Saturation Mutagenesis for Phenylalanine Ammonia Lyases of Enhanced Catalytic Properties.

ABSTRACT: Phenylalanine ammonia-lyases (PALs) are attractive biocatalysts for the stereoselective synthesis of non-natural phenylalanines. The rational design of PALs with extended substrate scope, highlighted the substrate specificity-modulator role of residue I460 of Petroselinum crispum PAL. Herein, saturation mutagenesis at key residue I460 was performed in order to identify PcPAL variants of enhanced activity or to validate the superior catalytic properties of the rationally explored I460V PcPAL compared with the other possible mutant variants. After optimizations, the saturation mutagenesis employing the NNK-degeneracy generated a high-quality transformant library. For high-throughput enzyme-activity screens of the mutant library, a PAL-activity assay was developed, allowing the identification

SUBMITTER: Tomoiaga RB 

PROVIDER: S-EPMC7355458 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature