Activating Compound | Comment | Organism | Structure |
---|---|---|---|
EDTA | 10 mM, 1.65fold activation | Thermus thermophilus |
Application | Comment | Organism |
---|---|---|
synthesis | enzyme catalyzes the following reactions with Prelog specificity: the reduction of acetophenone, 2,2,2-trifluoroacetophenone, alpha-tetralone, and alpha-methyl and alpha-ethyl benzoylformates to (S)-1-phenylethanol (>99% enantiomeric excess), (R)-alpha-(trifluoromethyl)benzyl alcohol (93% enantiomeric excess), (S)-alpha-tetralol (>99% enantiomeric excess), methyl (R)-mandelate (92% enantiomeric excess), and ethyl (R)-mandelate (95% enantiomeric excess), respectively, by way of an efficient in situ NADH-recycling system involving 2-propanol and a second thermophilic ADH | Thermus thermophilus |
Cloned (Comment) | Organism |
---|---|
overexpressed in Escherichia coli | Thermus thermophilus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
1-butyl-3-methylimidazolium tetrafluoroborate | 2 mM, 35% of initial activity; 2 mM, 65% inhibition | Thermus thermophilus | |
Ca2+ | 100 mM, 70% of initial activity | Thermus thermophilus | |
CaCl2 | 100 mM, 30% inhibition | Thermus thermophilus | |
Cu2+ | 100 mM, 76% of initial activity | Thermus thermophilus | |
CuCl2 | 1 mM, 24% inhibition | Thermus thermophilus | |
Hg2+ | 100 mM, 33% of initial activity | Thermus thermophilus | |
HgCl2 | 1 mM, 67% inhibition | Thermus thermophilus | |
Mg2+ | 100 mM, 67% of initial activity | Thermus thermophilus | |
MgCl2 | 100 mM, 33% inhibition | Thermus thermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0035 | - |
NADH | pH 10.5, 65°C | Thermus thermophilus | |
0.035 | - |
NADH | 65°C | Thermus thermophilus | |
0.24 | - |
NAD+ | 65°C | Thermus thermophilus | |
0.24 | - |
NAD+ | pH 10.5, 65°C | Thermus thermophilus | |
1 | - |
ethyl benzoylformate | 65°C | Thermus thermophilus | |
1 | - |
ethyl oxo(phenyl)acetate | pH 10.5, 65°C | Thermus thermophilus | |
2.7 | - |
methylbenzoylformate | 65°C | Thermus thermophilus | |
2.7 | - |
methyl oxo(phenyl)acetate | pH 10.5, 65°C | Thermus thermophilus | |
4.2 | - |
(S)-(+)-1-indanol | 65°C | Thermus thermophilus | |
4.2 | - |
(S)-(+)-alpha-tetraol | 65°C | Thermus thermophilus | |
4.2 | - |
(S)-1-indanol | pH 10.5, 65°C | Thermus thermophilus | |
4.2 | - |
(S)-alpha-tetralol | pH 10.5, 65°C | Thermus thermophilus | |
4.4 | - |
3-Methoxybenzaldehyde | 65°C | Thermus thermophilus | |
4.4 | - |
4-methoxybenzaldehyde | pH 6.0, 65°C | Thermus thermophilus | |
5.1 | - |
1-Indanol | 65°C | Thermus thermophilus | |
5.3 | - |
alpha-tetralol | 65°C | Thermus thermophilus | |
5.8 | - |
alpha-tetralone | 65°C | Thermus thermophilus | |
5.8 | - |
alpha-tetralone | pH 10.5, 65°C | Thermus thermophilus | |
5.9 | - |
1-phenyl-1,2-propanedione | 65°C | Thermus thermophilus | |
5.9 | - |
1-phenyl-1,2-propandione | pH 10.5, 65°C | Thermus thermophilus | |
11.2 | - |
2,2,2-trifluoroacetophenone | 65°C | Thermus thermophilus | |
11.2 | - |
2,2,2-trifluoroacetophenone | pH 10.5, 65°C | Thermus thermophilus | |
18.1 | - |
