Exopolysaccharides nanoparticles (EPS-NPs), especially from probiotics, became one of the most innovative approaches for intelligent cancer therapy due to their stability and biocompatibility. In this study, a previously identified sulphated hetero-polysacchraide was extracted from Weissella paramesenteroides MN2C2 and partially purified. Some parameters of the operational culture conditions of the produced EPS were optimized. Nano-forms of crude and partially purified EPS were synthesised by sonication and characterized by Transmission Electron Microscopy (TEM) and Dynamic Light Scattering (DLS). The particles were platelet-like and their size diameter ranged from 37 to 105 for crude EPS-NP and 45 to 204 nm for partially purified EPS-NP. Interestingly, both EPS-NPs were effective and highly selective against Colon Caco-2, Liver HepG-2 and Breast MCF-7 malignant cells. They were also found to have potent DPPH antioxidant activities dependent on their concentration. The producing probiotic strain showed high sensitivity towards Tetracycline, Fusidic acid and Ampiciline with no haemolytic activity. And it was tolerant to low pH, bile salts and pancreatic juices as well as it exerted antimicrobial activity against Pseudomonas aeruginosa and Bacillus subtilis. These findings suggest that nano-EPS from the probiotic W. paramesenteroides possess excellent characteristics for potential medicine applications.