AbstractThe structural arrangement of amino acid residues in native enzymes underlies their remarkable catalytic properties, thus providing a notable point of reference for designing potent yet simple biomimetic catalysts. Herein, we describe a minimalistic approach to construct a dipeptide‐based nano‐superstructure with enzyme‐like activity. The self‐assembled biocatalyst comprises one peptide as a single building block, readily synthesized from histidine. Through coordination with zinc ion, the peptide self‐assembly procedure allows the formation of supramolecular β‐sheet ordered nanocrystals, which can be used as basic units to further construct higher‐order superstructure. As a result, remarkable hydrolysis activity and enduring stability are demonstrated. Our work exemplifies the use of a bioinspired supramolecular assembly approach to develop next‐generation biocatalysts for biotechnological applications.