Biosynthesis of Conjugate Vaccines Using an O-Linked Glycosylation System

ABSTRACT Conjugate vaccines are known to be one of the most effective and safest types of vaccines against bacterial pathogens. Previously, vaccine biosynthesis has been performed by using N-linked glycosylation systems. However, the structural specificity of these systems for sugar substrates has...

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Autores principales:	Chao Pan, Peng Sun, Bo Liu, Haoyu Liang, Zhehui Peng, Yan Dong, Dongshu Wang, Xiankai Liu, Bin Wang, Ming Zeng, Jun Wu, Li Zhu, Hengliang Wang
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/c555ad5b8de74ce5a3d658427df08da0
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spelling	oai:doaj.org-article:c555ad5b8de74ce5a3d658427df08da02021-11-15T15:41:41ZBiosynthesis of Conjugate Vaccines Using an O-Linked Glycosylation System10.1128/mBio.00443-162150-7511https://doaj.org/article/c555ad5b8de74ce5a3d658427df08da02016-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00443-16https://doaj.org/toc/2150-7511ABSTRACT Conjugate vaccines are known to be one of the most effective and safest types of vaccines against bacterial pathogens. Previously, vaccine biosynthesis has been performed by using N-linked glycosylation systems. However, the structural specificity of these systems for sugar substrates has hindered their application. Here, we report a novel protein glycosylation system (O-linked glycosylation via Neisseria meningitidis) that can transfer virtually any glycan to produce a conjugate vaccine. We successfully established this system in Shigella spp., avoiding the construction of an expression vector for polysaccharide synthesis. We further found that different protein substrates can be glycosylated using this system and that the O-linked glycosylation system can also effectively function in other Gram-negative bacteria, including some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system. The results from a series of animal experiments show that the conjugate vaccine produced by this O-linked glycosylation system offered a potentially protective antibody response. Furthermore, we elucidated and optimized the recognition motif, named MOOR, for the O-glycosyltransferase PglL. Finally, we demonstrated that the fusion of other peptides recognized by major histocompatibility complex class II around MOOR had no adverse effects on substrate glycosylation, suggesting that this optimized system will be useful for future vaccine development. Our results expand the glycoengineering toolbox and provide a simpler and more robust strategy for producing bioconjugate vaccines against a variety of pathogens. IMPORTANCE Recently, the rapid development of synthetic biology has allowed bioconjugate vaccines with N-linked protein glycosylation to become a reality. However, the difficulty of reestablishing the exogenous polysaccharide synthetic pathway in Escherichia coli hinders their application. Here, we show that an O-linked protein glycosylation system from Neisseria meningitidis, which has a lower structure specificity for sugar substrates, could be engineered directly in attenuated pathogens to produce effective conjugate vaccines. To facilitate the further design of next-generation bioconjugate vaccines, we optimized a novel short motif consisting of 8 amino acids that is sufficient for glycosylation. Our results expand the application potential of O-linked protein glycosylation and demonstrate a simpler and more robust strategy for producing bioconjugate vaccines against different pathogens. In the future, bacterial antigenic polysaccharides could be attached to major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation.Chao PanPeng SunBo LiuHaoyu LiangZhehui PengYan DongDongshu WangXiankai LiuBin WangMing ZengJun WuLi ZhuHengliang WangAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 7, Iss 2 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Chao Pan Peng Sun Bo Liu Haoyu Liang Zhehui Peng Yan Dong Dongshu Wang Xiankai Liu Bin Wang Ming Zeng Jun Wu Li Zhu Hengliang Wang Biosynthesis of Conjugate Vaccines Using an O-Linked Glycosylation System
description	ABSTRACT Conjugate vaccines are known to be one of the most effective and safest types of vaccines against bacterial pathogens. Previously, vaccine biosynthesis has been performed by using N-linked glycosylation systems. However, the structural specificity of these systems for sugar substrates has hindered their application. Here, we report a novel protein glycosylation system (O-linked glycosylation via Neisseria meningitidis) that can transfer virtually any glycan to produce a conjugate vaccine. We successfully established this system in Shigella spp., avoiding the construction of an expression vector for polysaccharide synthesis. We further found that different protein substrates can be glycosylated using this system and that the O-linked glycosylation system can also effectively function in other Gram-negative bacteria, including some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system. The results from a series of animal experiments show that the conjugate vaccine produced by this O-linked glycosylation system offered a potentially protective antibody response. Furthermore, we elucidated and optimized the recognition motif, named MOOR, for the O-glycosyltransferase PglL. Finally, we demonstrated that the fusion of other peptides recognized by major histocompatibility complex class II around MOOR had no adverse effects on substrate glycosylation, suggesting that this optimized system will be useful for future vaccine development. Our results expand the glycoengineering toolbox and provide a simpler and more robust strategy for producing bioconjugate vaccines against a variety of pathogens. IMPORTANCE Recently, the rapid development of synthetic biology has allowed bioconjugate vaccines with N-linked protein glycosylation to become a reality. However, the difficulty of reestablishing the exogenous polysaccharide synthetic pathway in Escherichia coli hinders their application. Here, we show that an O-linked protein glycosylation system from Neisseria meningitidis, which has a lower structure specificity for sugar substrates, could be engineered directly in attenuated pathogens to produce effective conjugate vaccines. To facilitate the further design of next-generation bioconjugate vaccines, we optimized a novel short motif consisting of 8 amino acids that is sufficient for glycosylation. Our results expand the application potential of O-linked protein glycosylation and demonstrate a simpler and more robust strategy for producing bioconjugate vaccines against different pathogens. In the future, bacterial antigenic polysaccharides could be attached to major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation.
format	article
author	Chao Pan Peng Sun Bo Liu Haoyu Liang Zhehui Peng Yan Dong Dongshu Wang Xiankai Liu Bin Wang Ming Zeng Jun Wu Li Zhu Hengliang Wang
author_facet	Chao Pan Peng Sun Bo Liu Haoyu Liang Zhehui Peng Yan Dong Dongshu Wang Xiankai Liu Bin Wang Ming Zeng Jun Wu Li Zhu Hengliang Wang
author_sort	Chao Pan
title	Biosynthesis of Conjugate Vaccines Using an O-Linked Glycosylation System
title_short	Biosynthesis of Conjugate Vaccines Using an O-Linked Glycosylation System
title_full	Biosynthesis of Conjugate Vaccines Using an O-Linked Glycosylation System
title_fullStr	Biosynthesis of Conjugate Vaccines Using an O-Linked Glycosylation System
title_full_unstemmed	Biosynthesis of Conjugate Vaccines Using an O-Linked Glycosylation System
title_sort	biosynthesis of conjugate vaccines using an o-linked glycosylation system
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/c555ad5b8de74ce5a3d658427df08da0
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Biosynthesis of Conjugate Vaccines Using an O-Linked Glycosylation System

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