Information on EC 1.1.1.B4 - (R)-specific secondary alcohol dehydrogenase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea

EC NUMBER
COMMENTARY hide
1.1.1.B4
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
(R)-specific secondary alcohol dehydrogenase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(R)-R-CHOH-R' + NAD+ = R-CO-R' + NADH + H+
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
non-pathway related
-
-
SYSTEMATIC NAME
IUBMB Comments
secondary alcohol:NAD+ oxidoreductase (specific for (R)-configuration of alcohol)
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain AIU 652, growth on cholesterol or propanol as sole carbon source
-
-
Manually annotated by BRENDA team
strain AIU 652, growth on cholesterol or propanol as sole carbon source
-
-
Manually annotated by BRENDA team
strain PED, growth on 1-phenyl-1,2-ethanediol as sole carbon source
-
-
Manually annotated by BRENDA team
strain PED, growth on 1-phenyl-1,2-ethanediol as sole carbon source
-
-
Manually annotated by BRENDA team
strain Y379-50
-
-
Manually annotated by BRENDA team
strain Y379-50
-
-
Manually annotated by BRENDA team
gene adhB, mutant I86A
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
the mutation I86A allows large substituents to fit into the large pocket of I86ATeSADH, which corresponds to the small pocket in wild-type TeSADH, modeling of the stereopreference of TeSADH I86A
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(R)-(trifluoromethyl)benzyl alcohol + NADH + H+
?
show the reaction diagram
-
93% ethanoselective reaction
-
-
r
(R)-1-phenylethanol + NAD+
acetophenone + NADH + H+
show the reaction diagram
(R)-2-butanol + NAD+
2-butanone + NADH + H+
show the reaction diagram
(R)-2-pentanol + NAD+
2-pentanone + NADH + H+
show the reaction diagram
-
-
-
-
r
1,2-butanediol + NAD+
? + NADH
show the reaction diagram
-
20% of the activity with 2-propanol
-
-
r
1,2-propanediol + NAD+
? + NADH
show the reaction diagram
-
45% of the activity with 2-propanol
-
-
r
1,3-butanediol + NAD+
? + NADH
show the reaction diagram
-
126% of the activity with 2-propanol
-
-
r
1-(2-phenylcyclopropyl)ethanone + NADH
(1R)-1-(2-phenylcyclopropyl)ethanol + NAD+
show the reaction diagram
1-butanol + NAD+
butanal + NADH
show the reaction diagram
1-chloropentane-2,4-dione + NADH
(4R)-5-chloro-4-hydroxypentan-2-one + NAD+
show the reaction diagram
1-phenoxypropan-2-one + NADH
(2R)-1-phenoxypropan-2-ol + NAD+
show the reaction diagram
1-phenylethanone + NADH
(1R)-1-phenylethanol + NAD+
show the reaction diagram
1-phenylpropane-1,2-dione + NADH
(1S)-1-hydroxy-1-phenylpropan-2-one + NAD+
show the reaction diagram
1-phenylpropane-1,2-dione + NADH
? + NAD+
show the reaction diagram
-
-
-
-
r
1-propanol + NAD+
propanal + NADH
show the reaction diagram
-
9% of the activity with 2-propanol
-
-
r
2,2,2-trifluoro-1-phenylethanone + NADH
(1R)-2,2,2-trifluoro-1-phenylethanol + NAD+
show the reaction diagram
-
-
21% of the activity with 1-phenylpropane-1,2-dione
-
r
2,2,2-trifluoro-1-phenylethanone + NADH
(1S)-2,2,2-trifluoro-1-phenylethanol + NAD+
show the reaction diagram
-
-
37% yield, 92% enantiomeric excess
-
?
2,2,2-trifluoroacetophenone + NADH + H+
(1R)-2,2,2-trifluoro-1-phenylethanol + NAD+
show the reaction diagram
-
completely enantioselective reaction
-
-
r
2,3-butanediol + NAD+
? + NADH
show the reaction diagram
2-butanol + NAD+
2-butanone + NADH
show the reaction diagram
-
(R)-isomer is preferred over (S)-isomer
-
-
r
2-butanone + NAD+
2-butanol + NADH + H+
show the reaction diagram
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274% of the activity with beta-hydroxyacetophenone
-
-
?
