Creating high-performance artificial designer enzymes by unlocking design principles of their long-distance interactions

• Applicant: Dr. Minh Nguyen Trung
• Subject Area: Biological and Biomimetic Chemistry
• Term: since 2024
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 553951872

Biocatalysis represents a pivotal pillar of green, sustainable, contemporary synthetic chemistry providing an efficient tool to synthesize and modify intricate (bio)molecules. Despite the enormous efficiency of naturally evolved enzymes, they lack the versatility of the organic chemistry toolbox. Combining the best of both worlds, “designer enzymes” have been a growing field in recent years. Artificial designer enzymes enable mild, resource-efficient reaction conditions with a diverse array of chemical transformations. A bottleneck in the development of new designer enzymes is their inferior efficiency compared to natural enzymes, despite extensive optimization campaigns around the proteins’ catalytic pockets. The project proposed here-in aims to overcome this performance gap by exploiting structure-reactivity relationships, particularly with regards to long-distance allosteric interactions, which have not been explored in artificial enzymes before. Utilizing a multidisciplinary approach involving directed evolution, enzymology and protein-NMR spectroscopy, the project will focus on the designer enzyme LmrR_pAF (lactococcus multidrug resistance regulator with catalytic p-aminophenyl alanine side chains), which can catalyze several reactions through iminium-based organocatalysis. By targeting the dynamic elements of the LmrR_pAF scaffold for directed evolution and subsequent characterization of the underlying structural improvements the project will unravel the intricate interplay between enzyme performance and its structure and dynamics. The overarching goal is to establish universally applicable design principles for LmrR-derived designer enzymes to bring their performance to the same level as naturally evolved enzymes. Such principles will then facilitate the development of more innovative artificial designer enzymes and subsequently their use in chemoenzymatic synthesis.

• DFG Programme: WBP Fellowship
• International Connection: Netherlands
• Host: Professor Dr. Gerard Roelfes