The cell wall is highly conserved among bacteria and essential to their survival, that iswhy enzymes synthesizing it are a favoured target of antibiotics. Recent studies indicatethat the bacterial cell wall, also called peptidoglycan (PG), is synthesized by two distinctclasses of PG synthases: the bifunctional class A penicillin binding proteins (PBPs) thatpossess both the glycosyltransferase (GTase) and transpeptidase (TPase) activities andthe SEDS (shape, elongation, division, sporulation) GTases with their cognate class B PBPTPases. During division, the PG layer must grow inward to form a septum and bothclasses of PG synthases, i.e. the SEDS protein FtsW and the class A PBP1b, have beenassociated with the septal PG synthesis. It was recently demonstrated that FtsW, with itsassociated TPase, FtsI, purified as FtsWIQLB pentameric protein complex is active toproduce PG in vitro, however, the exact kinetics of the PG synthesis by this complexremains uncharacterized. Additionally, the interactions and crosstalk between these twoclasses of PG synthases remain poorly studied. We aim to determine the dynamics of PGsynthesis by FtsWIQLB complex reconstituted in polymer-supported lipid membranes.Additionally, we will characterize the composition of the PG product using a mass-spectrometry based pipeline. Lastly, by using this platform we will investigate thecrosstalk between the two classes of PG synthases. Overall, this project aims to enhanceour mechanistic understanding of the PG synthesis by the two classes of PG synthasesand their synergistic effects on PG assembly. This new platform can further be appliedfor the identification of novel inhibitors of these enzymes and thus facilitate theidentification of new antibiotics.