The purpose of this study is to optimize the extraction conditions (time, pH, and temperature) for water-soluble dietary fiber (SDF) and solids from Opuntia ficus-indica (OFI) stems and to evaluate the functionality of the optimized extract. The maximum extraction yield was obtained when 100% distilled water was used, indicating that ethanol hampered the extraction. The extraction yields were lower at higher temperatures and lower pHs. The maximum extraction of SDF (27.92%) and solids (5.6 g) was achieved with a 7-h extraction at 50oC and pH 7.0, with desirability being 0.958. The stem extract obtained under the optimum extraction conditions had an antioxidant capacity of 118.10 mg vitamin C equivalents (VCE) using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. In a medium containing 30% extract, the growth of E. coli was reduced by 34%, while that of B. cereus was reduced by 23%. These results suggest that OFI stems can be used as a functional resource.