Structural cracks pose a significant threat to their appearance and strength, potentially leading to catastrophic failures. These cracks facilitate moisture ingress, rendering concrete more permeable and accelerating rust in reinforcement. To address this issue and sustain concrete strength, friendly and long-lasting bacteria called Bacillus subtilis were employed. These bacteria thrive in concrete environments, absorbing moisture, and effectively repairing cracks. In this study, three different concentrations (104, 105, and 106 cells/ml) of B. subtilis were utilized to investigate their impact on mechanical properties, including compression, split tensile, impact strength, and flexural properties, as well as microstructural analysis through Scanning Electron Microscope (SEM) with Energy Dispersive X-Ray Analysis (EDAX) and X-ray Diffraction (XRD). Additionally, serviceability and durability studies were conducted to assess sorptivity and water absorption. From this study, the presence of calcium deposits in SEM images established a significant improvement in compressive strength and fracture healing. This study highlights the promising potential of Bacillus subtilis in enhancing structural integrity and durability.