Proteomic and in silico identification of potential allergenic proteins in cowpea (Vigna unguiculata L. Walp) seeds

Cowpea ( Vigna unguiculata L. Walp) seeds are rich in proteins (~24%), thus they have been considered as a viable dietary protein substitute. Despite its advantages, allergenicity risks of cowpea seeds need to be taken into account. Herein, the protein expression and potential allergens in cowpea seeds were analyzed using LC/timsTOF Pro 2, PEAK studio and multiple in silico analysis. Based on functional classification using STRING analysis, the result revealed response to stimuli, e.g. , oxidative stress and temperature, as the major cluster (187 proteins), followed by biosynthesis of secondary metabolite (117 proteins) and immune system (108 proteins). These suggest involvement of protein functions in maintaining homeostasis during seed development under stress conditions. By using webtool Allermatch™ and the Pfam database, 131 potential allergenic proteins were found from cowpea seeds. The findings revealed that cowpea seeds contain a number of recognized allergens, including endochitinase, beta-conglycinin, and vicilin, as well as numerous allergenic proteins not previously described, such as endochitinase 1B and 5-methyltetrahydropteroyltriglutamate–homocysteine methyltransferase (MetE). Additionally, use of the Kolaskar & Tongaonkar method predicted B-cell epitopes such as 30 VSGFGVI 36 , 152 VPVLVGP 158 and 153 PVLVGPV 159 , increasing the possibility of cowpea allergenicity. In conclusion, this study provides useful information on the potential allergens in cowpea seeds, providing a foundation for future cowpea allergenicity assessment including experimental IgE-binding or clinical validation. • Proteomics and in silico analyses identified 131 possible allergens in cowpea. • The Immune Epitope Database showed 696 epitopes that linked to allergenic responses. • This study revealed novel cowpea allergens such as endochitinase 1B. • 30 VSGFGVI 36 was the most potential B-cell epitope regions of endochitinase 1B.