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Literature summary extracted from

  • Rocha-Martin, J.; Vega, D.; Cabrera, Z.; Bolivar, J.; Fernandez-Lafuente, R.; Berenguer, J.; Guisan, J.
    Purification, immobilization and stabilization of a highly enantioselective alcohol dehydrogenase from Thermus thermophilus HB27 cloned in E. coli (2009), Process Biochem., 44, 1004-1012.
No PubMed abstract available

Application

EC Number Application Comment Organism
1.1.1.B3 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)

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

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.B3 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]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.B3 140000
-
 recombinant enzyme, gel filtration Thermus thermophilus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.B3 (S)-1-phenylethanol + NAD+ Thermus thermophilus
-
 acetophenone + NADH + H+
-
 r
1.1.1.B3 (S)-1-phenylethanol + NAD+ Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039
-
 acetophenone + NADH + H+
-
 r

Organism

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

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.B3 by heat treatment at 70°C and/or adsorption chromatography on an IDA-Cu2+ support, both methods combined result in 8.8fold purification 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.B3 1.5
-
 immobilized recombinant enzyme Thermus thermophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.B3 (S)-1-phenylethanol + NAD+
-
 Thermus thermophilus acetophenone + NADH + H+
-
 r
1.1.1.B3 (S)-1-phenylethanol + NAD+ asymmetric reduction of acetophenone to produce (S)-1-phenylethanol, with an enantiomeric excess of more than 99% Thermus thermophilus acetophenone + NADH + H+
-
 r
1.1.1.B3 (S)-1-phenylethanol + NAD+
-
 Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 acetophenone + NADH + H+
-
 r
1.1.1.B3 (S)-1-phenylethanol + NAD+ asymmetric reduction of acetophenone to produce (S)-1-phenylethanol, with an enantiomeric excess of more than 99% Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 acetophenone + NADH + H+
-
 r

Subunits

EC Number Subunits Comment Organism
1.1.1.B3 tetramer
-
 Thermus thermophilus

Synonyms

EC Number Synonyms Comment Organism
1.1.1.B3 More the enzyme belongs to the short-chain dehydrogenases/reductase superfamily Thermus thermophilus
1.1.1.B3 TtADH
-
 Thermus thermophilus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.B3 73
-
-
 Thermus thermophilus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.B3 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
1.1.1.B3 70
-
 after a thermal treatment at pH 7.0 and 70°C for 1 h, the remaining TtADH activity is over 90%, while the soluble protein concentration decreases by a 85% Thermus thermophilus
1.1.1.B3 90
-
 half-life is 30 min Thermus thermophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.B3 7
-
-
 Thermus thermophilus

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.1.1.B3 7
-
 after a thermal treatment at pH 7.0 and 70°C for 1 h, the remaining TtADH activity is over 90%, while the soluble protein concentration decreases by a 85% Thermus thermophilus

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.B3 NAD+
-
 Thermus thermophilus
1.1.1.B3 NADH
-
 Thermus thermophilus