Wall teichoic acid (WTA) is a Gram-positive bacterial cell wall polymer with roles in host cell adherence and coordination of essential physiological processes including cell division and peptidoglycan synthesis. Importantly, the biogenesis of WTA and peptidoglycan shares a limited pool of lipid precursors that must be continuously recycled for growth. As such, the WTA biogenesis pathway presents a subset of essential enzymes that can potentially be inhibited to treat Gram-positive bacterial infections, such as those arising from methicillin-resistant Staphylococcus aureus. To guide drug discovery and optimization of novel compounds targeting WTA biogenesis, we have selected two key S. aureus WTA biosynthetic enzymes for structural characterization by cryo-electron microscopy. Furthermore, structures of these targets are anticipated to provide novel mechanistic insights into related classes of proteins. Robust procedures for overexpression, purification and preparation of these drug targets for cryo-EM analysis have been established, and preliminary structures have been generated.