Ochratoxin A (OTA) poses a serious threat to human and animal health as well as the ecological environment due to its high toxicity, stable physicochemical properties, and widespread occurrence in food and feed. Because of their high efficiency, environmental friendliness and good specificity, biological enzymes have become an important research direction of OTA detoxification. This review systematically summarizes the OTA-degrading enzymes that have been reported, focusing on the strategies for optimizing their expression levels and enzymatic performance. The current research mainly focuses on carboxypeptidases and amidohydrolases, among which OTase and SaADH3 are representative in terms of their degradation efficiency and research depth. In view of the limited expression level and stability of natural OTA-degrading enzymes, the construction and optimization of efficient expression systems are crucial for their application. The expression level, catalytic activity, heat and acid tolerance of OTA-degrading enzymes can be significantly improved by gene element optimization, expression system selection, expression condition optimization, enzyme immobilization and rational design. Future research should further focus on the performance improvement and industrial application of OTA-degrading enzymes to promote their practical application in the field of food and feed safety.