A hemolysin secretion pathway-based novel secretory expression platform for efficient manufacturing of tag peptides and anti-microbial peptides in Escherichia coli

Abstract Background Although Escherichia coli has been widely used for the expression of exogenous proteins, the secretory expression in this system is still a big obstacle. As one of the most important secretion pathways, hemolysin A (HlyA) system of E. coli can transport substrates directly from t...

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Autores principales:	Wen Zhu, Lifu Hu, Yang Wang, Liangyin Lv, Hui Wang, Wenqiang Shi, Jianwei Zhu, Huili Lu
Formato:	article
Lenguaje:	EN
Publicado:	SpringerOpen 2021
Materias:	E. coli Hemolysin A Secretory expression Tag peptides Anti-microbial peptides Technology T Chemical technology TP1-1185 Biotechnology TP248.13-248.65
Acceso en línea:	https://doaj.org/article/e4897279f99f4e04a8695522cd8919ca
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spelling	oai:doaj.org-article:e4897279f99f4e04a8695522cd8919ca2021-11-28T12:03:55ZA hemolysin secretion pathway-based novel secretory expression platform for efficient manufacturing of tag peptides and anti-microbial peptides in Escherichia coli10.1186/s40643-021-00471-62197-4365https://doaj.org/article/e4897279f99f4e04a8695522cd8919ca2021-11-01T00:00:00Zhttps://doi.org/10.1186/s40643-021-00471-6https://doaj.org/toc/2197-4365Abstract Background Although Escherichia coli has been widely used for the expression of exogenous proteins, the secretory expression in this system is still a big obstacle. As one of the most important secretion pathways, hemolysin A (HlyA) system of E. coli can transport substrates directly from the cytoplasm to extracellular medium without the formation of any periplasmic intermediate, making it an ideal candidate for the development of the secretory production platform for exogenous proteins. Results In this work, we developed a novel production platform, THHly, based on the HlyA secretion system, and explored its applications in the efficient preparation and quick detection of tag peptides and anti-microbial peptides. In this novel platform the signal sequence of HlyA is fused to the C-terminal of target peptide, with Tobacco Etch Virus (TEV) protease cleavage site and 6*His tag between them. Five tag peptides displayed good secretory properties in E. coli BL21 (DE3), among which T7 tag and S tag were obtained by two rounds of purification steps and TEV cleavage, and maintained their intrinsic immunogenicity. Furthermore, Cecropin A and Melittin, two different types of widely explored anti-microbial peptides, were produced likewise and verified to possess anti-microbial/anti-tumor bioactivities. No significant bacterial growth inhibition was observed during the fusion protein expression, indicating that the fusion form not only mediated the secretion but also decreased the toxicity of anti-microbial peptides (AMPs) to the host bacteria. To the best of our knowledge, this is the first report to achieve the secretory expression of these two AMPs in E. coli with considerable potential for manufacturing and industrialization purposes. Conclusions The results demonstrate that the HlyA based novel production platform of E. coli allowed the efficient secretory production and purification of peptides, thus suggesting a promising strategy for the industrialized production of peptide pharmaceuticals or reagents. Graphical AbstractWen ZhuLifu HuYang WangLiangyin LvHui WangWenqiang ShiJianwei ZhuHuili LuSpringerOpenarticleE. coliHemolysin ASecretory expressionTag peptidesAnti-microbial peptidesTechnologyTChemical technologyTP1-1185BiotechnologyTP248.13-248.65ENBioresources and Bioprocessing, Vol 8, Iss 1, Pp 1-14 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	E. coli Hemolysin A Secretory expression Tag peptides Anti-microbial peptides Technology T Chemical technology TP1-1185 Biotechnology TP248.13-248.65
spellingShingle	E. coli Hemolysin A Secretory expression Tag peptides Anti-microbial peptides Technology T Chemical technology TP1-1185 Biotechnology TP248.13-248.65 Wen Zhu Lifu Hu Yang Wang Liangyin Lv Hui Wang Wenqiang Shi Jianwei Zhu Huili Lu A hemolysin secretion pathway-based novel secretory expression platform for efficient manufacturing of tag peptides and anti-microbial peptides in Escherichia coli
description	Abstract Background Although Escherichia coli has been widely used for the expression of exogenous proteins, the secretory expression in this system is still a big obstacle. As one of the most important secretion pathways, hemolysin A (HlyA) system of E. coli can transport substrates directly from the cytoplasm to extracellular medium without the formation of any periplasmic intermediate, making it an ideal candidate for the development of the secretory production platform for exogenous proteins. Results In this work, we developed a novel production platform, THHly, based on the HlyA secretion system, and explored its applications in the efficient preparation and quick detection of tag peptides and anti-microbial peptides. In this novel platform the signal sequence of HlyA is fused to the C-terminal of target peptide, with Tobacco Etch Virus (TEV) protease cleavage site and 6*His tag between them. Five tag peptides displayed good secretory properties in E. coli BL21 (DE3), among which T7 tag and S tag were obtained by two rounds of purification steps and TEV cleavage, and maintained their intrinsic immunogenicity. Furthermore, Cecropin A and Melittin, two different types of widely explored anti-microbial peptides, were produced likewise and verified to possess anti-microbial/anti-tumor bioactivities. No significant bacterial growth inhibition was observed during the fusion protein expression, indicating that the fusion form not only mediated the secretion but also decreased the toxicity of anti-microbial peptides (AMPs) to the host bacteria. To the best of our knowledge, this is the first report to achieve the secretory expression of these two AMPs in E. coli with considerable potential for manufacturing and industrialization purposes. Conclusions The results demonstrate that the HlyA based novel production platform of E. coli allowed the efficient secretory production and purification of peptides, thus suggesting a promising strategy for the industrialized production of peptide pharmaceuticals or reagents. Graphical Abstract
format	article
author	Wen Zhu Lifu Hu Yang Wang Liangyin Lv Hui Wang Wenqiang Shi Jianwei Zhu Huili Lu
author_facet	Wen Zhu Lifu Hu Yang Wang Liangyin Lv Hui Wang Wenqiang Shi Jianwei Zhu Huili Lu
author_sort	Wen Zhu
title	A hemolysin secretion pathway-based novel secretory expression platform for efficient manufacturing of tag peptides and anti-microbial peptides in Escherichia coli
title_short	A hemolysin secretion pathway-based novel secretory expression platform for efficient manufacturing of tag peptides and anti-microbial peptides in Escherichia coli
title_full	A hemolysin secretion pathway-based novel secretory expression platform for efficient manufacturing of tag peptides and anti-microbial peptides in Escherichia coli
title_fullStr	A hemolysin secretion pathway-based novel secretory expression platform for efficient manufacturing of tag peptides and anti-microbial peptides in Escherichia coli
title_full_unstemmed	A hemolysin secretion pathway-based novel secretory expression platform for efficient manufacturing of tag peptides and anti-microbial peptides in Escherichia coli
title_sort	hemolysin secretion pathway-based novel secretory expression platform for efficient manufacturing of tag peptides and anti-microbial peptides in escherichia coli
publisher	SpringerOpen
publishDate	2021
url	https://doaj.org/article/e4897279f99f4e04a8695522cd8919ca
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A hemolysin secretion pathway-based novel secretory expression platform for efficient manufacturing of tag peptides and anti-microbial peptides in Escherichia coli

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