Background <p>The <i>Mycobacterium tuberculosis</i> (<i>Mtb</i>) curli pili (MTP) adhesin has been reported as a significant target for TB diagnostic and intervention strategies. The precise contribution of MTP in modulating oxidative phosphorylation (OXPHOS) and central carbon metabolism (CCM) within host epithelial cells is currently unknown.</p> Objectives <p>This study aimed to investigate the impact of MTP in whole cell bioenergetics during early stages of infection.</p> Methods <p>Extracellular flux analysis was used to determine the role of MTP in modulating OXPHOS in A549 epithelial cells. ¹³C-metabolic flux analysis was performed on <i>Mtb mtp</i> proficient/deficient infected A549 epithelial cells to determine whether any specific changes in carbon flux through CCM are induced by the adhesin.</p> Results <p>The absence of MTP led to an increase in OXPHOS in infected A549 cells, thereby increasing ATP synthesis. The Δ<i>mtp</i>-infected A549 cells displayed a similar metabolic profile to the uninfected A549 cells. ¹³C-isotopomer metabolomic analysis of infected A549 cells suggested that MTP plays a role in decreasing glycolytic flux, enhancing flux through the pentose phosphate pathway (PPP), and modulating tricarboxylic acid (TCA) cycle intermediates by increasing flux through succinate.</p> Conclusions <p>The decreased basal respiration and flux through glycolysis and PPP of <i>Mtb</i>-infected A549 cells potentially decreased innate immune responses and production of signalling molecules to interact with immunocytes and activate adaptive immune responses. The similar metabolic profile of Δ<i>mtp</i>-infected A549 cells and uninfected A549 cells suggests that the absence of the adhesin decreases virulence of <i>Mtb</i>. These findings substantiate MTP as an eminent biomarker for TB diagnostics/intervention strategies, and a novel target for vaccine development.</p>