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Updated: Jul 26, 2025

Detection of Protease Activity by Fluorescent Peptide Zymography
Published on: January 20, 2019
Localization and identification of denatured antigenic sites of glycinin A3 subunit after using two processing
Yichao Wang1, Yida Gao1, Yuying Duan1
1College of Food Science and Engineering, Henan University of Technology, Zhengzhou 450001, Henan, China.
Abstract:
Glycinin is an important allergen in soybeans. In this study, molecular cloning and recombinant phage construction were performed to explore the antigenic sites of the glycinin A3 subunit that were denatured during processing. Next, the A-1-a fragment was located as the denatured antigenic sites by indirect ELISA. The combined UHP heat treatment showed better denaturation of this subunit than the single heat treatment assay. In addition, identification of the synthetic peptide showed that the A-1-a fragment was an amino acid sequence containing a conformational and linear IgE site, in which the first synthetic peptide (P1) being both an antigenic and allergenic site. The results of alanine-scanning showed that the key amino acids affecting antigenicity and allergenicity of A3 subunit were S28, K29, E32, L35 and N13. Our results could provide the basis for further development of more efficient methods to reduce the allergenicity of soybeans.
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