Activating Compound | Comment | Organism | Structure |
---|---|---|---|
o-phenanthroline | activates to 108% activity compared to the control activity at 0.1 mM and to 109% at 1 mM | Thermus thermophilus |
Application | Comment | Organism |
---|---|---|
synthesis | the enzyme is useful in production of chiral compounds for organic synthesis | Thermus thermophilus |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli strain BL21(DE3) | Thermus thermophilus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
1-butyl-3-methylimidazolium tetrafluoroborate | 65% inhibition at 2 mM | Thermus thermophilus | |
Ca2+ | 10% inhibition at 1 mM and 30% at 100 mM | Thermus thermophilus | |
Cu2+ | 23% inhibition at 1 mM | Thermus thermophilus | |
Hg2+ | 67% inhibition at 1 mM | Thermus thermophilus | |
iodoacetate | 5% inhibition at 1 mM | Thermus thermophilus | |
Mg2+ | 33% inhibition at 100 mM | Thermus thermophilus | |
additional information | no effects by 5 mM 2-mercaptoethanol | Thermus thermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics and substrate specificity, overview | Thermus thermophilus | |
4.2 | - |
(S)-1-indanol | pH 6.0, 65°C, recombinant enzyme, reduction reaction | Thermus thermophilus | |
5.1 | - |
rac-1-indanol | pH 6.0, 65°C, recombinant enzyme, reduction reaction | Thermus thermophilus | |
5.3 | - |
rac-alpha-tetralol | pH 6.0, 65°C, recombinant enzyme, reduction reaction | Thermus thermophilus | |
5.8 | - |
alpha-tetralone | pH 6.0, 65°C, recombinant enzyme, reduction reaction | Thermus thermophilus | |
27.6 | - |
1-Indanone | pH 6.0, 65°C, recombinant enzyme, reduction reaction | Thermus thermophilus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
EDTA | activates to 127% activity compared to the control activity at 1 mM and to 163% at 10 mM | Thermus thermophilus | |
K+ | activates to 167% activity compared to the control activity at 1 mM and to 193% at 100 mM | Thermus thermophilus | |
Li+ | activates to 127% activity compared to the control activity at 1 mM and to 178% at 100 mM | Thermus thermophilus | |
Na+ | activates to 179 at 100 mM | Thermus thermophilus | |
Zn2+ | activates to 114% activity compared to the control activity at 1 mM | Thermus thermophilus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
26000 | - |
4 * 26961, recombinant enzyme, mass spectrometry, 4 * 26000, recombinant enzyme, SDS-PAGE | Thermus thermophilus |
26961 | - |
4 * 26961, recombinant enzyme, mass spectrometry, 4 * 26000, recombinant enzyme, SDS-PAGE | Thermus thermophilus |
71000 | - |
about, recombinant enzyme, gel filtration in absence of NaCl | Thermus thermophilus |
105000 | - |
about, recombinant enzyme, gel filtration in presence of 0.15 M NaCl | Thermus thermophilus |
Organic Solvent | Comment | Organism |
---|---|---|
additional information | the enzyme shows a good tolerance to common organic solvents | Thermus thermophilus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermus thermophilus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme 11fold to homogeneity from Escherichia coli strain BL21(DE3) by heat denaturation of host proteins, anion exchange chromatography and gel filtration | Thermus thermophilus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
151.1 | - |
substrate: (S)-1-phenylethanol | Thermus thermophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-1-phenylethanol + NAD+ | 99% ethanoselective reaction | Thermus thermophilus | 1 phenylethanone + NADH + H+ | - |
r | |
(S)-alpha-tetralol + NAD+ | 99% ethanoselective reaction | Thermus thermophilus | alpha-tetralone + NADH + H+ | - |
r | |
1-indanone + NADH + H+ | - |
Thermus thermophilus | (S)-1-indanol + NAD+ | 99% enantiomeric excess after 6 h at 50°C | ? | |
