Biochemical and nutritional changes were studied during the ripening process of three Opuntia morphospecies with different ripening behavior: Naranjona (O. ficus-indica), Blanca Cristalina (Opuntia sp.), and Esmeralda (Opuntia sp.) of early, early-intermediate, and intermediate-late ripening, respectively. In loss of fresh weight, Naranjona showed the highest values, while in Blanca Cristalina and Esmeralda, a discrete weight loss was found. No significant differences were found among morphospecies in soluble solids, total titratable acidity and pH during the postharvest days. Blanca Cristalina and Esmeralda showed an increase in the content of carotenoids, while these diminished in Naranjona. The cell wall enzymes evaluated showed particular behaviors during the ripening of each morphospecies suggesting a fine biochemical control and not a clear relationship between fruit softening and enzyme activity. This study provides basic information on prickly pear ripening, in order to understand this process for its control and for improving shelf life.