Abstract
Gly m 5.0101, the alpha subunit of β-conglycinin, is one of the major allergens found in soybeans that has been identified as causing an allergic reaction. Here, we developed a quantification method of Gly m 5.0101 with multiple reaction monitoring using the synthetic peptide 194NPFLFGSNR202 as the external standard. Firstly, the ground soybean was defatted and extracted with a protein extraction buffer. Then the crude extract was on-filter digested by trypsin and analyzed by liquid chromatography-tandem mass spectrometry. The selected peptide exhibited a detection limit of 0.48 ng/mL and a linear relationship in a concentration range from 1.6 to 500 ng/mL (r2 > 0.99). The developed method was successfully applied to quantify the Gly m 5.0101 level in dozens of soybean varieties from different sources and soybean products derived from different processing techniques. The developed method could be used to further analyze β-conglycinin in soybean seeds combined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis.
Highlights
Glycine max, commonly known as soybean, is a major oilseed crop rich in protein and oil that is often used in food and pharmaceuticals [1]
The peptide is unique for the Gly m 5.0101
The results show that the contents of β-conglycinin in fresh soybean seeds ranged from 53.03 to 112.4 mg/g which was consistent with previous reports that β-conglycinin was detected by enzyme-linked immunosorbent assays (ELISA) methods in soybean seeds ranging from 51.1 to 131.2 mg/g [26,27,28]
Summary
Commonly known as soybean, is a major oilseed crop rich in protein and oil that is often used in food and pharmaceuticals [1]. Fat-free soybean meal is a significant and cheap source of protein for animal feeds. About 97% of soybean meal is used for animal feed and accounts for approximately 70% of all protein sources used in livestock diets. Β-conglycinin is the main allergen in soybean, which accounts for 10.0% to 12.7% of soybean seed or 24.7% to 45.3% of the total seed proteins. It contains three subunits: α subunit (MW, 57–76 kDa), α’ subunit (MW, 57-83 kDa) and β subunit (MW, 42–53 kDa). The percentage of each subunit varies among genotypes: α from 10.4% to
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