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Characteristics of antimicrobial peptide OaBac5mini and its bactericidal mechanism against Escherichia coli
INTRODUCTION: Antimicrobial peptides (AMPs) play an important role in defending against the attack of pathogenic microorganisms. Among them, the proline-rich antibacterial peptides (PrAMPs) have been attracting close attention due to their simple structure, strong antibacterial activity, and low cel...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2023
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9995905/ https://www.ncbi.nlm.nih.gov/pubmed/36908510 http://dx.doi.org/10.3389/fvets.2023.1123054 |
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author | Shen, Shanshan Sun, Yawei Ren, Fei Blair, Jessica M. A. Siasat, Pauline Fan, Shuaiqi Hu, Jianhe He, Junping |
author_facet | Shen, Shanshan Sun, Yawei Ren, Fei Blair, Jessica M. A. Siasat, Pauline Fan, Shuaiqi Hu, Jianhe He, Junping |
author_sort | Shen, Shanshan |
collection | PubMed |
description | INTRODUCTION: Antimicrobial peptides (AMPs) play an important role in defending against the attack of pathogenic microorganisms. Among them, the proline-rich antibacterial peptides (PrAMPs) have been attracting close attention due to their simple structure, strong antibacterial activity, and low cell toxicity. OaBac5mini is an active fragment of the sheep-derived OaBac5 belonging to the PrAMPs family. METHODS: In this study, the antibacterial activity of OaBac5mini was investigated by testing the MICs against different stains of E. coli and S. aureus as well as the time-kill curve. The bactericidal mechanism was explored by determining the effect of OaBac5mini on the cell membrane. The stability and biosafety were also evaluated. RESULTS: The susceptibility test demonstrated that OaBac5mini showed potent antibacterial activity against the multidrug-resistant (MDR) E. coli isolates. It is noticeable that the absence of inner membrane protein SbmA in E. coli ATCC 25922 caused the MIC of OaBac5mini to increase 4-fold, implying OaBac5mini can enter into the cytoplasm via SbmA and plays its antibacterial activity. Moreover, the antibacterial activity of OaBac5mini against E. coli ATCC 25922 was not remarkably affected by the serum salts except for CaCl(2) at a physiological concentration, pH, temperature, repeated freeze-thawing and proteases (trypsin < 20 μg/mL, pepsin or proteinase K). Time-kill curve analysis showed OaBac5mini at the concentration of 200 μg/mL (8 × MICs) could effectively kill E. coli ATCC 25922 after co-incubation for 12 h. In addition, OaBac5mini was not hemolytic against rabbit red blood cells and also was not cytotoxic to porcine small intestinal epithelial cells (IPEC-J2). Bioinformatic analysis indicated that OaBac5mini is a linear peptide with 8 net positive charges. Furthermore, OaBac5mini significantly increased the outer membrane permeability and impaired the inner membrane integrity and ultrastructure of E. coli ATCC25922. CONCLUSION: OaBac5mini is a stable and potent PrAMP that kills E. coli by two different modes of action - inhibiting intracellular target(s) and damaging cell membrane. |
format | Online Article Text |
id | pubmed-9995905 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-99959052023-03-10 Characteristics of antimicrobial peptide OaBac5mini and its bactericidal mechanism against Escherichia coli Shen, Shanshan Sun, Yawei Ren, Fei Blair, Jessica M. A. Siasat, Pauline Fan, Shuaiqi Hu, Jianhe He, Junping Front Vet Sci Veterinary Science INTRODUCTION: Antimicrobial peptides (AMPs) play an important role in defending against the attack of pathogenic microorganisms. Among them, the proline-rich antibacterial peptides (PrAMPs) have been attracting close attention due to their simple structure, strong antibacterial activity, and low cell toxicity. OaBac5mini is an active fragment of the sheep-derived OaBac5 belonging to the PrAMPs family. METHODS: In this study, the antibacterial activity of OaBac5mini was investigated by testing the MICs against different stains of E. coli and S. aureus as well as the time-kill curve. The bactericidal mechanism was explored by determining the effect of OaBac5mini on the cell membrane. The stability and biosafety were also evaluated. RESULTS: The susceptibility test demonstrated that OaBac5mini showed potent antibacterial activity against the multidrug-resistant (MDR) E. coli isolates. It is noticeable that the absence of inner membrane protein SbmA in E. coli ATCC 25922 caused the MIC of OaBac5mini to increase 4-fold, implying OaBac5mini can enter into the cytoplasm via SbmA and plays its antibacterial activity. Moreover, the antibacterial activity of OaBac5mini against E. coli ATCC 25922 was not remarkably affected by the serum salts except for CaCl(2) at a physiological concentration, pH, temperature, repeated freeze-thawing and proteases (trypsin < 20 μg/mL, pepsin or proteinase K). Time-kill curve analysis showed OaBac5mini at the concentration of 200 μg/mL (8 × MICs) could effectively kill E. coli ATCC 25922 after co-incubation for 12 h. In addition, OaBac5mini was not hemolytic against rabbit red blood cells and also was not cytotoxic to porcine small intestinal epithelial cells (IPEC-J2). Bioinformatic analysis indicated that OaBac5mini is a linear peptide with 8 net positive charges. Furthermore, OaBac5mini significantly increased the outer membrane permeability and impaired the inner membrane integrity and ultrastructure of E. coli ATCC25922. CONCLUSION: OaBac5mini is a stable and potent PrAMP that kills E. coli by two different modes of action - inhibiting intracellular target(s) and damaging cell membrane. Frontiers Media S.A. 2023-02-23 /pmc/articles/PMC9995905/ /pubmed/36908510 http://dx.doi.org/10.3389/fvets.2023.1123054 Text en Copyright © 2023 Shen, Sun, Ren, Blair, Siasat, Fan, Hu and He. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Veterinary Science Shen, Shanshan Sun, Yawei Ren, Fei Blair, Jessica M. A. Siasat, Pauline Fan, Shuaiqi Hu, Jianhe He, Junping Characteristics of antimicrobial peptide OaBac5mini and its bactericidal mechanism against Escherichia coli |
title | Characteristics of antimicrobial peptide OaBac5mini and its bactericidal mechanism against Escherichia coli |
title_full | Characteristics of antimicrobial peptide OaBac5mini and its bactericidal mechanism against Escherichia coli |
title_fullStr | Characteristics of antimicrobial peptide OaBac5mini and its bactericidal mechanism against Escherichia coli |
title_full_unstemmed | Characteristics of antimicrobial peptide OaBac5mini and its bactericidal mechanism against Escherichia coli |
title_short | Characteristics of antimicrobial peptide OaBac5mini and its bactericidal mechanism against Escherichia coli |
title_sort | characteristics of antimicrobial peptide oabac5mini and its bactericidal mechanism against escherichia coli |
topic | Veterinary Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9995905/ https://www.ncbi.nlm.nih.gov/pubmed/36908510 http://dx.doi.org/10.3389/fvets.2023.1123054 |
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