Mutagenesis‐Independent Stabilization of Class B Flavin Monooxygenases in Operation. Issue 12 (19th May 2017)

• Record Type: Journal Article
• Authors: Goncalves, Leticia C. P.; Kracher, Daniel; Milker, Sofia; Fink, Michael J.; Rudroff, Florian; Ludwig, Roland; Bommarius, Andreas S.; Mihovilovic, Marko D.
• Is Part Of: Advanced synthesis & catalysis. Volume 359:Issue 12(2017)
• Page Start: 2121
• Page End: 2131
• Publication Date: 2017-05-19
• Subjects: biocatalysis -- cofactors -- enzyme stabilization -- oxygenation -- reactive oxygen species; Catalysis -- Periodicals; Organic compounds -- Synthesis -- Periodicals; Chemistry -- Periodicals; Chemistry, Technical -- Periodicals; Chemistry -- Periodicals; Catalysis -- Periodicals; Technology, Pharmaceutical -- Periodicals; 547.2
• DOI: 10.1002/adsc.201700585
• Languages: English
• ISSNs: 1615-4150

Abstract

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 10 3 ‐ to 10 4 ‐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 :