Abstract
A recombinant E. coli ACV1003 releasing β-galactosidase by a SOS regulon system when it is exposed to a DNA-damaging compound, has been used to detect endocrine disruptors such as tributyltin (TBT) and triphenyltin (TPT). Maximum response ratio by E. coli ACV 1003 (recA::lacZ) – indicating the maximum ratio of enzyme produced against an environmental toxicant to that produced in the absence of a toxicant – was estimated as 6.3 with 1.0 μg TBT ml−1 at 37 °C, which was considerably higher than those with other strains. Extracellular β-galactosidase activity was 51 unit ml−1, which was 5% of that obtained by the conventional Miller's enzyme assay using solvents. Such a low enzyme activity can be rapidly determined, not by the usual time-consuming and tedious enzyme assay, but by an alternative interferometric biosensor. Heavily-doped porous silicon to apply to an interferometer was fabricated by etching to produce a Fabry–Pérot fringe pattern, which caused the change in the refractive index of the medium including β-galactosidase. The change in the effective optical thickness versus β-galactosidase activity showed a sigmoid increase up to the concentration of 250 unit β-galactosidase ml−1.
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Ryu, CS., Cho, S.M. & Kim, BW. Interferometeric sensing of β-galactosidase released by recombinant E. coli responding to an endocrine disruptor, tributyltin. Biotechnology Letters 23, 653–659 (2001). https://doi.org/10.1023/A:1010308126060
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DOI: https://doi.org/10.1023/A:1010308126060