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Literature summary for 1.1.1.71 extracted from

  • Kulig, J.; Frese, A.; Kroutil, W.; Pohl, M.; Rother, D.
    Biochemical characterization of an alcohol dehydrogenase from Ralstonia sp. (2013), Biotechnol. Bioeng., 110, 1838-1848.
    View publication on PubMed
Search for more literature for 1.1.1.71

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Ralstonia sp.

Inhibitors

Inhibitors Comment Organism Structure
(R)-2-hydroxypropiophenone
-
 Ralstonia sp.
Cyclohexanol
-
 Ralstonia sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.005
-
 NADPH with benzaldehyde as cosubstrate, at pH 7.5 and 30°C Ralstonia sp.
0.008
-
 NADPH with cyclohexanone as cosubstrate, at pH 7.5 and 30°C Ralstonia sp.
0.06
-
 NADP+ with benzyl alcohol as cosubstrate, at pH 9.0 and 30°C Ralstonia sp.
1.6
-
 (R)-2-hydroxypropiophenone at pH 7.5 and 30°C Ralstonia sp.
3.6
-
 (S)-2-Hydroxypropiophenone at pH 7.5 and 30°C Ralstonia sp.
5.2
-
 Cyclohexanol at pH 9.0 and 30°C Ralstonia sp.
10.8
-
 cyclohexanone at pH 7.5 and 30°C Ralstonia sp.
12.3
-
 benzyl alcohol at pH 9.0 and 30°C Ralstonia sp.
15.6
-
 benzaldehyde at pH 7.5 and 30°C Ralstonia sp.

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ Ca2+ has an stabilizing effect. With 1.0 mM CaCl2, the enzyme is completely stable at 0°C for 2 h and after 3.5 h almost 90% of the initial activity is retained Ralstonia sp.

Organism

Organism UniProt Comment Textmining
Ralstonia sp.
-
-
-
Ralstonia sp. DSM 6428
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Ralstonia sp.

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.01
-
 with acetone, at pH 7.5 and 30°C Ralstonia sp.
0.05
-
 with acetaldehyde, at pH 7.5 and 30°C Ralstonia sp.
0.08
-
 with 2,5-hexandione, at pH 7.5 and 30°C Ralstonia sp.
0.11
-
 with cyclohexanone, at pH 7.5 and 30°C Ralstonia sp.
0.56
-
 with 4-phenyl-2-butanone, at pH 7.5 and 30°C Ralstonia sp.
4.35
-
 with acetophenone, at pH 7.5 and 30°C Ralstonia sp.
4.89
-
 with benzaldehyde, at pH 7.5 and 30°C Ralstonia sp.
8.36
-
 with cyclohexanol, at pH 10.5 and 30°C Ralstonia sp.
9.78
-
 with cyclohexanone, at pH 7.5 and 30°C Ralstonia sp.
11.58
-
 with 3-methylcyclohexanol, at pH 10.5 and 30°C Ralstonia sp.
11.7
-
 with 3-fluorobenzaldehyde, at pH 7.5 and 30°C Ralstonia sp.
12.62
-
 with 3-methylcyclohexanone, at pH 7.5 and 30°C Ralstonia sp.
17.1
-
 with (S)-2-hydroxypropiophenone, at pH 7.5 and 30°C Ralstonia sp.
18.83
-
 with cyclohexenol, at pH 10.5 and 30°C Ralstonia sp.
23.31
-
 2-bromobenzaldehyde, at pH 7.5 and 30°C Ralstonia sp.
362.6
-
 with (R)-2-hydroxypropiophenone, at pH 7.5 and 30°C Ralstonia sp.

