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Literature summary extracted from

  • Pennacchio, A.; Pucci, B.; Secundo, F.; La Cara, F.; Rossi, M.; Raia, C.A.
    Purification and characterization of a novel recombinant highly enantioselective short-chain NAD(H)-dependent alcohol dehydrogenase from Thermus thermophilus (2008), Appl. Environ. Microbiol., 74, 3949-3958.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.1.1.1 EDTA 10 mM, 1.65fold activation Thermus thermophilus
1.1.1.B3 o-phenanthroline activates to 108% activity compared to the control activity at 0.1 mM and to 109% at 1 mM Thermus thermophilus
1.1.1.B4 o-phenanthroline activates to 108% activity compared to the control activity at 0.1 mM and to 109% at 1 mM Thermus thermophilus

Application

EC Number Application Comment Organism
1.1.1.1 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
1.1.1.B3 synthesis the enzyme is useful in production of chiral compounds for organic synthesis Thermus thermophilus
1.1.1.B4 synthesis the enzym is useful in production of chiral compounds for organic synthesis Thermus thermophilus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.1 overexpressed in Escherichia coli Thermus thermophilus
1.1.1.B3 expression in Escherichia coli strain BL21(DE3) Thermus thermophilus
1.1.1.B4 expression in Escherichia coli strain BL21(DE3) Thermus thermophilus

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.1 1-butyl-3-methylimidazolium tetrafluoroborate 2 mM, 35% of initial activity; 2 mM, 65% inhibition Thermus thermophilus
1.1.1.1 Ca2+ 100 mM, 70% of initial activity Thermus thermophilus
1.1.1.1 CaCl2 100 mM, 30% inhibition Thermus thermophilus
1.1.1.1 Cu2+ 100 mM, 76% of initial activity Thermus thermophilus
1.1.1.1 CuCl2 1 mM, 24% inhibition Thermus thermophilus
1.1.1.1 Hg2+ 100 mM, 33% of initial activity Thermus thermophilus
1.1.1.1 HgCl2 1 mM, 67% inhibition Thermus thermophilus
1.1.1.1 Mg2+ 100 mM, 67% of initial activity Thermus thermophilus
1.1.1.1 MgCl2 100 mM, 33% inhibition Thermus thermophilus
1.1.1.B3 1-butyl-3-methylimidazolium tetrafluoroborate 65% inhibition at 2 mM Thermus thermophilus
1.1.1.B3 Ca2+ 10% inhibition at 1 mM and 30% at 100 mM Thermus thermophilus
1.1.1.B3 Cu2+ 23% inhibition at 1 mM Thermus thermophilus
1.1.1.B3 Hg2+ 67% inhibition at 1 mM Thermus thermophilus
1.1.1.B3 iodoacetate 5% inhibition at 1 mM Thermus thermophilus
1.1.1.B3 Mg2+ 33% inhibition at 100 mM Thermus thermophilus
1.1.1.B3 additional information no effects by 5 mM 2-mercaptoethanol Thermus thermophilus
1.1.1.B4 1-butyl-3-methylimidazolium tetrafluoroborate 65% inhibition at 2 mM Thermus thermophilus
1.1.1.B4 Ca2+ 10% inhibition at 1 mM and 30% at 100 mM Thermus thermophilus
1.1.1.B4 Cu2+ 23% inhibition at 1 mM Thermus thermophilus
1.1.1.B4 Hg2+ 67% inhibition at 1 mM Thermus thermophilus
1.1.1.B4 iodoacetate 5% inhibition at 1 mM Thermus thermophilus
1.1.1.B4 Mg2+ 33% inhibition at 100 mM Thermus thermophilus
1.1.1.B4 additional information no effects by 5 mM 2-mercaptoethanol Thermus thermophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.1 0.0035
-
 NADH pH 10.5, 65°C Thermus thermophilus
1.1.1.1 0.035
-
 NADH 65°C Thermus thermophilus
1.1.1.1 0.24
-
 NAD+ 65°C Thermus thermophilus
1.1.1.1 0.24
-
 NAD+ pH 10.5, 65°C Thermus thermophilus
1.1.1.1 1
-
 ethyl benzoylformate 65°C Thermus thermophilus
1.1.1.1 1
-
