Biopolyme s such as polysaccharides and proteins have been vvidely used for the chiral separation of various components due to the intrinsic chirality of the polymers. Amyloidlike short peptides can also self-assemble into diverse chiral supramolecular nanostructures or polymers with precisely tailored architectures driving by noncovalent interactions. However, the use of such supramolecular nanostructures for the resolution and separation of chiral components remains largely unelplored. Here, we report that the self-assembled peptide supi-amolecular nanostructures can be used for the highly efficient chiral separation of various enantiomers. By rationally designing the constituent amino acid sequence of the peptides and the self-assembling environment, we can fabricate supramolecular polymers with distinct surface charges and architectures, including rianolielices, nanoriblaons, nanosheets, rianofibrils, and nanospheres. The various supramolecular nanostructures were then used to resolve the racemic mixtures of cr-methylbenzylamine, 2-lahenyliaropionic acid, and 1-phenylethanol. The results indicated that the self-assembled peptide polpners showed excellent enantioselective separation efficiency for different chiral molecules. The enantioselective separation efficiency of the peptide nanostructures can be tailored by changing their surface charges, morphology, and the constituent amino acid sequences of the peptides.