In the present study, Sulfated polysaccharides were extracted from macroalga Sargassum whitey collected from the Gulf of Mannar, Tamil Nadu, India. The maximum yield of 1.95g sulfated polysaccharide was extracted from Sargassum whitey and 0.81mg/ml of purified sulfated polysaccharide was obtained through DEAE anion exchange chromatography. The purified form was characterized using Fourier Transform Infrared Spectroscopy (FTIR), Energy Dispersive X-ray spectroscopy (EDX), and Field Emission Scanning Electron Microscopy (FESEM) analyses. The in vitro biological properties such as antibacterial, antioxidants activity (DPPH, Hydroxyl & Superoxide anion) of sulfated polysaccharides were evaluated and the results showed that 45μg mL-1 concentration of sulfated polysaccharide exhibited good antibacterial inhibitory activity against all the tested pathogenic bacteria strains, Escherichia coli, Klebsiella pneumoniae and Staphylococcus aureus. The in vitro antioxidant activity also found to be significant, where the exhibited IC<inf>50</inf> concentrations by the purified sulfated polysaccharide was 183.2µg/mL for DPPH, 204.2µg/mL for superoxide anion, and 163.8µg/mL for Hydroxyl scavenge free radicals activities. © 2021 Elsevier B.V., All rights reserved.