Mapping AMR Through Protein Libraries: Cell Surface Modification and the Role of Dlt and MprF Pathways in Bacillus subtilis

Abstract
This project aims to explore the mechanisms of antimicrobial resistance (AMR) in Bacillus subtilis, focusing on two key pathways: Dlt and MprF. These pathways modify the bacterial surface charge, reducing the binding efficiency of cationic antimicrobial peptides (AMPs). By investigating these pathways through mutagenesis, microfluidics, and single-cell microscopy, the study will characterize changes in surface properties like charge and membrane rigidity that contribute to resistance. The goal is to develop a genotype-phenotype map by generating protein libraries of Dlt proteins, and MprF mutants, linking specific mutations to AMP resistance. This map will provide critical insights into key residues and functional sites responsible for bacterial defence against AMPs. Real-time tracking of AMP binding and membrane penetration, along with various biochemical studies like lipidomics and mass spectrometry, will further clarify how these pathways alter bacterial membrane properties to resist AMP attacks. The knowledge gained from this study can be used to design small-molecule inhibitors targeting these pathways, offering a novel approach to restoring AMP efficacy. This project has the potential to develop innovative antimicrobial therapies that combine AMPs with targeted inhibitors, providing a much-needed alternative to combat the growing global threat of AMR.
BBSRC Priority Areas
Synthetic biology, technology development for biosciences, data-driven biology, understanding the rules of life, transformative technologies, Combatting antimicrobial resistance