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This study aimed to determine the effect of fermentation using an isolated Levilactobacillus brevis strain on the antinutrients and the protein quality of cassava leaves. Cassava leaves from tajirika and kibandameno variety which are considered bitter and sweet varieties, respectively, were fermented using the probiotic isolate and spontaneous fermentation was used as a control. The antinutrients of interest were cyanide and phytic acid whereas the protein quality of the leaves was analyzed in terms of the total crude protein and in-vitro digestibility. These were analyzed using standard procedures namely; picrate paper method for cyanide, spectrophotometry for phytic acid, Kjedahl method for the crude protein, and trypsin and pepsin enzymes simulation for the actual human digestion process for the in vitro protein digestibility. The data obtained was analyzed using the SPSS software and means separated at p 37% with the L. brevis isolate for both cassava varieties which was higher than 20–33% for the spontaneous fermentation (control). The crude protein in both the varieties ranged between 28% and 32% and improved up to 43% on fermentation. Upon digestion of the cassava leaves, the in vitro protein digestibility improved from 63% up to 94%. The findings of this study therefore indicates that the Levilactobacillus brevis isolate can reduce cyanide and phytic acid as well as improve the protein quality by increasing the crude protein content and enhancing the digestibility of protein in cassava leaves of kibandameno and tajirika varieties, hence can be used to detoxify cassava leaves and for food formulation.