title: Finding the switch: turning a baeyer-villiger monooxygenase into a NADPH oxidase.
abstract: By a targeted enzyme engineering approach, we were able to create an efficient NADPH oxidase from a monooxygenase. Intriguingly, replacement of only one specific single amino acid was sufficient for such a monooxygenase-to-oxidase switch-a complete transition in enzyme activity. Pre-steady-state kinetic analysis and elucidation of the crystal structure of the C65D PAMO mutant revealed that the mutation introduces small changes near the flavin cofactor, resulting in a rapid decay of the peroxyflavin intermediate. The engineered biocatalyst was shown to be a thermostable, solvent tolerant, and effective cofactor-regenerating biocatalyst. Therefore, it represents a valuable new biocatalytic tool.
authors.affiliation: Molecular Enzymology Group, University of Groningen , Nijenborgh 4, 9747AG Groningen, The Netherlands.
authors.forename: Patrícia B
authors.initials: PB
authors.lastname: Brondani
authors.forename: Hanna M
authors.initials: HM
authors.lastname: Dudek
authors.forename: Christian
authors.initials: C
authors.lastname: Martinoli
authors.forename: Andrea
authors.initials: A
authors.lastname: Mattevi
authors.forename: Marco W
authors.initials: MW
authors.lastname: Fraaije
mesh_terms: D000192:Actinomycetales / Q000201:enzymology; D055162:Biocatalysis; D009249:NADP / Q000737:chemistry / Q000378:metabolism*; D010105:Oxygenases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D009249:NADP; D010105:Oxygenases; C019961:cyclohexanone oxygenase