<p>Concrete is one of the most essential construction materials worldwide due to its universal availability and strong performance; however, it has significant limitations. One of the primary limitations of concrete is that it may develop microcracks for several reasons, including drying shrinkage, heavy loads, temperature fluctuations, and earthquakes. Conventional surface treatments, including epoxies, silanes, and pore-blocking agents, have been studied to enhance the durability of concrete. However, these methods are debated due to concerns over their long-term efficiency, high maintenance costs, accessibility, and environmental impact. Using microorganisms in concrete is becoming increasingly popular as a practical approach to increasing its durability and sustainability. This study aims to compare the outcomes of an experimental examination of the influence of nanomaterials produced by bacteria in concrete with those of conventional concrete on multiple concrete parameters. This research used four different proportions of bacteria (0.5, 1.5, 3.5, and 5%) and nutrients (0.25, 0.5, 0.75, and 1%) for the cement weight, respectively. At various time intervals, the samples were tested for their compressive strength, water absorption, chloride permeability, and microstructural characteristics using EDX, SEM, and XRD. The results showed that adding bacteria to concrete made it much stronger, more durable, and less porous and permeable. The most significant improvement occurred at a proportion of 1.5% bacteria, along with 0.5% calcium lactate in the cement weight, which led to a 31.3% increase in compressive strength, a 31.9% reduction in chloride permeability, a 62.3% decrease in water absorption, and a 32.8% w/w calcium content compared to conventional concrete. Furthermore, bacterial-incorporated concrete exhibits higher compressive strength, reduced water absorption, and lower chloride permeability compared to the control specimens.</p>