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Structural and functional features of a broad-spectrum prophage-encoded enzybiotic from Enterococcus faecium
Multidrug-resistant (MDR) bacteria have become a growing threat to public health. The gram-positive Enterococcus faecium is classified by WHO as a high-priority pathogen among the global priority list of antibiotic-resistant bacteria. Peptidoglycan-degrading enzymes (PDEs), also known as enzybiotics...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2023
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10167349/ https://www.ncbi.nlm.nih.gov/pubmed/37156923 http://dx.doi.org/10.1038/s41598-023-34309-2 |
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author | Premetis, Georgios E. Stathi, Angeliki Papageorgiou, Anastassios C. Labrou, Nikolaos E. |
author_facet | Premetis, Georgios E. Stathi, Angeliki Papageorgiou, Anastassios C. Labrou, Nikolaos E. |
author_sort | Premetis, Georgios E. |
collection | PubMed |
description | Multidrug-resistant (MDR) bacteria have become a growing threat to public health. The gram-positive Enterococcus faecium is classified by WHO as a high-priority pathogen among the global priority list of antibiotic-resistant bacteria. Peptidoglycan-degrading enzymes (PDEs), also known as enzybiotics, are useful bactericidal agents in the fight against resistant bacteria. In this work, a genome-based screening approach of the genome of E. faecium allowed the identification of a putative PDE gene with predictive amidase activity (EfAmi1; EC 3.5.1.28) in a prophage-integrated sequence. EfAmi1 is composed by two domains: a N-terminal Zn(2+)-dependent N-acetylmuramoyl-l-alanine amidase-2 (NALAA-2) domain and a C-terminal domain with unknown structure and function. The full-length gene of EfAmi1 was cloned and expressed as a 6xHis-tagged protein in E. coli. EfAmi1 was produced as a soluble protein, purified, and its lytic and antimicrobial activities were investigated using turbidity reduction and Kirby–Bauer disk-diffusion assays against clinically isolated bacterial pathogens. The crystal structure of the N-terminal amidase-2 domain was determined using X-ray crystallography at 1.97 Å resolution. It adopts a globular fold with several α-helices surrounding a central five-stranded β-sheet. Sequence comparison revealed a cluster of conserved amino acids that defines a putative binding site for a buried zinc ion. The results of the present study suggest that EfAmi1 displays high lytic and antimicrobial activity and may represent a promising new antimicrobial in the post-antibiotic era. |
format | Online Article Text |
id | pubmed-10167349 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-101673492023-05-10 Structural and functional features of a broad-spectrum prophage-encoded enzybiotic from Enterococcus faecium Premetis, Georgios E. Stathi, Angeliki Papageorgiou, Anastassios C. Labrou, Nikolaos E. Sci Rep Article Multidrug-resistant (MDR) bacteria have become a growing threat to public health. The gram-positive Enterococcus faecium is classified by WHO as a high-priority pathogen among the global priority list of antibiotic-resistant bacteria. Peptidoglycan-degrading enzymes (PDEs), also known as enzybiotics, are useful bactericidal agents in the fight against resistant bacteria. In this work, a genome-based screening approach of the genome of E. faecium allowed the identification of a putative PDE gene with predictive amidase activity (EfAmi1; EC 3.5.1.28) in a prophage-integrated sequence. EfAmi1 is composed by two domains: a N-terminal Zn(2+)-dependent N-acetylmuramoyl-l-alanine amidase-2 (NALAA-2) domain and a C-terminal domain with unknown structure and function. The full-length gene of EfAmi1 was cloned and expressed as a 6xHis-tagged protein in E. coli. EfAmi1 was produced as a soluble protein, purified, and its lytic and antimicrobial activities were investigated using turbidity reduction and Kirby–Bauer disk-diffusion assays against clinically isolated bacterial pathogens. The crystal structure of the N-terminal amidase-2 domain was determined using X-ray crystallography at 1.97 Å resolution. It adopts a globular fold with several α-helices surrounding a central five-stranded β-sheet. Sequence comparison revealed a cluster of conserved amino acids that defines a putative binding site for a buried zinc ion. The results of the present study suggest that EfAmi1 displays high lytic and antimicrobial activity and may represent a promising new antimicrobial in the post-antibiotic era. Nature Publishing Group UK 2023-05-08 /pmc/articles/PMC10167349/ /pubmed/37156923 http://dx.doi.org/10.1038/s41598-023-34309-2 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Premetis, Georgios E. Stathi, Angeliki Papageorgiou, Anastassios C. Labrou, Nikolaos E. Structural and functional features of a broad-spectrum prophage-encoded enzybiotic from Enterococcus faecium |
title | Structural and functional features of a broad-spectrum prophage-encoded enzybiotic from Enterococcus faecium |
title_full | Structural and functional features of a broad-spectrum prophage-encoded enzybiotic from Enterococcus faecium |
title_fullStr | Structural and functional features of a broad-spectrum prophage-encoded enzybiotic from Enterococcus faecium |
title_full_unstemmed | Structural and functional features of a broad-spectrum prophage-encoded enzybiotic from Enterococcus faecium |
title_short | Structural and functional features of a broad-spectrum prophage-encoded enzybiotic from Enterococcus faecium |
title_sort | structural and functional features of a broad-spectrum prophage-encoded enzybiotic from enterococcus faecium |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10167349/ https://www.ncbi.nlm.nih.gov/pubmed/37156923 http://dx.doi.org/10.1038/s41598-023-34309-2 |
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