The increasing incidence of antibiotic-resistant Mycobacterium tuberculosis infections is a global health threat necessitating the development of new antibiotics. Serine hydrolases (SHs) are a promising class of targets because of their importance for the synthesis of the mycobacterial cell envelope. We screen a library of small molecules containing serine-reactive electrophiles and identify narrow-spectrum inhibitors of M. tuberculosis growth. Using these lead molecules, we perform competitive activity-based protein profiling and identify multiple SH targets, including enzymes with uncharacterized functions. Lipidomic analyses of compound-treated cultures reveal an accumulation of free lipids and a substantial decrease in lipooligosaccharides, linking SH inhibition to defects in cell envelope biogenesis. Mutant analysis reveals a path to resistance via the synthesis of mycocerates, but not through mutations to SH targets. Our results suggest that simultaneous inhibition of multiple SH enzymes is likely to be an effective therapeutic strategy for the treatment of M. tuberculosis infections.

抗生素耐药性结核分枝杆菌感染的发病率不断上升是全球健康威胁，需要开发新的抗生素。丝氨酸水解酶 (SH) 是一类有前途的靶标，因为它们对分枝杆菌细胞包膜的合成很重要。我们筛选了一个包含丝氨酸反应性亲电子试剂的小分子库，并鉴定了结核分枝杆菌的窄谱抑制剂生长。使用这些先导分子，我们进行基于竞争活性的蛋白质分析并识别多个 SH 靶标，包括具有未表征功能的酶。化合物处理培养物的脂质组学分析显示游离脂质的积累和脂寡糖的显着减少，将 SH 抑制与细胞包膜生物发生的缺陷联系起来。突变体分析揭示了一条通过真菌角质酸合成产生耐药性的途径，而不是通过对 SH 靶点的突变。我们的结果表明，同时抑制多种 SH 酶可能是治疗结核分枝杆菌感染的有效治疗策略。