Aflatoxins, produced by fungi like Aspergillus flavus and Aspergillus parasiticus, pose significant threats to food safety, impacting human and animal health while causing economic losses. Detoxifying aflatoxins is crucial to reducing contamination in food and feed. Enzymes and microorganisms present an eco-friendly and efficient solution for this purpose. This review highlights their roles in detoxification, focusing on bacteria, yeasts, and fungi that can degrade or bind aflatoxins, thereby lowering toxicity. Enzymes such as laccase, peroxidase, and reductase facilitate detoxification through oxidative and hydrolytic degradation. The efficiency of these methods depends on factors like pH, substrate availability, and temperature. Understanding the interactions between enzymes, microorganisms, and aflatoxins is essential for optimizing detoxification strategies. While promising, further research is needed to enhance their application in food safety.