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

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.