Substrate expansion of Geotrichum candidum alcohol dehydrogenase towards diaryl ketones by mutation.

ABSTRACT: Chiral diaryl alcohols, such as (4-chlorophenyl)(pyridin-2-yl)methanol, are important intermediates for pharmaceutical synthesis. However, using alcohol dehydrogenases (ADHs) in the asymmetric reduction of diaryl ketones to produce the corresponding alcohols is challenging due to steric hindrance in the substrate binding pockets of the enzymes. In this study, the steric hindrance of the ADH from Geotrichum candidum NBRC 4597 (G. candidum acetophenone reductase, GcAPRD) was eliminated by simultaneous site-directed mutagenesis of Phe56 (in the large pocket) and Trp288 (in the small pocket). As a result, two double mutants, Phe56Ile/Trp288Ala, and Phe56Ala/Trp288Ala, exhibited much higher specific activities towards 2-(4'-chlorobenzoyl)pyridine (4.5 μmol/min/mg and 3.4 μmol/min/mg, respective

SUBMITTER: Tang Z 

PROVIDER: S-EPMC11680648 | biostudies-literature | 2024 Dec

REPOSITORIES: biostudies-literature