Bottom-Up Creation of an Enzyme for the Morita-Baylis-Hillman Reaction

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The combination of computational design and directed evolution is an attractive strategy to create enzymes with new function, that to date has delivered de novo biocatalysts for a small set of well-studied transformations. To establish 'bottom up' enzyme design as a reliable source of biocatalysts for practical applications, it is essential that we move beyond the realm of simple model reactions to demanding chemical transformations with complex mechanisms and high energy barriers. In this proposal, we will combine state-of-the-art methods in computational design, laboratory evolution and genetic code expansion to create efficient and selective enzymes for a bimolecular Morita-Baylis-Hillman (MBH) reaction, a synthetically valuable carbon-carbon bond forming reaction not observed in Nature. Successful implementation of the research program will expand the repertoire of biocatalysts for C-C bond formations and will highlight 'bottom-up' design as a viable strategy for creating enzymes for complex multi-step processes. The research will be carried out in close collaboration with GlaxoSmithKline (GSK) and aligns perfectly with the BBSRC strategic priorities in Industrial Biotechnology. This is a highly interdisciplinary project at the cutting edge of enzyme design and engineering research, offering diverse training opportunities for a post-graduate student in a state-of-the-art multidisciplinary setting. The student will gain broad expertise in organic synthesis, molecular and structural biology, protein expression and purification, laboratory evolution, biochemical assays and computational biology. Combined, the training program will equip the student with the necessary skills to pursue a future career in the chemical industry.