[Zurück]

F. Rudroff, C. P. Goncalves, D. Kracher, S. Milker, M. Fink, R. Ludwig, A. Bommarius, M.D. Mihovilovic:
"Mutagenesis-independent, 1000-fold stabilization of class B Flavin monooxygenases in operation";
Vortrag: ProtStab 2018, vilnius; 16.05.2018 - 18.05.2018; in: "The 12th Conference on Protein Stabilization", (2018), S. 1 - 59.

In this study we describe the stabilization of f flavin-dependent monooxygenases under reaction conditions, using an engineered formulation of additives (the natural cofactors NADPH, FAD, and superoxide dismutase and catalase as catalytic antixodans, Figure 1). This way, a 10^3 to 10^4-fold increase of the half-life under reaction conditions 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.