Fluorescent enzyme-coupled activity assay for phenylalanine ammonia-lyases.

ABSTRACT: Phenylalanine ammonia-lyases (PALs) catalyse the non-oxidative deamination of L-phenylalanine to trans-cinnamic acid, while in the presence of high ammonia concentration the reverse reaction occurs. PALs have been intensively studied, however, their industrial applications for amino acids synthesis remained limited, mainly due to their decreased operational stability or limited substrate specificity. The application of extensive directed evolution procedures to improve their stability, activity or selectivity, is hindered by the lack of reliable activity assays allowing facile screening of PAL-activity within large-sized mutant libraries. Herein, we describe the development of an enzyme-coupled fluorescent assay applicable for PAL-activity screens at whole cell level, involving decarboxyla

SUBMITTER: Moisa ME 

PROVIDER: S-EPMC7595223 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature