Mycobacterium tuberculosis is the leading cause of bacterial infectious disease deaths worldwide. Drug-resistant strains (MDR and XDR-TB) are an emerging problem and could lead to significant issues in the US because over half of the cases are in foreign born persons. Biotin biosynthesis has recently been identified as an attractive target for the development of new ant tubercular agents. In this proposal we describe the design, development, and optimization of a novel HTS-assay for BioA, which catalyzes the second step of biotin biosynthesis in Mtb. The objectives of this application are to screen BioA using our novel HTS assay and then perform secondary and tertiary screening of identified hits. In the first specific aim, we will prepare chemical and biochemical reagents for the primary and secondary assays. In the second specific aim, we will develop and screen hits using an orthogonal LC-MS based assay. In the third specific aim, we will perform tertiary screening using mutant strains of M. smegmatis that over- and under-express BioA to access biological activity and characterize on-target specificity.
. Tuberculosis (TB) caused by the slow growing bacillus Mycobacterium tuberculosis (Mtb) is the leading cause of infectious disease mortality in the world by a bacterial pathogen. M. tuberculosis and other atypical mycobacteria are now classified as opportunistic infections of AIDS patients. The proposed research is expected to identify new compounds that could be developed into potential new antitubercular agents. These compound could also be useful to understand the importance of biotin metabolism in Mtb.
|Park, Sae Woong; Casalena, Dominick E; Wilson, Daniel J et al. (2015) Target-based identification of whole-cell active inhibitors of biotin biosynthesis in Mycobacterium tuberculosis. Chem Biol 22:76-86|