Many natural materials exhibit a multilayer structure in which adjacent layers rotate in a helicoidal manner. The remarkable optical and mechanical properties of these materials have motivated research and development of man-made materials with similar morphology. Among them, composite materials by cellulose nanocrystals (CNCs) and polymers have attracted great interest; however, the relationship between the cholesteric structure and the material properties is not well understood. We used the composite CNC-polymer latex films with random, stratified, and cholesteric morphologies, all with the same compositions, to explore the effect of structure on the optical and mechanical properties of the composite films. Films with a cholesteric structure exhibited strong extinction, circular dichroism, and high stiffness; however, they had lower toughness than the films with the cholesteric stratified morphology. Films with disordered morphologies exhibited the highest toughness and the lowest stiffness. These trends were attributed to the confinement effects and the difference in polymer distribution in the films. These results provide guidance for the preparation of biomimetic cholesteric films with targeted optical and mechanical properties.