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show all sequences of 1.1.1.B4

Crystallization and preliminary X-ray diffraction analysis of the Thermoanaerobacter ethanolicus secondary alcohol dehydrogenase I86A mutant

Protsko, C.; Vieille, C.; Laivenieks, M.; Prasad, L.; Sanders, D.A.; Delbaere, L.T.; Acta Crystallogr. Sect. F 66, 831-833 (2010)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
gene adhB, expression of mutant I86A in Escherichia coli HB101 (DE3) as protein C-terminally tagged with two methionines followed by Leu-Glu-His6
Thermoanaerobacter ethanolicus
Crystallization (Commentary)
Crystallization
Organism
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
Thermoanaerobacter ethanolicus
Engineering
Amino acid exchange
Commentary
Organism
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
Thermoanaerobacter ethanolicus
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
37590
-
4 * 37590, sequence calculation, a tetramer formed by a dimer of dimers
Thermoanaerobacter ethanolicus
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
(R)-1-phenylethanol + NAD+
Thermoanaerobacter ethanolicus
activity of mutant I886A, the wild-type enzyme is (S)-specific forming (S)-1-phenylethanol
acetophenone + NADH + H+
-
-
r
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Thermoanaerobacter ethanolicus
P77990
gene adhB, mutant I86A
-
Purification (Commentary)
Commentary
Organism
recombinant C-terminally His-tagged mutant I86A from Escherichia coli HB101 (DE3) by nickel affinity chromatography
Thermoanaerobacter ethanolicus
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(R)-1-phenylethanol + NAD+
activity of mutant I886A, the wild-type enzyme is (S)-specific forming (S)-1-phenylethanol
723841
Thermoanaerobacter ethanolicus
acetophenone + NADH + H+
-
-
-
r
Subunits
Subunits
Commentary
Organism
tetramer
4 * 37590, sequence calculation, a tetramer formed by a dimer of dimers
Thermoanaerobacter ethanolicus
Cofactor
Cofactor
Commentary
Organism
Structure
NAD+
-
Thermoanaerobacter ethanolicus
NADH
-
Thermoanaerobacter ethanolicus
Cloned(Commentary) (protein specific)
Commentary
Organism
gene adhB, expression of mutant I86A in Escherichia coli HB101 (DE3) as protein C-terminally tagged with two methionines followed by Leu-Glu-His6
Thermoanaerobacter ethanolicus
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NAD+
-
Thermoanaerobacter ethanolicus
NADH
-
Thermoanaerobacter ethanolicus
Crystallization (Commentary) (protein specific)
Crystallization
Organism
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
Thermoanaerobacter ethanolicus
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
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
Thermoanaerobacter ethanolicus
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
37590
-
4 * 37590, sequence calculation, a tetramer formed by a dimer of dimers
Thermoanaerobacter ethanolicus
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
(R)-1-phenylethanol + NAD+
Thermoanaerobacter ethanolicus
activity of mutant I886A, the wild-type enzyme is (S)-specific forming (S)-1-phenylethanol
acetophenone + NADH + H+
-
-
r
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant C-terminally His-tagged mutant I86A from Escherichia coli HB101 (DE3) by nickel affinity chromatography
Thermoanaerobacter ethanolicus
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(R)-1-phenylethanol + NAD+
activity of mutant I886A, the wild-type enzyme is (S)-specific forming (S)-1-phenylethanol
723841
Thermoanaerobacter ethanolicus
acetophenone + NADH + H+
-
-
-
r
Subunits (protein specific)
Subunits
Commentary
Organism
tetramer
4 * 37590, sequence calculation, a tetramer formed by a dimer of dimers
Thermoanaerobacter ethanolicus
General Information
General Information
Commentary
Organism
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
Thermoanaerobacter ethanolicus
General Information (protein specific)
General Information
Commentary
Organism
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
Thermoanaerobacter ethanolicus
Other publictions for EC 1.1.1.B4
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
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Carboxydothermus hydrogenoformans, Carboxydothermus hydrogenoformans DSM 6008
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1
1
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1
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1
-
4
2
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15
1
1
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-
1
1
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2
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1
1
2
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1
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1
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4
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15
1
1
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1
1
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2010
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-
1
1
1
-
-
-
-
-
1
1
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1
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1
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1
1
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2
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1
2
1
1
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1
1
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1
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-
1
1
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-
-
-
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-
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1
1
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725726
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4
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2
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1
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2
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2
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4
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-
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697267
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178
16-23
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-
-
-
-
-
-
-
1
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3
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3
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2
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2
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1
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3
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-
697839
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4
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-
-
-
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1
-
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1
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-
2
-
-
-
-
4
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-
-
1
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-
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1
1
1
1
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4
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1
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2
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2
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2
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1
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1
1
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5
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4
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2
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2
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2
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4
4
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Thermus thermophilus
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1
1
1
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-
-
7
5
-
5
4
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1
1
-
-
1
-
-
-
-
-
7
1
2
1
4
4
2
2
-
3
-
-
-
1
1
1
3
-
-
-
-
7
-
5
-
5
4
-
1
-
-
1
-
-
-
-
7
1
2
1
4
4
2
2
-
-
-
-
-
-
-
-
688747
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Candida parapsilosis
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-
-
-
-
-
-
9
-
1
1
2
-
-
1
-
-
1
-
-
-
1
1
10
1
1
-
-
-
1
-
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2
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-
2
-
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-
-
9
-
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1
1
2
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-
1
-
-
1
1
10
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
687864
Isobe
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Burkholderia sp., Burkholderia sp. AIU 652
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96
387-393
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-
-
-
-
-
-
6
15
-
-
2
-
-
3
-
-
1
-
-
-
1
-
24
1
1
-
1
-
3
-
-
3
-
1
-
-
-
-
3
-
-
-
-
6
-
15
-
-
2
-
-
-
-
1
-
-
1
-
24
1
1
-
1
-
3
-
-
1
-
-
-
-
-
-
688571
Bradshaw
-
A Pseudomonas sp. alcohol dehy ...
Pseudomonas sp., Pseudomonas sp. PED
J. Org. Chem.
57
1526-1532
1992
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2
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2
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1
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1
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24
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2
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2
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1
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24
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2
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