2-butanone + NADH
(S)-2-hydroxybutane + NAD+
show the reaction diagram
2-butanone + NADH
2-butanol + NAD+
show the reaction diagram
-
83% of the activity with acetone
-
-
r
2-heptanone + NAD+
2-heptanol + NADH + H+
show the reaction diagram
-
281% of the activity with beta-hydroxyacetophenone
-
-
?
2-hexanone + NAD+
2-hexanol + NADH + H+
show the reaction diagram
-
259% of the activity with beta-hydroxyacetophenone
-
-
?
2-octanone + NAD+
2-octanol + NADH + H+
show the reaction diagram
-
350% of the activity with beta-hydroxyacetophenone
-
-
?
2-pentanol + NAD+
2-pentanone + NADH
show the reaction diagram
2-pentanone + NAD+
2-pentanol + NADH + H+
show the reaction diagram
-
249% of the activity with beta-hydroxyacetophenone
-
-
?
2-pentanone + NADH
(S)-2-hydroxypentane + NAD+
show the reaction diagram
-
6% of the activity with 2-butanone
-
-
r
2-pentanone + NADH
2-pentanol + NAD+
show the reaction diagram
-
44% of the activity with acetone
-
-
r
2-propanol + NAD+
acetone + NADH
show the reaction diagram
2-propanol + NAD+ + H+
acetone + NADH
show the reaction diagram
-
-
-
-
r
3-(dimethylamino)-1-phenylpropan-1-one + NADH
(1S)-3-dimethylamino-1-phenylpropan-1-ol + NAD+
show the reaction diagram
-
-
24% of the activity with 1-phenylpropane-1,2-dione
-
r
3-chloro-pentane-2,4-dione + NADH
(4R)-3-chloro-4-hydroxypentan-2-one + NAD+
show the reaction diagram
-
336% of the activity with 2-butanone
-
-
r
3-hexanone + NAD+
3-hexanol + NADH + H+
show the reaction diagram
-
29% of the activity with beta-hydroxyacetophenone
-
-
?
3-pentanol + NAD+
3-pentanone + NADH
show the reaction diagram
-
23% of the activity with 2-propanol
-
-
r
3-pentanone + NAD+
3-pentanol + NADH + H+
show the reaction diagram
-
35% of the activity with beta-hydroxyacetophenone
-
-
?
3-pentanone + NADH
3-pentanol + NAD+
show the reaction diagram
-
16% of the activity with acetone
-
-
r
4-hydroxy-2-butanone + NADH
? + NAD+
show the reaction diagram
-
95% of the activity with acetone
-
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r
4-methylpentan-2-one + NADH + H+
4-methylpentan-2-ol + NAD+
show the reaction diagram
4-phenylbutan-2-one + NADH
(2R)-4-phenylbutan-2-ol + NAD+
show the reaction diagram
-
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18% of the activity with 1-phenylpropane-1,2-dione
-
r
5-chloropentan-2-one + NADH
(2S)-5-chloropentan-2-ol + NAD+
show the reaction diagram
-
-
48% yield, 93% enantiomeric excess
-
?
6-methylhept-5-en-2-one + NADH
(2S)-6-methylhept-5-en-2-ol + NAD+
show the reaction diagram
-
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51% yield, 97% enantiomeric excess
-
?
acetone + NAD+
isopropanol + NADH + H+
show the reaction diagram
-
260% of the activity with beta-hydroxyacetophenone
-
-
?
acetone + NADH
2-propanol + NAD+
show the reaction diagram
-
-
-
-
r
acetophenone + NADH + H+
phenylethanol + NAD+
show the reaction diagram
acetophenone + NADPH + H+
phenylethanol + NADP+
show the reaction diagram
-
-
-
-
?
beta-hydroxyacetophenone + NAD+
(R)-1-phenyl-1,2-ethanediol + NADH + H+
show the reaction diagram
-
-
formation of (R)-enantiomner with 100% enantiomeric excess
-
?
