Purification and identification of an aflatoxin B1 degradation enzyme from Pantoea sp. T6

Aflatoxin B-1 (AFB(1)) is a secondary metabolite produced by Aspergillus fiavus and other fungi. Soil, crops, food, feed, etc. were susceptible to aflatoxin B-1 contamination, which caused adverse economic and health consequences. It is necessary to search for microorganisms or microbial enzymes that can degrade AFB(1). The degradation activity of AFB(1) by cell-free supernatant (68.30%) of isolate Pantoea sp. T6 was significantly higher (P < 0.05) than viable bacterial cells (4.87%) and intracellular cell extracts (3.68%). The supernatant's AFB(1) degradation activity was reduced from 68.30% to 5.33% in treatment with protease K and sodium dodecyl sulphate (SDS). An extracellular enzyme from the supernatant was secreted by Pantoea sp. T6 and was named as Pantoea aflatoxin degradation enzyme (PADE), which was obtained using Diethylaminoethanol (DEAE)Sepharose chromatography. PADE was further purified by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The PADE, outer membrane protein A, was identified by mass spectrometry and molecular mass was 38180.1Da. The optimum temperature and pH for the reaction of PADE with AFBI were 40 degrees C and 7.0, respectively. These finding showing that the PADE, which was isolated from the supernatant of Pantoea sp. T6, has the ability to degrade AFB(1), and may have potential application for aflatoxin B-1 reduction in the food and feed industry.