 ethyl oxo(phenyl)acetate pH 10.5, 65°C Thermus thermophilus
1.1.1.1 2.7
-
 methylbenzoylformate 65°C Thermus thermophilus
1.1.1.1 2.7
-
 methyl oxo(phenyl)acetate pH 10.5, 65°C Thermus thermophilus
1.1.1.1 4.2
-
 (S)-(+)-1-indanol 65°C Thermus thermophilus
1.1.1.1 4.2
-
 (S)-(+)-alpha-tetraol 65°C Thermus thermophilus
1.1.1.1 4.2
-
 (S)-1-indanol pH 10.5, 65°C Thermus thermophilus
1.1.1.1 4.2
-
 (S)-alpha-tetralol pH 10.5, 65°C Thermus thermophilus
1.1.1.1 4.4
-
 3-Methoxybenzaldehyde 65°C Thermus thermophilus
1.1.1.1 4.4
-
 4-methoxybenzaldehyde pH 6.0, 65°C Thermus thermophilus
1.1.1.1 5.1
-
 1-Indanol 65°C Thermus thermophilus
1.1.1.1 5.3
-
 alpha-tetralol 65°C Thermus thermophilus
1.1.1.1 5.8
-
 alpha-tetralone 65°C Thermus thermophilus
1.1.1.1 5.8
-
 alpha-tetralone pH 10.5, 65°C Thermus thermophilus
1.1.1.1 5.9
-
 1-phenyl-1,2-propanedione 65°C Thermus thermophilus
1.1.1.1 5.9
-
 1-phenyl-1,2-propandione pH 10.5, 65°C Thermus thermophilus
1.1.1.1 11.2
-
 2,2,2-trifluoroacetophenone 65°C Thermus thermophilus
1.1.1.1 11.2
-
 2,2,2-trifluoroacetophenone pH 10.5, 65°C Thermus thermophilus
1.1.1.1 18.1
-
 (S)-(-)-1-phenylethanol 65°C Thermus thermophilus
1.1.1.1 18.1
-
 (S)-1-phenylethanol pH 10.5, 65°C Thermus thermophilus
1.1.1.1 27.6
-
 1-Indanone 65°C Thermus thermophilus
1.1.1.1 27.6
-
 1-Indanone pH 10.5, 65°C Thermus thermophilus
1.1.1.1 61
-
 4-methoxybenzyl alcohol 65°C Thermus thermophilus
1.1.1.1 61
-
 4-methoxybenzyl alcohol pH 10.5, 65°C Thermus thermophilus
1.1.1.B3 additional information
-
 additional information kinetics and substrate specificity, overview Thermus thermophilus
1.1.1.B3 4.2
-
 (S)-1-indanol pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus
1.1.1.B3 5.1
-
 rac-1-indanol pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus
1.1.1.B3 5.3
-
 rac-alpha-tetralol pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus
1.1.1.B3 5.8
-
 alpha-tetralone pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus
1.1.1.B3 27.6
-
 1-Indanone pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus
1.1.1.B4 additional information
-
 additional information kinetics and substrate specificity, overview Thermus thermophilus
1.1.1.B4 0.035
-
 NADH pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus
1.1.1.B4 0.24
-
 NAD+ pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus
1.1.1.B4 1
-
 ethylbenzoylformate pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus
1.1.1.B4 11.2
-
 2,2,2-trifluoroacetophenone pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.1 K+ 100 mM KCl, 1.9fold activation Thermus thermophilus
1.1.1.1 K+ 100 mM, 193% of initial activity Thermus thermophilus
1.1.1.1 Li+ 100 mM LiCl, 1.8fold activation Thermus thermophilus
1.1.1.1 Li+ 100 mM, 178% of initial activity Thermus thermophilus
1.1.1.1 Na+ 100 mM NaCl, 1.8fold activation Thermus thermophilus
1.1.1.1 Na+ 100 mM, 179% of initial activity Thermus thermophilus
1.1.1.1 Zn2+ 1 mM ZnSO4, 1.14fold activation Thermus thermophilus
1.1.1.1 Zn2+ 1 mM, 114% of initial activity Thermus thermophilus
1.1.1.B3 EDTA activates to 127% activity compared to the control activity at 1 mM and to 163% at 10 mM Thermus thermophilus
1.1.1.B3 K+ activates to 167% activity compared to the control activity at 1 mM and to 193% at 100 mM Thermus thermophilus
1.1.1.B3 Li+ activates to 127% activity compared to the control activity at 1 mM and to 178% at 100 mM Thermus thermophilus
