Potential of a novel and thermostable recombinant phage endolysin of Escherichia phage KW1E_UTAR against Shigella spp.

Shigella causes millions of cases and hundreds of thousands of deaths annually, mainly affecting children in impoverished countries. Shigellosis outbreaks have been linked to waterborne transmission, and chlorination is commonly used to disinfect water. However, the emergence of chlorine-resistant S...

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Main Authors: Selvam Dharshini, Mohd Ismail Nor Ismaliza, Ng Michelle Y. T.
Format: Article
Language:English
Published: EDP Sciences 2025-01-01
Series:E3S Web of Conferences
Online Access:https://www.e3s-conferences.org/articles/e3sconf/pdf/2025/03/e3sconf_isgst2024_01015.pdf
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author Selvam Dharshini
Mohd Ismail Nor Ismaliza
Ng Michelle Y. T.
author_facet Selvam Dharshini
Mohd Ismail Nor Ismaliza
Ng Michelle Y. T.
author_sort Selvam Dharshini
collection DOAJ
description Shigella causes millions of cases and hundreds of thousands of deaths annually, mainly affecting children in impoverished countries. Shigellosis outbreaks have been linked to waterborne transmission, and chlorination is commonly used to disinfect water. However, the emergence of chlorine-resistant Shigella spp. is a concern. Therefore, it is imperative to develop an alternative antibacterial agent such as endolysin against the Shigella spp. In this study, the Escherichia phage KW1E_UTAR gene encoding for endolysin was successfully cloned and expressed in Escherichia coli pET-28a. The purified endolysin was characterized, and its antibacterial activity was determined. The purified endolysin remained stable between pH 4 and 9, and at a broad range of temperature (4°C–75°C). The endolysin's storage stability was tested for one week, one month, and three months at different temperatures (4°C, −20°C, and −80°C). Despite over 50% drop in the lytic activity, the purified endolysin outperformed the commercial lysozyme. Besides, all four Shigella spp. tested were susceptible to the purified endolysin, with Shigella flexneri being the most prominent. Subsequently, the purified endolysin’s effectiveness in removing S. flexneri from spiked water samples was evaluated. After an hour of treatment, the number of viable bacteria in the water samples was reduced by 99.9%. Therefore, endolysin could be effective at eradicating Shigella spp. in various water sources.
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spelling doaj-art-158a395d1dae4b7a922dac46302fe6472025-02-05T10:47:33ZengEDP SciencesE3S Web of Conferences2267-12422025-01-016030101510.1051/e3sconf/202560301015e3sconf_isgst2024_01015Potential of a novel and thermostable recombinant phage endolysin of Escherichia phage KW1E_UTAR against Shigella spp.Selvam Dharshini0Mohd Ismail Nor Ismaliza1Ng Michelle Y. T.2Department of Biological Science, Faculty of Science, Universiti Tunku Abdul Rahman, Jalan Universiti, Bandar Baru BaratDepartment of Biological Science, Faculty of Science, Universiti Tunku Abdul Rahman, Jalan Universiti, Bandar Baru BaratDepartment of Allied Health Sciences, Faculty of Science, Universiti Tunku Abdul Rahman, Jalan Universiti, Bandar Baru BaratShigella causes millions of cases and hundreds of thousands of deaths annually, mainly affecting children in impoverished countries. Shigellosis outbreaks have been linked to waterborne transmission, and chlorination is commonly used to disinfect water. However, the emergence of chlorine-resistant Shigella spp. is a concern. Therefore, it is imperative to develop an alternative antibacterial agent such as endolysin against the Shigella spp. In this study, the Escherichia phage KW1E_UTAR gene encoding for endolysin was successfully cloned and expressed in Escherichia coli pET-28a. The purified endolysin was characterized, and its antibacterial activity was determined. The purified endolysin remained stable between pH 4 and 9, and at a broad range of temperature (4°C–75°C). The endolysin's storage stability was tested for one week, one month, and three months at different temperatures (4°C, −20°C, and −80°C). Despite over 50% drop in the lytic activity, the purified endolysin outperformed the commercial lysozyme. Besides, all four Shigella spp. tested were susceptible to the purified endolysin, with Shigella flexneri being the most prominent. Subsequently, the purified endolysin’s effectiveness in removing S. flexneri from spiked water samples was evaluated. After an hour of treatment, the number of viable bacteria in the water samples was reduced by 99.9%. Therefore, endolysin could be effective at eradicating Shigella spp. in various water sources.https://www.e3s-conferences.org/articles/e3sconf/pdf/2025/03/e3sconf_isgst2024_01015.pdf
spellingShingle Selvam Dharshini
Mohd Ismail Nor Ismaliza
Ng Michelle Y. T.
Potential of a novel and thermostable recombinant phage endolysin of Escherichia phage KW1E_UTAR against Shigella spp.
E3S Web of Conferences
title Potential of a novel and thermostable recombinant phage endolysin of Escherichia phage KW1E_UTAR against Shigella spp.
title_full Potential of a novel and thermostable recombinant phage endolysin of Escherichia phage KW1E_UTAR against Shigella spp.
title_fullStr Potential of a novel and thermostable recombinant phage endolysin of Escherichia phage KW1E_UTAR against Shigella spp.
title_full_unstemmed Potential of a novel and thermostable recombinant phage endolysin of Escherichia phage KW1E_UTAR against Shigella spp.
title_short Potential of a novel and thermostable recombinant phage endolysin of Escherichia phage KW1E_UTAR against Shigella spp.
title_sort potential of a novel and thermostable recombinant phage endolysin of escherichia phage kw1e utar against shigella spp
url https://www.e3s-conferences.org/articles/e3sconf/pdf/2025/03/e3sconf_isgst2024_01015.pdf
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