Gracilaria fisheri is a red seaweed that possesses several biological activities. The major important compounds comprising this seaweed are sulfated polysaccharides. However, limited data exists on the optimal extraction conditions for this seaweed using eco-friendly techniques like microwave-assisted extraction (MAE). This work aimed to optimize the MAE conditions for extracting sulfated polysaccharides from G. fisheri. Three factors influencing MAE were investigated: microwave time, solvent-to-solid ratio, and microwave power. These factors were initially screened by varying one factor at a time to identify levels leading to high extraction yields. These levels were then included in a Box-Behnken design. Microwave time was varied from 20 to 40 s, solvent-to-solid ratio from 8:1 to 10:1 mL/g, and microwave power from 300 to 600 W. Two responses were monitored: extraction yield and sulfate content. The optimal condition for both high extraction yield and sulfate content was found to be 30 s of microwave time, a solvent-to-solid ratio of 9.5:1 mL/g, and 450 W of microwave power. This condition yielded 20.32-20.93% extraction yield and 48.28-49.95% sulfate content. The computer program's predictions were accurate and reliable, with a very low percentage error of less than 3%. Therefore, the Box-Behnken design proved to be an effective tool for optimizing the extraction of sulfated polysaccharides from G. fisheri using the MAE method.