The aim of this work was to select suitable fermentation treatments for the efficient bioconversion of cactus (Opuntia humifusa Raf.) bioactive components with an improved radical scavenging activity for use as a nutraceutical. To obtain microorganisms for the microbial conversion of cactus, Leuconostoc mesenteroides ATCC8294, Lactobacillus plantarum KCTC 3099, Lactobacillus plantarum KERI 236 and Monascus pilosus KCCM 60029 (ATCC 22080) were used for fermentation. Fermentation by Lac. plantarum KCTC 3099 was the most effective at scavenging 1,1-diphenyl-2-picrylhydrazyl hydrate (DPPH) and 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals and reducing iron ( ). In particular, uronic acid levels showed a remarkable increase in fermentation. The polyphenol and quercetin content of the fermented cactus showed large increases from 108.65 μg/mL and 2.71 μg/mL to 227.83 μg/mL and 9.73 μg/mL, respectively, showing a maximum level at 36 h of fermentation with Lac. plantarum KCTC 3099. Thus, cactus fermentation with Lac. plantarum is an useful process for the enhancement of antioxidant contents and activity of fresh cactus.