cycloheptanone + NADH + H+
cycloheptanol + NAD+
show the reaction diagram
ethanol + NAD+
ethanal + NADH
show the reaction diagram
ethyl (R)-mandelate + NADH + H+
ethyl benzoylformate + NAD+
show the reaction diagram
-
95% ethanoselective reaction
-
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r
ethyl 2-oxo-3-phenylpropanoate + NADH + H+
ethyl 2-hydroxy-3-phenylpropanoate + NAD+
show the reaction diagram
ethyl 2-oxo-4-phenylbutanoate + NADH + H+
ethyl 2-hydroxy-4-phenylbutanoate + NAD+
show the reaction diagram
135% of the activity with ethyl 2-oxopropanoate
chirality of the product not determined. The enzyme prouces ethyl (R)-mandelate from ethal benzoylformate
-
?
ethyl 2-oxopropanoate + NADH + H+
ethyl 2-hydroxypropanoate + NAD+
show the reaction diagram
-
chirality of the product not determined. The enzyme prouces ethyl (R)-mandelate from ethal benzoylformate
-
?
ethyl 3-oxohexanoate + NADH + H+
ethyl 3-hydroxyhexanoate + NAD+
show the reaction diagram
105% of the activity with ethyl 2-oxopropanoate
chirality of the product not determined. The enzyme prouces ethyl (R)-mandelate from ethal benzoylformate
-
?
ethyl 4-chloro-3-oxobutanoate + NADH + H+
ethyl 4-chloro-3-hydroxybutanoate + NAD+
show the reaction diagram
ethyl benzoylformate + NADH + H+
ethyl (R)-mandelate + NAD+
show the reaction diagram
-
completely enantioselective reaction
95% enantiomeric excess after 6 h at 50C
-
r
ethyl benzoylformate + NADH + H+
ethyl (R)-mandelate+ NAD+
show the reaction diagram
270% of the activity with ethyl 2-oxopropanoate
95% conversion with 99.9% enantiomeric excess
-
?
ethyl oxo(phenyl)acetate + NADH + H+
ethyl hydroxy(phenyl)acetate + NAD+
show the reaction diagram
270% of the activity with ethyl 2-oxopropanoate
chirality of the product not determined. The enzyme prouces ethyl (R)-mandelate from ethal benzoylformate
-
?
hydroxyacetone + NADH
? + NAD+
show the reaction diagram
-
92% of the activity with acetone
-
-
r
methyl (R)-mandelate + NADH + H+
methyl benzoylformate + NAD+
show the reaction diagram
-
92% ethanoselective reaction
-
-
r
methyl 3-oxobutanoate + NADH
methyl (3S)-3-hydroxybutanoate + NAD+
show the reaction diagram
-
135% of the activity with 2-butanone
-
-
r
methyl benzoylformate + NADH + H+
methyl (R)-mandelate + NAD+
show the reaction diagram
-
completely enantioselective reaction
92% enantiomeric excess after 6 h at 50C
-
r
methyl oxo(phenyl)acetate + NADH
methyl (2S)-hydroxy(phenyl)ethanoate + NAD+
show the reaction diagram
-
-
79% yield, 98% enantiomeric excess
-
?
