| Bacteria propagation requires precise control of cell wall synthesis driven by concerted enzyme activities. Peptidoglycan, the main component of the cell wall, serves to dictate the morphology of nearly all bacteria and protect them from environmental stress. In this document, I present the use of molecular probes, microscopy, quantitative analysis and genetic engineering to elucidate peptidoglycan biosynthesis during cell division. My collaborators and I reported the direct visualization of septal peptidoglycan formation, with high spatial and temporal resolution, during Bacillus subtilis septation. We showed how peptidoglycan synthesis enzymes move in concert to drive division septum formation. Next, I report the design and synthesis of novel probes for visualizing peptidoglycan formation in real-time. These new probes enable, for the first time, time-lapse monitoring of peptidoglycan biosynthesis in live cells as well as high-throughput transpeptidase assays in vitro. Finally, I provide physical and photochemical characterization of the probes used in my research. This information could serve as a guideline for other researchers to utilize the probes, maximizing their application in various research and advancing our understanding of peptidoglycan biosynthesis. |
