Oppositely Charged Single Enzyme Nanogels Form Versatile Coacervates for Efficient Enzyme Cascade Catalysis
Abstract
ABSTRACT Enzymatic cascade systems enhance the efficiency of biocatalysis by mimicking natural metabolic pathways. However, conventional platforms face challenges such as ineffective enzyme distribution, uncontrolled stoichiometry, limited substrate accessibility, and a microenvironment that is suboptimal for enzyme performance. Inspired by cellular biomolecular condensates, we here present a design based on oppositely charged single enzyme nanogels (SENs) that phase separate into denser microdroplet coacervates, which thereby serve both as an immobilization scaffold and functional unit. These coacervates, the first fabricated exclusively from oppositely charged nanogels, form under broad environmental conditions and enable specific microenvironments by adjusting SEN ratios to match the requirements of coupled enzymes. Selective substrate enrichment and efficient diffusion of intermediates enhance cascade productivity. The applicability of the platform is demonstrated with pharmaceutically relevant enzymes, highlighting its potential for biosynthetic applications. This work establishes a new paradigm that integrates protein‐polymer engineering with phase separation chemistry to overcome key limitations in cascade enzyme catalysis.
Article Details
Authors (12)
Andoni Rodriguez‐Abetxuko
Department of Biomedical Engineering and Chemical Engineering and Chemistry Institute for Complex Molecular Systems Eindhoven University of Technology Helix, P. O. Box 513 Eindhoven 5600 MB The Netherlands
Nadia A. Erkamp
Neshat Moslehi
Yannick H. A. Leurs
Laboratory of Chemical Biology, Department of Biomedical Engineering
Lars Paffen
Department of Biomedical Engineering and Chemical Engineering and Chemistry Institute for Complex Molecular Systems Eindhoven University of Technology Helix, P. O. Box 513 Eindhoven 5600 MB The Netherlands
Işil Yeşil Gür
Department of Chemical Engineering and Chemistry and Institute for Complex Molecular Systems (ICMS) Eindhoven University of Technology Eindhoven The Netherlands
Jana Stojanović
Department of Chemical Engineering and Chemistry and Institute for Complex Molecular Systems (ICMS) Eindhoven University of Technology Eindhoven The Netherlands
Hugo Brasselet
Department of Biotechnology Delft University of Technology Delft The Netherlands
Laura van der Weel
Department of Biotechnology Delft University of Technology Delft The Netherlands
Ulf Hanefeld
Ilja K. Voets
Self-Organizing Soft Matter, Department of Chemical Engineering and Chemistry & Institute of Complex Molecular Systems
Jan C. M. van Hest
Bio-Organic Chemistry, Departments of Biomedical Engineering and Chemical Engineering and Chemistry, Institute for Complex Molecular Systems