1. Finding the Switch: Turning a Baeyer-Villiger Monooxygenase into a NADPH Oxidase.
- Author
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Brondani, Patrícia B., Dudek, Hanna M., Martinoli, Christian, Mattevi, Andrea, and Fraaije, Marco W.
- Subjects
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ENZYMES , *CHEMICAL synthesis , *NADPH oxidase , *BAEYER-Villiger rearrangement , *MONOOXYGENASES , *FLAVINS , *INTERMEDIATES (Chemistry) - 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. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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