Project Title: A structure-based approached to biocatalysis
Supervisor: Prof. Andrea Mattevi, University of Pavia, Italy
Objectives: There are several urgent issues to be resolved for preparing industrial applicable flavin-dependent monooxygenases. This project will focus on improving the rate of catalysis, understanding the molecular basis for formation and stabilisation of the oxidising enzyme intermediate in Flavin Monooxygenases (FMO) and boosting enzyme robustness by establishing (1) efficient expression systems for soluble and membrane-bound eukaryotic FMOs; (2) protocols for protein purification as to provide the material for structural and biochemical studies; and, (3) providing structure determination of enzymes and enzyme-ligand complexes including analysis of valuable mutants.
Methodology: The work will involve the production, crystallization and structure elucidation of human FMOs, as no structure is available yet for these flavoproteins, in spite of them playing a crucial role in metabolism of drugs and metabolites. The FMOs will also be studied regarding their mechanistic features.
Expected Results: (i) Expressed and purified eukaryotic FMOs; (ii) structure-based mechanisms for enzyme function and substrate selectivity.