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bioRxiv · 10.1101/2025.10.18.683236

Dynamic and Catalytic Multiphase Coacervates

Booth, R.·Pei, J.·Shaffer, J.·Keating, C. D.
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Abstract

Subcellular organization is critical for myriad cellular functions, including spatiotemporal regulation of biocatalysts. Liquid-liquid phase separation contributes to intracellular organization by providing microenvironments that sequester biomolecules and control their activities. Although catalytic reactions in multiphase coacervate systems are important for living cells, provide functionality to artificial systems, and may aid understanding of how metabolism first arose, they are not well understood. Here we report an artificial multiphase system in which catalytically active droplets containing three distinct coexisting coacervate phases undergo reorganization in response to pH, enabling control over chemical reactions. Catalysis is performed by an inner, polyhistidine-dense coacervate phase with inherent nonenzymatic activity. We show how phase composition, in addition to pH, affects catalytic activity, and how the addition of enzymes that alter local pH provides a means to regulate composition and catalysis. By exploiting competitive interplay between intramolecular self-association and intermolecular complexation among polymers within multiphase coacervate systems, it was possible to (re)organize and tune the reactivity of dynamic synthetic biological systems beyond what is possible for single-phase coacervates. Such approaches could prove valuable in designing artificial cells and related microreactor systems capable of catalyzing and controlling complex chemical and biochemical processes.

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Booth, R., Pei, J., Shaffer, J., Keating, C. D.. 2025-10-18. Dynamic and Catalytic Multiphase Coacervates. https://doi.org/10.1101/2025.10.18.683236

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