Rational assembly of multi-enzyme cascades: A paradigm shift from stochastic immobilization to precision assembly...

Area of Science:

• Biocatalysis and green chemistry.
• Enzyme engineering and synthetic biology.

Background:

• Multi-enzyme cascade catalysis mimics natural pathways for efficient synthesis.
• Current systems face challenges in inter-enzyme synergy and operational stability.
• Precise spatial organization is key to overcoming these limitations.

Purpose of the Study:

• To systematically review multi-enzyme cascade reaction types and assembly materials.
• To examine enzyme assembly strategies and engineering methodologies.
• To introduce "ordered assembly" as a novel paradigm for advanced multi-enzyme systems.

Main Methods:

• Categorization of cascade reactions into five modular types (linear, parallel, orthogonal, cyclic, triangular).
• Classification of assembly materials into non-biological (polymers, inorganic) and biological (polysaccharides, nucleic acids, proteins, organisms).
• Systematic literature review of enzyme assembly strategies and engineering approaches.

Main Results:

• Diverse enzyme assembly strategies and state-of-the-art engineering methods are examined.
• "Ordered assembly" is proposed as a novel paradigm for constructing multi-enzyme systems.
• Design principles for customized assembly strategies based on cascade modules are elaborated.

Conclusions:

• Multi-enzyme assembly technology holds significant potential for biocatalysis and sustainable fine chemical manufacturing.
• Further development of ordered assembly strategies is crucial for overcoming current bottlenecks.
• This review provides a theoretical foundation for advancing multi-enzyme assembly applications.