Microalgae are primary producers of aquatic ecosystems, securing biodiversity and health of the ecosystem and contributing to reducing the impact of climate change through carbon dioxide fixation. Also, they are useful biomass that can be used as biological resources for producing valuable industrial products. However, harvesting process, which is the separation of microalgal biomass from mixed liquor, is an important bottleneck in use of valorization of microalgae as a bioresource accounting for 20 to 30% of the total production cost. This study investigates the applicability of sewage sludge-derived extracellular polymeric substance (EPS) as bioflucculant for harvesting microalgae. We compared the flocculation characteristics of microalgae using EPSs extracted from sewage sludge by three methods. The flocculation efficiency of microalgae is closely related to the carbohydrate and protein concentrations of EPS. Heat-extracted EPS contains the highest carbohydrate and protein concentrations and can be a best-suited bioflocculant for microalgae recovery with 87.2% flocculation efficiency. Injection of bioflocculant improved the flocculation efficiency of all three different algal strains, Chlorella Vulgaris, Chlamydomonas Asymmetrica, Scenedesmus sp., however the improvement was more significant when it was used for flocculation of Chlamydomonas Asymmetrica with flagella.