Abstract
In the present study, recombinant organophosphorus hydrolase OPHC2 was successfully produced by Yarrowia lipolytica and purified. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analyses showed a major polypeptide band of 36 kDa. The purified enzyme was optimally active at 65°C and pH 8.5 and also displayed good thermal and pH stability using methyl parathion (O,O-dimethyl-O-4-p-nitrophenyl phosphorothioate) as a substrate. Moreover, as Y. lipolytica is a non-pathogenic, generally regarded as safe (GRAS) yeast, the cell culture supernatant can be used directly on vegetables and fruits that are contaminated by organophosphorus pesticides.
Acknowledgments
We thank Professor Catherine Madzak from the Institut National Agronomique Paris-Grignon for the kind gifts of the expression vector pINA1296 and Y. lipolytica Polg. Meng Li and Xiaolan Yu contributed equally to this work.
Funding
This study was supported by the National 973 Program of China (No. 2013CB910801), National Natural Sciences Foundation of China (No. 31340036, 31172320), the Nature Science Fund for Creative Research Groups of Hubei Province of China (No. 2012FFA034), and the Foundation for High and New Technology Industrial Innovative Research Groups of the Wuhan Science and Technology Bureau (No. 2014070504020239).