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Beyond Compartmentalization: Deciphering Reaction Kinetics in Liquid-Liquid Phase Separation for Rational

Kang Zhang1, Yuyao Wan1, Xiao Jia1

  • 1School of Life Sciences and Engineering, Sichuan Engineering Research Center for Biomimetic Synthesis of Natural Drugs, Southwest Jiaotong University, Chengdu, Sichuan 610031, P. R. China.

ACS Synthetic Biology
|June 25, 2026
PubMed
Summary
This summary is machine-generated.

Liquid-liquid phase separation (LLPS) organizes biochemical processes for synthetic biology. Understanding LLPS mechanisms is key to optimizing reaction kinetics for advanced applications.

Keywords:
bioreactionsenrichment effectenzyme activity modulationliquid−liquid phase separationmicroenvironmentreaction kineticsreaction–diffusion coupling

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Area of Science:

  • Biochemistry
  • Synthetic Biology
  • Biotechnology

Background:

  • Liquid-liquid phase separation (LLPS) enables membrane-less compartmentalization for biochemical processes.
  • LLPS applications in synthetic biology, metabolic engineering, and artificial cells offer novel solutions.
  • Current limitations in LLPS applications stem from an incomplete understanding of its impact on reaction kinetics.

Purpose of the Study:

  • To provide a comprehensive review of LLPS mechanisms governing reaction kinetics.
  • To integrate protein and non-protein mediated LLPS.
  • To establish mechanistic insights and design guidelines for LLPS-driven synthetic biology.

Main Methods:

  • Systematic dissection of LLPS's role in orchestrating reaction kinetics.
  • Analysis of mechanisms including reactant concentration, reaction-diffusion coupling, microenvironment engineering, and enzyme activity modulation.
  • Review of thermodynamic foundations and classifications of LLPS.

Main Results:

  • LLPS dictates bioreaction outcomes through various kinetic regulatory mechanisms.
  • Identified key factors: reactant concentration, reaction-diffusion coupling, microenvironment, and enzyme activity.
  • Summarized diverse applications in biocatalysis, metabolic engineering, diagnostics, therapeutics, and artificial cell construction.

Conclusions:

  • LLPS is a powerful tool for controlling biochemical processes in synthetic biology.
  • Bridging the knowledge gap in LLPS mechanistic understanding is crucial for rational design.
  • This review provides essential insights for developing next-generation LLPS-driven technologies.