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Due to drug addiction and the emergence of antibiotic resistance in pathogens, the disease load and medication intake have risen worldwide. The alternative treatment for drug-resistant infections is Nano formulation-based antimicrobial agents. The plant extract of Conocarpus Lancifolius fruits was used to synthesize silver nanoparticles in the current study, and it was further employed as an antimicrobial and anticancer agent. Nanoparticles have been characterized by UV–visible spectrometer revealed the notable peak of λ max = 410–442 nm, which confirms the reduction of silver ion to elemental silver nanoparticles, and the biological moieties in the synthesis were further confirmed by FTIR analysis. The stability and crystalline nature of materials were approved by XRD analysis and expected the size of the nanomaterials of 21 to 173 nm analyzed by a nanophox particle-size analyzer. In vitro, synthesized materials act as an antibacterial agent against Streptococcus pneumonia and Staphylococcus aureus. The inhibition zones of 18 and 24 mm have been estimated to be antibacterial activity against both bacteria. The potency of up to 100% of AgNPs for bacterial strains was incubated overnight at 60 μg/ml. Based on our results, biogenic AgNPs reveal significant activity against fungal pathogen Rhizopusus stolonifera and Aspergillus flavus that cause leading infectious diseases. Additionally, nanomaterials were biocompatible and demonstrated the potential anticancer activities against MDA MB-231 cells after 24-hour exposure.