Mutagenesis‐Independent Stabilization of Class B Flavin Monooxygenases in Operation

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‐...

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Bibliographic Details
Published in:Advanced synthesis & catalysis 2017-06, Vol.359 (12), p.2121-2131
Main Authors: Goncalves, Leticia C. P., Kracher, Daniel, Milker, Sofia, Fink, Michael J., Rudroff, Florian, Ludwig, Roland, Bommarius, Andreas S., Mihovilovic, Marko D.
Format: Article
Language:English
Subjects:
Additives
Antioxidants
biocatalysis
Biotechnology
Catalase
Catalysis
cofactors
enzyme stabilization
Enzymes
Fragility
Metabolism
Mutagenesis
oxygenation
reactive oxygen species
Stabilization
Sulfur
Superoxide dismutase
Technology utilization
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Summary: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.
ISSN:1615-4150
1615-4169
DOI:10.1002/adsc.201700585