2,2,2-trifluoroacetophenone + NADH + H+ | completely enantioselective reaction | Thermus thermophilus | ? | - |
r | |
acetophenone + NADH + H+ | completely enantioselective reaction | Thermus thermophilus | (S)-1-phenylethanol + NAD+ | 99% enantiomeric excess after 6 h at 50°C | r | |
alpha-tetralone + NADH + H+ | completely enantioselective reaction | Thermus thermophilus | (S)-alpha-tetralol + NAD+ | 99% enantiomeric excess after 6 h at 50°C | r | |
additional information | substrate specificity of ADHTt in the oxidation and the reduction reactions depends on the substrate, cf. (R)-specific secondary alcohol dehydrogenase, detailed overview. The enzyme shows a high reduction rate with halogenated aryl ketones, such as 2,2,2-trifluoroacetophenone, 2-chloroacetophenone, and 4-chlorobutyrophenone, and with aryl diketones, such as 1-phenyl-1,2-propanedione, although it is not active on benzil, i.e. diphenylethanedione. ADHTt proves to be very effective in reducing aryl alpha-keto esters, although it is not active on aliphatic alpha-keto esters and aryl beta-keto ester. Critical role of the D37 residue in discriminating NAD(H) from NADP(H) | Thermus thermophilus | ? | - |
? | |
rac-1-indanol + NAD+ | 3900% activity compared to (S)-1-phenylethanol | Thermus thermophilus | 1-indanone + NADH + H+ | - |
r | |
rac-alpha-tetralol + NAD+ | 2700% activity compared to (S)-1-phenylethanol | Thermus thermophilus | alpha-tetralone + NADH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
tetramer | 4 * 26961, recombinant enzyme, mass spectrometry, 4 * 26000, recombinant enzyme, SDS-PAGE | Thermus thermophilus |
Synonyms | Comment | Organism |
---|---|---|
ADHTt | - |
Thermus thermophilus |
More | the enzyme belongs to the short-chain dehydrogenase/reductase, SDR, superfamily | Thermus thermophilus |
short-chain NAD(H)-dependent alcohol dehydrogenase | - |
Thermus thermophilus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
oxidation reaction | Thermus thermophilus |
65 | - |
reduction reaction | Thermus thermophilus |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 73 | the enzyme displays activity at temperatures up to 73°C | Thermus thermophilus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
24 h, purified recombinant enzyme, 142% inactivation | Thermus thermophilus |
60 | - |
24 h, purified recombinant enzyme, 134% inactivation | Thermus thermophilus |
70 | - |
24 h, purified recombinant enzyme, 107% inactivation | Thermus thermophilus |
90 | - |
30 min, purified recombinant enzyme, 50% inactivation | Thermus thermophilus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
7.7 | - |
alpha-tetralone | pH 6.0, 65°C, recombinant enzyme, reduction reaction | Thermus thermophilus | |
8.3 | - |
1-Indanone | pH 6.0, 65°C, recombinant enzyme, reduction reaction | Thermus thermophilus | |
45.7 | - |
rac-1-indanol | pH 6.0, 65°C, recombinant enzyme, reduction reaction | Thermus thermophilus | |
48.1 | - |
rac-alpha-tetralol | pH 6.0, 65°C, recombinant enzyme, reduction reaction | Thermus thermophilus | |
61.4 | - |
(S)-1-indanol | pH 6.0, 65°C, recombinant enzyme, reduction reaction | Thermus thermophilus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
reduction reaction | Thermus thermophilus |
10 | - |
oxidation reaction | Thermus thermophilus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 7.7 | reduction reaction, high activity | Thermus thermophilus |
9 | 10 | oxidation reaction, high activity | Thermus thermophilus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | the enzyme shows no activity with NADP(H) | Thermus thermophilus | |
NAD+ | critical role of the D37 residue in discriminating NAD(H) from NADP(H) | Thermus thermophilus | |
NADH | critical role of the D37 residue in discriminating NAD(H) from NADP(H) | Thermus thermophilus |