The approval of bedaquiline (BDQ) for the treatment of tuberculosis has generated substantial interest in inhibiting energy metabolism as a therapeutic paradigm. However, it is not known precisely how BDQ triggers cell death in Mycobacterium tuberculosis (Mtb). Using ¹³C isotopomer analysis, we show that BDQ-treated Mtb redirects central carbon metabolism to induce a metabolically vulnerable state susceptible to genetic disruption of glycolysis and gluconeogenesis. Metabolic flux profiles indicate that BDQ-treated Mtb is dependent on glycolysis for ATP production, operates a bifurcated TCA cycle by increasing flux through the glyoxylate shunt, and requires enzymes of the anaplerotic node and methylcitrate cycle. Targeting oxidative phosphorylation (OXPHOS) with BDQ and simultaneously inhibiting substrate level phosphorylation via genetic disruption of glycolysis leads to rapid sterilization. Our findings provide insight into the metabolic mechanism of BDQ-induced cell death and establish a paradigm for the development of combination therapies that target OXPHOS and glycolysis.

苯达喹啉（BDQ）用于治疗肺结核的批准引起了人们对抑制能量代谢作为治疗范例的浓厚兴趣。然而，确切地不清楚BDQ如何触发结核分枝杆菌（Mtb）中的细胞死亡。使用¹³ C同位素分析，我们显示BDQ处理的Mtb重定向中央碳代谢，以诱导易受糖酵解和糖异生的遗传破坏的代谢脆弱状态。代谢通量曲线表明BDQ处理的Mtb它依赖于糖酵解来产生ATP，通过增加通过乙醛酸分流器的通量来操作分叉的TCA循环，并且需要无节肢节和柠檬酸甲酯循环的酶。用BDQ靶向氧化磷酸化（OXPHOS）并同时通过糖酵解的遗传破坏抑制底物水平的磷酸化导致快速灭菌。我们的发现为BDQ诱导的细胞死亡的代谢机制提供了见识，并为针对OXPHOS和糖酵解的联合疗法的开发建立了范例。