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  • Open Access

Enzyme-Driven Phase Separation

Damiano Andreghetti1,2, Alfredo Braunstein1,3,2, Luca Dall'Asta1,2,3, and Andrea Gamba1,2,3,*

  • *Contact author: andrea.gamba@polito.it
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PRX Life 4, 033004 – Published 14 July, 2026

DOI: https://doi.org/10.1103/jmcc-mvrs

Abstract

The formation of polarized signaling domains on cell membranes is a fundamental example of biological pattern formation. While such patterns resemble structures from equilibrium phase separation, they are intrinsically nonequilibrium, driven by energy-consuming enzymatic cycles that switch molecules like phosphoinositides or small GTPases between distinct states. Here, we develop a minimal model of this enzyme-driven phase-ordering process. Starting from microscopic reaction kinetics, we derive a mesoscopic theory that belongs to the class of active model A with a global constraint. This framework yields an explicit mean-field phase diagram and closed-form expressions for key observables, such as interfacial tension, domain fractions, and phase-coexistence boundaries, in terms of kinetic rates. In this context, phase coexistence is controlled by nonequilibrium parameters such as- catalytic rates and enzymatic asymmetry, rather than equilibrium parameters such as saturation concentrations. The resulting phase-separated domains rapidly exchange material with their surroundings. Their maintenance requires a continuous power input determined by enzymatic kinetics. The predicted phenomenology is consistent with experimental observations on reconstituted systems of phosphoinositide and Rab5 membrane patterning. We further study how metastable uniform states decay via nucleation of minority-phase domains and subsequent coarsening, driven by an effective interfacial tension. Using largedeviation theory, we derive the critical nucleation radius under the action of the intrinsic, multiplicative chemical noise. The analytical results are quantitatively confirmed by stochastic simulations of the process. Our work provides a theoretical framework identifying key biochemical parameters controlling active phase separation on membrane scaffolds, offering testable predictions for experiments.

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PRX Life 4, 033004– Published 14 July, 2026
  • Received 7 December 2025
  • Accepted 12 June 2026
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