DNAzyme-based biosensors have emerged as a promising tool for ensuring food safety due to their high sensitivity, specificity, and potential for rapid, cost-effective detection of hazardous substances. These biosensors leverage DNAzymes-catalytically active DNA molecules-to detect a range of contaminants, including metal ions, fungal toxins, pesticides, and pathogens. While DNAzyme-based biosensors show significant advantages over conventional techniques, challenges such as nuclease degradation, interference from complex sample matrices, and the high costs associated with DNAzyme synthesis still hinder their widespread application. Recent advancements in the stability of DNAzymes, their immobilization strategies, and integration with nanomaterials are progressively addressing these limitations, enhancing the performance and reliability of DNAzyme-based sensors. This review highlights the structural and catalytic characteristics of DNAzymes, assesses their current applications in food safety, and discusses innovative strategies to overcome existing challenges. The continuous evolution of DNAzyme-based biosensors, particularly in design and device integration, holds great promise for their future role in routine, reliable food analysis.