Mutations Increasing Cofactor Affinity, Improve Stability and Activity of a Baeyer–Villiger Monooxygenase
Hamid R. Mansouri, Oriol Gracia Carmona, Julia Jodlbauer, Lorenz Schweiger, M. Fink, Erik Breslmayr, Christophe V. F. P. Laurent, Saima Feroz, Letícia Christina Pires Gonçalves, Daniela V. Rial, Marko D. Mihovilovič, Andreas S. Bommarius, Roland Ludwig, Chris Oostenbrink, Florian Rudroff
Abstract
) and kinetic stability (8-fold). Our analysis revealed a crucial position in the cofactor binding domain, responsible for an 11-fold increase in affinity to the flavin cofactor, and explained using MD simulations. This gain in affinity was compatible with other mutations. While our study focused on a particular model enzyme, previous studies indicate that these findings are plausibly applicable to other BVMOs, and possibly to other flavin-dependent monooxygenases. These new design principles can inform the development of industrially robust, flavin-dependent biocatalysts for various oxidations.