1.1.1.B3 Na+ activates to 179 at 100 mM Thermus thermophilus
1.1.1.B3 Zn2+ activates to 114% activity compared to the control activity at 1 mM Thermus thermophilus
1.1.1.B4 EDTA activates to 127% activity compared to the control activity at 1 mM and to 163% at 10 mM Thermus thermophilus
1.1.1.B4 K+ activates to 167% activity compared to the control activity at 1 mM and to 193% at 100 mM Thermus thermophilus
1.1.1.B4 Li+ activates to 127% activity compared to the control activity at 1 mM and to 178% at 100 mM Thermus thermophilus
1.1.1.B4 Na+ activates to 179 at 100 mM Thermus thermophilus
1.1.1.B4 Zn2+ activates to 114% activity compared to the control activity at 1 mM Thermus thermophilus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.1 26961
-
 4 * 26961, calculated from sequence Thermus thermophilus
1.1.1.1 105000
-
 buffer containing 25 mM NaCl, gel filtration Thermus thermophilus
1.1.1.B3 26000
-
 4 * 26961, recombinant enzyme, mass spectrometry, 4 * 26000, recombinant enzyme, SDS-PAGE Thermus thermophilus
1.1.1.B3 26961
-
 4 * 26961, recombinant enzyme, mass spectrometry, 4 * 26000, recombinant enzyme, SDS-PAGE Thermus thermophilus
1.1.1.B3 71000
-
 about, recombinant enzyme, gel filtration in absence of NaCl Thermus thermophilus
1.1.1.B3 105000
-
 about, recombinant enzyme, gel filtration in presence of 0.15 M NaCl Thermus thermophilus
1.1.1.B4 26000
-
 4 * 26961, recombinant enzyme, mass spectrometry, 4 * 26000, recombinant enzyme, SDS-PAGE Thermus thermophilus
1.1.1.B4 26961
-
 4 * 26961, recombinant enzyme, mass spectrometry, 4 * 26000, recombinant enzyme, SDS-PAGE Thermus thermophilus
1.1.1.B4 71000
-
 about, recombinant enzyme, gel filtration in absence of NaCl Thermus thermophilus
1.1.1.B4 105000
-
 about, recombinant enzyme, gel filtration in presence of 0.15 M NaCl Thermus thermophilus

Organic Solvent Stability

EC Number Organic Solvent Comment Organism
1.1.1.B3 additional information the enzyme shows a good tolerance to common organic solvents Thermus thermophilus
1.1.1.B4 additional information the enzyme shows a good tolerance to common organic solvents Thermus thermophilus

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.1 Thermus thermophilus
-
-
-
1.1.1.1 Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039
-
-
-
1.1.1.B3 Thermus thermophilus
-
-
-
1.1.1.B4 Thermus thermophilus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.1
-
 Thermus thermophilus
1.1.1.B3 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
1.1.1.B4 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 [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.1 151.1
-
-
 Thermus thermophilus
1.1.1.B3 151.1
-
 substrate: (S)-1-phenylethanol Thermus thermophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.1 (S)-(+)-1-indanol + NAD+
-
 Thermus thermophilus indanone + NADH + H+
-
 ?
1.1.1.1 (S)-1-indanol + NAD+ 3900% of the activity with (S)-1-phenylethanol Thermus thermophilus 1-indanone + NADH + H+
-
 r
1.1.1.1 (S)-1-phenylethanol + NAD+
-
 Thermus thermophilus 1-phenylethanone + NADH + H+
-
 r
1.1.1.1 (S)-alpha-tetralol + NAD+ 2700% of the activity with (S)-1-phenylethanol Thermus thermophilus alpha-tetralone + NADH + H+
-
 r
1.1.1.1 1-(4'-chlorophenyl)ethanol + NAD+ 26% of the activity with (S)-(-)-1-phenylethanol Thermus thermophilus 1-(4'-chlorophenyl)ethanone + NADH + H+
-
 ?
