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Abstract Currently, Nano-materials have been explored for their abundant biomedical applications. In the present study the green synthesized (SC-AgNPs) by root extract of Smilax Chenensis have been characterized by UV–visible spectroscopy revealed SPR peak at 432 nm and FT-IR data reveals that the bioactive components of root extract have been actively involved in the reduction and stabilization of SC-AgNPs. TEM and AFM results revealed that SC-AgNPs were roughly spherical in shape. Further, the particle size of SC-AgNPs was also carried out by Dynamic Light Scattering method by aqueous colloidal solution and the results reveals that the SC-AgNPs are poly-dispersed in nature with an average size 45.6 nm with a Z average of 39.5 nm. The stability of colloidal SC-AgNPs was further confirmed by negative zeta potential value of − 21.0 mV. The SC-AgNPs showed good antibacterial activity against both gram –ve and gram + ve bacteria, whereas, SC-AgNPs coupled with antibiotic reveals excellent and enhanced antibacterial activity. The gram –ve E.coli and gram + ve S.aureus revealed highest zone of inhibition when compared to other two bacterial species. So, SC-AgNPs coupled with antibiotics can be excellent alternative to treat antibiotic resistant bacteria. The SC-AgNPs also reveals excellent antioxidant activity among them DPPH method revealed superior activity with an IC50 value76.22. The SC-AgNPs also reveals superior anticancer activity against MDA-MB-231 with IC50 value of 33.98 µg/mL and photo-catalytic activity the optical density of reduced from 1.861 to 0.135 OD within 30 min. The green SC-AgNPs detected to have multiple therapeutic applications.