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C. P. Goncalves, S. Milker, D. Kracher, M. Fink, F. Rudroff, R. Ludwig, A. Bommarius, M.D. Mihovilovic:
"More than 1000-fold stabilization of class B flavin monooxygenases in operandi without mutagenesis";
Advanced Synthesis & Catalysis, 359 (2017), 2121 - 2131.

This paper describesthe stabilizationof flavin-dependent monooxygenases under reaction conditions,using an engineered formulation of addi-tives (the natural cofactors NADPH andFAD,and superoxide dismutase and catalase as catalytic anti-oxidants). This way,a103-to104-foldincrease of the half-life was reached without resource-intensive di-rected evolution or structure-dependent protein en-gineering methods. Thestabilized enzymes are highly valuedfor theirsynthetic potential in biotech-nology and medicinalchemistry (enantioselective sulfur, nitrogen and Baeyer-Villiger oxidations;oxi-dative human metabolism), but widespread applica-tion was so far hindered by their notoriousfragility. Our technology immediately enables their use,does not require structural knowledge of the biocatalyst, and creates astrong basis for the targeted develop-ment of improvedvariants by mutagenesis.

biocatalysis;cofactors;enzyme stabiliza-tion;oxygenation;reactive oxygen species

http://dx.doi.org/10.1002/adsc.201700585

http://publik.tuwien.ac.at/files/publik_266584.pdf