1.1.1.1 1-(4'-fluorophenyl)ethanol + NAD+ 45% of the activity with (S)-1-phenylethanol Thermus thermophilus 1-(4'-fluorophenyl)ethanone + NADH + H+
-
 r
1.1.1.1 1-(4-fluorophenyl)ethanol + NAD+ 45% of the activity with (S)-(-)-1-phenylethanol Thermus thermophilus 1-(4-fluorophenyl)ethanone + NADH + H+
-
 ?
1.1.1.1 1-indanol + NAD+
-
 Thermus thermophilus 1-indanone + NADH + H+
-
 ?
1.1.1.1 1-indanone + NADH + H+
-
 Thermus thermophilus 1-indanol + NAD+
-
 r
1.1.1.1 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
1.1.1.1 1-phenyl-1,2-propanedione + NADH + H+ activity is 1.46fold higher than with 2,2,2-trifluoroacetophenone Thermus thermophilus ?
-
 ?
1.1.1.1 1-phenyl-1-propanol + NAD+ 59% of the activity with (S)-(-)-1-phenylethanol Thermus thermophilus 1-phenyl-1-propanone + NADH + H+
-
 ?
1.1.1.1 1-phenyl-1-propanol + NAD+ 59% of the activity with (S)-1-phenylethanol Thermus thermophilus 1-phenylpropan-1-one + NADH + H+
-
 r
1.1.1.1 1-phenyl-2-propanol + NAD+ 16% of the activity with (S)-(-)-1-phenylethanol Thermus thermophilus 1-phenyl-2-propanone + NADH + H+
-
 ?
1.1.1.1 2,2,2-trifluoroacetophenone + NADH + H+
-
 Thermus thermophilus (R)-alpha-(trifluoromethyl)benzyl alcohol + NAD+ 93% enantiomeric excess r
1.1.1.1 2,2,2-trifluoroacetophenone + NADH + H+ 93% enantiomeric excess Thermus thermophilus (R)-alpha-(trifluoromethyl)benzyl alcohol + NAD+
-
 ?
1.1.1.1 2,2,2-trifluoroacetophenone + NADH + H+
-
 Thermus thermophilus 2,2,2-trifluoro-1-phenylethanol + NAD+
-
 r
1.1.1.1 2,2-dichloroacetophenone + NADH + H+ 32% of the activity with 2,2,2-trifluoroacetophenone Thermus thermophilus 2,2-dichloro-1-phenylethanol + NAD+
-
 ?
1.1.1.1 2-methoxybenzaldehyde + NADH + H+ 13% of the activity with 2,2,2-trifluoroacetophenone Thermus thermophilus 2-methoxybenzylalcohol + NAD+
-
 ?
1.1.1.1 2-methoxybenzyl alcohol + NAD+ 25% of the activity with (S)-(-)-1-phenylethanol Thermus thermophilus 2-methoxybenzaldehyde + NADH + H+
-
 ?
1.1.1.1 2-propanol + NAD+
-
 Thermus thermophilus acetone + NADH + H+
-
 ?
1.1.1.1 2-propanol + NAD+
-
 Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 acetone + NADH + H+
-
 ?
1.1.1.1 3-methoxybenzaldehyde + NADH + H+ 14% of the activity with 2,2,2-trifluoroacetophenone Thermus thermophilus 3-methoxybenzylalcohol + NAD+
-
 ?
1.1.1.1 3-methoxybenzaldehyde + NADH + H+
-
 Thermus thermophilus 3-methoxybenzyl alcohol + NAD+
-
 r
1.1.1.1 3-methoxybenzyl alcohol + NAD+ 13% of the activity with (S)-(-)-1-phenylethanol Thermus thermophilus 3-methoxybenzaldehyde + NADH + H+
-
 ?
1.1.1.1 3-methoxybenzyl alcohol + NAD+ 13% of the activity with (S)-(-)-1-phenylethanol Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 3-methoxybenzaldehyde + NADH + H+
-
 ?
1.1.1.1 4'-chlorobutyrophenone + NADH + H+ 10% of the activity with 2,2,2-trifluoroacetophenone Thermus thermophilus 1-(4-chlorophenyl)butan-1-ol + NAD+
-
 ?
