Developing efficient antibacterial nanomaterials has potential across diverse fields, but it requires a deeper understanding of material‐bacteria interactions. In this study, a novel 2D core‐shell MoAlB@MBene structure is synthesized using a mild wet‐chemical etching approach. The growth of E. coli, S. aureus, and B. subtilis bacteria in the presence of MoAlB@MBene decreased in a concentration‐dependent manner, with a prolonged lag phase in the initial 6 h of incubation. Even under dark conditions, MoAlB@MBene triggered the formation of intercellular reactive oxygen species (ROS) and singlet oxygen (1O2) in bacteria, while the bacteria protected themselves by forming biofilm and altering cell morphology. The MoAlB@MBene shows consistent light absorption across the visible range, along with a distinctive UV absorption edge. Two types of band gaps are identified: direct (1.67 eV) and indirect (0.74 eV), which facilitate complex light interactions with MoAlB@MBene. Exposure to simulated white light led to decreased viability rates of E. coli (20.6%), S. aureus (22.9%), and B. subtilis (21.4%). Altogether, the presented study enhances the understanding of bacteria responses in the presence of light‐activated 2D nanomaterials.