Storage Stability

Storage Stability Organism
-20°C, 10 mM TEA-HCl buffer pH 7.5, at least 80 days, no loss of activity Ralstonia sp.
8°C, 50 mM TEA-HCl buffer pH 7.5, 1 day, 91% loss of activity Ralstonia sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(1R)-1-hydroxy-1-phenylpropan-2-one + NADPH + H+
-
 Ralstonia sp. (1R)-1-phenylpropane-1,2-diol + NADP+
-
 ?
(2R)-2-hydroxy-1-(4-methoxyphenyl)propan-1-one + NADPH + H+
-
 Ralstonia sp. (2R)-1-(4-methoxyphenyl)propane-1,2-diol + NADP+
-
 ?
(2S)-2-hydroxy-1-(4-methoxyphenyl)propan-1-one + NADPH + H+
-
 Ralstonia sp. (2S)-1-(4-methoxyphenyl)propane-1,2-diol + NADP+
-
 ?
(R)-2-hydroxypropiophenone + NADPH + H+
-
 Ralstonia sp. ?
-
 r
(S)-2-hydroxypropiophenone + NADPH + H+
-
 Ralstonia sp. ?
-
 r
2,5-hexandione + NADH + H+
-
 Ralstonia sp. ?
-
 r
2-bromobenzaldehyde + NADPH
-
 Ralstonia sp. 2-bromobenzyl alcohol + NADP+
-
 r
2-bromobenzaldehyde + NADPH + H+
-
 Ralstonia sp. 2-bromobenzyl alcohol + NADP+
-
 ?
2-cyclohexenol + NADP+
-
 Ralstonia sp. 2-cyclohexenone + NADH + H+
-
 r
2-cyclohexenone + NADH + H+
-
 Ralstonia sp. 2-cyclohexenol + NADP+
-
 r
2-fluorobenzaldehyde + NADPH + H+
-
 Ralstonia sp. 2-fluorobenzyl alcohol + NADP+
-
 ?
2-hydroxy-2-methyl-1-phenylpropan-1-one + NADPH + H+
-
 Ralstonia sp. 2-methyl-1-phenylpropan-1,2-diol + NADP+
-
 ?
2-methoxybenzaldehyde + NADPH + H+
-
 Ralstonia sp. 2-methoxybenzyl alcohol + NADP+
-
 ?
2-methoxybenzaldehyde + NADPH + H+
-
 Ralstonia sp. DSM 6428 2-methoxybenzyl alcohol + NADP+
-
 ?
2-methylbenzaldehyde + NADPH + H+
-
 Ralstonia sp. 2-methylbenzyl alcohol + NADP+
-
 ?
3-fluorobenzaldehyde + NADPH
-
 Ralstonia sp. 3-fluorobenzyl alcohol + NADP+
-
 r
3-methoxybenzaldehyde + NADPH + H+
-
 Ralstonia sp. 3-methoxybenzyl alcohol + NADP+
-
 ?
3-methylbenzaldehyde + NADPH + H+
-
 Ralstonia sp. 3-methylbenzyl alcohol + NADP+
-
 ?
3-methylcyclohexanol + NADP+
-
 Ralstonia sp. ?
-
 r
3-methylcyclohexanone + NADH + H+
-
 Ralstonia sp. 3-methylcyclohexanol + NAD+
-
 r
4-bromobenzaldehyde + NADPH + H+
-
 Ralstonia sp. 4-bromobenzyl alcohol + NADP+
-
 ?
4-fluorobenzaldehyde + NADPH + H+
-
 Ralstonia sp. 4-fluorobenzyl alcohol + NADP+
-
 ?
4-methoxybenzaldehyde + NADPH + H+
-
 Ralstonia sp. 4-methoxybenzyl alcohol + NADP+
-
 ?
4-methoxybenzaldehyde + NADPH + H+
-
 Ralstonia sp. DSM 6428 4-methoxybenzyl alcohol + NADP+
-
 ?
4-methylbenzaldehyde + NADPH + H+
-
 Ralstonia sp. 4-methylbenzyl alcohol + NADP+
-
 ?
4-methylpentan-2-one + NADPH + H+
-
 Ralstonia sp. 4-methylpentan-2-ol + NADP+
-
 ?
4-phenyl-2-butanone + NADH + H+
-
 Ralstonia sp. 4-phenyl-2-butanol + NAD+
-
 r
acetaldehyde + NADPH
-
 Ralstonia sp. ethanol + NADP+
-
