Peter-Leon Hagedoorn’s group at Delft University of Technology, focuses on the elucidation of reaction mechanisms of (metallo-)enzymes containing heme, Fe-S clusters and/or W/Mo cofactors. We use microsecond timescale rapid mixing techniques, unique in the world, employed in a continuous flow UV-vis spectrophotometric device called Nanospec, and a rapid freeze hyperquench device called MHQ.
These techniques are circa 100 times faster than commercial instruments, and both techniques have been very powerful in the discovery of novel transient intermediates in the heme enzyme chlorite dismutase and Cu2+ binding to ATCUN/NTS motif peptides. In recent years Hagedoorn ventured together with prof. Hanefeld into enzyme immobilization and flow biocatalysis using Mn2+ dependent aldolases and hydroxynitrile lyases as well as thermophilic glycosyl transferases and hydratases (including FeS cluster containing enzymes).
Within the W-BioCat project, our focus is on the recombinant expression and characterization of W-containing aldehyde oxidoreductases (AORs). Heterologous expression of W-enzymes is challenging due to the requirement of an efficient W-cofactor biosynthesis pathway. The W-BioCat strains developed in this project will enable expression of new W-enzymes from genetic databases, and facilitate production of new engineered W-enzymes.
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