Structural and Functional Characterization of a Novel Recombinant Antimicrobial Peptide from <i>Hermetia illucens</i>
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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| Format: | Article |
| Language: | English |
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MDPI AG
2021-12-01
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| Series: | Current Issues in Molecular Biology |
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| Online Access: | https://www.mdpi.com/1467-3045/44/1/1 |
| _version_ | 1811183601932304384 |
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| author | Angela Di Somma Antonio Moretta Carolina Cané Carmen Scieuzo Rosanna Salvia Patrizia Falabella Angela Duilio |
| author_facet | Angela Di Somma Antonio Moretta Carolina Cané Carmen Scieuzo Rosanna Salvia Patrizia Falabella Angela Duilio |
| author_sort | Angela Di Somma |
| collection | DOAJ |
| 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 <i>Hermetia illucens</i> 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. |
| first_indexed | 2024-04-11T09:49:44Z |
| format | Article |
| id | doaj.art-76c056f23e2d4c9d8a8a15c4e7f48bb9 |
| institution | Directory Open Access Journal |
| issn | 1467-3037 1467-3045 |
| language | English |
| last_indexed | 2024-04-11T09:49:44Z |
| publishDate | 2021-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Current Issues in Molecular Biology |
| spelling | doaj.art-76c056f23e2d4c9d8a8a15c4e7f48bb92022-12-22T04:30:50ZengMDPI AGCurrent Issues in Molecular Biology1467-30371467-30452021-12-0144111310.3390/cimb44010001Structural and Functional Characterization of a Novel Recombinant Antimicrobial Peptide from <i>Hermetia illucens</i>Angela Di Somma0Antonio Moretta1Carolina Cané2Carmen Scieuzo3Rosanna Salvia4Patrizia Falabella5Angela Duilio6Department of Chemical Sciences, University of Naples “Federico II”, Via Cinthia 4, 80126 Napoli, ItalyDepartment of Sciences, University of Basilicata, 85100 Potenza, ItalyDepartment of Chemical Sciences, University of Naples “Federico II”, Via Cinthia 4, 80126 Napoli, ItalyDepartment of Sciences, University of Basilicata, 85100 Potenza, ItalyDepartment of Sciences, University of Basilicata, 85100 Potenza, ItalyDepartment of Sciences, University of Basilicata, 85100 Potenza, ItalyDepartment of Chemical Sciences, University of Naples “Federico II”, Via Cinthia 4, 80126 Napoli, ItalyAntibiotics 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 <i>Hermetia illucens</i> 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.https://www.mdpi.com/1467-3045/44/1/1antimicrobial peptidesdefensinsinsects<i>Hermetia illucens</i>antibacterial activity |
| spellingShingle | Angela Di Somma Antonio Moretta Carolina Cané Carmen Scieuzo Rosanna Salvia Patrizia Falabella Angela Duilio Structural and Functional Characterization of a Novel Recombinant Antimicrobial Peptide from <i>Hermetia illucens</i> Current Issues in Molecular Biology antimicrobial peptides defensins insects <i>Hermetia illucens</i> antibacterial activity |
| title | Structural and Functional Characterization of a Novel Recombinant Antimicrobial Peptide from <i>Hermetia illucens</i> |
| title_full | Structural and Functional Characterization of a Novel Recombinant Antimicrobial Peptide from <i>Hermetia illucens</i> |
| title_fullStr | Structural and Functional Characterization of a Novel Recombinant Antimicrobial Peptide from <i>Hermetia illucens</i> |
| title_full_unstemmed | Structural and Functional Characterization of a Novel Recombinant Antimicrobial Peptide from <i>Hermetia illucens</i> |
| title_short | Structural and Functional Characterization of a Novel Recombinant Antimicrobial Peptide from <i>Hermetia illucens</i> |
| title_sort | structural and functional characterization of a novel recombinant antimicrobial peptide from i hermetia illucens i |
| topic | antimicrobial peptides defensins insects <i>Hermetia illucens</i> antibacterial activity |
| url | https://www.mdpi.com/1467-3045/44/1/1 |
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