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Paramylon from Euglena gracilis is an insoluble crystalline β-1,3-glucan which have pharmaceutical and nutraceuticals applications. The present study aims to check the prebiotic potential of paramylon derived from heterotrophically grown E. gracilis in bioreactor. The Paramylon was extracted using sodium dodecyl sulfate from E. gracilis biomass. The Fourier Transform-Infra Red spectroscopy and scanning electron microscopy demonstrated the isolated paramylon to be equivalent to that of analytical standard. The prebiotic activity of E. gracilis cell extract and isolated paramylon was studied. E. gracilis cell extract as well as isolated paramylon led to cell number enhancement of Lacfid (Lactobacillus) strain exhibiting the prebiotic activities.