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Literature summary for 1.1.1.1 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

Activating Compound Comment Organism Structure
EDTA 10 mM, 1.65fold activation Thermus thermophilus

Application

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(Commentary)

Cloned (Comment) Organism
overexpressed in Escherichia coli Thermus thermophilus

Inhibitors

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 [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

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 [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

Organism UniProt Comment Textmining
Thermus thermophilus
-
-
-
Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Thermus thermophilus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
151.1
-
-
Thermus thermophilus

Substrates and Products (Substrate)

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

Subunits Comment Organism
tetramer 4 * 26961, calculated from sequence Thermus thermophilus

Synonyms

Synonyms Comment Organism
NAD(H)-dependent alcohol dehydrogenase
-
Thermus thermophilus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
73
-
-
Thermus thermophilus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
73
-
inactivation above Thermus thermophilus
90
-
half-life 30 min Thermus thermophilus

Turnover Number [1/s]

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

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

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 [mM/s]

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