Development and characterization of LipoCatch: a bacterial lipoprotein-based biomaterial that self-assembles into...

Area of Science:

• Biomaterials Engineering
• Synthetic Biology
• Nanotechnology

Background:

• Genetically encoded nanomaterials offer precise control over molecular composition and function.
• The bacterial lipidation pathway has not been previously utilized for constructing hybrid protein/lipid nanostructures.

Purpose of the Study:

• To design and characterize a versatile bacterial lipoprotein, LipoCatch, for modular nanostructure formation.
• To establish a genetically encoded platform for creating customizable bacterial lipoprotein-based biomaterials.

Main Methods:

• Engineered a bacterial lipoprotein (LipoCatch) by appending a signal peptide for lipidation.
• Biosynthetically produced LipoCatch in *E. coli* and purified it.
• Utilized LC-MS, SEC, DLS, and TEM for characterization.
• Investigated two functionalization strategies: SpyCatcher/SpyTag covalent modification and phospholipid incorporation.

Main Results:

• Confirmed site-specific lipidation and formation of 13 nm nanoparticles.
• Demonstrated LipoCatch's ability to undergo covalent modification and non-covalent phospholipid incorporation.
• Showcased enhanced stability of LipoCatch and hybrid nanostructures, including tolerance to lyophilization compared to liposomes.

Conclusions:

• Established LipoCatch as an engineered bacterial lipoprotein platform for customizable biomaterial development.
• Highlighted the potential of LipoCatch for creating stable, hybrid protein/lipid nanostructures.
• Provided a proof-of-principle for utilizing bacterial lipidation pathways in nanomaterial synthesis.