Cancer is a major leading cause of death worldwide. However, lactic acid bacteria (LAB) were found to produce different anticancer agents, such as l- asparaginase (ASNase), which effectively contribute against the cancer cells growth. This study aimed for the production of ASNase from Weissella paramesenteroides MN2C2, as well as the evaluation of its anticancer and antioxidant properties. The maximal ASNase production conditions were found to be reached when 10% (v/v) of the strain was inoculated in MRS medium of pH 6.5, then statically incubated for 48h at 37°C. The partially purified ASNase showed a final specific activity of 8.75 nmol/min/mg proteins, and a molecular weight of 36 kDal which was determined by SDSPAGE investigation. The anticancer effect of the partially purified ASNase was tested against different cell lines. The most optimum IC50 of 0.135 mg/mL was recorded against Caco-2 cells, while higher IC50 values of 1.034, 4.5 and 1.636 mg/mL were also obtained when tested against HepG-2, MCF-7 andA549 respectively. The partially purified ASNase showed excellent antioxidant activity with an IC 50 of 235.8 µg/ml. The tested strain was also able to participate in the production of yoghurt with enhanced quality, which consequently comprises a number of health beneficial bioactive compounds. These results demonstrated that the probiotic W. paramesenteroides MN2C2 is a very promising microorganism for both pharmaceutical and nutritional uses.