Ordered Coimmobilization of Multimeric Enzyme Arrays with Enhanced Biocatalytic Cascade Performance

Area of Science:

• Biotechnology
• Enzyme Engineering
• Synthetic Biology

Background:

• Enzyme coimmobilization mimics natural cascade pathways, offering broad applications.
• Efficient regeneration of expensive sugar donors like UDP-glucose is crucial for biosynthesis.
• Existing methods for enzyme assembly often lack order and efficiency.

Purpose of the Study:

• To develop a strategy for orderly coimmobilizing multiple enzymes.
• To construct an ordered coimmobilization array for ginsenoside Rh2 biosynthesis.
• To enhance the efficiency and stability of biocatalytic cascade reactions.

Main Methods:

• Hierarchically self-assembled multimeric enzymes combined with polyhistidine tag affinity-mediated immobilization.
• Ordered coimmobilization of glycosyltransferase UGT51 mutant and sucrose synthase.
• Comparison with random coimmobilization and free enzyme systems.

Main Results:

• The ordered coimmobilization array significantly boosted immobilization and catalysis efficiency.
• Improved UDP-glucose regeneration, storage stability, and reusability were observed.
• The system demonstrated enhanced performance compared to random coimmobilization and free enzyme systems.

Conclusions:

• The developed strategy enables orderly coimmobilization of multienzymes.
• This approach promises advancements in fabricating enzyme arrays for biocatalysis.
• Nanocomplexes offer synergistic benefits for enhancing biocatalytic cascade performance.