Sauer Group, Metabolomics, Antibiotics, More of action
Identifying the target of the antimicrobial peptide Sublancin
Tutor: Dr. Ruben Mars ()
Duration: 3 months
Background
Knowing the mode of action of naturally occurring antimicrobial compounds can help to design new targeted antibiotics. Since metabolomics provides a direct and global read-out of the state of the cell, it could help to unravel the mode of action of antimicrobial compounds. Sublancin is a bacteriocin produced by the Gram-positive soil bacterium Bacillus subtilis 168. Sublancin is capable of killing several species of Gram-positive bacteria, such as Staphylococcus aureus, including methicillin resistant S. aureus (MRSA). Sublancin is post-translationally modified with two internal disulfide bridges, making it extraordinarily stable. In a second post-translational modification, a glucose molecule is added to sublancin. The role of this glucose and the mode of action of sublancin is not known, but central metabolism and the phosphotransferase system may be involved.
Your project
In this semester project you will apply untargeted metabolomics technologies to screen for clues to the mode of action of sublancin. The project consists of the following parts:
1) Perform time-course metabolite sampling on multiple sublancin-sensitive and resistant strains to look for conserved patterns in intra- and extracellular metabolites.
2) Compare the identified patterns with experimental outcome for antimicrobials with a known mode of action.
3) Quantifying the observed metabolomic phenotypes with targeted metabolomics.
4) Try to test sublancin uptake in multiple genetic backgrounds of B. subtilis.
The project will be a combination of experimental design, lab work, data analysis, and hypothesis generation, and will have a strong practical component.