Engineering of phenylalanine ammonia lyase from Rhodotorula graminis for the enhanced synthesis of unnatural l-amino acids. Issue 46 (17th November 2016)

• Record Type: Journal Article
• Authors: Rowles, Ian; Groenendaal, Bas; Binay, Baris; Malone, Kirk J.; Willies, Simon C.; Turner, Nicholas J.
• Is Part Of: Tetrahedron. Volume 72:Issue 46(2016)
• Page Start: 7343
• Page End: 7347
• Publication Date: 2016-11-17
• Subjects: Rhodotorula graminis Phenylalanine ammonia lyase -- Unnatural l-amino acids -- Directed evolution -- Modeling; Chemistry, Organic -- Periodicals; 547.005
• DOI: 10.1016/j.tet.2016.06.026
• Languages: English
• ISSNs: 0040-4020

Abstract

Abstract: Phenylalanine ammonia lyase (PAL) catalyses the reversible non-oxidative deamination of phenylalanine to trans -cinnamic acid and ammonia. Analogues ofl -phenylalanine are incorporated as pharmacophores in several peptidomimetic drug molecules and are therefore of particular interest to the fine chemical industry. PAL from Rhodotorula graminis ( Rgr PAL) has shown an ability to accept analogues ofl -phenylalanine. Our aim was to increase enzymatic activity with directed evolution towards a specific non-natural substrate through the cloning and over-production of PAL in Escherichia coli . The identified variants of Rgr PAL with significantly showed more catalytic efficient compared to the wild-type enzyme. These variants were used in a preparative scale biotransformation resulting in a 94% conversion to L-4-Br-phenylalanine (>99% ee). Graphical abstract: