The constant golden tides of Sargassum spp., identified to be a mixture of Sargassum natans and Sargassum fluitans, observed recently in the Mexican Caribbean have affected the marine ecosystem and the local economy and have created the need for solutions for their management and use. The Sargassum arrivals have thus been considered as third-generation feedstock for biofuel. Their potential for energetic conversion to biomethane was investigated, with hydrolysis as the limiting step due to its complex composition; therefore, in the present study, different physical, chemical, and enzymatic pretreatments and a combination of them have been evaluated, with the additional use of granular activated carbon, to determine the best yield and methane quality. The combined pretreatments of 2.5% hydrogen peroxide, followed by an enzymatic pretreatment (enzymatic extract from Trametes hirsuta isolated from decomposing wood in the Yucatán Peninsula-Mexico), was the best option, reaching a biodegradability of 95% and maximum methane yield of 387 ± 3.09 L CH4/kg volatile solid. The use of a conductive material, such as granular activated carbon, did not generate significant changes in performance and methane concentration.
