Enzyme catalysis in biomembranes - from biotechnological application to biomedical research

Membrane-bound enzymes are involved in essential biological processes, such as metabolising difficult-to-convert substrates and activating fatty acids. Among them, membrane-bound oxidoreductases have high potential for biotechnological application and are of biomedical relevance. However, their requirement for a membrane scaffold makes their functional characterization and engineering very challenging. This BioTechMed YRG aims to characterize and engineer nonheme diiron enzymes in a cell-free system. We will investigate industrially relevant alkane monooxygenases, used to synthesize monomers for biobased polymers, and fatty acid hydroxylases, associated with deficiencies that lead to neurodegenerative diseases. Functional characterization of membrane enzymes will be achieved by combining reconstitution in nanodiscs, in vitro functional studies, and the development of high-throughput assays for protein engineering. Reconstitution of membrane enzymes into the robust nanodisc system will enable their study in a physiologically relevant reaction environment, free from interference from host enzymes. The results of the functional characterization will advance our mechanistic understanding of these enzymes. The gained knowledge can then be applied to improve biotechnologically relevant enzymes for the sustainable synthesis of renewable chemicals on the one hand and to investigate the effects of mutations in medicinally relevant enzymes on the other. 

Membrane-bound oxidoreductases have high potential for biotechnological application and are of biomedical relevance. The dependency of these enzymes on a membrane scaffold represents a major obstacle for their functional characterization. This BioTechMed YRG aims to develop a toolbox for the analysis of structure-function relationships of these enzymes and advance our mechanistic understanding of membrane-bound oxidoreductases.

Project funding

Funding EUR 500.000,-