methylpyruvate + NADH
? + NAD+
show the reaction diagram
-
64% of the activity with acetone
-
-
r
pentane-2,4-dione + NADH
(4R)-4-hydroxypentan-2-one + NAD+
show the reaction diagram
-
32% of the activity with 2-butanone
-
-
r
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(R)-1-phenylethanol + NAD+
acetophenone + NADH + H+
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADPH
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
EDTA
-
activates to 127% activity compared to the control activity at 1 mM and to 163% at 10 mM
K+
-
activates to 167% activity compared to the control activity at 1 mM and to 193% at 100 mM
Li+
-
activates to 127% activity compared to the control activity at 1 mM and to 178% at 100 mM
Na+
-
activates to 179 at 100 mM
Zn2+
-
activates to 114% activity compared to the control activity at 1 mM
ZnSO4
-
1mM, 135% of initial activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-butyl-3-methylimidazolium tetrafluoroborate
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65% inhibition at 2 mM
8-hydroxyquinoline
-
1 mM, 55% residual activity
alpha,alpha'-dipyridyl
-
1 71, 43% residual activity
Ca2+
-
10% inhibition at 1 mM and 30% at 100 mM
CaCl2
-
1 mM, 21% residual activity
CoCl2
-
1 mM, 57% residual activity
Cu2+
-
23% inhibition at 1 mM
CuSO4
-
1 mM, complete loss of activity
EDTA
-
1 mM, 68% residual activity
FeCl2
-
1 mM, 81% residual activity
FeCl3
-
1 mM, 65% residual activity
FeSO4
-
1 mM, complete loss of activity
Hg2+
-
67% inhibition at 1 mM
iodoacetate
-
5% inhibition at 1 mM
Mg2+
-
33% inhibition at 100 mM
MgSO4
-
1 mM, 94% residual activity
MnSO4
-
1 mM, 69% residual activity
NiCl2
-
1 mM, 96% residual activity
o-phenanthroline
-
1 mM, 43% residual activity
p-chloromercuribenzoate
-
1 mM, 4% residual activity
ZnCl2
-
1 mM, 73% residual activity
additional information
-
no effects by 5 mM 2-mercaptoethanol
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
o-phenanthroline
-
activates to 108% activity compared to the control activity at 0.1 mM and to 109% at 1 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5.4
(R)-1,3-butandiol
-
pH 8.5, 30C
0.045 - 37.4
(R)-2-butanol
0.22 - 11.7
(R)-2-pentanol
52
(S)-1,3-butandiol
-
pH 8.5, 30C
0.35
(S)-2-butanol
-
pH 8.5, 30C
10
1-butanol
-
pH 8.5, 30C
4
1-propanol
-
pH 8.5, 30C
11.2
2,2,2-trifluoroacetophenone
-
pH 6.0, 65C, recombinant enzyme, reduction reaction
0.065
2-butanol
-
pH 8.5, 30C
0.04
2-butanone
-
pH 6.0, 30C
0.84
2-Pentanol
-
pH 8.5, 30C
0.075 - 0.11
2-propanol
0.76
3-Pentanol
-
pH 8.5, 30C
0.065
acetone
-
pH 6.0, 30C
6.5
ethanol
-
pH 8.5, 30C
5.3
ethyl 2-oxopropanoate
pH 6.5, 50C
-
1
ethylbenzoylformate
-
pH 6.0, 65C, recombinant enzyme, reduction reaction
0.15 - 0.525
NAD+
0.01 - 0.13
NADH
0.017 - 0.028
NADPH
additional information
additional information
-
kinetics and substrate specificity, overview
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1 - 1.7
(R)-2-butanol
1.1 - 1.9
(R)-2-pentanol
25.5
2,2,2-trifluoroacetophenone
Thermus thermophilus
-
pH 6.0, 65C, recombinant enzyme, reduction reaction
3.97
ethyl 2-oxopropanoate
Carboxydothermus hydrogenoformans
Q3ACV3
pH 6.5, 50C
-
50.1
ethylbenzoylformate
Thermus thermophilus
-
pH 6.0, 65C, recombinant enzyme, reduction reaction
0.84
NAD+
Thermus thermophilus
-
pH 6.0, 65C, recombinant enzyme, reduction reaction
52.4
NADH
Thermus thermophilus
-
pH 6.0, 65C, recombinant enzyme, reduction reaction
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
75 - 300
NADH
8
5100 - 9300
NADPH
5
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
14
-
pH 6.0, 30C
36
-
pH 8.5, 30C, substrate 2-propanol
104
-
pH 8.5, 30C, substrate 2-propanol
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5
-
wild-type, cofactor NADH. Mutant R38P, both cofactors NADH and NADPH
6 - 6.5
-
wild-type, cofactor NADPH
6
-
reduction reaction
7.1
-
reduction
8
-
reduction
9
-
and above, oxidation, Tris buffer
10
-
oxidation reaction
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 7.7
-
reduction reaction, high activity
9 - 10
