Microalgae, particularly Spirulina sp., have garnered significant attention due to their high protein content, abundance of pigments, and bioactive compounds. In recent years, research efforts have expanded to include the exploration of microbial exopolysaccharide (EPS) producers as potential sources of biopolymers with unique chemical structures and functional properties. This work investigates various extraction protocols for EPS from Spirulina platensis with promising results obtained through treatment with ethanol 95%. The obtained dry EPS extract was subjected to biochemical characterization, revealing the presence of sulphated deoxyhexose fucose sugars with multiple functional groups. Additionally, structural analysis indicated a complex polysaccharide backbone, which might be contributed to its broad range of bioactivities. The analysis demonstrated an increase in DPPH free radical scavenging activity with EPS concentration, highlighting its potential antioxidant properties. Furthermore, the EPS exhibited antibacterial activity against opportunistic pathogenic bacteria, showed a 23 mm zone of inhibition against S. aureus and approximately 22 mm against E. coli. The minimum inhibitory concentration (MIC) value for S. aureus (80.17 µg.mL−1) was lower than those for E. coli (88 µg.mL−1initially and 91.95 µg.mL−1 later). Both strains exhibited substantial reductions in biofilm formation upon EPS treatment, with E. coli showing a 78% decrease and S. aureus a 75% decrease, underscoring the EPS’s potent antibiofilm properties. Preliminary rheological studies also indicated favorable gel-forming properties, broadening its applicability in food and pharmaceutical formulations. These findings underscored the potential of S. platensis derived EPS as a valuable resource for various applications, including functional food ingredients and wound-healing agents.