(S)-(-)-1-phenylethanol | 65°C | Thermus thermophilus | |
18.1 | - |
(S)-1-phenylethanol | pH 10.5, 65°C | Thermus thermophilus | |
27.6 | - |
1-Indanone | 65°C | Thermus thermophilus | |
27.6 | - |
1-Indanone | pH 10.5, 65°C | Thermus thermophilus | |
61 | - |
4-methoxybenzyl alcohol | 65°C | Thermus thermophilus | |
61 | - |
4-methoxybenzyl alcohol | pH 10.5, 65°C | Thermus thermophilus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
K+ | 100 mM KCl, 1.9fold activation | Thermus thermophilus | |
K+ | 100 mM, 193% of initial activity | Thermus thermophilus | |
Li+ | 100 mM LiCl, 1.8fold activation | Thermus thermophilus | |
Li+ | 100 mM, 178% of initial activity | Thermus thermophilus | |
Na+ | 100 mM NaCl, 1.8fold activation | Thermus thermophilus | |
Na+ | 100 mM, 179% of initial activity | Thermus thermophilus | |
Zn2+ | 1 mM ZnSO4, 1.14fold activation | Thermus thermophilus | |
Zn2+ | 1 mM, 114% of initial activity | Thermus thermophilus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
26961 | - |
4 * 26961, calculated from sequence | Thermus thermophilus |
105000 | - |
buffer containing 25 mM NaCl, gel filtration | Thermus thermophilus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermus thermophilus | - |
- |
- |
Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Thermus thermophilus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
151.1 | - |
- |
Thermus thermophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-(+)-1-indanol + NAD+ | - |
Thermus thermophilus | indanone + NADH + H+ | - |
? | |
(S)-1-indanol + NAD+ | 3900% of the activity with (S)-1-phenylethanol | Thermus thermophilus | 1-indanone + NADH + H+ | - |
r | |
(S)-1-phenylethanol + NAD+ | - |
Thermus thermophilus | 1-phenylethanone + NADH + H+ | - |
r | |
(S)-alpha-tetralol + NAD+ | 2700% of the activity with (S)-1-phenylethanol | Thermus thermophilus | alpha-tetralone + NADH + H+ | - |
r | |
1-(4'-chlorophenyl)ethanol + NAD+ | 26% of the activity with (S)-(-)-1-phenylethanol | Thermus thermophilus | 1-(4'-chlorophenyl)ethanone + NADH + H+ | - |
? | |
1-(4'-fluorophenyl)ethanol + NAD+ | 45% of the activity with (S)-1-phenylethanol | Thermus thermophilus | 1-(4'-fluorophenyl)ethanone + NADH + H+ | - |
r | |
1-(4-fluorophenyl)ethanol + NAD+ | 45% of the activity with (S)-(-)-1-phenylethanol | Thermus thermophilus | 1-(4-fluorophenyl)ethanone + NADH + H+ | - |
? | |
1-indanol + NAD+ | - |
Thermus thermophilus | 1-indanone + NADH + H+ | - |
? | |
1-indanone + NADH + H+ | - |
Thermus thermophilus | 1-indanol + NAD+ | - |
r | |
1-phenyl-1,2-propandione + NADH + H+ | 146% of the activity with 2,2,2-trifluoroacetophenone | Thermus thermophilus | 1-phenyl-2-hydroxy-1-propanone + NAD+ | - |
r | |
1-phenyl-1,2-propanedione + NADH + H+ | activity is 1.46fold higher than with 2,2,2-trifluoroacetophenone | Thermus thermophilus | ? | - |
? | |
1-phenyl-1-propanol + NAD+ | 59% of the activity with (S)-(-)-1-phenylethanol | Thermus thermophilus | 1-phenyl-1-propanone + NADH + H+ | - |
? | |
