(1) Background: Probiotication is an important method in the food industry, and the use of probiotic dairy products has prevented lactose intolerant patients and vegetarians from their consumption. Hence, there is a need to incorporate probiotics in fruit juice without lactose; (2) Method: Probiotic viability, and physicochemical and sensory evaluation of stored probioticated pineapple juice using lactic acid bacteria (LAB) (Pediococcus pentosaceus LaG1, Lactobacillus rhamnosus GG, Pediococcus pentosaceus LBF2) as a single and mixed starter was investigated; (3) Results: There was an increase in the lactic acid production, and reduction in pH, vitamin C content, and colour during storage. At weeks 3 and 4, Propp2 and Pcontrol samples had the highest lactic acid content (317.9 mg/L and 160.34 mg/L). The vitamin C content ranged from 2.91-7.10 mg/100 g. There was a general reduction in total soluble solids during storage. The probiotic LAB were viable throughout the storage time (1.05-1.10 × 10 9 cfu/mL) in the juice samples. There was no significant difference in terms of taste, aroma, colour, or appearance during the time of storage; (4) Conclusion: The pineapple juice supported the viability, lactic acid production, vitamin C development, and the antagonistic potential of the probiotic candidate. This result is useful for the development of probiotic fruit juice as functional foods and nutraceuticals with health beneficial effect.