1.1.1.1 4-methoxybenzaldehyde + NADH + H+ 13% of the activity with 2,2,2-trifluoroacetophenone Thermus thermophilus 4-methoxybenzylalcohol + NAD+
-
 ?
1.1.1.1 4-methoxybenzyl alcohol + NAD+ 99% of the activity with (S)-(-)-1-phenylethanol Thermus thermophilus 4-methoxybenzaldehyde + NADH + H+
-
 ?
1.1.1.1 4-methoxybenzyl alcohol + NAD+ 99% of the activity with (S)-1-phenylethanol Thermus thermophilus 4-methoxybenzaldehyde + NADH + H+
-
 r
1.1.1.1 acetophenone + NADH + H+ more than 99% enantiomeric excess Thermus thermophilus (S)-(-)-1-phenylethanol + NAD+
-
 r
1.1.1.1 acetophenone + NADH + H+
-
 Thermus thermophilus (S)-1-phenylethanol + NAD+ 99% enantiomeric excess r
1.1.1.1 alpha-ethyl benzoylformate + NADH + H+
-
 Thermus thermophilus ethyl (R)-(-)-mandelate + NAD+ 95% enantiomeric excess r
1.1.1.1 alpha-methyl benzoylformate + NADH + H+
-
 Thermus thermophilus methyl (R)-mandelate + NAD+ 92% enantiomeric excess r
1.1.1.1 alpha-tetralone + NADH + H+
-
 Thermus thermophilus (S)-alpha-tetralol + NAD+ 99% enantiomeric excess r
1.1.1.1 alpha-tetralone + NADH + H+ more than 99% enantiomeric excess Thermus thermophilus (S)-alpha-tetralol + NAD+
-
 ?
1.1.1.1 alpha-tetralone + NADH + H+
-
 Thermus thermophilus alpha-tetralol + NAD+
-
 r
1.1.1.1 benzaldehyde + NADH + H+ 14% of the activity with 2,2,2-trifluoroacetophenone Thermus thermophilus benzyl alcohol + NAD+
-
 ?
1.1.1.1 benzaldehyde + NADH + H+ 14% of the activity with 2,2,2-trifluoroacetophenone Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 benzyl alcohol + NAD+
-
 ?
1.1.1.1 cyclohexanol + NAD+ 13% of the activity with (S)-(-)-1-phenylethanol Thermus thermophilus cyclohexanone + NADH + H+
-
 ?
1.1.1.1 cyclohexanol + NAD+ 13% of the activity with (S)-(-)-1-phenylethanol Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 cyclohexanone + NADH + H+
-
 ?
1.1.1.1 ethyl benzoylformate + NADH + H+ 95% enantiomeric excess Thermus thermophilus ethyl (R)-(-)-mandelate + NAD+
-
 ?
1.1.1.1 ethyl oxo(phenyl)acetate + NADH + H+ 100% of the activity with 2,2,2-trifluoroacetophenone Thermus thermophilus ethyl hydroxy(phenyl)acetate + NAD+
-
 r
1.1.1.1 methyl benzoylformate + NADH + H+ 92% enantiomeric excess Thermus thermophilus methyl (R)-(-)-mandelate + NAD+
-
 ?
1.1.1.1 methyl oxo(phenyl)acetate + NADH + H+ 57% of the activity with 2,2,2-trifluoroacetophenone Thermus thermophilus methyl hydroxy(phenyl)acetate + NAD+
-
 r
1.1.1.1 methylbenzoylformate + NADH + H+ 13% of the activity with 2,2,2-trifluoroacetophenone Thermus thermophilus ?
-
 ?
1.1.1.1 additional information preference for reduction of aromatic ketones and alpha-keto esters, and poor activity on aromatic alcohols and aldehydes Thermus thermophilus ?
-
 ?
1.1.1.1 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 ?
-
 ?
1.1.1.1 trans-cinnamyl alcohol + NAD+ activity is 3.9fold higher than with (S)-(-)-1-phenylethanol Thermus thermophilus (2E)-3-phenylprop-2-enal + NADH + H+
-
 ?
