Directed Evolution of an Amine Oxidase for the Preparative Deracemisation of Cyclic Secondary Amines

• Published in: Chembiochem : a European journal of chemical biology Vol. 6; no. 4; pp. 637 - 639
• Main Authors: Carr, Reuben, Alexeeva, Marina, Dawson, Michael J, Gotor-Fernández, Vicente, Humphrey, Cara E, Turner, Nicholas J
• Format: Journal Article
• Language: English
• Published: Weinheim Wiley-VCH Verlag 01.04.2005; WILEY-VCH Verlag; WILEY‐VCH Verlag
• Subjects: amine oxidase; amines; Amines - chemistry; Amines - metabolism; Amino Acid Substitution - genetics; Aspergillus niger - enzymology; biocatalysis; Catalytic Domain - genetics; deracemisation; directed evolution; Directed Molecular Evolution; enantioselectivity; Enzymes, Immobilized - chemistry; Enzymes, Immobilized - metabolism; Humans; Kinetics; Models, Molecular; Monoamine Oxidase - chemistry; Monoamine Oxidase - genetics; Monoamine Oxidase - metabolism; Pargyline - chemistry; Phenethylamines - chemistry; Phenethylamines - metabolism; Protein Engineering; Sequence Homology, Amino Acid; Stereoisomerism; Tetrahydroisoquinolines - chemistry; Tetrahydroisoquinolines - metabolism
• ISSN: 1439-4227; 1439-7633
• DOI: 10.1002/cbic.200400329

Off the rac. Directed evolution of the amine oxidase from Aspergillus niger (MAO‐N), by using 1‐methyltetrahydroisoquinoline as the target substrate, led to the identification of a variant enzyme (Asn336Ser/Ile246Met) that possessed a greater kcat value (5.5‐fold) than the parent as well as high enantioselectivity (see scheme). This new variant was used for the deracemisation of cyclic secondary amines on a preparative scale.