Molecular weight and degree of substitution of extracted sodium carboxymethyl cellulose (CMC) from barley stem wastes were assessed. A novel bio-nanocomposite, CMC/Cu@RS, was developed in the presence of CMC, a copper complex, and Rhazya stricta. This bio-nanocomposite was characterized by techniques such as X-ray photoelectron spectroscopy (XPS), dynamic light scattering (DLS), zeta potential analysis, nuclear magnetic resonance (NMR), scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), UV–visible spectroscopy, and fourier-transform infrared spectroscopy (FT-IR). The anticancer properties of Rhazya stricta, Paclitaxel, and the CMC/Cu@RS bio-nanocomposite were evaluated against the human breast cancer cell line MCF-7 using the MTT assay. The cytotoxicity of the CMC/Cu@RS bionanocomposite was significantly lower compared to Rhazya stricta. Additionally, the cellular uptake of the CMC/Cu@RS bio-nanocomposite by MCF-7 cells was investigated through fluorescence microscopy. It seems that the nanocomposite was absorbed and penetrated the cancer cells. The catalytic activity of bio-nanocomposite was considered as a nano-catalyst for the oxidation of alcohols using hydrogen peroxide in aqueous conditions. The excellent conversions and selectivities were gained in this green strategy. This straightforward synthesis of the nanocomposite from agricultural waste, coupled with its significant anticancer and catalytic activities, underscored its potential for both the environmentally friendly processes and the sustainability initiatives.

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