Abstract
Cytochrome P450 enzymes (CYP) are a group of monooxygenases able to biotransform several kinds of xenobiotics including aflatoxin B1 (AFB1), a highly toxic mycotoxin. These enzymes have been widely studied in humans and others mammals, but there is not enough information in commercial poultry species about their biochemical characteristics or substrate specificity. The aim of the present study was to identify CYPs from avian liver microsomes with the use of prototype substrates specific for human CYP enzymes and AFB1. Biochemical characterization was carried out in vitro and biotransformation products were detected by high-performance liquid chromatography (HPLC). Enzymatic constants were calculated and comparisons between turkey, duck, quail and chicken activities were done. The results demonstrate the presence of four avian ortholog enzyme activities possibly related with a CYP1A1, CYP1A2, CYP2A6 (activity not previously identified) and CYP3A4 poultry orthologs, respectively. Large differences in enzyme kinetics specific for prototype substrates were found among the poultry species studied. Turkey liver microsomes had the highest affinity and catalytic rate for AFB1 whereas chicken enzymes had the lowest affinity and catalytic rate for the same substrate. Quail and duck microsomes showed intermediate values. These results correlate well with the known in vivo sensitivity for AFB1 except for the duck. A high correlation coefficient between 7-ethoxyresorufin-Odeethylase (EROD) and 7-methoxyresorufin- O-deethylase (MROD) activities was found in the four poultry species, suggesting that these two enzymatic activities might be carried out by the same enzyme. The results of the present study indicate that four prototype enzyme activities are present in poultry liver microsomes, possibly related with the presence of three CYP avian orthologs. More studies are needed in order to further characterize these enzymes.
Highlights
Turkey liver microsomes had of Cytochrome P450 enzymes (CYP) genes and protein information in other phosphate sodium salt, glucose 6-phosphate the highest affinity and catalytic rate for aflatoxin B1 (AFB1) avian species
Quail and duck microsomes showed such as aflatoxin B1 (AFB1), a toxic compound hydrate, glycerol, bovine serum albumin, intermediate values
A high correlation bioactivated by CYP enzymes to a more toxic purchased from Sigma Chemical Co
Summary
Samples it is necessary to search for specific Key words: CYP prototype substrates, avian CYP, Sandra Milena Cepeda. Laboratorio de Toxicología, Facultad de Medicina Veterinaria y de Zootecnia, Universidad Nacional de Colombia, Bogotá, D.C., Colombia substrates metabolized by a specific enzyme (or group of enzymes). Only few prototype substrates have been found and most of them are intended for human CYPs.[1] mammalian and avian CYPs are not strictly orthologs, some substrates, inhibitors and aflatoxin B1, enzymatic parameters, avian CYP2A6 ortholog
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