Abstract Macroalgae have not met their full potential to date as biomass for the production of energy. One reason is the high cost associated with the pretreatment which breaks the biomass's crystalline structure and better exposes the fermentable sugars to anaerobes. In the attempt to overcome this technological barrier, the performance of a Hollander beater mechanical pretreatment is assessed in this paper. This pretreatment has been applied to a batch of Laminariaceae biomass and inoculated with sludge from a wastewater treatment plant. The derived biogas and methane yields were used as the responses of a complex system in order to identify the optimal system input variables by using the response surface methodology (RSM). The system's inputs considered are the mechanical pretreatment time (5–15 min range), the machine's chopping gap (76–836 μm) and the mesophilic to thermophilic range of temperatures (30–50 °C). The mechanical pretreatment was carried out with the purpose of enhancing the biodegradability of the macroalgal feedstock by increasing the specific surface area available during the anaerobic co-digestion. The pretreatment effects on the two considered responses are estimated, discussed and optimized using the tools provided by the statistical software Design-Expert v.8. The best biogas yield of treated macroalgae was found at 50 °C after 10 min of treatment, providing 52% extra biogas and 53% extra methane yield when compared to untreated samples at the same temperature conditions. The highest biogas rate achieved by treating the biomass was 685 cc gTS−1, which is 430 cc gTS−1 in terms of CH4 yield.