This research investigated the impact of calcium carbonate precipitation induced by the activity of four microbial strains known as Sporosarcina pasteurii, Bacillus megaterium, Sporosarcina ureae, and Bacillus licheniformis at cell concentrations of 107 cells/ml on lightweight fiber concrete (LWFC). Three mixed designs with different fiber percentages (0%, 0.5%, and 1%) were considered for evaluation of the effects of these bacteria and polypropylene fiber on the mechanical properties and durability of LWC, which contained Leca aggregate. The experimental results indicated that the simultaneous addition of bacteria and fiber was effective and improved compressive, tensile, and flexural strength. It also reduced concrete water absorption and permeability in the specimens. In specimens with B. licheniformis and 1% fiber, a maximum increase of approximately 32.65% was observed in tensile strength and a 28.39% decrease was observed in water absorption with respect to the control specimens. A comparison of the bacterial specimens revealed that B. licheniformis and B. megateterium performed better in the induction of calcium carbonate precipitation and distribution of fiber in the concrete matrix than S. pasteurii and S. ureae to reduce the balling phenomenon. In addition, SEM-EDX and XRD analyses were conducted to verify the formation of calcium carbonate in the concrete matrix and characterize the crystalline phases of the calcite.