How To Break the Janus Effect of H2O2 in Biocatalysis? Understanding Inactivation Mechanisms To Generate more Robust Enzymes

H2O2, is an attractive oxidant for synthetic chemistry, especially if activated as percarboxylic acid. H2O2, however, is also a potent inactivator of enzymes. Protein engineering efforts to improve enzyme resistance against H2O2 in the past have mostly focused on tedious probabilistic directed evolution approaches. Here we demonstrate that a rational approach combining multiscale MD simulations and Born–Oppenheimer ab initio QM/MM MD simulations is an efficient approach to rapidly identify improved enzyme variants. Thus, the lipase from Penicillium camembertii was redesigned with a single mutation (I260R), leading to drastic improvements in H2O2 resistance while maintaining the catalytic activity. Also the extension of this methodology to other enzymes is demonstrated.