In this work, hierarchical ZnO particles were prepared using a biomineralization strategy at room temperature in the presence of peptides acidified from spider silk proteins. A mechanism of the mineralization of the ZnO particles was that the affinity of original ZnO nanoparticles and zinc ions in the peptide chains played an important role in controlling the biocrystallizing formation of the pore ZnO particles. The intensity of their visible green luminescence was enhanced with increases of the mineralization time due to the porous surface defects. The hierarchical ZnO materials with biomolecules will facilitate their photoluminescence spectra applications as biosensors or optoelectronic nanodevices in the future, when covalently coupled with peptides or other biomolecules to achieve patterned growth over large areas of substrate.