Liquid‐Liquid Phase Separation Modulated Thylakoid–Enzyme Interfacial Coupling for Enhanced Light‐Powered Biocatalysis
Year: 2026
Type: article
Abstract: ABSTRACT Thylakoid (TK) membranes serve as promising photosynthesis platforms for light‐driven biocatalysis, yet their catalytic efficiency is largely restricted by poor coupling between photo‐dependent NADPH regeneration and enzymatic conversion. Herein, we report a facile TK‐anchored enzyme condensate strategy to overcome this bottleneck. Specifically, the UTX‐derived intrinsically disordered region (IDR) fused with target enzymes mediates liquid–liquid phase separation to form enzyme‐rich condensates, which then stably anchor on TK membranes and spatially confine catalysis at the NADPH‐generating membrane interface. Taking carbonyl reductase Sm CR V4 and ene‐reductase Ss ER A314S as model enzymes, the TK–enzyme hybrid condensates exhibit prominently improved catalytic performance, especially under NADPH regeneration‐limited conditions, relative to freely dispersed enzyme systems. The TK–UTX‐ Sm CR V4 hybrid improves the total product yield up to 3.1‐fold, and the TK–UTX‐ Ss ER A314S hybrid reaches a 2.9‐fold higher conversion within 20 min. Coarse‐grained molecular dynamics simulations validate a dynamic multivalent interfacial anchoring mode: the IDR domain mediates stable membrane binding via high‐frequency residue–lipid interactions dominated by short‐range Lennard–Jones forces (with auxiliary electrostatic contributions), while the enzyme domain remains solvent‐exposed. This work establishes a feasible strategy that physically links light‐driven NADPH recycling with enzymatic catalysis, offering a general approach for constructing high‐efficiency artificial photosynthesis systems.
DOI: 10.1002/anie.7079393
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: bronze
OpenAlex ID: W7223475604