Background: Traditional fermented foods are a typical niche for probiotic Lactic Acid Bacteria (LAB) and are involved with many therapeutic attributes. Among LAB, Lactobacillus plantarum is one of the most versatile members. Several L. plantarum strains have been documented for their probiotic attributes. There are limited reports on the evaluation of probiotic potential along with the therapeutic properties such as antioxidative ability. With this background, the present study was carried out to screen ‘Sannas’ which is collected from the Coorg region of Karnataka state, India, in the presence of potential probiotic L. plantarum strains for their therapeutic potential.Methods: Sannas was screened for the potential probiotic strains and identified using physiological, biochemical and molecular methods. A series of probiotic attributes were assessed for the selection of potential probiotic strains. Further, the potential probiotic strains were assessed for the in vitro antibiofilm activity against Pseudomonas aeruginosa MTCC 7903. In addition, the cholesterol lowering ability of the potential probiotic strain using four-percent commercial fresh hen egg yolk as the cholesterol source. On the other hand, the in vitro antioxidant activity was also determined by a series of assays. Furthermore, the toxicity assessment was also done by the mitotic index using onion root tips as the source of chromosomes. Results: L. plantarum MYS14 isolated from an unexploited traditional cereal-based fermented food had good functional probiotic attributes and the antibiotic susceptibility was an intrinsic feature, thus consumption of this strain does not possess any health risk to humans and animals. In addition, the strain exhibited strong radical scavenging activity which might be useful in controlling or slowing the progress of several oxidative stress related disorders. On the other hand, the strain also exhibited better cholesterol assimilation, antibiofilm ability and the mitotic index revealed the non-toxic effect of the CFS. Conclusion: The probiotic therapy is gaining significant research interest in relation to gut microbiota in an attempt to better understand the therapeutic potential of probiotic strains isolated from unexploited traditional cereal based foods. It is confirmed that this L. plantarum MYS14 strain possesses several characteristics suitable for the production of various antioxidant probiotic products. Collectively, the results suggest that L. plantarum MYS14 may be used as probiotic strain for therapeutic applications.