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Literature summary for 1.1.1.2 extracted from

  • Ying, X.; Grunden, A.M.; Nie, L.; Adams, M.W.; Ma, K.
    Molecular characterization of the recombinant iron-containing alcohol dehydrogenase from the hyperthermophilic Archaeon, Thermococcus strain ES1 (2009), Extremophiles, 13, 299-311.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21 cells Thermococcus sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.006
-
NADPH recombinant enzyme, with 90 mM ethanol as cosubstrate, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.
0.007
-
NADPH native enzyme, with 90 mM ethanol as cosubstrate, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.
0.84
-
acetaldehyde recombinant enzyme, with 0.4 mM NADP+ as cosubstrate, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.
1
-
acetaldehyde native enzyme, with 0.4 mM NADP+ as cosubstrate, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.
9.7
-
ethanol recombinant enzyme, with 0.4 mM NADP+ as cosubstrate, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.
9.7
-
NADP+ recombinant enzyme, with 90 mM ethanol as cosubstrate, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.
10.4
-
ethanol native enzyme, with 0.4 mM NADP+ as cosubstrate, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.
10.4
-
NADP+ native enzyme, with 90 mM ethanol as cosubstrate, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ contains 1.0 g-atom iron per subunit Thermococcus sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
45000
-
4 * 45000, SDS-PAGE Thermococcus sp.
176000
-
recombinant enzyme, gel filtration Thermococcus sp.
179200
-
calculated from amino acid sequence Thermococcus sp.
180000
-
SDS-PAGE Thermococcus sp.
182000
-
native enzyme, gel filtration Thermococcus sp.

Organism

Organism UniProt Comment Textmining
Thermococcus sp. C1IWT4
-
-
Thermococcus sp. ES1 C1IWT4
-
-

Oxidation Stability

Oxidation Stability Organism
the enzyme is sensitive to oxygen with a half-life upon exposure to air of about 4 min Thermococcus sp.

Purification (Commentary)

Purification (Comment) Organism
DEAE-Sepharose column chromatography, hydroxyapatite column chromatography, and Superdex 200 gel filtration Thermococcus sp.

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
5.6
-
native enzyme from cell-free extract, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.
6.8
-
recombinant enzyme from cell-free extract, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.
52.8
-
recombinant enzyme after 7.8fold purification, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.
57
-
native enzyme after 10.1fold purification, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1-butanol + NADP+ the native enzyme shows 154.6% activity with 1-butanol compared to ethanol Thermococcus sp. butanal + NADPH + H+
-
?
1-butanol + NADP+ the native enzyme shows 154.6% activity with 1-butanol compared to ethanol Thermococcus sp. ES1 butanal + NADPH + H+
-
?
1-heptanol + NADP+ the native enzyme shows 94% activity with 1-heptanol compared to ethanol Thermococcus sp. heptanal + NADPH + H+
-
?
1-heptanol + NADP+ the native enzyme shows 94% activity with 1-heptanol compared to ethanol Thermococcus sp. ES1 heptanal + NADPH + H+
-
?
1-octanol + NADP+ the native enzyme shows 68.8% activity with 1-octanol compared to ethanol Thermococcus sp. octanal + NADPH + H+
-
?
1-octanol + NADP+ the native enzyme shows 68.8% activity with 1-octanol compared to ethanol Thermococcus sp. ES1 octanal + NADPH + H+
-
?
1-pentanol + NADP+ the native enzyme shows 184% activity with 1-pentanol compared to ethanol Thermococcus sp. pentanal + NADPH + H+
-
?
1-pentanol + NADP+ the native enzyme shows 184% activity with 1-pentanol compared to ethanol Thermococcus sp. ES1 pentanal + NADPH + H+
-
?
1-propanol + NADP+ the native enzyme shows 120.6% activity with 1-propanol compared to ethanol Thermococcus sp. propanal + NADPH + H+
-
?
2-phenylethanol + NADP+ the native enzyme shows 40.2% activity with 2-phenylethanol compared to ethanol Thermococcus sp. 2-phenylacetaldehyde + NADPH + H+
-
?
ethanol + NADP+ the native enzyme shows 100% activity with ethanol Thermococcus sp. acetaldehyde + NADPH + H+
-
?
ethanol + NADP+ the native enzyme shows 100% activity with ethanol Thermococcus sp. ES1 acetaldehyde + NADPH + H+
-
?
hexyl alcohol + NADP+ the native enzyme shows 137.8% activity with hexyl alcohol compared to ethanol Thermococcus sp. ?
-
?

Subunits

Subunits Comment Organism
homotetramer 4 * 45000, SDS-PAGE Thermococcus sp.

Synonyms

Synonyms Comment Organism
ADH
-
Thermococcus sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
22.9
-
acetaldehyde recombinant enzyme, with 0.4 mM NADP+ as cosubstrate, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.
23.6
-
acetaldehyde native enzyme, with 0.4 mM NADP+ as cosubstrate, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.
24
-
NADPH native enzyme, with 90 mM ethanol as cosubstrate, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.
24
-
NADPH recombinant enzyme, with 90 mM ethanol as cosubstrate, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.
81
-
ethanol recombinant enzyme, with 0.4 mM NADP+ as cosubstrate, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.
81
-
NADP+ recombinant enzyme, with 90 mM ethanol as cosubstrate, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.
84
-
ethanol native enzyme, with 0.4 mM NADP+ as cosubstrate, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.
84
-
NADP+ native enzyme, with 90 mM ethanol as cosubstrate, in 100 mM EPPS buffer (pH 8.8) Thermococcus sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
for acetaldehyde reduction Thermococcus sp.
10.3 10.5 for ethanol oxidation Thermococcus sp.

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Thermococcus sp.

pI Value

Organism Comment pI Value Maximum pI Value
Thermococcus sp. calculated from amino acid sequence
-
5.5