1.1.1.B3 (S)-1-phenylethanol + NAD+ 99% ethanoselective reaction Thermus thermophilus 1 phenylethanone + NADH + H+
-
 r
1.1.1.B3 (S)-alpha-tetralol + NAD+ 99% ethanoselective reaction Thermus thermophilus alpha-tetralone + NADH + H+
-
 r
1.1.1.B3 1-indanone + NADH + H+
-
 Thermus thermophilus (S)-1-indanol + NAD+ 99% enantiomeric excess after 6 h at 50°C ?
1.1.1.B3 2,2,2-trifluoroacetophenone + NADH + H+ completely enantioselective reaction Thermus thermophilus ?
-
 r
1.1.1.B3 acetophenone + NADH + H+ completely enantioselective reaction Thermus thermophilus (S)-1-phenylethanol + NAD+ 99% enantiomeric excess after 6 h at 50°C r
1.1.1.B3 alpha-tetralone + NADH + H+ completely enantioselective reaction Thermus thermophilus (S)-alpha-tetralol + NAD+ 99% enantiomeric excess after 6 h at 50°C r
1.1.1.B3 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 ?
-
 ?
1.1.1.B3 rac-1-indanol + NAD+ 3900% activity compared to (S)-1-phenylethanol Thermus thermophilus 1-indanone + NADH + H+
-
 r
1.1.1.B3 rac-alpha-tetralol + NAD+ 2700% activity compared to (S)-1-phenylethanol Thermus thermophilus alpha-tetralone + NADH + H+
-
 r
1.1.1.B4 (R)-(trifluoromethyl)benzyl alcohol + NADH + H+ 93% ethanoselective reaction Thermus thermophilus ?
-
 r
1.1.1.B4 2,2,2-trifluoroacetophenone + NADH + H+ completely enantioselective reaction Thermus thermophilus (1R)-2,2,2-trifluoro-1-phenylethanol + NAD+
-
 r
1.1.1.B4 ethyl (R)-mandelate + NADH + H+ 95% ethanoselective reaction Thermus thermophilus ethyl benzoylformate + NAD+
-
 r
1.1.1.B4 ethyl benzoylformate + NADH + H+ completely enantioselective reaction Thermus thermophilus ethyl (R)-mandelate + NAD+ 95% enantiomeric excess after 6 h at 50°C r
1.1.1.B4 methyl (R)-mandelate + NADH + H+ 92% ethanoselective reaction Thermus thermophilus methyl benzoylformate + NAD+
-
 r
1.1.1.B4 methyl benzoylformate + NADH + H+ completely enantioselective reaction Thermus thermophilus methyl (R)-mandelate + NAD+ 92% enantiomeric excess after 6 h at 50°C r
1.1.1.B4 additional information substrate specificity of ADHTt in the oxidation and the reduction reactions depends on the substrate, cf. (S)-1-phenylethanol 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 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.1 tetramer 4 * 26961, calculated from sequence Thermus thermophilus
1.1.1.B3 tetramer 4 * 26961, recombinant enzyme, mass spectrometry, 4 * 26000, recombinant enzyme, SDS-PAGE Thermus thermophilus
1.1.1.B4 tetramer 4 * 26961, recombinant enzyme, mass spectrometry, 4 * 26000, recombinant enzyme, SDS-PAGE Thermus thermophilus

Synonyms

EC Number Synonyms Comment Organism
1.1.1.1 NAD(H)-dependent alcohol dehydrogenase
-
 Thermus thermophilus
1.1.1.B3 ADHTt
-
 Thermus thermophilus
1.1.1.B3 More the enzyme belongs to the short-chain dehydrogenase/reductase, SDR, superfamily Thermus thermophilus
1.1.1.B3 short-chain NAD(H)-dependent alcohol dehydrogenase
-
 Thermus thermophilus
1.1.1.B4 ADHTt
-
 Thermus thermophilus
1.1.1.B4 More the enzyme belongs to the short-chain dehydrogenase/reductase, SDR, superfamily Thermus thermophilus
1.1.1.B4 short-chain NAD(H)-dependent alcohol dehydrogenase
-
 Thermus thermophilus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.1 73
-
-
 Thermus thermophilus
1.1.1.B3 60
-
 oxidation reaction Thermus thermophilus