1-phenyl-1-propanol + NAD+ | 59% of the activity with (S)-1-phenylethanol | Thermus thermophilus | 1-phenylpropan-1-one + NADH + H+ | - |
r | |
1-phenyl-2-propanol + NAD+ | 16% of the activity with (S)-(-)-1-phenylethanol | Thermus thermophilus | 1-phenyl-2-propanone + NADH + H+ | - |
? | |
2,2,2-trifluoroacetophenone + NADH + H+ | - |
Thermus thermophilus | (R)-alpha-(trifluoromethyl)benzyl alcohol + NAD+ | 93% enantiomeric excess | r | |
2,2,2-trifluoroacetophenone + NADH + H+ | 93% enantiomeric excess | Thermus thermophilus | (R)-alpha-(trifluoromethyl)benzyl alcohol + NAD+ | - |
? | |
2,2,2-trifluoroacetophenone + NADH + H+ | - |
Thermus thermophilus | 2,2,2-trifluoro-1-phenylethanol + NAD+ | - |
r | |
2,2-dichloroacetophenone + NADH + H+ | 32% of the activity with 2,2,2-trifluoroacetophenone | Thermus thermophilus | 2,2-dichloro-1-phenylethanol + NAD+ | - |
? | |
2-methoxybenzaldehyde + NADH + H+ | 13% of the activity with 2,2,2-trifluoroacetophenone | Thermus thermophilus | 2-methoxybenzylalcohol + NAD+ | - |
? | |
2-methoxybenzyl alcohol + NAD+ | 25% of the activity with (S)-(-)-1-phenylethanol | Thermus thermophilus | 2-methoxybenzaldehyde + NADH + H+ | - |
? | |
2-propanol + NAD+ | - |
Thermus thermophilus | acetone + NADH + H+ | - |
? | |
2-propanol + NAD+ | - |
Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 | acetone + NADH + H+ | - |
? | |
3-methoxybenzaldehyde + NADH + H+ | 14% of the activity with 2,2,2-trifluoroacetophenone | Thermus thermophilus | 3-methoxybenzylalcohol + NAD+ | - |
? | |
3-methoxybenzaldehyde + NADH + H+ | - |
Thermus thermophilus | 3-methoxybenzyl alcohol + NAD+ | - |
r | |
3-methoxybenzyl alcohol + NAD+ | 13% of the activity with (S)-(-)-1-phenylethanol | Thermus thermophilus | 3-methoxybenzaldehyde + NADH + H+ | - |
? | |
3-methoxybenzyl alcohol + NAD+ | 13% of the activity with (S)-(-)-1-phenylethanol | Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 | 3-methoxybenzaldehyde + NADH + H+ | - |
? | |
4'-chlorobutyrophenone + NADH + H+ | 10% of the activity with 2,2,2-trifluoroacetophenone | Thermus thermophilus | 1-(4-chlorophenyl)butan-1-ol + NAD+ | - |
? | |
4-methoxybenzaldehyde + NADH + H+ | 13% of the activity with 2,2,2-trifluoroacetophenone | Thermus thermophilus | 4-methoxybenzylalcohol + NAD+ | - |
? | |
4-methoxybenzyl alcohol + NAD+ | 99% of the activity with (S)-(-)-1-phenylethanol | Thermus thermophilus | 4-methoxybenzaldehyde + NADH + H+ | - |
? | |
4-methoxybenzyl alcohol + NAD+ | 99% of the activity with (S)-1-phenylethanol | Thermus thermophilus | 4-methoxybenzaldehyde + NADH + H+ | - |
r | |
acetophenone + NADH + H+ | more than 99% enantiomeric excess | Thermus thermophilus | (S)-(-)-1-phenylethanol + NAD+ | - |
r | |
acetophenone + NADH + H+ | - |
Thermus thermophilus | (S)-1-phenylethanol + NAD+ | 99% enantiomeric excess | r | |
alpha-ethyl benzoylformate + NADH + H+ | - |
Thermus thermophilus | ethyl (R)-(-)-mandelate + NAD+ | 95% enantiomeric excess | r | |
alpha-methyl benzoylformate + NADH + H+ | - |
Thermus thermophilus | methyl (R)-mandelate + NAD+ | 92% enantiomeric excess | r | |
alpha-tetralone + NADH + H+ | - |
