Tuberculosis (TB) remains an insidious scourge of civilization. The causative agent, Mycobacterium tuberculosis (Mtb), is a global health crisis, and TB ranks as the second leading cause of death from an infectious disease worldwide after the human immunodeficiency virus (HIV). In 2020, there were approximately 1.5 million deaths reported from TB and an estimated 10 million new cases of TB. Consequently, there is an extremely urgent need for the discovery of new therapeutics effective against both drug resistant and nondrug resistant strains of TB. The product of the Rv2224c gene, Hip1 (hydrolase important for pathogenesis), is a Mtb cell-wall associated serine hydrolase that plays an important role in the pathogenic strategies of Mtb. It plays a role in cell envelop maintenance and in the dampening of host cell proinflammatory responses. Functional studies indicate that Hip1 is a promising target for drug discovery. In fact, mice infected with a Hip1 mutant strain survive significantly longer than wild-type Mtb-infected mice and exhibit mild lung immunopathology despite high bacterial burdens. Here we present the expression, purification, and cocrystallization of Hip1 with a novel, potent (Ki = 309 + 15 pM), reversible, covalent inhibitor, NS-049-2. The 2.9 Å crystal structure is the first ligand-bound structure of Hip1 and will be useful for the development of novel TB antibiotics and diagnostic assays.