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High Level Expression and Purification of Cecropin-like Antimicrobial Peptides in Escherichia coli
Cecropins are a family of antimicrobial peptides (AMPs) that are widely found in the innate immune system of Cecropia moths. Cecropins exhibit a broad spectrum of antimicrobial and anticancer activities. The structures of Cecropins are composed of 34–39 amino acids with an N-terminal amphipathic α-h...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9220022/ https://www.ncbi.nlm.nih.gov/pubmed/35740373 http://dx.doi.org/10.3390/biomedicines10061351 |
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author | Wu, Chih-Lung Chih, Ya-Han Hsieh, Hsin-Ying Peng, Kuang-Li Lee, Yi-Zong Yip, Bak-Sau Sue, Shih-Che Cheng, Jya-Wei |
author_facet | Wu, Chih-Lung Chih, Ya-Han Hsieh, Hsin-Ying Peng, Kuang-Li Lee, Yi-Zong Yip, Bak-Sau Sue, Shih-Che Cheng, Jya-Wei |
author_sort | Wu, Chih-Lung |
collection | PubMed |
description | Cecropins are a family of antimicrobial peptides (AMPs) that are widely found in the innate immune system of Cecropia moths. Cecropins exhibit a broad spectrum of antimicrobial and anticancer activities. The structures of Cecropins are composed of 34–39 amino acids with an N-terminal amphipathic α-helix, an AGP hinge and a hydrophobic C-terminal α-helix. KR12AGPWR6 was designed based on the Cecropin-like structural feature. In addition to its antimicrobial activities, KR12AGPWR6 also possesses enhanced salt resistance, antiendotoxin and anticancer properties. Herein, we have developed a strategy to produce recombinant KR12AGPWR6 through a salt-sensitive, pH and temperature dependent intein self-cleavage system. The His6-Intein-KR12AGPWR6 was expressed by E. coli and KR12AGPWR6 was released by the self-cleavage of intein under optimized ionic strength, pH and temperature conditions. The molecular weight and structural feature of the recombinant KR12AGPWR6 was determined by MALDI-TOF mass, CD, and NMR spectroscopy. The recombinant KR12AGPWR6 exhibited similar antimicrobial activities compared to the chemically synthesized KR12AGPWR6. Our results provide a potential strategy to obtain large quantities of AMPs and this method is feasible and easy to scale up for commercial production. |
format | Online Article Text |
id | pubmed-9220022 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-92200222022-06-24 High Level Expression and Purification of Cecropin-like Antimicrobial Peptides in Escherichia coli Wu, Chih-Lung Chih, Ya-Han Hsieh, Hsin-Ying Peng, Kuang-Li Lee, Yi-Zong Yip, Bak-Sau Sue, Shih-Che Cheng, Jya-Wei Biomedicines Article Cecropins are a family of antimicrobial peptides (AMPs) that are widely found in the innate immune system of Cecropia moths. Cecropins exhibit a broad spectrum of antimicrobial and anticancer activities. The structures of Cecropins are composed of 34–39 amino acids with an N-terminal amphipathic α-helix, an AGP hinge and a hydrophobic C-terminal α-helix. KR12AGPWR6 was designed based on the Cecropin-like structural feature. In addition to its antimicrobial activities, KR12AGPWR6 also possesses enhanced salt resistance, antiendotoxin and anticancer properties. Herein, we have developed a strategy to produce recombinant KR12AGPWR6 through a salt-sensitive, pH and temperature dependent intein self-cleavage system. The His6-Intein-KR12AGPWR6 was expressed by E. coli and KR12AGPWR6 was released by the self-cleavage of intein under optimized ionic strength, pH and temperature conditions. The molecular weight and structural feature of the recombinant KR12AGPWR6 was determined by MALDI-TOF mass, CD, and NMR spectroscopy. The recombinant KR12AGPWR6 exhibited similar antimicrobial activities compared to the chemically synthesized KR12AGPWR6. Our results provide a potential strategy to obtain large quantities of AMPs and this method is feasible and easy to scale up for commercial production. MDPI 2022-06-08 /pmc/articles/PMC9220022/ /pubmed/35740373 http://dx.doi.org/10.3390/biomedicines10061351 Text en © 2022 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 Wu, Chih-Lung Chih, Ya-Han Hsieh, Hsin-Ying Peng, Kuang-Li Lee, Yi-Zong Yip, Bak-Sau Sue, Shih-Che Cheng, Jya-Wei High Level Expression and Purification of Cecropin-like Antimicrobial Peptides in Escherichia coli |
title | High Level Expression and Purification of Cecropin-like Antimicrobial Peptides in Escherichia coli |
title_full | High Level Expression and Purification of Cecropin-like Antimicrobial Peptides in Escherichia coli |
title_fullStr | High Level Expression and Purification of Cecropin-like Antimicrobial Peptides in Escherichia coli |
title_full_unstemmed | High Level Expression and Purification of Cecropin-like Antimicrobial Peptides in Escherichia coli |
title_short | High Level Expression and Purification of Cecropin-like Antimicrobial Peptides in Escherichia coli |
title_sort | high level expression and purification of cecropin-like antimicrobial peptides in escherichia coli |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9220022/ https://www.ncbi.nlm.nih.gov/pubmed/35740373 http://dx.doi.org/10.3390/biomedicines10061351 |
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