1.1.1.B3 65
-
 reduction reaction Thermus thermophilus
1.1.1.B4 60
-
 oxidation reaction Thermus thermophilus
1.1.1.B4 65
-
 reduction reaction Thermus thermophilus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.1.1.B3 25 73 the enzyme displays activity at temperatures up to 73°C Thermus thermophilus
1.1.1.B4 25 73 the enzyme displays activity at temperatures up to 73°C Thermus thermophilus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.1 73
-
 inactivation above Thermus thermophilus
1.1.1.1 90
-
 half-life 30 min Thermus thermophilus
1.1.1.B3 50
-
 24 h, purified recombinant enzyme, 142% inactivation Thermus thermophilus
1.1.1.B3 60
-
 24 h, purified recombinant enzyme, 134% inactivation Thermus thermophilus
1.1.1.B3 70
-
 24 h, purified recombinant enzyme, 107% inactivation Thermus thermophilus
1.1.1.B3 90
-
 30 min, purified recombinant enzyme, 50% inactivation Thermus thermophilus
1.1.1.B4 50
-
 24 h, purified recombinant enzyme, 142% inactivation Thermus thermophilus
1.1.1.B4 60
-
 24 h, purified recombinant enzyme, 134% inactivation Thermus thermophilus
1.1.1.B4 70
-
 24 h, purified recombinant enzyme, 107% inactivation Thermus thermophilus
1.1.1.B4 90
-
 30 min, purified recombinant enzyme, 50% inactivation Thermus thermophilus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.1 0.84
-
 NAD+ 65°C Thermus thermophilus
1.1.1.1 0.84
-
 NAD+ pH 10.5, 65°C Thermus thermophilus
1.1.1.1 1.1
-
 (S)-(-)-1-phenylethanol 65°C Thermus thermophilus
1.1.1.1 1.1
-
 (S)-1-phenylethanol pH 10.5, 65°C Thermus thermophilus
1.1.1.1 1.6
-
 4-methoxybenzyl alcohol 65°C Thermus thermophilus
1.1.1.1 1.6
-
 4-methoxybenzyl alcohol pH 10.5, 65°C Thermus thermophilus
1.1.1.1 3.1
-
 4-methoxybenzaldehyde pH 6.0, 65°C Thermus thermophilus
1.1.1.1 3.1
-
 3-Methoxybenzaldehyde 65°C Thermus thermophilus
1.1.1.1 7.7
-
 alpha-tetralone 65°C Thermus thermophilus
1.1.1.1 7.7
-
 alpha-tetralone pH 10.5, 65°C Thermus thermophilus
1.1.1.1 8.3
-
 1-Indanone 65°C Thermus thermophilus
1.1.1.1 8.3
-
 1-Indanone pH 10.5, 65°C Thermus thermophilus
1.1.1.1 17.1
-
 1-phenyl-1,2-propanedione 65°C Thermus thermophilus
1.1.1.1 17.1
-
 1-phenyl-1,2-propandione pH 10.5, 65°C Thermus thermophilus
1.1.1.1 25.5
-
 2,2,2-trifluoroacetophenone 65°C Thermus thermophilus
1.1.1.1 25.5
-
 2,2,2-trifluoroacetophenone pH 10.5, 65°C Thermus thermophilus
1.1.1.1 38.1
-
 methylbenzoylformate 65°C Thermus thermophilus
1.1.1.1 38.1
-
 methyl oxo(phenyl)acetate pH 10.5, 65°C Thermus thermophilus
1.1.1.1 45.7
-
 1-Indanol 65°C Thermus thermophilus
1.1.1.1 48.1
-
 alpha-tetralol 65°C Thermus thermophilus
1.1.1.1 50.1
-
 ethyl benzoylformate 65°C Thermus thermophilus
1.1.1.1 50.1
-
 ethyl oxo(phenyl)acetate pH 10.5, 65°C Thermus thermophilus
1.1.1.1 52.4
-
 NADH 65°C Thermus thermophilus
1.1.1.1 52.4
-
 NADH pH 10.5, 65°C Thermus thermophilus
1.1.1.1 57
-
 (S)-(+)-alpha-tetraol 65°C Thermus thermophilus
1.1.1.1 57
-
 (S)-alpha-tetralol pH 10.5, 65°C Thermus thermophilus
1.1.1.1 61.4
-
 (S)-(+)-1-indanol 65°C Thermus thermophilus
1.1.1.1 61.4
-
 (S)-1-indanol pH 10.5, 65°C Thermus thermophilus
1.1.1.B3 7.7
-
 alpha-tetralone pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus
1.1.1.B3 8.3
-
 1-Indanone pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus
1.1.1.B3 45.7
-
