1. Mutagenesis-Independent Stabilization of Class B Flavin Monooxygenases in Operation.
- Author
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Goncalves, Leticia C. P., Kracher, Daniel, Milker, Sofia, Fink, Michael J., Rudroff, Florian, Ludwig, Roland, Bommarius, Andreas S., and Mihovilovic, Marko D.
- Subjects
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FLAVINS , *MUTAGENESIS , *MONOOXYGENASES , *PHARMACEUTICAL chemistry , *BIOTECHNOLOGY - Abstract
This paper describes the stabilization of flavin-dependent monooxygenases under reaction conditions, using an engineered formulation of additives (the natural cofactors NADPH and FAD, and superoxide dismutase and catalase as catalytic antioxidants). This way, a 103- to 104-fold increase of the half-life was reached without resource-intensive directed evolution or structure-dependent protein engineering methods. The stabilized enzymes are highly valued for their synthetic potential in biotechnology and medicinal chemistry (enantioselective sulfur, nitrogen and Baeyer-Villiger oxidations; oxidative human metabolism), but widespread application was so far hindered by their notorious fragility. Our technology immediately enables their use, does not require structural knowledge of the biocatalyst, and creates a strong basis for the targeted development of improved variants by mutagenesis. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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