Expression and application of aflatoxin degrading enzyme gene in Pichia pastoris

Abstract

In this study, three aflatoxin degrading enzyme genes, tv-adtz, arm-adtz and cu-adtz, were heterologously expressed in Pichia pastoris. The protein expression of the enzyme solution was detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and the results showed that specific protein bands were detected and the target genes were successfully integrated into Pichia pastoris. The enzyme activities and detoxification efficiency of TV-ADTZ, Arm-ADTZ and Cu-ADTZ crude enzyme solutions were detected, and the highest enzyme activities were up to 3.57, 4.30, and 2.41 U mL⁻¹, and the highest degradation rates were up to 45.58%, 60.0% and 34.21%, respectively. Arm-ADTZ with the best degradation effect was selected and designed for detoxification application experiments to test its detoxification efficiency of AFB₁ in aqueous phase and in the process of moldy ground corn and preparation of DDGS, respectively, and the degradation rates reached 78.94%, 56.48%, and 24.31% after 24 h of reaction, respectively. Thus, it can be seen that the aflatoxin-degrading enzyme gene was successfully integrated into Pichia pastoris and secreted for expression, and the expressed product could effectively degrade AFB₁.