Current Research on Reducing Pre‐ and Post‐harvest Aflatoxin Contamination of U.S. Almond, Pistachio, and Walnut

In a search for an antifungal peptide with a high activity against Aspergillus flavus, Bacillus subtilis AU195 was selected from a collection of isolates with antagonistic activity against A. flavus. To identify the antifungal peptides, a protein purification scheme was developed based on the detection of the antifungal activity in purified fractions against A. flavus. Two lipopeptides were purified with anion exchange and gel filtration chromatography. Their masses were determined to be 1045 and 1059 m/z with mass spectrometry, and their peptide moiety was identical to bacillomycin D. AU195 synthesized a mixture of two antifungal bacillomycin D analogues with masses of 1045 and 1059, the 14 mass unit difference representing the difference between a C15 and a C16 lipid chain. Both bacillomycin D analogues were active at the same concentration against A. flavus, but the different lipid chain length apparently affected the activity of the lipopeptide against other fungi.

Approximately half of hepatocellular carcinoma (HCC) from regions in the world with high contamination of food with the mycotoxin aflatoxin B1 (AFB1) contain a mutation in codon 249 of the p53 tumor suppressor gene. The mutation almost exclusively consists of a G-->T transversion in the third position of this codon, resulting in the insertion of serine at position 249 in the mutant protein. To gain insight into the mechanism of formation of this striking mutational hot spot in hepatocarcinogenesis, we studied the mutagenesis of codons 247-250 of p53 by rat liver microsome-activated AFB1 in human HCC cells HepG2 by restriction fragment length polymorphism/polymerase chain reaction genotypic analysis. AFB1 preferentially induced the transversion of G-->T in the third position of codon 249. However, AFB1 also induced G-->T and C-->A transversions into adjacent codons, albeit at lower frequencies. Since the latter mutations are not observed in HCC it follows that both mutability on the DNA level and altered function of the mutant serine 249 p53 protein are responsible for the observed mutational hot spot in p53 in HCC from AFB1-contaminated areas. Our results are in agreement with an etiological role of AFB1 in hepatocarcinogenesis in regions of the world with AFB1-contaminated food.