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Characterization of Three Different Endolysins Effective against Gram-Negative Bacteria

Genes encoding endolysins were identified and cloned from three different Escherichia coli bacteriophages, 10-24(13), PBEC30, and PBEC56. Putative antimicrobial peptide (AMP)-like C-terminal alpha helix structures with amphipathic natures were predicted from the three endolysins. Each gene was clone...

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Autores principales: Jeong, Tae-Hwan, Hong, Hye-Won, Kim, Min Soo, Song, Miryoung, Myung, Heejoon
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10053066/
https://www.ncbi.nlm.nih.gov/pubmed/36992387
http://dx.doi.org/10.3390/v15030679
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author Jeong, Tae-Hwan
Hong, Hye-Won
Kim, Min Soo
Song, Miryoung
Myung, Heejoon
author_facet Jeong, Tae-Hwan
Hong, Hye-Won
Kim, Min Soo
Song, Miryoung
Myung, Heejoon
author_sort Jeong, Tae-Hwan
collection PubMed
description Genes encoding endolysins were identified and cloned from three different Escherichia coli bacteriophages, 10-24(13), PBEC30, and PBEC56. Putative antimicrobial peptide (AMP)-like C-terminal alpha helix structures with amphipathic natures were predicted from the three endolysins. Each gene was cloned and expressed as hexahistidine-tagged forms, and the products were purified and characterized. The purified endolysins exhibited antibacterial activities against a variety of Gram-negative bacteria including Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, and Klebsiella pneumonia. Their antibacterial activities were improved by N-terminal fusion with an antimicrobial peptide, cecropin A. Minimum inhibitory concentrations (MIC) were as low as 4 μg/mL, depending on the targeted strain. The endolysins’ enzymatic activities were not affected by changes in pH at ranges from 5 to 10 and were stable at temperatures between 4 and 65 °C. The in vivo efficacies of the three endolysins were also demonstrated using Galleria melonella for infection models.
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spelling pubmed-100530662023-03-30 Characterization of Three Different Endolysins Effective against Gram-Negative Bacteria Jeong, Tae-Hwan Hong, Hye-Won Kim, Min Soo Song, Miryoung Myung, Heejoon Viruses Article Genes encoding endolysins were identified and cloned from three different Escherichia coli bacteriophages, 10-24(13), PBEC30, and PBEC56. Putative antimicrobial peptide (AMP)-like C-terminal alpha helix structures with amphipathic natures were predicted from the three endolysins. Each gene was cloned and expressed as hexahistidine-tagged forms, and the products were purified and characterized. The purified endolysins exhibited antibacterial activities against a variety of Gram-negative bacteria including Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, and Klebsiella pneumonia. Their antibacterial activities were improved by N-terminal fusion with an antimicrobial peptide, cecropin A. Minimum inhibitory concentrations (MIC) were as low as 4 μg/mL, depending on the targeted strain. The endolysins’ enzymatic activities were not affected by changes in pH at ranges from 5 to 10 and were stable at temperatures between 4 and 65 °C. The in vivo efficacies of the three endolysins were also demonstrated using Galleria melonella for infection models. MDPI 2023-03-04 /pmc/articles/PMC10053066/ /pubmed/36992387 http://dx.doi.org/10.3390/v15030679 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Jeong, Tae-Hwan
Hong, Hye-Won
Kim, Min Soo
Song, Miryoung
Myung, Heejoon
Characterization of Three Different Endolysins Effective against Gram-Negative Bacteria
title Characterization of Three Different Endolysins Effective against Gram-Negative Bacteria
title_full Characterization of Three Different Endolysins Effective against Gram-Negative Bacteria
title_fullStr Characterization of Three Different Endolysins Effective against Gram-Negative Bacteria
title_full_unstemmed Characterization of Three Different Endolysins Effective against Gram-Negative Bacteria
title_short Characterization of Three Different Endolysins Effective against Gram-Negative Bacteria
title_sort characterization of three different endolysins effective against gram-negative bacteria
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10053066/
https://www.ncbi.nlm.nih.gov/pubmed/36992387
http://dx.doi.org/10.3390/v15030679
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