Herein, we describe a detailed protocol to extract the mucilage from different species of the genus Opuntia spp. (i.e., Opuntia Ficus (OFi), Opuntia Dillenii (ODi) and Opuntia Robusta (ORo)). The extracted mucilage was characterized by NMR, FTIR-ATR, HPLC, and TGA. OFi was found to have the highest phenolic content, 7.84 ± 1.93 mg catechol/g mucilage. The mucilage from the three species were characterized by having a high content of monosaccharides, being mannose and glucose the most abundant components (ca. 48-73 % and 23-35 %, respectively). In the context of biomass revalorization, the mucilage was proven to serve as a reducing and stabilizing agent in the synthesis of gold nanoparticles (AuNP/mucilage). The synthesis was optimized with a mucilage concentration of 2 mg/mL using 12.5 μL of KAuCl4 and was carried out at 80 °C for 90 min. This protocol afforded spherical nanoparticles with an average size of 9.7 ± 4.0 nm that were stable for at least 14 days, as demonstrated by TEM. Synthesized AuNP/mucilage was evaluated as a plasmonic catalyst for the reduction of 4-nitrophenol as model reaction, showing a considerable enhancement in its kapp of 97 % under white light and a decrease of 24.8 % in its activation energy.