Engineering of Thermostable ß-Hydroxyacid Dehydrogenase for the Asymmetric Reduction of Imines.
Chembiochem
; 21(24): 3511-3514, 2020 12 11.
Article
em En
| MEDLINE
| ID: mdl-32939899
ABSTRACT
The ß-hydroxyacid dehydrogenase from Thermocrinus albus (Ta-ßHAD), which catalyzes the NADP+ -dependent oxidation of ß-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 (N192â
L, N193â
L). Engineering the putative alternative proton donor (D258S) and the gate-keeping residue (F250â
A) led to a switched substrate specificity as compared to the single and triple variants. The two most active Ta-ßHAD variants were applied to biocatalytic asymmetric reductions of imines at elevated temperatures and enabled enhanced product formation at a reaction temperature of 50 °C.
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Base de dados:
MEDLINE
Assunto principal:
Temperatura
/
Desidrogenases de Carboidrato
/
Engenharia de Proteínas
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Iminas
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article