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A Whole Cell Bioreporter Approach to Assess Transport and Bioavailability of Organic Contaminants in Water Unsaturated Systems
Published on: December 24, 2014
Internal arsenite bioassay calibration using multiple bioreporter cell lines.
Anke Wackwitz1, Hauke Harms, Antonis Chatzinotas
1UFZ, Helmholtz Centre for Environmental Research, Department of Environmental Microbiology, 04318 Leipzig, Germany. anke.wackwitz@ufz.de
Microbial Biotechnology
|January 26, 2011
Summary
This study introduces a novel internal calibration method for bioreporter bacteria assays using multiple cell lines. This approach enhances accuracy and ease of use for detecting contaminants like arsenite in water.
Area of Science:
- Environmental microbiology
- Biosensor technology
- Analytical chemistry
Background:
- Bioreporter bacteria assays require calibration due to variable outputs influenced by incubation time and cell physiology.
- Field comparisons with standardized colorimetric methods are challenging and prone to errors.
- Existing methods lack robust internal calibration for reliable field applications.
Purpose of the Study:
- To develop and validate a new internal calibration strategy for bioreporter assays using multiple cell lines with distinct reporter outputs.
- To improve the accuracy and user-friendliness of bioreporter assays for environmental monitoring.
- To establish a method for detecting specific analyte concentrations, such as arsenite, independent of assay variations.
Main Methods:
- Construction of Escherichia coli-based bioreporter strains expressing cytochrome c peroxidase (CCP) or β-galactosidase.
- Engineered variations in reporter protein catalytic activity (CCP) or synthesis rates (β-galactosidase) to achieve differential signal intensities.
- Utilized combinations of these engineered cell lines for internal calibration and analyte concentration range discrimination.
Main Results:
- Constructed bioreporter strains exhibited distinct signal intensities at identical arsenite concentrations.
- Combinations of cell lines enabled the definition of arsenite concentration ranges with qualitative (yes/no) signals.
- The developed method demonstrated relative independence from incubation time and bioreporter cell activity.
- Discriminated concentration ranges aligned with permissive levels of arsenite in drinking water (e.g., WHO guidelines).
Conclusions:
- The proposed internal calibration method using multiple bioreporter cell lines effectively controls assay variations.
- This approach enhances the reliability and simplifies the application of bioreporter assays for environmental monitoring.
- The method shows promise for accurate detection of contaminants like arsenite within relevant environmental concentration ranges.
