Detoxification of ochratoxin A by Lysobacter sp. CW239 and characteristics of a novel degrading gene carboxypeptidase cp4

Ochratoxin A (OTA) is a potent mycotoxin that frequently contaminates agro-products and threatens food safety. A highly efficient OTA degrading strain Lysobacter sp. CW239 was isolated, and the OTA degradation characteristics were investigated. A novel OTA degrading gene carboxypeptidase cp4 was successfully cloned and characterized from CW239. The heterologous recombinant was constructed by gene cp4 and expression vector pET-32a((+)) and overexpressed by E. coli BL21 CodonPlus (TM) (DE3). The recombinant protein rCP4 was purified, and the OTA-degrading activity was evaluated. Although OTA was efficiently degraded by CW239 (24-h degradation ratio of 86.2%), the 24-h OTA degradation ratio for rCP4 was only 36.8% at fairly high concentration (0.25 mg/mL) protein. The degraded product was obtained by immune affinity column (IAC) and determined by mass spectrometry (MS), and the degraded product was the less toxic ochratoxin alpha (OT alpha). Based on the serial investigations of this study, OTA might be simultaneously co-degraded by CP4 and another unknown degrading agent in that degrading strain. (C) 2019 Elsevier Ltd. All rights reserved.