Prokaryotic Expression, Purification, and Antibacterial Activity of the Hepcidin Peptide of Crescent Sweetlips (Plectorhinchus cinctus)
The hepcidin peptide of crescent sweetlips (Plectorhinchus cinctus) is a cysteine-rich, cationic antimicrobial peptide that plays a crucial role in the innate immune system’s defense against invading microbes. The aim of this study was to identify the optimal parameters for prokaryotic expression an...
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ftmdpi:oai:mdpi.com:/1467-3045/45/9/456/ 2023-10-01T03:54:10+02:00 Prokaryotic Expression, Purification, and Antibacterial Activity of the Hepcidin Peptide of Crescent Sweetlips (Plectorhinchus cinctus) Peixin Wang Zhongjing Lin Shaoling Lin Baodong Zheng Yi Zhang Jiamiao Hu agris 2023-08-31 application/pdf https://doi.org/10.3390/cimb45090456 eng eng Multidisciplinary Digital Publishing Institute Molecular Pharmacology https://dx.doi.org/10.3390/cimb45090456 https://creativecommons.org/licenses/by/4.0/ Current Issues in Molecular Biology Volume 45 Issue 9 Pages: 7212-7227 crescent sweetlips hepcidin soluble expression optimization antibacterial activity Text 2023 ftmdpi https://doi.org/10.3390/cimb45090456 2023-09-03T23:54:56Z The hepcidin peptide of crescent sweetlips (Plectorhinchus cinctus) is a cysteine-rich, cationic antimicrobial peptide that plays a crucial role in the innate immune system’s defense against invading microbes. The aim of this study was to identify the optimal parameters for prokaryotic expression and purification of this hepcidin peptide and characterize its antibacterial activity. The recombinant hepcidin peptides were expressed in Escherichia coli strain Arctic Express (DE3), with culture and induction conditions optimized using response surface methodology (RSM). The obtained hepcidin peptides were then purified before tag cleavage, and their antibacterial activity was determined. The obtained results revealed that induction temperature had the most significant impact on the production of soluble recombinant peptides. The optimum induction conditions were determined to be an isopropylthio-β-galactoside (IPTG) concentration of 0.21 mmol/L, induction temperature of 18.81 °C, and an induction time of 16.01 h. Subsequently, the recombinant hepcidin peptide was successfully purified using Ni-IDA affinity chromatography followed by SUMO protease cleavage. The obtained hepcidin peptide (without His-SUMO tag) demonstrated strong antimicrobial activity in vitro against V. parahaemolyticus, E. coli, and S. aureus. The results showed prokaryotic (E. coli) expression is a feasible way to produce the hepcidin peptide of crescent sweetlips in a cost-effective way, which has great potential to be used as an antimicrobial agent in aquaculture. Text Arctic MDPI Open Access Publishing Arctic Ida ENVELOPE(170.483,170.483,-83.583,-83.583) Current Issues in Molecular Biology 45 9 7212 7227 |
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English |
topic |
crescent sweetlips hepcidin soluble expression optimization antibacterial activity |
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crescent sweetlips hepcidin soluble expression optimization antibacterial activity Peixin Wang Zhongjing Lin Shaoling Lin Baodong Zheng Yi Zhang Jiamiao Hu Prokaryotic Expression, Purification, and Antibacterial Activity of the Hepcidin Peptide of Crescent Sweetlips (Plectorhinchus cinctus) |
topic_facet |
crescent sweetlips hepcidin soluble expression optimization antibacterial activity |
description |
The hepcidin peptide of crescent sweetlips (Plectorhinchus cinctus) is a cysteine-rich, cationic antimicrobial peptide that plays a crucial role in the innate immune system’s defense against invading microbes. The aim of this study was to identify the optimal parameters for prokaryotic expression and purification of this hepcidin peptide and characterize its antibacterial activity. The recombinant hepcidin peptides were expressed in Escherichia coli strain Arctic Express (DE3), with culture and induction conditions optimized using response surface methodology (RSM). The obtained hepcidin peptides were then purified before tag cleavage, and their antibacterial activity was determined. The obtained results revealed that induction temperature had the most significant impact on the production of soluble recombinant peptides. The optimum induction conditions were determined to be an isopropylthio-β-galactoside (IPTG) concentration of 0.21 mmol/L, induction temperature of 18.81 °C, and an induction time of 16.01 h. Subsequently, the recombinant hepcidin peptide was successfully purified using Ni-IDA affinity chromatography followed by SUMO protease cleavage. The obtained hepcidin peptide (without His-SUMO tag) demonstrated strong antimicrobial activity in vitro against V. parahaemolyticus, E. coli, and S. aureus. The results showed prokaryotic (E. coli) expression is a feasible way to produce the hepcidin peptide of crescent sweetlips in a cost-effective way, which has great potential to be used as an antimicrobial agent in aquaculture. |
format |
Text |
author |
Peixin Wang Zhongjing Lin Shaoling Lin Baodong Zheng Yi Zhang Jiamiao Hu |
author_facet |
Peixin Wang Zhongjing Lin Shaoling Lin Baodong Zheng Yi Zhang Jiamiao Hu |
author_sort |
Peixin Wang |
title |
Prokaryotic Expression, Purification, and Antibacterial Activity of the Hepcidin Peptide of Crescent Sweetlips (Plectorhinchus cinctus) |
title_short |
Prokaryotic Expression, Purification, and Antibacterial Activity of the Hepcidin Peptide of Crescent Sweetlips (Plectorhinchus cinctus) |
title_full |
Prokaryotic Expression, Purification, and Antibacterial Activity of the Hepcidin Peptide of Crescent Sweetlips (Plectorhinchus cinctus) |
title_fullStr |
Prokaryotic Expression, Purification, and Antibacterial Activity of the Hepcidin Peptide of Crescent Sweetlips (Plectorhinchus cinctus) |
title_full_unstemmed |
Prokaryotic Expression, Purification, and Antibacterial Activity of the Hepcidin Peptide of Crescent Sweetlips (Plectorhinchus cinctus) |
title_sort |
prokaryotic expression, purification, and antibacterial activity of the hepcidin peptide of crescent sweetlips (plectorhinchus cinctus) |
publisher |
Multidisciplinary Digital Publishing Institute |
publishDate |
2023 |
url |
https://doi.org/10.3390/cimb45090456 |
op_coverage |
agris |
long_lat |
ENVELOPE(170.483,170.483,-83.583,-83.583) |
geographic |
Arctic Ida |
geographic_facet |
Arctic Ida |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Current Issues in Molecular Biology Volume 45 Issue 9 Pages: 7212-7227 |
op_relation |
Molecular Pharmacology https://dx.doi.org/10.3390/cimb45090456 |
op_rights |
https://creativecommons.org/licenses/by/4.0/ |
op_doi |
https://doi.org/10.3390/cimb45090456 |
container_title |
Current Issues in Molecular Biology |
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45 |
container_issue |
9 |
container_start_page |
7212 |
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7227 |
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