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oxidation reaction, high activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
60
-
oxidation reaction
65
-
reduction reaction
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 73
-
the enzyme displays activity at temperatures up to 73C
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.4
-
isoelectric focusing
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
26000
-
4 * 26961, recombinant enzyme, mass spectrometry, 4 * 26000, recombinant enzyme, SDS-PAGE
26961
-
4 * 26961, recombinant enzyme, mass spectrometry, 4 * 26000, recombinant enzyme, SDS-PAGE
30000
x * 30000, SDS-PAGE and calculated
35000
-
gel filtration
37000
-
4 * 37000, SDS-PAGE
37500
-
x * 37500, SDS-PAGE
37590
4 * 37590, sequence calculation, a tetramer formed by a dimer of dimers
71000
-
about, recombinant enzyme, gel filtration in absence of NaCl
105000
-
about, recombinant enzyme, gel filtration in presence of 0.15 M NaCl
150000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
-
x * 37500, SDS-PAGE
tetramer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant His-tagged mutant I86A, hanging drop vapour diffusion, mixing of 0.002 ml of 8 mg/ml protein in 50 mM 2-(N-morpholino)ethanesulfonic acid, pH 6.6, 20 mM NaCl, 0.1 mM dithiothreitol, 0.1 mM ethylenediaminetetraacetic acid, and 2 mM (R)-1-phenylethanol, with 0.2% D-fructose 1,6-disphosphate trisodium salt octahydrate, 0.2% glycerol phosphate disodium salt hydrate, 0.2% O-phospho-L-serine, 0.2% O-phosphol-L-styrosine, 0.2% phytic acid sodium salt hydrate, 0.02 M HEPES sodium pH 6.8, and 0.002 ml reservoir solution containing 20% v/v PEG 400, 0.1 M Na HEPES, pH 7.0, room temperature, 6-12 h, X-ray diffraction structure determination and analysis, molecular replacement, modeling
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
wild-type, half-life 160 h, mutant R38P, half-life 220 h
42
-
wild-type, half-life 1.25 h, mutant R38P, half-life 3 h
50
-
24 h, purified recombinant enzyme, 142% inactivation
60
-
24 h, purified recombinant enzyme, 134% inactivation
90
-
30 min, purified recombinant enzyme, 50% inactivation
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Acetone
Ethanol
tetrahydrofuran
additional information
-
the enzyme shows a good tolerance to common organic solvents
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4C, pH 6.0-8.0, 24 h, more than 90% residual activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant C-terminally His-tagged mutant I86A from Escherichia coli HB101 (DE3) by nickel affinity chromatography
recombinant enzyme 11fold to homogeneity from Escherichia coli strain BL21(DE3) by heat denaturation of host proteins, anion exchange chromatography and gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli RB791 cells
-
expression in Escherichia coli
expression in Escherichia coli strain BL21(DE3)
-
gene adhB, expression of mutant I86A in Escherichia coli HB101 (DE3) as protein C-terminally tagged with two methionines followed by Leu-Glu-His6
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A90S
-
activity with NADPH as cofactor decreased to about 50% of wild-type, activity with NADH strongly decreased
G37D/R38P
-
activity with NADH is decreased relative to Arg38Pro alone, but is higher than that of the wild-type enzyme
M140I
-
no activity with NADPH as cofactor
R38P
-
no activity with NADPH as cofactor, fourfold increase in activity with NADH
V112D
-
no activity with NADPH as cofactor, strongly decreased activity with NADH
W95L/N249Y
-
the mutant exhibits higher activity but decreased affinity toward aliphatic alcohols, aldehydes as well as NAD+ and NADH compared to the wild type enzyme, optimum pH is at about pH 8.6
I86A
site-directed mutagensis, the secondary alcohol dehydrogenase I86A mutant is stereospecific for (R)-alcohols instead of (S)-alcohols, in contrast to the wild-type enzyme, the mutation I86A allows large substituents to fit into the large pocket of I86ATeSADH, which corresponds to the small pocket in wild-type TeSADH, modeling of the stereopreference of TeSADH I86A
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis