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show all sequences of 1.1.1.B3

Purification, immobilization and stabilization of a highly enantioselective alcohol dehydrogenase from Thermus thermophilus HB27 cloned in E. coli

Rocha-Martin, J.; Vega, D.; Cabrera, Z.; Bolivar, J.; Fernandez-Lafuente, R.; Berenguer, J.; Guisan, J.; Process Biochem. 44, 1004-1012 (2009)
No PubMed abstract available

Data extracted from this reference:

Application
Application
Commentary
Organism
synthesis
the immobilized enzyme is utilized in the asymmetric reduction of acetophenone to produce (S)-1-phenylethanol, with an enantiomeric excess of more than 99%
Thermus thermophilus
Cloned(Commentary)
Commentary
Organism
expression in Escherichia coli strain BL21(DE3)
Thermus thermophilus
Engineering
Amino acid exchange
Commentary
Organism
additional information
covalent immobilization of the purified recombinant enzyme on different supports, i.e. on glyoxyl agarose, amino epoxy agarose, CNBr-activated sepharose, monoaminoethyl-N-ethylagarose-glutaraldehyde, monoaminoethyl-N-ethyl agarose, or polyethyleneimine agarose, immobilized enzyme activities, overview
Thermus thermophilus
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
140000
-
recombinant enzyme, gel filtration
Thermus thermophilus
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
(S)-1-phenylethanol + NAD+
Thermus thermophilus
-
acetophenone + NADH + H+
-
-
r
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Thermus thermophilus
-
strain HB27, DSM 7039
-
Purification (Commentary)
Commentary
Organism
by heat treatment at 70C and/or adsorption chromatography on an IDA-Cu2+ support, both methods combined result in 8.8fold purification
Thermus thermophilus
Specific Activity [micromol/min/mg]
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
1.5
-
immobilized recombinant enzyme
Thermus thermophilus
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(S)-1-phenylethanol + NAD+
-
701031
Thermus thermophilus
acetophenone + NADH + H+
-
-
-
r
(S)-1-phenylethanol + NAD+
asymmetric reduction of acetophenone to produce (S)-1-phenylethanol, with an enantiomeric excess of more than 99%
701031
Thermus thermophilus
acetophenone + NADH + H+
-
-
-
r
Subunits
Subunits
Commentary
Organism
tetramer
-
Thermus thermophilus
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
73
-
-
Thermus thermophilus
Temperature Stability [C]
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
additional information
-
the thermal stability of the enzyme adsorbed by ionic exchange is very similar to the stability of the covalently immobilized enzymes in the pH range 5-9, thermal inactivation kinetics, overview
Thermus thermophilus
70
-
after a thermal treatment at pH 7.0 and 70C for 1 h, the remaining TtADH activity is over 90%, while the soluble protein concentration decreases by a 85%
Thermus thermophilus
90
-
half-life is 30 min
Thermus thermophilus
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
-
Thermus thermophilus
pH Stability
pH Stability
pH Stability Maximum
Commentary
Organism
7
-
after a thermal treatment at pH 7.0 and 70C for 1 h, the remaining TtADH activity is over 90%, while the soluble protein concentration decreases by a 85%
Thermus thermophilus
Cofactor
Cofactor
Commentary
Organism
Structure
NAD+
-
Thermus thermophilus
NADH
-
Thermus thermophilus
Application (protein specific)
Application
Commentary
Organism
synthesis
the immobilized enzyme is utilized in the asymmetric reduction of acetophenone to produce (S)-1-phenylethanol, with an enantiomeric excess of more than 99%
Thermus thermophilus
Cloned(Commentary) (protein specific)
Commentary
Organism
expression in Escherichia coli strain BL21(DE3)
Thermus thermophilus
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NAD+
-
Thermus thermophilus
NADH
-
Thermus thermophilus
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
additional information
covalent immobilization of the purified recombinant enzyme on different supports, i.e. on glyoxyl agarose, amino epoxy agarose, CNBr-activated sepharose, monoaminoethyl-N-ethylagarose-glutaraldehyde, monoaminoethyl-N-ethyl agarose, or polyethyleneimine agarose, immobilized enzyme activities, overview
Thermus thermophilus
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
140000
-
recombinant enzyme, gel filtration
Thermus thermophilus
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
(S)-1-phenylethanol + NAD+
Thermus thermophilus
-
acetophenone + NADH + H+
-
-
r
Purification (Commentary) (protein specific)
Commentary
Organism
by heat treatment at 70C and/or adsorption chromatography on an IDA-Cu2+ support, both methods combined result in 8.8fold purification
Thermus thermophilus
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
1.5
-
immobilized recombinant enzyme
Thermus thermophilus
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(S)-1-phenylethanol + NAD+
-
701031
Thermus thermophilus
acetophenone + NADH + H+
-
-
-
r
(S)-1-phenylethanol + NAD+
asymmetric reduction of acetophenone to produce (S)-1-phenylethanol, with an enantiomeric excess of more than 99%
701031
Thermus thermophilus
acetophenone + NADH + H+
-
-
-
r
Subunits (protein specific)
Subunits
Commentary
Organism
tetramer
-
Thermus thermophilus
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
73
-
-
Thermus thermophilus
Temperature Stability [C] (protein specific)
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
additional information
-
the thermal stability of the enzyme adsorbed by ionic exchange is very similar to the stability of the covalently immobilized enzymes in the pH range 5-9, thermal inactivation kinetics, overview
Thermus thermophilus
70
-
after a thermal treatment at pH 7.0 and 70C for 1 h, the remaining TtADH activity is over 90%, while the soluble protein concentration decreases by a 85%
Thermus thermophilus
90
-
half-life is 30 min
Thermus thermophilus
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
-
Thermus thermophilus
pH Stability (protein specific)
pH Stability
pH Stability Maximum
Commentary
Organism
7
-
after a thermal treatment at pH 7.0 and 70C for 1 h, the remaining TtADH activity is over 90%, while the soluble protein concentration decreases by a 85%
Thermus thermophilus
Other publictions for EC 1.1.1.B3
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)
724568
Cao
Purification and characterizat ...
Scheffersomyces stipitis
Biores. Technol.
102
1733-1739
2011
-
1
1
-
-
-
3
1
-
-
-
-
3
1
-
-
1
-
-
-
1
-
5
-
1
2
-
-
1
1
2
2
-
-
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-
1
1
2
-
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3
-
1
-
-
-
-
3
-
-
1
-
-
1
-
5
-
1
2
-
-
1
1
2
-
-
-
-
-
-
-
723841
Protsko
Crystallization and preliminar ...
Thermoanaerobacter ethanolicus
Acta Crystallogr. Sect. F
66
831-833
2010
-
-
-
-
1
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
1
-
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2
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2
-
1
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1
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-
-
-
1
-
-
-
-
-
-
-
-
-
-
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-
-
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-
725726
Li
-
Deracemization of aryl seconda ...
Rhodotorula sp., Rhodotorula sp. AS2.2241
J. Mol. Catal. B
64
48-52
2010
-
1
-
-
-
-
-
-
-
-
-
2
-
2
-
-
-
-
-
-
-
-
4
-
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1
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1
-
1
-
-
-
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-
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-
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-
2
-
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-
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-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
697267
Pal
Activity of yeast alcohol dehy ...
Saccharomyces carlsbergensis, Saccharomyces carlsbergensis Y379-50, Saccharomyces cerevisiae
Chem. Biol. Interact.
178
16-23
2009
-
-
-
-
-
-
-
-
1
-
-
-
-
3
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
2
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-
2
-
-
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-
-
-
-
1
-
-
-
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-
-
-
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-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
697839
Pennacchio
Role of Tryptophan 95 in subst ...
Sulfolobus solfataricus
Extremophiles
13
751-761
2009
-
-
1
1
1
-
-
4
-
-
-
-
-
1
-
-
1
-
-
-
-
-
2
-
-
-
-
4
-
-
-
1
-
-
-
-
-
1
1
1
1
-
-
-
-
4
-
-
-
-
-
-
-
1
-
-
-
-
2
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
701031
Rocha-Martin
-
Purification, immobilization a ...
Thermus thermophilus
Process Biochem.
44
1004-1012
2009
-
1
1
-
1
-
-
-
-
-
1
1
-
1
-
-
1
-
-
-
1
-
2
1
1
-
3
-
1
-
1
2
-
-
-
-
1
1
2
-
1
-
-
-
-
-
-
-
1
1
-
-
-
1
-
-
1
-
2
1
1
-
3
-
1
-
1
-
-
-
-
-
-
-
684583
Pennacchio
Purification and characterizat ...
Thermus thermophilus
Appl. Environ. Microbiol.
74
3949-3958
2008
1
1
1
-
-
-
7
6
-
5
4
-
1
5
-
-
1
-
-
-
1
-
7
1
2
1
4
5
2
2
-
3
-
-
-
1
1
1
3
-
-
-
-
7
-
6
-
5
4
-
1
-
-
1
-
-
1
-
7
1
2
1
4
5
2
2
-
-
-
-
-
-
-
-
689823
Soni
-
Purification and characterizat ...
Candida viswanathii, Candida viswanathii MTCC 5158
Process Biochem.
42
1632-1640
2007
1
-
-
-
-
-
30
2
-
-
2
-
7
3
-
-
1
-
-
-
1
1
14
1
1
2
1
-
1
1
-
2
-
-
-
1
-
-
2
-
-
-
-
30
-
2
-
-
2
-
7
-
-
1
-
-
1
1
14
1
1
2
1
-
1
1
-
-
-
-
-
-
-
-
685078
Hoeffken
Crystal structure and enzyme k ...
uncultured bacterium, uncultured bacterium EbN1
Biochemistry
45
82-93
2006
-
-
1
1
-
-
3
3
-
-
3
-
-
2
-
-
1
-
-
-
-
-
4
1
-
-
-
-
-
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-
-
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-
1
-
1
-
-
-
3
-
3
-
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3
-
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-
1
-
-
-
-
4
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
684588
Abokitse
Cloning, sequence analysis, an ...
Rhodococcus erythropolis
Appl. Microbiol. Biotechnol.
62
380-386
2003
-
-
1
-
-
-
-
-
-
1
2
-
-
1
-
-
1
-
-
-
1
-
5
1
-
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-
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-
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-
1
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1
2
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-
1
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1
-
5
1
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-
-
-
-
-
-
-
-
-
-
-
-
-
685641
Yamamoto
-
Synthesis of (R)-1,3-butanedio ...
Candida parapsilosis
Biosci. Biotechnol. Biochem.
4
925-927
2002
-
1
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
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1
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2
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1
1
2
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-
1
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-
-
-
-
-
-
-
-
-
-
-
-
-
685644
Yamamoto
Synthesis of ethyl (R)-4-chlor ...
Candida parapsilosis
Biosci. Biotechnol. Biochem.
66
481-483
2002
-
1
1
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
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-
1
-
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-
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1
1
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-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
685643
Xie
NAD+-Dependent (S)-specific se ...
Nocardia fusca AKU 2123
Biosci. Biotechnol. Biochem.
63
1721-1729
1999
-
-
-
-
-
-
7
3
-
-
2
-
-
4
-
-
1
-
-
-
1
-
24
1
2
-
2
-
-
2
1
3
-
-
-
-
-
-
3
-
-
-
-
7
-
3
-
-
2
-
-
-
-
1
-
-
1
-
24
1
2
-
2
-
-
2
1
-
-
-
-
-
-
-