Enteromorpha prolifera, the marine algae frequently causing green tide in the littoral areas in recent years, was investigated as a feasible biosorbent to remove azo dye Direct Fast Scarlet 4BS (DFS-4BS) from aqueous solution. The FTIR spectra of the acid-treated Enteromorpha prolifera (ATEP) biomass before and after dye sorption were used to identify the main functionalities involved in the biosorption process. Results of batch experiments showed that the adsorption capacity of ATEP was dependent on solution pH, ATEP dosage and temperature. The adsorption kinetics followed the pseudo-second order rate equation, while the adsorption isotherm could be well described by the Langmuir model with maximum sorption capacity of 318.87mg·g-1, demonstrating the excellent performance of ATEP as a biosorbent to remove DFS-4BS from aqueous solution. The negative values of free energy change (ΔG°) and positive value of enthalpy change (ΔH°) confirmed that the sorption of DFS-4BS onto ATEP was spontaneous and endothermic in the temperature range of 303-333K. The isosteric heat of adsorption increased with the increase of surface loading, suggesting that the ATEP biomass had heterogeneous surface and lateral interactions might exist among adsorbed molecules. According to the sorption results and the FTIR spectra, it was deduced that the adsorption mechanism involved hydrogen bonding, electrostatic attraction and bonding, and hydrophobic and van der Waals interaction.