Lead contamination has posed a potential threat to the environment and food safety, arousing extensive concern. In this work, we fabricated a novel fluorescent sensing platform based on zeolitic imidazolate framework-8 (ZIF-8) and DNAzyme for monitoring Pb2+ in water and fish samples. ZIF-8 was proposed as a fluorescence quencher with the advantages of simple synthesis, low cost, and high quenching efficiency. The Pb2+-dependent GR5 DNAzyme containing the large ssDNA loop can be adsorbed onto ZIF-8 accompanied by fluorescence quenching. Upon binding with Pb2+, GR5 DNAzyme was activated and cleaved, leading to the release of FAM-labeled 5-base ssDNA, which restored the fluorescence. The “turn-on” assay can detect Pb2+ through the one-pot procedure in the range of 0.01–10.0 nM with a detection limit of 7.1 pM. The platform is promising for on-site monitoring of Pb2+ owing to the excellent performance of high sensitivity, low background, strong anti-interference ability, and simple operation.