Using CO2 as reagent: FDCA and other commodity chemical bioproduction through enzymatic carboxylation

Background: The Leys and Hay groups has been at the forefront of characterizing, understanding and application of the ubiquitous microbial ubiD family of (de)carboxylases. We have demonstrated at proof-of-principle level that furan carboxylic acid can be converted to FDCA using a UbiD enzyme (called HmfF). However, the catalyst properties, yield and reaction conditions are far removed from what is needed to support industrial application. Objectives: We seek to answer following questions: how can we explore the wider ubiD enzyme family to select the best chassis for future studies and directed evolution, aimed at producing high specific activity and stability under relevant industrial conditions? Can HmfF activity be installed in the more tractable fungal UbiDs, that are smaller and oxygen tolerant? How can we further evolve the selected enzyme(s) to match the required properties for industrial use? Can we build upon literature reports describing UbiD activity in supercritical CO2, and operate HmfF under such conditions? What are the most efficient routes to control HmfF activity and isolate the FDCA product? Can we alter substrate profile to include carboxylation of furfural? To what extend can these approaches be applied to other commodity chemical production, either through carboxylation or decarboxylation (for production of hydrocarbons). The student will work in the groups of David Leys and Sam Hay at the Manchester Institute of Biotechnology (MIB) and undertake protein biochemistry, enzymology, structural biology, chemical engineering and computational studies aimed at improved understanding of the ubiD enzymes and their properties. They will explore directed evolution of ubiD as well as perform the necessarily high-throughput analytical chemistry aimed at generating evolved variant with suitable substrate selectivity, stability and oxygen-tolerance amongst other properties. The project falls squarely within TRL1, seeking to transition basic research on UbiD enzymology towards applied research on how to optimize FDCA/commodity chemical production from biomass. It will make use of analytical science, biophysics, catalysis and chemical mechanisms, include computational chemistry and contribute to carbon capture.