title: Tailoring D-amino acid oxidase from the pig kidney to R-stereoselective amine oxidase and its use in the deracemization of α-methylbenzylamine.
abstract: The deracemization of racemic amines to yield enantioenriched amines using S-stereoselective amine oxidases (AOx) has recently been attracting attention. However, R-stereoselective AOx that are suitable for deracemization have not yet been identified. An R-stereoselective AOx was now evolved from porcine kidney D-amino acid oxidase (pkDAO) and subsequently use for the deracemization of racemic amines. The engineered pkDAO, which was obtained by directed evolution, displayed a markedly changed substrate specificity towards R amines. The mutant enzyme exhibited a high preference towards the substrate α-methylbenzylamine and was used to synthesize the S amine through deracemization. The findings of this study indicate that further investigations on the structure-activity relationship of AOx are warranted and also provide a new method for biotransformations in organic synthesis.
authors.affiliation: Biotechnology Research Center and Department of Biotechnology, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama 939-0398 (Japan); Asano Active Enzyme Molecule Project, ERATO, JST, 5180 Kurokawa, Imizu, Toyama 939-0398 (Japan).
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Yasukawa
authors.forename: Shogo
authors.initials: S
authors.lastname: Nakano
authors.forename: Yasuhisa
authors.initials: Y
authors.lastname: Asano
mesh_terms: D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D055162:Biocatalysis; D020134:Catalytic Domain; D007668:Kidney / Q000201:enzymology*; D008958:Models, Molecular; D015394:Molecular Structure; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism*; D010627:Phenethylamines / Q000737:chemistry*; D011487:Protein Conformation; D013237:Stereoisomerism; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D013552:Swine
chemical_list: D050505:Mutant Proteins; D010627:Phenethylamines; D000581:Amidohydrolases; C407355:D-amino acid amidase; C033198:1-phenethylamine
keywords: amine oxidases; biocatalysis; chiral amines; directed evolution; oxidation