Enzymes in the D-Alanine Branch of Bacterial Cell Wall Peptidoglycan Assembly

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This renewal proposal deals with the enzymes involved in providing and incorporating D-alanine (D-ala) and the dipeptide D-Ala-D-Ala into the peptoglycan layer of cell walls in bacteria. Three enzymes comprise the D-Alanine branch, alanine racemase, D-Ala-D-Ala ligase and the D-Ala-D-Ala adding enzyme. In the past grant period (s) all three enzymes from E.coli and Salmonella typhimurium were purified to homogeneity for the first time. In the new grant period will be devoted to the study of the mechanism, structure/function of each of the genes and the encolded enzymes. In addition, the first enzyme in the peptoglycan pathway, the UDP-Nacetylglucosamine enolpyruvyltransferase will be purified and characterized. The results will greatly enchance our understanding of hw bacteria grow and build their cell walls. They may also lead to new strategies for designing effective chemotherapeutic agents against bacterial infections.