The effect of pediocin-assisted thermosonication (PAT) treatment on physicochemical properties and shelf-life enhancement of blood fruit juice was investigated. A comparison of PAT treatment with pasteurization was done based on antioxidant activity, phytochemical retention, polyphenol oxidase residual activity, and microbial count of the juice. The optimized condition for PAT treatment was found at 50.92°C, 10.28 min, and 155.53 ppm of pediocin concentration. A comparative study of sensory attributes showed pasteurized juice to be significantly different (p < .05) from PAT and fresh juice characteristics. Storage study showed maximum quality changes in pasteurized juice than PAT juice. Weibull model satisfactorily represented the survival kinetics of Leuconostoc mesenteroides under PAT treatment with R2 and RMSE values of 0.98 and 0.24 respectively. PAT treatment successfully retarded bacterial proliferation including spores of Alicyclobacillus acidoterrestris and, therefore, potentially be opted as an alternative to pasteurization by commercial organizations in fruit juice processing. Practical applications The non-thermal technologies have emerged intending to apply in the food processing industries. Ultrasound-assisted with a mild heat and bacteriocin is one of the novel hurdle technologies, which can potentially improve the shelf life of fruit juice by inhibiting common spoilage microorganisms including spore-forming bacteria. It also maintains fresh product characteristics up to a certain extent by keeping valuable phytonutrients and physiochemical properties intact. Therefore, the present study demonstrates the viability of a bacteriocin-assisted thermosonication treatment on blood fruit juice which can be opted by commercial organizations as it delivers products with superior quality than the thermally processed ones.