Construction of a sodA::luxCDABE fusion Escherichia coli: comparison with a katG fusion strain through their responses to oxidative stresses

Abstract. A recombinant bioluminescent Escherichia coli strain, EBHJ, (sodA::luxCDABE), containing the promoter for the manganese superoxide dismutase (sodA) gene fused to the Vibrio fischeriluxCDABE operon, was successfully constructed and characterized. Redox-cycling agents, such as paraquat and chromium, strongly induced a sodA- regulated response in dose-dependent manners, resulting in an increase of the bioluminescence. In a comparison with an existing oxidative stress responsive strain, DPD2511 (katG::luxCDABE), which is sensitive to H2O2, the mechanism of chemicals that cause oxidative damage was elucidated via the key transcriptional factors involved in induction of the sodA and katG promoters, i.e. SoxRS and OxyR, respectively. It was found that responses from the katG- and sodA-based strains were significantly different dependent upon the chemicals being tested. Therefore, EBHJ, alone or in parallel with DPD2511, can be used to characterize and monitor chemicals that cause oxidative damage.

[1]  K B Konstantinov,et al.  Characterization of the Stress Response of a Bioluminescent Biological Sensor in Batch and Continuous Cultures , 1996, Biotechnology progress.

[2]  D. Touati,et al.  Interaction of six global transcription regulators in expression of manganese superoxide dismutase in Escherichia coli K-12 , 1993, Journal of bacteriology.

[3]  I. Fridovich,et al.  Assay of metabolic superoxide production in Escherichia coli. , 1991, The Journal of biological chemistry.

[4]  D. Bagchi,et al.  Oxidative mechanisms in the toxicity of chromium and cadmium ions. , 2001, Journal of environmental pathology, toxicology and oncology : official organ of the International Society for Environmental Toxicology and Cancer.

[5]  B Demple,et al.  Positive control of a global antioxidant defense regulon activated by superoxide-generating agents in Escherichia coli. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[6]  H. A. Leon,et al.  Effects of ethidium bromide on development and aging of Drosophila: Implications for the free radical theory of aging , 1981, Experimental Gerontology.

[7]  R. Larossa,et al.  Distinct responses of a recA::luxCDABE Escherichia coli strain to direct and indirect DNA damaging agents. , 1999, Mutation research.

[8]  R. Larossa,et al.  Oxidative stress detection with Escherichia coli harboring a katG'::lux fusion. , 1996, Applied and environmental microbiology.

[9]  I. Fridovich,et al.  The role of O2.- in the production of HO.: in vitro and in vivo. , 1994, Free radical biology & medicine.

[10]  R. Menzel,et al.  A microtiter plate-based system for the semiautomated growth and assay of bacterial cells for beta-galactosidase activity. , 1989, Analytical biochemistry.

[11]  S. Kubota,et al.  Formation of hydroxy radicals by environmental estrogen-like chemicals in rat striatum , 2000, Neuroscience Letters.

[12]  B Demple,et al.  Redox-operated genetic switches: the SoxR and OxyR transcription factors. , 2001, Trends in biotechnology.

[13]  T. Close,et al.  Regulation of the vir genes of Agrobacterium tumefaciens plasmid pTiC58 , 1987, Journal of bacteriology.

[14]  G. Storz,et al.  Oxidative stress. , 1999, Current opinion in microbiology.

[15]  D. Touati,et al.  Iron and oxygen regulation of Escherichia coli MnSOD expression: competition between the global regulators Fur and ArcA for binding to DNA , 1993, Molecular microbiology.

[16]  J. Lobos,et al.  Novel pathway for bacterial metabolism of bisphenol A. Rearrangements and stilbene cleavage in bisphenol A metabolism. , 1994, The Journal of biological chemistry.