Thermus thermophilus | (S)-alpha-tetralol + NAD+ | 99% enantiomeric excess | r | |
alpha-tetralone + NADH + H+ | more than 99% enantiomeric excess | Thermus thermophilus | (S)-alpha-tetralol + NAD+ | - |
? | |
alpha-tetralone + NADH + H+ | - |
Thermus thermophilus | alpha-tetralol + NAD+ | - |
r | |
benzaldehyde + NADH + H+ | 14% of the activity with 2,2,2-trifluoroacetophenone | Thermus thermophilus | benzyl alcohol + NAD+ | - |
? | |
benzaldehyde + NADH + H+ | 14% of the activity with 2,2,2-trifluoroacetophenone | Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 | benzyl alcohol + NAD+ | - |
? | |
cyclohexanol + NAD+ | 13% of the activity with (S)-(-)-1-phenylethanol | Thermus thermophilus | cyclohexanone + NADH + H+ | - |
? | |
cyclohexanol + NAD+ | 13% of the activity with (S)-(-)-1-phenylethanol | Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 | cyclohexanone + NADH + H+ | - |
? | |
ethyl benzoylformate + NADH + H+ | 95% enantiomeric excess | Thermus thermophilus | ethyl (R)-(-)-mandelate + NAD+ | - |
? | |
ethyl oxo(phenyl)acetate + NADH + H+ | 100% of the activity with 2,2,2-trifluoroacetophenone | Thermus thermophilus | ethyl hydroxy(phenyl)acetate + NAD+ | - |
r | |
methyl benzoylformate + NADH + H+ | 92% enantiomeric excess | Thermus thermophilus | methyl (R)-(-)-mandelate + NAD+ | - |
? | |
methyl oxo(phenyl)acetate + NADH + H+ | 57% of the activity with 2,2,2-trifluoroacetophenone | Thermus thermophilus | methyl hydroxy(phenyl)acetate + NAD+ | - |
r | |
methylbenzoylformate + NADH + H+ | 13% of the activity with 2,2,2-trifluoroacetophenone | Thermus thermophilus | ? | - |
? | |
additional information | preference for reduction of aromatic ketones and alpha-keto esters, and poor activity on aromatic alcohols and aldehydes | Thermus thermophilus | ? | - |
? | |
additional information | preference for reduction of aromatic ketones and alpha-keto esters, and poor activity on aromatic alcohols and aldehydes | Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 | ? | - |
? | |
trans-cinnamyl alcohol + NAD+ | activity is 3.9fold higher than with (S)-(-)-1-phenylethanol | Thermus thermophilus | (2E)-3-phenylprop-2-enal + NADH + H+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
tetramer | 4 * 26961, calculated from sequence | Thermus thermophilus |
Synonyms | Comment | Organism |
---|---|---|
NAD(H)-dependent alcohol dehydrogenase | - |
Thermus thermophilus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
73 | - |
- |
Thermus thermophilus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
73 | - |
inactivation above | Thermus thermophilus |
90 | - |
half-life 30 min | Thermus thermophilus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.84 | - |
NAD+ | 65°C | Thermus thermophilus | |
0.84 | - |
NAD+ | pH 10.5, 65°C | Thermus thermophilus | |
1.1 | - |
(S)-(-)-1-phenylethanol | 65°C | Thermus thermophilus | |
1.1 | - |
(S)-1-phenylethanol | pH 10.5, 65°C | Thermus thermophilus | |
1.6 | - |
4-methoxybenzyl alcohol | 65°C | Thermus thermophilus | |
1.6 | - |
