Plant Activation of Bicyclic Aromatic Amines, Benzidine and 4-Aminobiphenyl

Bicyclic aromatic amines, benzidine and 4-aminobiphenyl (4-ABP) have been known as human bladder carcinogens and used in industrial manufacturing. Using the plant cell/microbe coincubation assay, this study showed that benzidine and 4-ABP were activated by plant peroxidase into frameshift- and base pair substitution-mutagens by plant-activation. Bacterial acetyl O-acetyltransferase as well as human N-acetyltransferase play a key role in the activation of benzidine and 4-ABP, and bacterial nitroreductase is partially involved in the mutagenic activation in Salmonella typhimurium. The mutagenic plant-activated benzidine and 4-ABP metabolites were fractionated using ultrafiltration membrane separation. The mutagenic metabolites were recovered in a retentate fraction with a molecular weight of $\ge$100 kDa and stable for more than 7 months when stored at 4$\sp\circ$C. To define the mutagenic specificity of the plant-activated aromatic amine metabolite, mutant spectra of spontaneous revertants and revertants induced by the plant- and mammalian-activated of aromatic amines were analyzed at the hisG46 allele in S. typhimurium strain YG1029. The plant-activated aromatic amines induced different types of base pair substitutions from the mammalian S9-activated aromatic amines. To isolate and characterize the plant-activated benzidine product(s), we generated polyclonal anti-benzidine antibodies. The anti-benzidine antibody did not recognize the plant-activated benzidine metabolites. This suggests that the plant-activated benzidine product is structurally different from benzidine parent compound. To define the molecular effects of plant- and mammalain-activated benzidine DNA adducts were induced from plant-activated benzidine in S. typhimurium and analyzed using $\sp{32}$P-postlabelmg and HPLC. Mammalian- and plant-activate benzidine metabolites induced DNA adducts in bacteria. To determine the genotoxicity of plant-activated benzidine in Chinese hamster lung cells a rapid single cell gel electrophoresis assay was employed. The plant-activated benzidine did not directly induce any DNA damage in mammalian cells. However, the plant-activated benzidine after treatment with pectinase was a potent genotoxin in mammalian cells. These data suggest that plant-activated xenobiotics may be converted into stable forms, perhaps in the pectin component in the plant cell wall. This plant activation may be a process to transport xenobiotics laterally through the environment and vertically through the food chain.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Agronomy

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2015

Author dc:creator

• Ju, Young Hwa

Contributors dc:contributor

• Plewa, Michael J.

• Biology, Plant Physiology

Language dc:language

eng

Identifier

(MiAaPQ)AAI9904495

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/87670