Engineering of Thermostable β‐Hydroxyacid Dehydrogenase for the Asymmetric Reduction of Imines

The β‐hydroxyacid dehydrogenase from Thermocrinus albus (Ta‐βHAD), which catalyzes the NADP+‐dependent oxidation of β‐hydroxyacids, was engineered to accept imines as substrates. The catalytic activity of the proton‐donor variant K189D was further increased by the introduction of two nonpolar flanki...

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Bibliographic Details
Published in:Chembiochem : a European journal of chemical biology 2020-12, Vol.21 (24), p.3511-3514
Main Authors: Stockinger, Peter, Schelle, Luca, Schober, Benedikt, Buchholz, Patrick C. F., Pleiss, Jürgen, Nestl, Bettina M.
Format: Article
Language:English
Subjects:
Asymmetry
beta-HAD
Catalytic activity
Communication
Communications
Dehydrogenase
Dehydrogenases
High temperature
imine reductases
Imines
NADP
Oxidation
promiscuity
Protons
Residues
Substrate specificity
Substrates
thermostability
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Summary:The β‐hydroxyacid dehydrogenase from Thermocrinus albus (Ta‐βHAD), which catalyzes the NADP+‐dependent oxidation of β‐hydroxyacids, was engineered to accept imines as substrates. The catalytic activity of the proton‐donor variant K189D was further increased by the introduction of two nonpolar flanking residues (N192 L, N193 L). Engineering the putative alternative proton donor (D258S) and the gate‐keeping residue (F250 A) led to a switched substrate specificity as compared to the single and triple variants. The two most active Ta‐βHAD variants were applied to biocatalytic asymmetric reductions of imines at elevated temperatures and enabled enhanced product formation at a reaction temperature of 50 °C. Get more from your enzyme: A β‐hydroxyacid dehydrogenase from Thermocrinus albus was engineered to accept two different imine compounds as substrates. The variants were applied to biocatalytic asymmetric reductions at elevated temperatures leading to an enhanced product formation.
ISSN:1439-4227
1439-7633
DOI:10.1002/cbic.202000526