Bacterial biofilm functionalization through Bap amyloid engineering.

Area of Science:

• Microbial Engineering
• Synthetic Biology
• Biomaterials Science

Background:

• Biofilm engineering enables the creation of living structures with programmable functions.
• Amyloidogenic proteins in biofilms can be engineered for self-assembling, functionalized surfaces.
• Facultative amyloids, like Bap, act as adhesins and matrix scaffolds, making them ideal for biofilm modification.

Purpose of the Study:

• To utilize the facultative amyloid-like Bap protein from Staphylococcus aureus for decorating bacterial biofilms and cell surfaces.
• To demonstrate pH-dependent control over the localization of functional tags on biofilms.
• To develop a system for molecular immobilization on bacterial surfaces and biofilm matrices.

Main Methods:

• Engineering the Bap protein from Staphylococcus aureus.
• Utilizing the SpyTag/SpyCatcher system for molecular trapping and immobilization.
• Modulating medium pH to control functional tag localization.
• Functionalizing the cell wall of Gram-positive bacteria with engineered Bap domains.

Main Results:

• The Bap protein successfully decorated extracellular biofilm matrices and bacterial cell surfaces with functional domains.
• Functional tag localization was controllable by altering the pH of the medium.
• A tool for covalent molecular immobilization using Bap and the SpyTag/SpyCatcher system was established.
• Recombinant engineered Bap-amyloid domains functionalized the cell walls of various Gram-positive bacteria.

Conclusions:

• The Bap protein provides a simple and tunable platform for biofilm functionalization.
• This approach allows for programmable modification of bacterial surfaces and matrices.
• The developed system has potential applications in biotechnology and materials science.