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Research Article | Open Access

An innovative method used for the identification of N-glycans on soybean allergen β-conglycinin

Cheng Lia,1, Yang Tiana,1, Jianli Hanb,1, Yu Lub, Meiyi Zoub, Yue Jiab, Chengjian Wanga, Linjuan Huanga,b( ), Zhongfu Wanga,b( )
Key Laboratory of Glycobiology and Glycoengineering of Xi'an, College of Food Science and Technology, Northwest University, Xi'an 710069, China
College of Life Sciences, Northwest University, Xi'an 710069, China

1Co-first authors.Peer review under responsibility of KeAi Communications Co., Ltd.]]>

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Abstract

β-Conglycinin is one of the major allergens existed in soybean. N-Glycans attached to the β-conglycinin influenced the immunoreactivity and antigen presenting efficiency of β-conglycinin. In this study, we described a new method used to release and collect the N-glycans from β-conglycinin, and the N-glycans existed in linear epitopes of β-conglycinin were identified. Glycopeptides hydrolyzed from β-conglycinin were purified by cotton hydrophilic chromatography. Trifluoromethylsulfonic acid was then used to release glycans from glycopeptides, and new glycopeptides containing one single N-acetyl-D-glucosamine (GlcNAc) moiety were then utilized for mass spectrometry. Five glycosylation sites (Asn-199, Asn-455, Asn-215, Asn-489 and Asn-326) and 22 kinds of glycopeptides were identified. It is noteworthy that the peptide VVN#ATSNL (where # represents for the glycosylation site) was analyzed to be both glycopeptide and linear epitope. Our results provided a new method for the N-glycoform analysis of food allergens, and laid a foundation for understanding the relationship between glycosylation and food allergy.

Keywords

Soybean allergen β-conglycininGlycopeptideMass spectrometryN-GlycanGlycosylation site

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Food Science and Human Wellness
Volume 12 Issue 3,
May 2023
Pages 842-850
DOI: 10.1016/j.fshw.2022.09.025
Cite this article:
Li C, Tian Y, Han J, et al. An innovative method used for the identification of N-glycans on soybean allergen β-conglycinin. Food Science and Human Wellness, 2023, 12(3): 842-850. https://doi.org/10.1016/j.fshw.2022.09.025

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Received: 28 May 2021
Revised: 12 July 2021
Accepted: 29 August 2021
Published: 15 October 2022
© 2023 Beijing Academy of Food Sciences. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co., Ltd.

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
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