Mycobacterium tuberculosis is the causative agent of human tuberculosis, which remains the major cause of death from a single infectious agent, killing nearly 1.7 million people each year and infecting around one-third of the entire human population. Tuberculosis has increased in incidence in recent years due to co-infection with HIV and the appearance of multi-drug resistance, which presents an increasing threat worldwide. In our efforts to discover new treatments for tuberculosis we selected for our target the Mycobacterium cell wall synthesis enzyme PimA. PimA has been demonstrated to be an essential enzyme in mycobacteria, both by saturation mutagenesis studies and construction of bacteria containing conditional knockouts. Importantly, the synthetic step catalysed by PimA does not occur in humans, which suggests that a high degree of selective toxicity may be attainable by inhibitors. We screened our in-house fragment library by NMR spectroscopy against PimA. The fragment hits derived from the initial screen were subsequently validated using surface plasmon resonance. A set of compounds with sub-millimolar affinity were identified and subsequently shown to have commensurate inhibitory activity in a PimA enzymatic assay. The success of our fragment-based screen supports the contention that PimA is a tractable drug target for the future development of novel tuberculosis therapies. Moreover, the relatively favourable ligand efficiencies of the top ranking compounds encourage their further optimization towards the establishment of a lead compound.