Professor Roberto A. Chica - Manchester Institute of Biotechnology

Professor Roberto A. Chica 

"Building Catalysis from Scratch: Scaffold Customization and Multistate Reaction Optimization in De Novo Enzyme Design"

Natural enzymes have evolved to catalyze reactions with remarkable efficiency by precisely positioning catalytic groups while retaining the flexibility to reshape their active sites for stabilization of multiple intermediates along a reaction coordinate. Yet, de novo enzyme design has struggled to reproduce these features, limiting the catalytic efficiency of designed biocatalysts. In this presentation, I will describe our development of computational methods that recapitulate these principles of natural enzyme catalysis and how we used them to build efficient enzymes from scratch. First, I will introduce CANVAS, a workflow that customizes minimal de novo TIM barrels with tailored structural lids that precisely position catalytic residues and create functional active sites. In a single round of design, CANVAS produced Kemp elimination catalysts with catalytic efficiencies up to 21,000 M⁻¹ s⁻¹, 10–100-fold higher than previous de novo designs. I will then describe multistate enzyme design, which optimizes active sites across an entire reaction coordinate rather than a single transition state. Applied to a de novo Morita–Baylis–Hillmanase, this approach yielded biocatalysts with up to 100-fold higher catalytic efficiency than conventional single-state design, with the most active variant surpassing an extensively optimized enzyme in both efficiency and enantioselectivity. Together, these approaches provide a blueprint for designing tailored enzymes for complex, multistep reactions by combining the precise catalytic geometry and adaptive flexibility that make natural enzymes so powerful.