Enhancing the mechanical, thermal, and degradation properties of a poly(lactic acid) (PLA) blend without deteriorating its other useful features was the goal of this work. The isolation of cellulose nanocrystals (CNCs) from Agave angustifolia fibers was carried out, and the properties of the bio-nanocomposites comprising these CNCs were evaluated, which included PLA, natural rubber (NR), and liquid NR (LNR). Transmission electron microscopy and zeta potential analysis confirmed the successful isolation of CNCs from agave fibers after several chemical treatment steps. The effects of different CNC loadings on the properties of the bio-nanocomposites were investigated using tensile tests, thermal analysis, morphological analysis, and water absorption tests. Bio-nanocomposites containing 5 wt% and 7.5 wt% CNC had the optimal tensile modulus and strength, respectively. Different levels of CNC did not noticeably affect the thermal stability of the bio-nanocomposites, although the thermogram curves increased slightly as CNC content increased. The addition of CNC at different loadings affects the crystallization rate of PLA blend. The water absorption capacity increased as CNC level increased, and 5 wt% CNC gave rise to the highest water absorption. The four-component bio-nanocomposites created in this study provided an alternative for producing new green materials with tunable physical, mechanical, and thermal properties.