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Expression, Purification, and Characterization of a Novel Hybrid Peptide with Potent Antibacterial Activity
The hybrid peptide cecropin A (1–8)–LL37 (17–30) (C–L), derived from the sequence of cecropin A (C) and LL-37 (L), showed significantly increased antibacterial activity and minimized hemolytic activity than C and L alone. To obtain high-level production of C–L, the deoxyribonucleic acid sequence enc...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6099547/ https://www.ncbi.nlm.nih.gov/pubmed/29925795 http://dx.doi.org/10.3390/molecules23061491 |
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author | Wei, Xubiao Wu, Rujuan Zhang, Lulu Ahmad, Baseer Si, Dayong Zhang, Rijun |
author_facet | Wei, Xubiao Wu, Rujuan Zhang, Lulu Ahmad, Baseer Si, Dayong Zhang, Rijun |
author_sort | Wei, Xubiao |
collection | PubMed |
description | The hybrid peptide cecropin A (1–8)–LL37 (17–30) (C–L), derived from the sequence of cecropin A (C) and LL-37 (L), showed significantly increased antibacterial activity and minimized hemolytic activity than C and L alone. To obtain high-level production of C–L, the deoxyribonucleic acid sequence encoding C–L with preferred codons was cloned into pET-SUMO to construct a fusion expression vector, and overexpressed in Escherichia coli (E. coli) BL21 (DE3). The maximum fusion protein (92% purity) was obtained with the yield of 89.14 mg/L fermentation culture after purification with Ni-NTA Sepharose column. The hybrid C–L was cleaved from the fusion protein by SUMO-protease, and 17.54 mg/L pure active C–L was obtained. Furthermore, the purified C–L showed identical antibacterial and hemolytic activity to synthesized C–L. Stability analysis results exhibited that the activity of C–L changed little below 80 °C for 20 min, but when the temperature exceeded 80 °C, a significant decrease was observed. Varying the pH from 5.0 to 10.0 did not appear to influence the activity of C–L, however, pH below 4.0 decreased the antibacterial activity of C–L rapidly. Under the challenge of several proteases (pepsin, trypsin, and proteinase K), the functional activity of C–L was maintained over 50%. In summary, this study not only supplied an effective approach for high-level production of hybrid peptide C–L, but paved the way for its further exploration in controlling infectious diseases of farm animals or even humans. |
format | Online Article Text |
id | pubmed-6099547 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-60995472018-11-13 Expression, Purification, and Characterization of a Novel Hybrid Peptide with Potent Antibacterial Activity Wei, Xubiao Wu, Rujuan Zhang, Lulu Ahmad, Baseer Si, Dayong Zhang, Rijun Molecules Article The hybrid peptide cecropin A (1–8)–LL37 (17–30) (C–L), derived from the sequence of cecropin A (C) and LL-37 (L), showed significantly increased antibacterial activity and minimized hemolytic activity than C and L alone. To obtain high-level production of C–L, the deoxyribonucleic acid sequence encoding C–L with preferred codons was cloned into pET-SUMO to construct a fusion expression vector, and overexpressed in Escherichia coli (E. coli) BL21 (DE3). The maximum fusion protein (92% purity) was obtained with the yield of 89.14 mg/L fermentation culture after purification with Ni-NTA Sepharose column. The hybrid C–L was cleaved from the fusion protein by SUMO-protease, and 17.54 mg/L pure active C–L was obtained. Furthermore, the purified C–L showed identical antibacterial and hemolytic activity to synthesized C–L. Stability analysis results exhibited that the activity of C–L changed little below 80 °C for 20 min, but when the temperature exceeded 80 °C, a significant decrease was observed. Varying the pH from 5.0 to 10.0 did not appear to influence the activity of C–L, however, pH below 4.0 decreased the antibacterial activity of C–L rapidly. Under the challenge of several proteases (pepsin, trypsin, and proteinase K), the functional activity of C–L was maintained over 50%. In summary, this study not only supplied an effective approach for high-level production of hybrid peptide C–L, but paved the way for its further exploration in controlling infectious diseases of farm animals or even humans. MDPI 2018-06-20 /pmc/articles/PMC6099547/ /pubmed/29925795 http://dx.doi.org/10.3390/molecules23061491 Text en © 2018 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Wei, Xubiao Wu, Rujuan Zhang, Lulu Ahmad, Baseer Si, Dayong Zhang, Rijun Expression, Purification, and Characterization of a Novel Hybrid Peptide with Potent Antibacterial Activity |
title | Expression, Purification, and Characterization of a Novel Hybrid Peptide with Potent Antibacterial Activity |
title_full | Expression, Purification, and Characterization of a Novel Hybrid Peptide with Potent Antibacterial Activity |
title_fullStr | Expression, Purification, and Characterization of a Novel Hybrid Peptide with Potent Antibacterial Activity |
title_full_unstemmed | Expression, Purification, and Characterization of a Novel Hybrid Peptide with Potent Antibacterial Activity |
title_short | Expression, Purification, and Characterization of a Novel Hybrid Peptide with Potent Antibacterial Activity |
title_sort | expression, purification, and characterization of a novel hybrid peptide with potent antibacterial activity |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6099547/ https://www.ncbi.nlm.nih.gov/pubmed/29925795 http://dx.doi.org/10.3390/molecules23061491 |
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