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Eradication of drug-resistant Acinetobacter baumannii by cell-penetrating peptide fused endolysin
Antimicrobial agents targeting peptidoglycan have shown successful results in eliminating bacteria with high selective toxicity. Bacteriophage encoded endolysin as an alternative antibiotics is a peptidoglycan degrading enzyme with a low rate of resistance. Here, the engineered endolysin was develop...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Microbiological Society of Korea
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9132170/ https://www.ncbi.nlm.nih.gov/pubmed/35614377 http://dx.doi.org/10.1007/s12275-022-2107-y |
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author | Lim, Jeonghyun Jang, Jaeyeon Myung, Heejoon Song, Miryoung |
author_facet | Lim, Jeonghyun Jang, Jaeyeon Myung, Heejoon Song, Miryoung |
author_sort | Lim, Jeonghyun |
collection | PubMed |
description | Antimicrobial agents targeting peptidoglycan have shown successful results in eliminating bacteria with high selective toxicity. Bacteriophage encoded endolysin as an alternative antibiotics is a peptidoglycan degrading enzyme with a low rate of resistance. Here, the engineered endolysin was developed to defeat multiple drug-resistant (MDR) Acinetobacter baumannii. First, putative endolysin PA90 was predicted by genome analysis of isolated Pseudomonas phage PBPA. The His-tagged PA90 was purified from BL21(DE3) pLysS and tested for the enzymatic activity using Gram-negative pathogens known for having a high antibiotic resistance rate including A. baumannii. Since the measured activity of PA90 was low, probably due to the outer membrane, cell-penetrating peptide (CPP) DS4.3 was introduced at the N-terminus of PA90 to aid access to its substrate. This engineered endolysin, DS-PA90, completely killed A. baumannii at 0.25 µM, at which concentration PA90 could only eliminate less than one log in CFU/ml. Additionally, DS-PA90 has tolerance to NaCl, where the ∼50% of activity could be maintained in the presence of 150 mM NaCl, and stable activity was also observed with changes in pH or temperature. Even MDR A. baumannii strains were highly susceptible to DS-PA90 treatment: five out of nine strains were entirely killed and four strains were reduced by 3–4 log in CFU/ml. Consequently, DS-PA90 could protect waxworm from A. baumannii-induced death by ∼70% for ATCC 17978 or ∼44% for MDR strain 1656-2 infection. Collectively, our data suggest that CPP-fused endolysin can be an effective antibacterial agent against Gram-negative pathogens regardless of antibiotics resistance mechanisms. ELECTRONIC SUPPLEMENTARY MATERIAL: Supplemental material for this article may be found at 10.1007/s12275-022-2107-y. |
format | Online Article Text |
id | pubmed-9132170 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | The Microbiological Society of Korea |
record_format | MEDLINE/PubMed |
spelling | pubmed-91321702022-05-26 Eradication of drug-resistant Acinetobacter baumannii by cell-penetrating peptide fused endolysin Lim, Jeonghyun Jang, Jaeyeon Myung, Heejoon Song, Miryoung J Microbiol Virology Antimicrobial agents targeting peptidoglycan have shown successful results in eliminating bacteria with high selective toxicity. Bacteriophage encoded endolysin as an alternative antibiotics is a peptidoglycan degrading enzyme with a low rate of resistance. Here, the engineered endolysin was developed to defeat multiple drug-resistant (MDR) Acinetobacter baumannii. First, putative endolysin PA90 was predicted by genome analysis of isolated Pseudomonas phage PBPA. The His-tagged PA90 was purified from BL21(DE3) pLysS and tested for the enzymatic activity using Gram-negative pathogens known for having a high antibiotic resistance rate including A. baumannii. Since the measured activity of PA90 was low, probably due to the outer membrane, cell-penetrating peptide (CPP) DS4.3 was introduced at the N-terminus of PA90 to aid access to its substrate. This engineered endolysin, DS-PA90, completely killed A. baumannii at 0.25 µM, at which concentration PA90 could only eliminate less than one log in CFU/ml. Additionally, DS-PA90 has tolerance to NaCl, where the ∼50% of activity could be maintained in the presence of 150 mM NaCl, and stable activity was also observed with changes in pH or temperature. Even MDR A. baumannii strains were highly susceptible to DS-PA90 treatment: five out of nine strains were entirely killed and four strains were reduced by 3–4 log in CFU/ml. Consequently, DS-PA90 could protect waxworm from A. baumannii-induced death by ∼70% for ATCC 17978 or ∼44% for MDR strain 1656-2 infection. Collectively, our data suggest that CPP-fused endolysin can be an effective antibacterial agent against Gram-negative pathogens regardless of antibiotics resistance mechanisms. ELECTRONIC SUPPLEMENTARY MATERIAL: Supplemental material for this article may be found at 10.1007/s12275-022-2107-y. The Microbiological Society of Korea 2022-05-25 2022 /pmc/articles/PMC9132170/ /pubmed/35614377 http://dx.doi.org/10.1007/s12275-022-2107-y Text en © Author(s) 2022, under the exclusive license with the Microbiological Society of Korea This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic. |
spellingShingle | Virology Lim, Jeonghyun Jang, Jaeyeon Myung, Heejoon Song, Miryoung Eradication of drug-resistant Acinetobacter baumannii by cell-penetrating peptide fused endolysin |
title | Eradication of drug-resistant Acinetobacter baumannii by cell-penetrating peptide fused endolysin |
title_full | Eradication of drug-resistant Acinetobacter baumannii by cell-penetrating peptide fused endolysin |
title_fullStr | Eradication of drug-resistant Acinetobacter baumannii by cell-penetrating peptide fused endolysin |
title_full_unstemmed | Eradication of drug-resistant Acinetobacter baumannii by cell-penetrating peptide fused endolysin |
title_short | Eradication of drug-resistant Acinetobacter baumannii by cell-penetrating peptide fused endolysin |
title_sort | eradication of drug-resistant acinetobacter baumannii by cell-penetrating peptide fused endolysin |
topic | Virology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9132170/ https://www.ncbi.nlm.nih.gov/pubmed/35614377 http://dx.doi.org/10.1007/s12275-022-2107-y |
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