Chemoselective reagents for the stabilization and modular coupling of enzymes

• Applicant: Professor Dr. Tom N. Großmann
• Subject Area: Biological and Biomimetic Chemistry ; Biochemistry
• Term: since 2026
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 576988756

The selective chemical modification of expressed proteins is essential for many biotechnological and therapeutic applications such as the generation of antibody-drug conjugates, the labeling of proteins in their biological environment, and for protein immobilization. Surprisingly, covalent modification strategies have rarely been used to enhance the stability and activity of enzymes. In particular, biocatalytic processes, which use enzymes to perform chemical transformations on organic molecules, would greatly benefit from such features. So far, biocatalytic applications have mainly focused on the use of fairly stable enzymes that typically perform one-step reactions. This severely limits their applicability. In cells, different enzymes are often spatially aligned, allowing the substrate to pass from one to the next, enabling efficient ‘cascade’ reactions. Such a setup is also desirable for biocatalytic applications in vitro, but the majority of biocatalytic processes employ an individual enzyme that catalyzes a single-step transformation. Multi-enzyme processes could greatly enhance productivity and efficiency, but general methods required for the modular and spatial organization of enzymes are currently lacking. In the proposed project, a novel approach for the generation of stable enzyme assemblies will be developed than can then be applied to provide more efficient biocatalytic processes. Previously, we have reported a protein ‘clamp’ that covalently modifies and stabilizes enzymes. We now aim to utilize this discovery to go beyond the current state-of-the-art and develop a general stabilization strategy for enzymes which facilitates the defined assembly of biocatalytic modules. Building upon our expertise in bioorganic synthesis as well as protein engineering, the aim is to develop a set of molecular clamps and to generate stable biocatalytic modules. These modules will then be used for the generation of multi-functional biocatalysts for cascade reactions. The proposed project will provide a general method towards robust and efficient biocatalysts that can be applied to provide sustainable production processes.

• DFG Programme: Research Grants
• International Connection: Netherlands
• Cooperation Partners: Dr. Ivana Drienovska; Dr. Francesco Mutti