title: Aflatoxin detoxification by manganese peroxidase purified from Pleurotus ostreatus.
abstract: Manganese peroxidase (MnP) was produced from white rot edible mushroom Pleurotus ostreatus on the culture filtrate. The enzyme was purified to homogeneity using (NH4)2SO4 precipitation, DEAE-Sepharose and Sephadex G-100 column chromatography. The final enzyme activity achieved 81 U mL(-1), specific activity 78 U mg(-1) with purification fold of 130 and recovery 1.2% of the crude enzyme. SDS-PAGE indicated that the pure enzyme have a molecular mass of approximately 42 kDa. The optimum pH was between 4-5 and the optimum temperature was 25 °C. The pure MnP activity was enhanced by Mn(2+), Cu(2+), Ca(2+) and K(+) and inhibited by Hg(+2) and Cd(+2). H2O2 at 5 mM enhanced MnP activity while at 10 mM inhibited it significantly. The MnP-cDNA encoding gene was sequenced and determined (GenBank accession no. AB698450.1). The MnP-cDNA was found to consist of 497 bp in an Open Reading Frame (ORF) encoding 165 amino acids. MnP from P. ostreatus could detoxify aflatoxin B1 (AFB1) depending on enzyme concentration and incubation period. The highest detoxification power (90%) was observed after 48 h incubation at 1.5 U mL(-1) enzyme activities.
authors.affiliation: Department of Botany Faculty of Science Cairo University Giza Egypt.
authors.forename: Ramy Sayed
authors.initials: RS
authors.lastname: Yehia
mesh_terms: D000348:Aflatoxins / Q000378:metabolism*; D001711:Biotransformation; D011232:Chemical Precipitation; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D020536:Enzyme Activators / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008670:Metals / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D010544:Peroxidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D020076:Pleurotus / Q000201:enzymology*; D017422:Sequence Analysis, DNA; D013696:Temperature
chemical_list: D000348:Aflatoxins; D004271:DNA, Fungal; D020536:Enzyme Activators; D004791:Enzyme Inhibitors; D008670:Metals; D010544:Peroxidases; C051129:manganese peroxidase
keywords: Pleurotus ostreatus; aflatoxin B1; manganese peroxidase; purification