Cloned (Comment) | Organism |
---|---|
gene adh3, sequence comparison, expression of N-terminally His-tagged enzyme in Escherichia coli strains NovaBlue Singles and M15[pREP4] | Oenococcus oeni |
Crystallization (Comment) | Organism |
---|---|
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 |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
EDTA | - |
Oenococcus oeni | |
Zn2+ | complete inhibition | Oenococcus oeni |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.05 | - |
NAD+ | pH 9.0, 60°C, recombinant enzyme | Oenococcus oeni | |
0.2 | - |
NADH | pH 8.0, 60°C, recombinant enzyme | Oenococcus oeni | |
2.6 | - |
acetaldehyde | pH 8.0, 60°C, recombinant enzyme | Oenococcus oeni | |
10.4 | - |
ethanol | pH 9.0, 60°C, recombinant enzyme | Oenococcus oeni | |
11.5 | - |
1,2-propanediol | pH 8.0, 60°C, recombinant enzyme | Oenococcus oeni | |
26.7 | - |
1,3-Propanediol | pH 9.0, 60°C, recombinant enzyme | Oenococcus oeni |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | 40% activation at 10 mM | Oenococcus oeni | |
additional information | Adh3 exhibits tolerance to several metal ions | Oenococcus oeni | |
Na+ | 40% activation at 1 mM | Oenococcus oeni | |
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 |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
74000 | - |
recombinant His-tagged enzyme, gel filtration | Oenococcus oeni |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ethanol + NAD+ | Oenococcus oeni | - |
acetaldehyde + NADH + H+ | - |
r |
Organic Solvent | Comment | Organism |
---|---|---|
additional information | Adh3 exhibits tolerance to several organic solvents | Oenococcus oeni |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Oenococcus oeni | - |
gene adh3 | - |
Oenococcus oeni DSM 20252 | - |
gene adh3 | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.2 | - |
substrates 1,3-propanediol and NAD+, pH 9.0, 60°C, recombinant enzyme | Oenococcus oeni |
0.2 | - |
substrates acetaldehyde and NADH, pH 8.0, 60°C, recombinant enzyme | Oenococcus oeni |
0.3 | - |
substrates 1,3-propanediol and NAD+, pH 9.0, 60°C, recombinant enzyme, with NiCl2 addition | Oenococcus oeni |
1.1 | - |
substrates ethanol and NAD+, pH 9.0, 60°C, recombinant enzyme | Oenococcus oeni |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1,2-propanediol + NAD+ | - |
Oenococcus oeni | ? | - |
? | |
1,2-propanediol + NAD+ | - |
Oenococcus oeni DSM 20252 | ? | - |
? | |
1,3-propanediol + NAD+ | - |
Oenococcus oeni | ? | - |
? | |
1,3-propanediol + NAD+ | - |
Oenococcus oeni DSM 20252 | ? | - |
? | |
1-butanol + NAD+ | - |
Oenococcus oeni | butanal + NADH + H+ | - |
r | |
1-butanol + NAD+ | - |
Oenococcus oeni DSM 20252 | butanal + NADH + H+ | - |
r | |
1-propanol + NAD+ | - |
Oenococcus oeni | propanaldehyde + NADH + H+ | - |
r | |
2-mercaptoethanol + NAD+ | - |
Oenococcus oeni | ? | - |
? | |
acetaldehyde + NADH + H+ | - |
Oenococcus oeni | ethanol + NAD+ | - |
r | |
acetaldehyde + NADH + H+ | - |
Oenococcus oeni DSM 20252 | ethanol + NAD+ | - |
r | |
ethanol + NAD+ | - |
Oenococcus oeni | acetaldehyde + NADH + H+ | - |
r | |
ethanol + NAD+ | best substrate | Oenococcus oeni | acetaldehyde + NADH + H+ | - |
r | |
additional information | no activity with methanol, 1-butanol, glycerol or 2-propanol | Oenococcus oeni | ? | - |
? | |
additional information | no activity with methanol, 1-butanol, glycerol or 2-propanol | Oenococcus oeni DSM 20252 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 42200-43000, recombinant His-tagged enzyme, SDS-PAGE | Oenococcus oeni |
More | Adh3 forms a Ni2+-containing homodimer in its active form, crystal structure analysis, larger aggregates are inactive | Oenococcus oeni |
Synonyms | Comment | Organism |
---|---|---|
NAD+-dependent alcohol dehydrogenase | - |
Oenococcus oeni |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | 60 | - |
Oenococcus oeni |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
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 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
aldehyde reduction | Oenococcus oeni |
9 | - |
ethanol oxidation | Oenococcus oeni |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | Adh3 is strictly dependent on NAD+/NADH, and shows no activity with NADP+/NADPH as cofactor | Oenococcus oeni | |
NADH | Adh3 is strictly dependent on NAD+/NADH, and shows no activity with NADP+/NADPH as cofactor | Oenococcus oeni |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Oenococcus oeni | sequence calculation | - |
5.23 |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme ADH3 belongs to the group III alcohol dehydrogenases | Oenococcus oeni |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.01 | - |
1,2-propanediol | pH 8.0, 60°C, recombinant enzyme | Oenococcus oeni | |
0.01 | - |
1,3-Propanediol | pH 9.0, 60°C, recombinant enzyme | Oenococcus oeni | |
0.5 | - |
ethanol | pH 9.0, 60°C, recombinant enzyme | Oenococcus oeni | |
0.9 | - |
acetaldehyde | pH 8.0, 60°C, recombinant enzyme | Oenococcus oeni | |
4.9 | - |
NAD+ | pH 9.0, 60°C, recombinant enzyme | Oenococcus oeni | |
12.9 | - |
NADH | pH 8.0, 60°C, recombinant enzyme | Oenococcus oeni |