Dados do Trabalho

Título

SHOTGUN PROTEOMICS AS A TOOL TO UNDERSTAND THE ALLERGENIC PROFILE OF BRAZILIAN WHEAT GENOTYPES

Introdução

Despite being widely grown and consumed worldwide, wheat can provoke sensitivities or inflammatory responses in susceptible individuals due to the immune system's reaction to specific proteins. This includes conditions such as celiac disease, non-celiac gluten sensitivity, wheat allergy, and related disorders. This study aimed to elucidate the impact of different processing methods on the expression of allergenic proteins in wheat flours with distinct technological applications.

Material e Métodos

Products derived from two Brazilian wheat genotypes (Triticum aestivum), ORS Agile (bread/improver wheat) [1] and ORS Vintecinco (biscuit wheat) [2], were analyzed: refined flours, thermoplastic extrudates, raw and cooked pasta, and breads formulated with varying ingredients. Sequential protein extraction was performed, considering Osborne fractions: salt-soluble (albumins/globulins), ethanol-soluble (gliadins), and urea-soluble with reduction (glutenins) proteins, followed by quantification using RP-HPLC. Peptides were identified using nanoLC-MS/MS on a Q Exactive Plus Orbitrap in data-dependent acquisition mode, processed with MaxQuant against a custom Triticum database from UniprotKB, and further analyzed against a curated wheat allergen database.

Resultados e Discussão

Among the 380 proteins evaluated, approximately 12% (48 proteins) were confirmed as allergens, covering the albumins/globulins, gliadins, and glutenins fractions. Well-established wheat allergens, as HMW-GS Ax2, Bx7, Dx5 and Dy10, gamma-gliadins and alpha-amylase/trypsin inhibitors could be found in all analyzed samples.

Conclusão

ORS Agile flour showed the highest allergen expression intensity, contrasting with the lowest observed in enriched-formulation bread from ORS Vintecinco. Notably, ORS Agile flour exhibited over threefold higher allergen expression compared to ORS Vintecinco, likely attributable to its higher gluten content typical of bread/improver wheat. The study underscores the significant impact of processing on allergen levels, particularly pronounced in ORS Agile samples. However, additional investigations, including targeted proteomics to quantify specific allergenic peptides and epitopes, are essential to delineate the precise influence of technological quality and processing methods on altering the allergenic potential of wheat products.