS-PAGE of recombinant His-tagged protein | Oenococcus oeni |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.1.1.1 | NAD+-dependent alcohol dehydrogenase | - |
Oenococcus oeni |
1.1.1.202 | ADH3 | - |
Oenococcus oeni |
1.1.1.202 | DhaT | - |
Oenococcus oeni |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.1.1.1 | 50 | 60 | - |
Oenococcus oeni |
1.1.1.202 | 50 | 60 | - |
Oenococcus oeni |
EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.1.1.1 | 10 | 80 | more than 40% of activity between 40°C and 70°C with all substrates. The enzyme is not capable of reducing acetaldehyde at 80°C, while remaining activity for alcohol oxidation, temperature optimum lies between 50°C to 60°C | Oenococcus oeni |
EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.1.1.202 | 50 | - |
60 min, inactivation | Oenococcus oeni |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.1.1.1 | 8 | - |
aldehyde reduction | Oenococcus oeni |
1.1.1.1 | 9 | - |
ethanol oxidation | Oenococcus oeni |
1.1.1.202 | 8 | - |
reduction of aldehydes | Oenococcus oeni |
1.1.1.202 | 9 | - |
oxidation of alcohols | Oenococcus oeni |
EC Number | pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|---|
1.1.1.202 | 6 | 7 | most stable within this range | Oenococcus oeni |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.1 | NAD+ | Adh3 is strictly dependent on NAD+/NADH, and shows no activity with NADP+/NADPH as cofactor | Oenococcus oeni | |
1.1.1.1 | NADH | Adh3 is strictly dependent on NAD+/NADH, and shows no activity with NADP+/NADPH as cofactor | Oenococcus oeni | |
1.1.1.202 | additional information | no cofactor: NADPH | Oenococcus oeni | |
1.1.1.202 | NAD+ | - |
Oenococcus oeni | |
1.1.1.202 | NADH | - |
Oenococcus oeni |
EC Number | Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|---|
1.1.1.1 | Oenococcus oeni | sequence calculation | - |
5.23 |
1.1.1.202 | Oenococcus oeni | calculated | - |
5.2 |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.1.1.1 | evolution | the enzyme ADH3 belongs to the group III alcohol dehydrogenases | Oenococcus oeni |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.1 | 0.01 | - |
1,2-propanediol | pH 8.0, 60°C, recombinant enzyme | Oenococcus oeni | |
1.1.1.1 | 0.01 | - |
1,3-Propanediol | pH 9.0, 60°C, recombinant enzyme | Oenococcus oeni | |
1.1.1.1 | 0.5 | - |
ethanol | pH 9.0, 60°C, recombinant enzyme | Oenococcus oeni | |
1.1.1.1 | 0.9 | - |
acetaldehyde | pH 8.0, 60°C, recombinant enzyme | Oenococcus oeni | |
1.1.1.1 | 4.9 | - |
NAD+ | pH 9.0, 60°C, recombinant enzyme | Oenococcus oeni | |
1.1.1.1 | 12.9 | - |
NADH | pH 8.0, 60°C, recombinant enzyme | Oenococcus oeni | |
1.1.1.202 | 0.01 | - |
1,2-propanediol | pH 9.0, 60°C | Oenococcus oeni | |
1.1.1.202 | 0.01 | - |
1,3-Propanediol | pH 9.0, 60°C | Oenococcus oeni | |
1.1.1.202 | 4.9 | - |
NAD+ | pH 9.0, 60°C | Oenococcus oeni | |
1.1.1.202 | 12.9 | - |
NADH | pH 8.0, 60°C | Oenococcus oeni |