Aflatoxin B1 activation to a plasmid mutagen by a chemical model of cytochrome P-450.
Aflatoxin B1 (AFB1) was oxidised by a chemical model of cytochrome P-450 and the products obtained analysed by reversed-phase hplc. The oxidation system employs a water-insoluble iron(III)porphyrin catalyst (tetraphenylporphinatoiron(III) chloride; FeTPPCl) and an oxygen donor, iodosylbenzene (PhIO). The two AFB1 products obtained from this reaction were derived by initial oxidation across the 8-9 double bond of AFB1 and subsequent breakdown of AFB1-8,9-epoxide, the compound postulated to be the ultimate carcinogenic and mutagenic derivative of AFB1. Oxidation of AFB1 by the porphyrin-catalysed system in the presence of calf-thymus DNA, resulted in the formation of AFB1-DNA adducts identical to those formed in vivo and in vitro after liver mono-oxygenase activation. The chemical model system was therefore used on a microscale to react AFB1 with plasmid pHR1800, a plasmid containing the genes for beta-lactamase (amp-r) and galactokinase (galK). AFB1-modified plasmid DNA was then transformed into E. coli AB1886 (uvrA-) and the effects of the AFB1-DNA adducts on plasmid survival and the ability of pHR1800-transformed bacteria to metabolise galactose, were then studied. The results showed an AFB1 dose-dependent decrease in plasmid survival and increase in the mutation frequency of the galK gene only when the complete oxidation system was used to generate the reactive AFB1 metabolite.
D016604:Aflatoxin B1; D000348:Aflatoxins / Q000378:metabolism / Q000494:pharmacology*; D003577:Cytochrome P-450 Enzyme System / Q000494:pharmacology*; D004247:DNA / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D008956:Models, Chemical; D009153:Mutagens; D010084:Oxidation-Reduction; D010957:Plasmids / Q000187:drug effects*; D014169:Transformation, Bacterial
D000348:Aflatoxins; D009153:Mutagens; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; D016604:Aflatoxin B1