Evaluation of a high throughput toxicity biosensor and comparison with a Daphnia magna bioassay.
A high throughput toxicity biosensor has been designed and constructed using recombinant Escherichia coli cells, containing stress specific promoters (recA, fabA, or katG) or constitutive promoters (lac) fused to luciferase genes originating from Vibrio fisheri. These genetically engineered cells were immobilized in 96 well plates. By optimizing cell immobilization conditions and the strains' response specificity to toxic chemicals, bioluminescent outputs decreased or increased dose-dependently upon adding test chemicals. However, to date the toxicity data obtained using this biosensor have not been compared with the results of other toxicity tests. Phenolics were chosen to evaluate the correlation between the LD50 and the EC50 (GC2) or EC120 (DPD2540) of Daphnia magna and E. coli, respectively. Toxicity data obtained from constitutive strains by bioluminescent level decrements were compared with the results from D. magna as a standard. LD50 values were used as parameters of D. magna toxicity and EC50 of EC120 values were used for the immobilized biosensor. In the DPD2540 test, phenolics, membrane damaging toxic chemicals, for testing immobilized stress specific bacterial strains trigger dose-dependant bioluminescence increase within specific concentration. Although the stress specific responsiveness from the strains could not be compared with D. magna's LD50 values, these responses offer additional information, such as upon the mode of toxic action in the sample, in addition to the cellular toxicity results as indicated by the EC50. This novel high throughput toxicity biosensor can be implemented to investigate the toxicity of any other soluble materials, and can be used as a standardization tool for the evaluation of toxicity.
Kim
Byoung Chan
BC
National Research Laboratory on Environmental Biotechnology and Department of Environmental Science and Engineering, Kwangju Institute of Science and Technology, 1, Oryong-dong, Puk-gu, 500-712, Kwangju, South Korea.
Park
Kyeong Seo
KS
Kim
Sang Don
SD
Gu
Man Bock
MB
D000818:Animals; D001681:Biological Assay / Q000379:methods*; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D002470:Cell Survival / Q000187:drug effects; D003621:Daphnia / Q000187:drug effects* / Q000254:growth & development; D004305:Dose-Response Relationship, Drug; D004867:Equipment Design; D019544:Equipment Failure Analysis; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics* / Q000254:growth & development; D005470:Fluorometry / Q000379:methods; D007928:Lethal Dose 50; D008163:Luminescent Measurements; D010636:Phenols / Q000633:toxicity*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D016019:Survival Analysis; D018675:Toxicity Tests / Q000295:instrumentation* / Q000379:methods
D010636:Phenols