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Central Composite Design Optimization of Biological Dye Removal in the Presence of Macroalgae Chara sp.Response surface methodology (RSM) was employed to investigate the effects of different operational parameters on the biological decolorization of a dye solution containing malachite green (MG) in the presence of macroalgae Chara sp. The investigated variables were the initial pH, initial dye concentration, algae amount, and reaction time. Central composite design (CCD) was used for the optimization of biological decolorization process. Predicted values were found to be in good agreement with experimental values (R 2 ¼ 0.982 and Adj-R 2 ¼ 0.966), which indicated suitability of the employed model and the success of RSM. The results of optimization predicted by the model showed that maximum decolorization efficiency was achieved at the optimum condition of the initial pH 6.8, initial dye concentration 9.7 mg/L, algae amount 3.9 g, and reaction time 75 min. UV-VIS spectra and FT-IR analysis showed degradation of MG.