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Literature summary for 1.1.1.1 extracted from

  • Elleuche, S.; Fodor, K.; Klippel, B.; von der Heyde, A.; Wilmanns, M.; Antranikian, G.
    Structural and biochemical characterisation of a NAD+-dependent alcohol dehydrogenase from Oenococcus oeni as a new model molecule for industrial biotechnology applications (2013), Appl. Microbiol. Biotechnol., 97, 8963-8975.
    View publication on PubMed

Cloned(Commentary)

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 (Commentary)

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

Inhibitors Comment Organism Structure
EDTA
-
Oenococcus oeni
Zn2+ complete inhibition Oenococcus oeni

KM Value [mM]

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

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 [Da] Molecular Weight Maximum [Da] Comment Organism
74000
-
recombinant His-tagged enzyme, gel filtration Oenococcus oeni

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ethanol + NAD+ Oenococcus oeni
-
acetaldehyde + NADH + H+
-
r

Organic Solvent Stability

Organic Solvent Comment Organism
additional information Adh3 exhibits tolerance to several organic solvents Oenococcus oeni

Organism

Organism UniProt Comment Textmining
Oenococcus oeni
-
gene adh3
-
Oenococcus oeni DSM 20252
-
gene adh3
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [┬Ámol/min/mg] Specific Activity Maximum [┬Ámol/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 and Products (Substrate)

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

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

Synonyms Comment Organism
NAD+-dependent alcohol dehydrogenase
-
Oenococcus oeni

Temperature Optimum [┬░C]

Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
50 60
-
Oenococcus oeni

Temperature Range [┬░C]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
aldehyde reduction Oenococcus oeni
9
-
ethanol oxidation Oenococcus oeni

Cofactor

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

pI Value

Organism Comment pI Value Maximum pI Value
Oenococcus oeni sequence calculation
-
5.23

General Information

General Information Comment Organism
evolution the enzyme ADH3 belongs to the group III alcohol dehydrogenases Oenococcus oeni

kcat/KM [mM/s]

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