Transforming Crowded Coacervates into Multi-Compartmental Hybrid Microreactors for Practical Enzymatic Catalysis.

Area of Science:

• Synthetic biology and bioengineering
• Biomimetic materials science
• Chemical engineering

Background:

• Designing artificial protocells with complex organization and functionality is a key challenge in synthetic biology.
• Existing methods often struggle to achieve robust, stable, and highly functional protocell architectures.
• There is a need for versatile platforms to create advanced microreactors for in vitro applications.

Purpose of the Study:

• To develop a versatile Pickering emulsion-based encapsulation approach for creating robust multicompartmental hybrid microreactors.
• To integrate hierarchical compartmentalization, molecular crowding, selective permeability, and mechanical stability into artificial protocells.
• To demonstrate the enhanced catalytic efficiency and stability of these hybrid microreactors for biocatalysis and cascade reactions.

Main Methods:

• Utilizing Pickering emulsions to encapsulate membraneless coacervate compartments.
• Creating multicompartmental hybrid microreactors with spatially sequestered catalytic species.
• Characterizing the structural features, including molecular crowding and confinement effects.
• Testing the performance of the microreactors in enzymatic kinetic resolution and chemo-enzymatic cascade reactions.

Main Results:

• Achieved a 1.9-9.2 fold enhancement in catalytic activity for lipase-driven reactions.
• Demonstrated strengthened thermostability up to 100 °C and long-term durability (1600 h).
• Successfully showcased superior performance in complex chemo-enzymatic and multi-enzymatic cascade reactions.
• Identified macromolecular crowding and confinement as key factors promoting catalytic functions.

Conclusions:

• The Pickering emulsion-based approach provides a versatile platform for constructing advanced artificial protocells.
• The resulting hybrid microreactors exhibit enhanced stability, compartmentalization, and catalytic efficiency.
• This work offers a promising strategy for designing functional microreactors in synthetic biology and bioengineering.