Bacterial pathogens confer virulence by resisting the inhibitory nature of antimicrobial agents during infection. Membrane transporters of the major facilitator superfamily (MFS) mediate the active efflux of structurally distinctive antimicrobial agents. Treatment failures in patients with bacterial infectious disease frequently involve multidrug efflux systems of the MFS. These multidrug transporters are energized by secondary active transport mechanisms involving conformational changes tied to drug transport catalysis to mediate efflux across the bacterial membrane. This review article briefly summarizes recent developments regarding the molecular nature of the MFS multidrug efflux in bacterial pathogens. These active transporter systems represent promising targets for biotechnological approaches to reduce the conditions that foster infections that are recalcitrant to chemotherapy.