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[Display omitted] • Novel chitosan based bioactive biocomposite was successfully synthesized. • Nano crystalline hydroxyapatite material improves the mechanical properties. • Opuntiaficus-indica fruit gum directly increases the thermal stability of composites. • Cs-Lg/ n -Hap/Fg biocomposite owns higher antioxidant and antibacterial properties. • It provides substantial role against bone cancer cells beneficial for cancer therapy. In this paper, a novel bioactive Chitosan-Lignin/ n -Hydroxyapatite/ Opuntia ficus-indica -Fruit gum (Cs-Lg/ n -Hap/OFI-Fg) biocomposite was fabricated. The as-obtained biocomposite properties are analyzed by optical and structural investigation. The biological properties like antibacterial, antioxidant, and anticancer ability were evaluated for the biocomposites. The FT-IR spectrum result shows the biocomposites possess significant intermolecular interaction with biomolecules. The powder XRD result indicated the as-obtained Cs-Lg/ n -Hap/OFI-Fg biocomposites are crystalline in nature. The microhardness test shows the addition of fruit gum increases the mechanical strength of the biocomposite. The thermal stabilities of the as-obtained Cs-Lg/ n -Hap/OFI-Fg biocomposite were higher than to Cs-Lg/ n -Hap composite as revealed by TGA analysis. The SEM images show an addition of fruit gum leads to the formation of spherical-shaped discrete particle morphology evidenced by TEM analysis. The antioxidant and antibacterial activity of bioactive biocomposite shows better activity due to the presence of numerous biomolecules. The MTT assay result revealed significant activity in as-obtained Cs-Lg/ n -Hap/OFI-Fg biocomposite compared to Cs-Lg/ n -Hap composite against MG63 bone cells and L929 cells due to the active biomolecules extant in fruit gum. The anticancer results revealed that 5 µg/ml concentration biocomposite is a significant role against bone cancer cells which proves that the as-obtained composites could be more valuable for the treatment of bone deficiency. [ABSTRACT FROM AUTHOR]