Biopolymer based metal oxide nanoparticles, prepared by eco-friendly approach, are gaining interest owing to their wide range of applications. In this study, aqueous extract of Trianthema portulacastrum was used for the green synthesis of chitosan base copper oxide (CH-CuO) nanoparticles. The nanoparticles were characterized through UV-Vis Spectrophotometry, SEM, TEM, FTIR and XRD analysis. These techniques provided evidence for the successful synthesis of the nanoparticles, having poly-dispersed spherical shaped morphology with average crystallite size of 17.37 nm. The antibacterial activity for the CH-CuO nanoparticles was determined against multi-drug resistant (MDR), Escherichia coli, Pseudomonas aeruginosa (gram-negative), Enterococcus faecium and Staphylococcus aureus (gram-positive). Maximum activity was obtained against Escherichia coli (24 ± 1.99 mm) while least activity was observed against Staphylococcus aureus (17 ± 1.54 mm). In-vitro analysis for biofilm inhibition, EPS and cell surface hydrophobicity showed >60 % inhibitions for all the bacterial isolates. Antioxidant and photocatalytic assays for the nanoparticles showed significant activities of radical scavenging (81 ± 4.32 %) and dye degradation (88 %), respectively. Antidiabetic activity for the nanoparticles, determined by in-vitro analysis of alpha amylase inhibition, showed enzyme inhibition of 47 ± 3.29 %. The study signifies the potential of CH-CuO nanoparticle as an effective antimicrobial agent against MDR bacteria along with the antidiabetic and photocatalytic activities.