4-methoxybenzyl alcohol | pH 10.5, 65°C | Thermus thermophilus | |
3.1 | - |
4-methoxybenzaldehyde | pH 6.0, 65°C | Thermus thermophilus | |
3.1 | - |
3-Methoxybenzaldehyde | 65°C | Thermus thermophilus | |
7.7 | - |
alpha-tetralone | 65°C | Thermus thermophilus | |
7.7 | - |
alpha-tetralone | pH 10.5, 65°C | Thermus thermophilus | |
8.3 | - |
1-Indanone | 65°C | Thermus thermophilus | |
8.3 | - |
1-Indanone | pH 10.5, 65°C | Thermus thermophilus | |
17.1 | - |
1-phenyl-1,2-propanedione | 65°C | Thermus thermophilus | |
17.1 | - |
1-phenyl-1,2-propandione | pH 10.5, 65°C | Thermus thermophilus | |
25.5 | - |
2,2,2-trifluoroacetophenone | 65°C | Thermus thermophilus | |
25.5 | - |
2,2,2-trifluoroacetophenone | pH 10.5, 65°C | Thermus thermophilus | |
38.1 | - |
methylbenzoylformate | 65°C | Thermus thermophilus | |
38.1 | - |
methyl oxo(phenyl)acetate | pH 10.5, 65°C | Thermus thermophilus | |
45.7 | - |
1-Indanol | 65°C | Thermus thermophilus | |
48.1 | - |
alpha-tetralol | 65°C | Thermus thermophilus | |
50.1 | - |
ethyl benzoylformate | 65°C | Thermus thermophilus | |
50.1 | - |
ethyl oxo(phenyl)acetate | pH 10.5, 65°C | Thermus thermophilus | |
52.4 | - |
NADH | 65°C | Thermus thermophilus | |
52.4 | - |
NADH | pH 10.5, 65°C | Thermus thermophilus | |
57 | - |
(S)-(+)-alpha-tetraol | 65°C | Thermus thermophilus | |
57 | - |
(S)-alpha-tetralol | pH 10.5, 65°C | Thermus thermophilus | |
61.4 | - |
(S)-(+)-1-indanol | 65°C | Thermus thermophilus | |
61.4 | - |
(S)-1-indanol | pH 10.5, 65°C | Thermus thermophilus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | 10 | substrate: (S)-(-)-1-phenylethanol and 3 mM NAD+ or methyl benzoylformate + NADH | Thermus thermophilus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Thermus thermophilus | |
NAD+ | strict requirement for NAD(H) as the coenzyme. Critical role of the D37 residue in discriminating NAD(H) from NADP(H) | Thermus thermophilus | |
NADH | - |
Thermus thermophilus | |
NADH | strict requirement for NAD(H) as the coenzyme. Critical role of the D37 residue in discriminating NAD(H) from NADP(H) | Thermus thermophilus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.026 | - |
4-methoxybenzyl alcohol | pH 10.5, 65°C | Thermus thermophilus | |
0.06 | - |
(S)-1-phenylethanol | pH 10.5, 65°C | Thermus thermophilus | |
0.3 | - |
1-Indanone | pH 10.5, 65°C | Thermus thermophilus | |
0.7 | - |
4-methoxybenzaldehyde | pH 6.0, 65°C | Thermus thermophilus | |
1.3 | - |
alpha-tetralone | pH 10.5, 65°C | Thermus thermophilus | |
2.3 | - |
2,2,2-trifluoroacetophenone | pH 10.5, 65°C | Thermus thermophilus | |
2.9 | - |
1-phenyl-1,2-propandione | pH 10.5, 65°C | Thermus thermophilus | |
3.5 | - |
NAD+ | pH 10.5, 65°C | Thermus thermophilus | |
13.6 | - |
(S)-alpha-tetralol | pH 10.5, 65°C | Thermus thermophilus | |
14.1 | - |
methyl oxo(phenyl)acetate | pH 10.5, 65°C | Thermus thermophilus | |
14.6 | - |
(S)-1-indanol | pH 10.5, 65°C | Thermus thermophilus | |
50.1 | - |
ethyl oxo(phenyl)acetate | pH 10.5, 65°C | Thermus thermophilus | |
1490 | - |
NADH | pH 10.5, 65°C | Thermus thermophilus |