Designed Protein Cages as Scaffolds for Building Multienzyme Materials

Area of Science:

• Biochemistry
• Synthetic Biology
• Protein Engineering

Background:

• Enzyme function is influenced by spatial organization.
• Developing methods to control enzyme arrangement is crucial for biocatalysis and synthetic biology.

Purpose of the Study:

• To create a novel platform for organizing multiple enzymes using designer protein cages and sortase-based ligation.
• To demonstrate enhanced enzymatic activity through controlled enzyme spatial arrangement.

Main Methods:

• Utilized a 24-subunit designer protein cage scaffold with outwardly exposed termini.
• Employed sortase-mediated covalent attachment of cellulase enzymes, tagged with polyglycine, to sort-tag modified cage subunits.
• Assessed cellulose degradation activity using the engineered enzyme-cage construct.

Main Results:

• Successfully attached multiple cellulase enzymes to the protein cage scaffold with high reactivity and copy number.
• Demonstrated significantly enhanced cellulose degradation activity compared to free enzymes or singly modified cages.
• Validated the efficacy of sortase-based enzymatic attachments for enzyme organization.

Conclusions:

• The developed platform enables precise spatial organization of enzymes.
• Engineered enzyme-cage constructs exhibit superior catalytic performance.
• This strategy offers broad applicability in enzymatic materials and synthetic biology.