WP1 – Flavin-dependent monooxygenases (RUG – ESR1, UNIPV – ESR2, ISKO – ESR3)
The research will focus on improving the rate of catalysis, understanding the molecular basis for formation and stabilisation of the oxidising enzyme intermediate and boosting enzyme robustness. The Fraaije group (RUG – ESR1) will focus on the engineering of flavin-containing monooxygenases (FMOs) from microbial sources. By dedicated enzyme redesign, through creating and screening focused mutant libraries using the unique SeSaM methodology (SSB) and assisted by computational predictions for library design, improved enzymes will be produced and studied. The generated enzymes will be evaluated as biocatalysts for among others the conversion of indole derivatives for dye synthesis (ISKO). The Mattevi group (UNIPV – ESR2) will focus on the production, crystallization and structure elucidation of FMOs. Emphasis will be on the structural elucidation of human FMOs as no structure is available yet for these flavoproteins while they play a crucial role in metabolism of drugs and metabolites. The FMOs will also be studied regarding their mechanistic features. The ESR at ISKO (ESR3) will work on the application of FMOs for the production of indigoid dyes. FMOs made available through the Fraaije group will be used for this. In addition to synthesis of indigoid dyes, ISKO will also focus on i) isolation of dye molecules for vat dyeing bath preparation as well as ii) in-situ dyeing of textile materials.