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In vitro study of DNA damage induced by acid orange 52 and its biodegradation derivatives
Hedi Ben Mansour1, Daniel Barillier, David Corroler
1Equipe de Recherche en Physico-Chimie et Biotechnologie (ERPCB-EA3914), Institut Universitaire de Technologie-Unité de Formation et de Recherche des Sciences, Unviersité de Caen-Basse Normandie, France.
Abstract:
Mutagenicity of acid orange 52 (AO52) and its degradation products by Pseudomonas putida mt-2 was evaluated with the use of Salmonella Typhimurium TA102 and TA104 with and without the metabolic activation system (S9). No mutagenicity was observed in the absence of S9 and in the presence of S9 for biodegradation under shaking conditions, but it increased significantly in the presence of S9 after biodegradation under static conditions. In addition, the ability of tested compounds to induce DNA damage in vitro was evaluated with the DNA strand scission assay. The toxicity generated by the pure azo dye and the corresponding azoreduction products (4-aminobenzenesulfonic acid and N,N'-dimethyl-p-phenylenediamine) were compared. We suggest that the mutagenicity mechanism of these molecules occurs through free radical generation processes. In this study, we demonstrate that P. putida mt-2 incubated under aerobic conditions undergoes catabolism that enables it to degrade AO52 completely and, especially, to detoxify the dye mixtures.
Insights
Pseudomonas putida mt-2 degrades acid orange 52 (AO52) and its byproducts. Mutagenicity increased under static conditions with S9, suggesting free radical generation, but P. putida mt-2 detoxified dye mixtures.
Area of Science:
- Environmental microbiology
- Toxicology
- Bioremediation
Background:
- Acid orange 52 (AO52) is an azo dye with potential mutagenic concerns.
- Microbial degradation is a key strategy for detoxifying industrial pollutants.
- Understanding the mutagenicity of dye degradation products is crucial for risk assessment.
Purpose of the Study:
- To evaluate the mutagenicity of AO52 and its degradation products by Pseudomonas putida mt-2.
- To investigate the role of the metabolic activation system (S9) in AO52 mutagenicity.
- To assess the DNA damaging potential and compare the toxicity of AO52 and its azoreduction products.
Main Methods:
- Bacterial strains: Salmonella Typhimurium TA102 and TA104.
- Mutagenicity testing with and without the metabolic activation system (S9).
- Biodegradation under shaking and static conditions.
- DNA strand scission assay for in vitro DNA damage.
- Comparison of toxicity of pure AO52 and its azoreduction products.
Main Results:
- No mutagenicity of AO52 was observed without S9 or with S9 under shaking conditions.
- Mutagenicity significantly increased with S9 after biodegradation under static conditions.
- P. putida mt-2 completely degraded AO52 and detoxified dye mixtures under aerobic conditions.
- Azoreduction products showed varying toxicity compared to the parent dye.
Conclusions:
- The mutagenicity of AO52 and its products may involve free radical generation.
- Biodegradation conditions (static vs. shaking) influence the mutagenic potential of AO52.
- P. putida mt-2 demonstrates effective AO52 degradation and detoxification capabilities.
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