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Open AccessArticle

The Mechanism of Chlorine Damage Using Enhanced Green Fluorescent Protein-Expressing Escherichia coli

1
Department of Human Centered Engineering, Ochanomizu University, Tokyo 112-8610, Japan
2
Faculty of Core Research, Natural Science Division, Ochanomizu University, Tokyo 112-8610, Japan
*
Author to whom correspondence should be addressed.
Water 2019, 11(10), 2156; https://doi.org/10.3390/w11102156
Received: 12 September 2019 / Revised: 12 October 2019 / Accepted: 14 October 2019 / Published: 16 October 2019
(This article belongs to the Section Wastewater Treatment and Reuse)
This study investigated how chlorine inactivates and damages Escherichia coli cells. E. coli that had transformed to express enhanced green fluorescent protein (EGFP) at the cytoplasm was treated with chlorine. Damage to the cell membrane and cell wall was analyzed by measuring the fluorescence intensity of the leaked EGFP, then accounting for the fluorescence deterioration. At pH 7, E. coli was lethally damaged after treatment with chlorine, but significant leakage of EGFP was not observed. In contrast, significant leakage of EGFP was observed at pH 9, even though E. coli was not as inactivated as it was at pH 7. Flow cytometry was used to confirm the fluorescence intensity of the remaining EGFP inside the cells. No significant fluorescence loss was observed in the cells at pH 7. However, at pH 9, the fluorescence intensity in the cells decreased, indicating leakage of EGFP. These results suggest that hypochlorous acid inactivates E. coli without damaging its cell membrane and cell wall, whereas the hypochlorite ion inactivates E. coli by damaging its cell membrane and cell wall. It was possible to confirm the chlorine damage mechanism on E. coli by measuring the fluorescence intensity of the leaked EGFP. View Full-Text
Keywords: Escherichia coli; damage mechanism; chlorine; EGFP; flow cytometry Escherichia coli; damage mechanism; chlorine; EGFP; flow cytometry
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Mizozoe, M.; Otaki, M.; Aikawa, K. The Mechanism of Chlorine Damage Using Enhanced Green Fluorescent Protein-Expressing Escherichia coli. Water 2019, 11, 2156.

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