Engineering artificial metalloenzymes for detoxification of organophosphorous nerve agents

The objective of this project is to engineer new artificial metalloenzymes for the degradation of persistent organophosphorous (OP) neurotoxics. We will focus on the development of original directed evolution strategies to find new proteins able to reduce toxicity of chosen OP molecules used as models to target hydrolysis and/or oxidation activities. A first directed evolution approach will be explored to change the catalytic properties of a natural human metalloenzyme for which the metal is directly coordinated by the protein. Based on a structural comparison, a diverse library of protein variants will be designed. Catalytic activities will be screened by innovative high throughput screening assays on OP surrogates and toxics. In a second approach, biohybrid metalloenzymes will be obtained by covalent grafting of synthetic organometallic complexes on a protein scaffold. An alphaRep issued from a new family of modular artificial repeat proteins by will be chosen for its high stability (pH, T, solvent).