 r
acetone + NADH + H+
-
 Ralstonia sp. propan-2-ol + NAD+
-
 r
acetophenone + NADH + H+
-
 Ralstonia sp. 1-phenylethan-1-ol + NAD+
-
 r
benzaldehyde + NADPH
-
 Ralstonia sp. benzyl alcohol + NADP+
-
 r
benzaldehyde + NADPH
-
 Ralstonia sp. DSM 6428 benzyl alcohol + NADP+
-
 r
benzyl alcohol + NADP+
-
 Ralstonia sp. benzaldehyde + NADPH
-
 r
benzyl alcohol + NADP+
-
 Ralstonia sp. DSM 6428 benzaldehyde + NADPH
-
 r
butan-2-ol + NADP+
-
 Ralstonia sp. butan-2-one + NADPH + H+
-
 ?
butan-2-one + NADPH + H+
-
 Ralstonia sp. butan-2-ol + NADP+
-
 ?
cyclohexanol + NADP+
-
 Ralstonia sp. cyclohexanone + NADH + H+
-
 r
cyclohexanone + NADH + H+
-
 Ralstonia sp. cyclohexanol + NAD+
-
 r
cyclohexanone + NADH + H+
-
 Ralstonia sp. cyclohexanol + NADP+
-
 r
formaldehyde + NADPH + H+
-
 Ralstonia sp. methanol + NADP+
-
 ?
furan-2-carbaldehyde + NADPH + H+
-
 Ralstonia sp. (furan-2-yl)methanol + NADP+
-
 ?
heptan-4-one + NADPH + H+
-
 Ralstonia sp. heptan-4-ol + NADP+
-
 ?
hexan-1-ol + NADP+
-
 Ralstonia sp. hexaldehyde + NADPH + H+
-
 ?
hexan-2-one + NADPH + H+
-
 Ralstonia sp. hexan-2-ol + NADP+
-
 ?
hexanal + NADPH + H+
-
 Ralstonia sp. hexan-1-ol + NADP+
-
 ?
additional information no activity with methanol and ethanol Ralstonia sp. ?
-
 ?
additional information no activity with methanol and ethanol Ralstonia sp. DSM 6428 ?
-
 ?
octan-1-ol + NADP+
-
 Ralstonia sp. octaldehyde + NADPH + H+
-
 ?
octan-2-one + NADPH + H+
-
 Ralstonia sp. octan-2-ol + NADP+
-
 ?
pentan-1-ol + NADP+
-
 Ralstonia sp. pentaldehyde + NADPH + H+
-
 ?
propan-1-ol + NADP+
-
 Ralstonia sp. propionaldehyde + NADPH + H+
-
 ?
propan-2-ol + NADP+
-
 Ralstonia sp. propan-2-one + NADPH + H+
-
 ?
pyridine-2-carbaldehyde + NADPH + H+
-
 Ralstonia sp. (pyridin-2-yl)methanol + NADP+
-
 ?

Synonyms

Synonyms Comment Organism
NADPH-dependent alcohol dehydrogenase
-
 Ralstonia sp.
RADH
-
 Ralstonia sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
8 25 the enzyme has a half-life of 80 h at 25°C and stability optimum between 8 and 15°C in the presence of 0.8 mM CaCl2 (half-life of 130 h) Ralstonia sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6 9.5 reduction reaction Ralstonia sp.
10 11.5 oxidation reaction Ralstonia sp.

pH Stability

pH Stability pH Stability Maximum Comment Organism
5.5 8 the enzyme shows half-lives of 60-70 h between pH 5.5 and 8.0 at room temperature Ralstonia sp.

Cofactor

Cofactor Comment Organism Structure
NADP+
-
 Ralstonia sp.
NADPH
-
 Ralstonia sp.

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
11.4
-
 Cyclohexanol at pH 9.0 and 30°C Ralstonia sp.
16.4
-
 (R)-2-hydroxypropiophenone at pH 7.5 and 30°C Ralstonia sp.