A bioluminescent sensor for high throughput toxicity classification.
A high throughput toxicity monitoring and classification biosensor system has been successfully developed using four immobilized bioluminescent Escherichia coli strains, DPD2511, DPD2540, DPD2794 and TV1061, which have plasmids bearing a fusion of a specific promoter to the luxCDABE operon. The bioluminescence of DPD2511 increases in the presence of oxidative damage, DPD2540 by membrane damage, DPD2794 by DNA damage and TV1061 by protein damage. In the developed biosensor these strains are immobilized in a single 96 well plate using an LB-agar matrix, and are able to detect the toxicities of hydrogen peroxide, phenol and mitomycin C in water samples. As the concentration of each chemical was increased, the bioluminescence levels from the corresponding wells, containing either DPD2511, DPD2540, DPD2794 or TV1061, increased. This increase in bioluminescence followed a dose dependent response to the toxic chemicals within a specific concentration range. In particular, each test requires only 4 h to give clear bioluminescent response signature. Storage of the biosensor at 4 degrees C for 2 weeks caused no change in its dose-dependent response. The fast and easy detection of oxidative, membrane, protein and DNA damaging agents in aqueous environments is possible due to the high throughput capability of this biosensor.
Kim
Byoung Chan
BC
National Research Laboratory on Environmental Biotechnology, and Department of Environmental Science and Engineering, Kwangju Institute of Science and Technology, 1 Oryoung-dong, Puk-gu, 500-712 Kwangju, South Korea.
Gu
Man Bock
MB
D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000145:classification / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000032:analysis / Q000633:toxicity*; D008163:Luminescent Measurements; D016685:Mitomycin / Q000032:analysis / Q000633:toxicity*; D019800:Phenol / Q000032:analysis / Q000633:toxicity*; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods; D018675:Toxicity Tests / Q000295:instrumentation* / Q000379:methods; D014874:Water Pollutants, Chemical / Q000032:analysis / Q000145:classification* / Q000633:toxicity
D014874:Water Pollutants, Chemical; D019800:Phenol; D016685:Mitomycin; D006861:Hydrogen Peroxide