Biosynthesis of Conjugate Vaccines Using an O-Linked Glycosylation System

mBio. 2016;7(2):e00443-16 DOI 10.1128/mBio.00443-16

 

Journal Homepage

Journal Title: mBio

ISSN: 2150-7511 (Online)

Publisher: American Society for Microbiology

LCC Subject Category: Science: Microbiology

Country of publisher: United States

Language of fulltext: English

Full-text formats available: PDF, HTML

 

AUTHORS

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

EDITORIAL INFORMATION

Blind peer review

Editorial Board

Instructions for authors

Time From Submission to Publication: 12 weeks

 

Abstract | Full Text

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.