Remazol brilliant blue (RBB) is an anthraquinone anionic dye that has several commercial uses, especially in the textile industries and is well-known for its detrimental impacts on marine life and the surrounding ecosystem. Mercury (Hg2+) is also one of the most severe hazardous environmental contaminants due to its bioaccumulation through the food chain and high toxicity to the human embryo and fetus. The biosorption potential of Gelidium corneum biomass for bioremoval of Hg2+ and RBB dye simultaneously from binary mixture was assessed. The effects of initial pH, contact time, Hg2+, RBB, and biomass concentrations on the biosorption process were investigated in 50 batch experiments using a Face-centered central composite design. The maximum removal percentage of Hg2+ (98.25%) was achieved in the run no. 14, under optimum experimental conditions: 200 mg/L Hg2+, 75 mg/L RBB, pH 5. At 30 °C, 4 g/L algal biomass was used, with a contact time of 180 min. Whereas, the maximum removal percentage of RBB (89.18%) was obtained in the run no. 49 using 200 mg/L Hg2+, 100 mg/L RBB, pH 5, 4 g/L algal biomass and 180 min of contact time. FTIR analysis of Gelidium corneum biomass surface demonstrated the presence of many functional groups that are important binding sites responsible for Hg2+ and RBB biosorption. SEM analysis showed apparent morphological alterations including surface shrinkage and the appearance of new shiny adsorbate ion particles on the Gelidium corneum biomass surface after the biosorption process. The EDX study reveals an additional optical absorption peak for Hg2+, confirming the role of Gelidium corneum biomass in Hg2+ biosorption. In conclusion, Gelidium corneum biomass has been shown to be an eco-friendly, sustainable, promising, cost-effective and biodegradable biosorbent to simultaneously biosorb Hg2+ and RBB dye from aquatic ecosystems.