 rac-1-indanol pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus
1.1.1.B3 48.1
-
 rac-alpha-tetralol pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus
1.1.1.B3 61.4
-
 (S)-1-indanol pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus
1.1.1.B4 0.84
-
 NAD+ pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus
1.1.1.B4 25.5
-
 2,2,2-trifluoroacetophenone pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus
1.1.1.B4 50.1
-
 ethylbenzoylformate pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus
1.1.1.B4 52.4
-
 NADH pH 6.0, 65°C, recombinant enzyme, reduction reaction Thermus thermophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.1 9 10 substrate: (S)-(-)-1-phenylethanol and 3 mM NAD+ or methyl benzoylformate + NADH Thermus thermophilus
1.1.1.B3 6
-
 reduction reaction Thermus thermophilus
1.1.1.B3 10
-
 oxidation reaction Thermus thermophilus
1.1.1.B4 6
-
 reduction reaction Thermus thermophilus
1.1.1.B4 10
-
 oxidation reaction Thermus thermophilus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.B3 6 7.7 reduction reaction, high activity Thermus thermophilus
1.1.1.B3 9 10 oxidation reaction, high activity Thermus thermophilus
1.1.1.B4 6 7.7 reduction reaction, high activity Thermus thermophilus
1.1.1.B4 9 10 oxidation reaction, high activity Thermus thermophilus

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.1 NAD+
-
 Thermus thermophilus
1.1.1.1 NAD+ strict requirement for NAD(H) as the coenzyme. Critical role of the D37 residue in discriminating NAD(H) from NADP(H) Thermus thermophilus
1.1.1.1 NADH
-
 Thermus thermophilus
1.1.1.1 NADH strict requirement for NAD(H) as the coenzyme. Critical role of the D37 residue in discriminating NAD(H) from NADP(H) Thermus thermophilus
1.1.1.B3 additional information the enzyme shows no activity with NADP(H) Thermus thermophilus
1.1.1.B3 NAD+ critical role of the D37 residue in discriminating NAD(H) from NADP(H) Thermus thermophilus
1.1.1.B3 NADH critical role of the D37 residue in discriminating NAD(H) from NADP(H) Thermus thermophilus
1.1.1.B4 additional information the enzyme shows no activity with NADP(H) Thermus thermophilus
1.1.1.B4 NAD+ critical role of the D37 residue in discriminating NAD(H) from NADP(H) Thermus thermophilus
1.1.1.B4 NADH critical role of the D37 residue in discriminating NAD(H) from NADP(H) Thermus thermophilus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.1 0.026
-
 4-methoxybenzyl alcohol pH 10.5, 65°C Thermus thermophilus
1.1.1.1 0.06
-
 (S)-1-phenylethanol pH 10.5, 65°C Thermus thermophilus
1.1.1.1 0.3
-
 1-Indanone pH 10.5, 65°C Thermus thermophilus
1.1.1.1 0.7
-
 4-methoxybenzaldehyde pH 6.0, 65°C Thermus thermophilus
1.1.1.1 1.3
-
 alpha-tetralone pH 10.5, 65°C Thermus thermophilus
1.1.1.1 2.3
-
 2,2,2-trifluoroacetophenone pH 10.5, 65°C Thermus thermophilus
1.1.1.1 2.9
-
 1-phenyl-1,2-propandione pH 10.5, 65°C Thermus thermophilus
1.1.1.1 3.5
-
 NAD+ pH 10.5, 65°C Thermus thermophilus
1.1.1.1 13.6
-
 (S)-alpha-tetralol pH 10.5, 65°C Thermus thermophilus
1.1.1.1 14.1
-
 methyl oxo(phenyl)acetate pH 10.5, 65°C Thermus thermophilus
1.1.1.1 14.6
-
 (S)-1-indanol pH 10.5, 65°C Thermus thermophilus
1.1.1.1 50.1
-
 ethyl oxo(phenyl)acetate pH 10.5, 65°C Thermus thermophilus
1.1.1.1 1490
-
 NADH pH 10.5, 65°C Thermus thermophilus