Mechanistic insights into ochratoxin A biodetoxification by Pediococcus acidilactici NX8 involving a multi-hydrolase toolbox.

This study aimed to develop efficient and environmentally friendly detoxification strategies for ochratoxin A (OTA). Pediococcus acidilactici NX8, isolated from traditional fermented dairy products, completely degraded 1 μg/mL OTA within 24 h. By integrating degradation pathway analysis and multi-omics approaches, the molecular mechanism underlying OTA degradation was elucidated, and three novel OTA-hydrolyzing enzymes (XlyAB, AmiE, and Pbp4b) were identified. The recombinant variants completely degraded 2 μg/mL OTA within 5 min and remained stable at pH 4-7 and 4-50 °C. Molecular simulations revealed the specific binding mode between the enzymes and OTA at the atomic level. This study is the first to reveal the multienzyme mechanism of OTA degradation in the genus Pediococcus, providing a theoretical basis and genetic resources for probiotic- and enzyme-based bioremediation technologies. The efficient degradation of OTA in wine and feed highlights the potential of these enzymes for applications in the food and feed industries.