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Silver nanoparticles (AgNPs) have become a research focus due to its antimicrobial, anticancer applications and cost-effective properties. In this study the effectiveness of green synthesis of NPs using biological macromolecule chitosan which is acting as a reducing cum stabilizing agent is carried out. An in-depth analysis of the synthesized AgNPs was conducted using a variety of sophisticated characterization techniques such as UV-visible spectroscopy, particle size analysis, zeta potential measurements, transmission electron microscopy (TEM), photoluminescence, and Fourier transform infrared (FTIR) spectroscopy. The antimicrobial activity of the formulated AgNPs were inspected against two human pathogenic strains.In the antimicrobial activity, the synthesized AgNPs exhibited a reduction in the growth of both the microbes. The minimum inhibitory concentration (MIC) of 1.22 𝜇M was observed for both Candida albicansand Mycobacterium smegmatis.Consequently, AgNPs may be used as an opportunity for modern-day antibiotics to treat infections due to human pathogens. Antiproliferative analysis revealed that AgNPs showed antiproliferative characteristics against MDA-MB-231 cells compared to the control. Such AgNPs have an anticancer effect and are likely to be used as smart drug delivery mediators to treat late-stage cancer.