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Structural and Functional Characterization of a Novel Recombinant Antimicrobial Peptide from Hermetia illucens

Antibiotics are commonly used to treat pathogenic bacteria, but their prolonged use contributes to the development and spread of drug-resistant microorganisms raising the challenge to find new alternative drugs. Antimicrobial peptides (AMPs) are small/medium molecules ranging 10–60 residues synthesi...

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Autores principales: Di Somma, Angela, Moretta, Antonio, Cané, Carolina, Scieuzo, Carmen, Salvia, Rosanna, Falabella, Patrizia, Duilio, Angela
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8929087/
https://www.ncbi.nlm.nih.gov/pubmed/35723380
http://dx.doi.org/10.3390/cimb44010001
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author Di Somma, Angela
Moretta, Antonio
Cané, Carolina
Scieuzo, Carmen
Salvia, Rosanna
Falabella, Patrizia
Duilio, Angela
author_facet Di Somma, Angela
Moretta, Antonio
Cané, Carolina
Scieuzo, Carmen
Salvia, Rosanna
Falabella, Patrizia
Duilio, Angela
author_sort Di Somma, Angela
collection PubMed
description Antibiotics are commonly used to treat pathogenic bacteria, but their prolonged use contributes to the development and spread of drug-resistant microorganisms raising the challenge to find new alternative drugs. Antimicrobial peptides (AMPs) are small/medium molecules ranging 10–60 residues synthesized by all living organisms and playing important roles in the defense systems. These features, together with the inability of microorganisms to develop resistance against the majority of AMPs, suggest that these molecules might represent effective alternatives to classical antibiotics. Because of their high biodiversity, with over one million described species, and their ability to live in hostile environments, insects represent the largest source of these molecules. However, production of insect AMPs in native forms is challenging. In this work we investigate a defensin-like antimicrobial peptide identified in the Hermetia illucens insect through a combination of transcriptomics and bioinformatics approaches. The C-15867 AMP was produced by recombinant DNA technology as a glutathione S-transferase (GST) fusion peptide and purified by affinity chromatography. The free peptide was then obtained by thrombin proteolysis and structurally characterized by mass spectrometry and circular dichroism analyses. The antibacterial activity of the C-15867 peptide was evaluated in vivo by determination of the minimum inhibitory concentration (MIC). Finally, crystal violet assays and SEM analyses suggested disruption of the cell membrane architecture and pore formation with leaking of cytosolic material.
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spelling pubmed-89290872022-06-04 Structural and Functional Characterization of a Novel Recombinant Antimicrobial Peptide from Hermetia illucens Di Somma, Angela Moretta, Antonio Cané, Carolina Scieuzo, Carmen Salvia, Rosanna Falabella, Patrizia Duilio, Angela Curr Issues Mol Biol Article Antibiotics are commonly used to treat pathogenic bacteria, but their prolonged use contributes to the development and spread of drug-resistant microorganisms raising the challenge to find new alternative drugs. Antimicrobial peptides (AMPs) are small/medium molecules ranging 10–60 residues synthesized by all living organisms and playing important roles in the defense systems. These features, together with the inability of microorganisms to develop resistance against the majority of AMPs, suggest that these molecules might represent effective alternatives to classical antibiotics. Because of their high biodiversity, with over one million described species, and their ability to live in hostile environments, insects represent the largest source of these molecules. However, production of insect AMPs in native forms is challenging. In this work we investigate a defensin-like antimicrobial peptide identified in the Hermetia illucens insect through a combination of transcriptomics and bioinformatics approaches. The C-15867 AMP was produced by recombinant DNA technology as a glutathione S-transferase (GST) fusion peptide and purified by affinity chromatography. The free peptide was then obtained by thrombin proteolysis and structurally characterized by mass spectrometry and circular dichroism analyses. The antibacterial activity of the C-15867 peptide was evaluated in vivo by determination of the minimum inhibitory concentration (MIC). Finally, crystal violet assays and SEM analyses suggested disruption of the cell membrane architecture and pore formation with leaking of cytosolic material. MDPI 2021-12-21 /pmc/articles/PMC8929087/ /pubmed/35723380 http://dx.doi.org/10.3390/cimb44010001 Text en © 2021 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
Di Somma, Angela
Moretta, Antonio
Cané, Carolina
Scieuzo, Carmen
Salvia, Rosanna
Falabella, Patrizia
Duilio, Angela
Structural and Functional Characterization of a Novel Recombinant Antimicrobial Peptide from Hermetia illucens
title Structural and Functional Characterization of a Novel Recombinant Antimicrobial Peptide from Hermetia illucens
title_full Structural and Functional Characterization of a Novel Recombinant Antimicrobial Peptide from Hermetia illucens
title_fullStr Structural and Functional Characterization of a Novel Recombinant Antimicrobial Peptide from Hermetia illucens
title_full_unstemmed Structural and Functional Characterization of a Novel Recombinant Antimicrobial Peptide from Hermetia illucens
title_short Structural and Functional Characterization of a Novel Recombinant Antimicrobial Peptide from Hermetia illucens
title_sort structural and functional characterization of a novel recombinant antimicrobial peptide from hermetia illucens
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8929087/
https://www.ncbi.nlm.nih.gov/pubmed/35723380
http://dx.doi.org/10.3390/cimb44010001
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