Cultivating microalgae for the remediation of aquaculture wastewater provides a promising solution for pollution control. However, the economic viability of this approach faces challenges due to the high costs associated with microalgal biomass harvesting. This study aims to address this issue by immobilizing microalgae onto coral velvet carriers, enhancing the efficiency of biomass recovery. Four types of microalgae were screened: Chlorella sp., Isochrysis galbana, Chaetoceros sp., and Nannochloropsis sp. Among them, Isochrysis galbana exhibited the best self sedimentation rate, achieving a self sedimentation rate of 94.36%. Chlorella sp. demonstrated the best denitrification rate, with a nitrate removal rate of 100% and an inorganic nitrogen removal rate of 79.13%. In addition, this study found that extracellular polymeric substances(EPS) affects the self-settling performance of microalgae, and the results emphasize the key role of tightly-bound EPS(TB-EPS) content in determining self settling efficiency. Furthermore,the assessments of the purification of simulated aquaculture wastewater were conducted, comparing the outcomes of co-cultivation with mono-culture. The co-cultivation strategy showed exceptional efficacy, achieving a 100% removal rate for NO-N by the 5th day. In contrast, mono-cultures of Chlorella sp. and Isochrysis galbana showed removal rates of 77.76% and 45.72%, respectively, at the same interval. Applying of the co-cultivation microalgal biofilm to treat the actual aquaculture wastewater showed remarkable denitrification performance, attaining a 100% removal rate for NO-N by the 7th day. The study proposes the co-cultivation of Chlorella sp. and Isochrysis galbana for treating aquaculture wastewater and explores the potential application of immobilization technology to remove nitrogen-containing pollutants.