This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Antimicrobial therapy was been an integral part of healthcare since its inception;however, antibiotic resistance is becoming increasingly dangerous as it claims numerous lives every year. The number of deaths in the United States attributable to antibiotic resistance is projected to surpass deaths due to HIV/AIDS;therefore, a way to combat or circumvent antibiotic resistance is critical to advancement in antimicrobial therapy. Anhydromuramic acid kinase (AnmK) is a peptidoglycan recycling enzyme that has recently been isolated, expressed and purified. Inhibiting the enzymes that control peptidoglycan recycling may hinder bacterial growth in a novel way. The goals of this project are multifacted and include both native and substrate-bound crystal structures of AnmK, the synthesis of its substrate (anhydromuramic acid) and potential substrate mimics (anhydroglucosamine, levoglucosan) and a full kinetic characterization of the enzyme. This enzyme has novel activity as it is a simultaneous hydrolase and kinase, so structural characterization of the enzyme with a product-like complex would also be desirable in order complete the picture of structural changes that may occur upon binding and release of substrates. Currently, there is not a structure of AnmK published in the literature nor is there an enzyme mechanism known for this type of kinase. Structural work with AnmK could reveal a novel family of kinases, help discover the solution to a new enzyme mechanism, and further advance the understanding of enzymes involved in bacterial cell wall biosynthesis so that this knowledge can be utilized to combat emerging antibiotic resistance. Future plans also include mutagenesis of potential key residues to investigate the effect on enzyme kinetics and structure-based inhibitor design.