Tuberculosis (TB) is the leading cause of death from infectious diseases in the world, affecting more than ten million patients each year. However, multi-drug resistance (MDR-TB) threatens progress achieved in TB care and control, and there are few drugs available to treat MDR-TB. Our overall aim was to identify anti-TB natural products from microbes sourced from unique environments. This thesis presents efforts to achieve an effective approach to identify anti-TB microbial natural products with the combination of one strain many compounds (OSMAC) strategy, NMR fingerprint and principal component analysis. The thesis begins with an introduction of TB and the current anti-TB drugs and candidates. It also covers a review on anti-TB natural products from marine microbe and endophyte origin and analysis of their physicochemical properties using Lipinski’s rule of five as well as the ChemGPS tool. As part of a research program aiming to identify anti-TB microbial constituents, a cell-based screening assay was developed to screen 2562 crude extracts. Among the active hits, 46 actinomyces isolated from marine, desert or Traditional Chinese Medicines were selected for further chemical investigation according to their chemical profiles or anti-TB activities. The results are presented in chapters 2 to 7.