Non-peptide guided auto-secretion of recombinant proteins by super-folder green fluorescent protein in Escherichia coli

Abstract Protein secretion in Escherichia coli is usually led by a signal peptide that targets the protein to specific secretory pathways. In this study, we demonstrated that the superfolder green fluorescent protein (sfGFP) could be served as a non-signal peptide to guide protein auto-secretion in...

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Autores principales:	Zhen Zhang, Rongxing Tang, Dewu Zhu, Wenfeng Wang, Li Yi, Lixin Ma
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2a3fb9ce3a5f42dfb77153b471291d87
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spelling	oai:doaj.org-article:2a3fb9ce3a5f42dfb77153b471291d872021-12-02T16:06:51ZNon-peptide guided auto-secretion of recombinant proteins by super-folder green fluorescent protein in Escherichia coli10.1038/s41598-017-07421-32045-2322https://doaj.org/article/2a3fb9ce3a5f42dfb77153b471291d872017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-07421-3https://doaj.org/toc/2045-2322Abstract Protein secretion in Escherichia coli is usually led by a signal peptide that targets the protein to specific secretory pathways. In this study, we demonstrated that the superfolder green fluorescent protein (sfGFP) could be served as a non-signal peptide to guide protein auto-secretion in E. coli. This auto-secretion was characterized as a three-step process through the sub-cellular localization analysis: inner membrane trans-location followed by anchoring at outer membrane, and then being released into culture media. We further determined that the beta-barrel structure and net negative charges of sfGFP played important roles in its auto-extracellular secretion property. Using sfGFP as a carrier, heterologous proteins ranging from peptide to complex protein, including antibacterial peptide PG4, endo-beta-N-acethylglucosamindase H (Endo H), human arginase-1 (ARG1), and glutamate decarboxylase (GAD) were all successfully expressed and secreted extracellularly when fused to the carboxyl end of sfGFP. Besides facilitating the extracellular secretion, sfGFP fusion proteins can also be correctly folded and formed the active complex protein structure, including the trimetric human ARG1 and homo-hexametric GAD. This is the first report that sfGFP can guide the secretion of recombinant proteins out of the cells from cytoplasm in E. coli without affecting their conformation and function.Zhen ZhangRongxing TangDewu ZhuWenfeng WangLi YiLixin MaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Zhen Zhang Rongxing Tang Dewu Zhu Wenfeng Wang Li Yi Lixin Ma Non-peptide guided auto-secretion of recombinant proteins by super-folder green fluorescent protein in Escherichia coli
description	Abstract Protein secretion in Escherichia coli is usually led by a signal peptide that targets the protein to specific secretory pathways. In this study, we demonstrated that the superfolder green fluorescent protein (sfGFP) could be served as a non-signal peptide to guide protein auto-secretion in E. coli. This auto-secretion was characterized as a three-step process through the sub-cellular localization analysis: inner membrane trans-location followed by anchoring at outer membrane, and then being released into culture media. We further determined that the beta-barrel structure and net negative charges of sfGFP played important roles in its auto-extracellular secretion property. Using sfGFP as a carrier, heterologous proteins ranging from peptide to complex protein, including antibacterial peptide PG4, endo-beta-N-acethylglucosamindase H (Endo H), human arginase-1 (ARG1), and glutamate decarboxylase (GAD) were all successfully expressed and secreted extracellularly when fused to the carboxyl end of sfGFP. Besides facilitating the extracellular secretion, sfGFP fusion proteins can also be correctly folded and formed the active complex protein structure, including the trimetric human ARG1 and homo-hexametric GAD. This is the first report that sfGFP can guide the secretion of recombinant proteins out of the cells from cytoplasm in E. coli without affecting their conformation and function.
format	article
author	Zhen Zhang Rongxing Tang Dewu Zhu Wenfeng Wang Li Yi Lixin Ma
author_facet	Zhen Zhang Rongxing Tang Dewu Zhu Wenfeng Wang Li Yi Lixin Ma
author_sort	Zhen Zhang
title	Non-peptide guided auto-secretion of recombinant proteins by super-folder green fluorescent protein in Escherichia coli
title_short	Non-peptide guided auto-secretion of recombinant proteins by super-folder green fluorescent protein in Escherichia coli
title_full	Non-peptide guided auto-secretion of recombinant proteins by super-folder green fluorescent protein in Escherichia coli
title_fullStr	Non-peptide guided auto-secretion of recombinant proteins by super-folder green fluorescent protein in Escherichia coli
title_full_unstemmed	Non-peptide guided auto-secretion of recombinant proteins by super-folder green fluorescent protein in Escherichia coli
title_sort	non-peptide guided auto-secretion of recombinant proteins by super-folder green fluorescent protein in escherichia coli
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/2a3fb9ce3a5f42dfb77153b471291d87
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Non-peptide guided auto-secretion of recombinant proteins by super-folder green fluorescent protein in Escherichia coli

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