ThermaCORE: Thermus thermophilus Accelerated Co-evolution of Robust Enzymes for industrial applications

"Applied biocatalysis faces a critical limitation: insufficient robust enzymes capable of enduring harsh industrial conditions, impeding circular economy development from waste materials. Current directed evolution methods are slow to screen large mutant libraries (10⁶-10¹²) and encounter significant stability-activity trade-offs where activity enhancement destabilise enzymes.This interdisciplinary project leverages Thermus thermophilus, a thermophilic bacterium, as a living factory for directed evolution. Producing enzymes within this thermostable organism enables selection for inherently stable enzymes. The approach targets engineering robust enzymes for plastic degradation (PET, PBS, PLA), where high temperatures are essential for efficient breakdown and recycling.The methodology employs comprehensive Design-Build-Test-Learn cycles combining cutting-edge biotechnology with computational approaches. Microfluidic droplet technology enables rapid, high-throughput screening of millions of enzyme variants through fluorometric/colorimetric assays. Deep mutational scanning on sorted variants provides detailed sequence-function relationship insights by revealing effects of thousands of amino-acid substitutions on stability and activity. Experimental deep mutational scanning data integrates with machine learning models to approximate protein fitness landscapes and design ""smart libraries"" for subsequent evolution rounds.This fellowship will develop my expertise across biotechnology, computational biology, and biocatalyst engineering, fostering interdisciplinary skills essential for modern R&I careers. The project benefits the host organization by strenghting its capacity in emerging biotechnologies, and biocatalyst discovery that contributes to European leadership in sustainable bioeconomy solutions, and delivers robust biocatalysts that accelerate plastic recycling, reducing environmental impact while creating economic value in the circular economy sector."