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<p>Red kidney beans (<i>Phaseolus vulgaris</i> L.), a nutrient-dense legume, face significant limitations in food applications due to persistent allergenicity. The dual effects of industrial autoclaving (121&#xa0;°C, 15&#xa0;min) on allergenicity modulation were evaluated through in vitro gastrointestinal digestion models in this study, integrated with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), immunoblotting, and computational epitope prediction. Electrophoretic profiling demonstrated partial degradation of major allergens for 43&#xa0;kDa phaseollin-like protein, lectin subunits (31/25&#xa0;kDa). Legumin <i>α</i>-subunits exhibited pepsin resistance (74.8% stability), contrasting with rapid degradation of <i>α</i>-amylase inhibitor (<i>α</i>-AI) and group 3 late embryogenesis abundance (G3LEA) proteins. Chromatographic immunoreactivity screening revealed autoclaved samples generated 8 antigenic fractions, including novel hydrophobic peptides with enhanced immunoglobulin E (IgE)-binding potential. Mass spectrometry identified 849 and 1 516 unique peptides in raw and autoclaved digests, respectively, with phaseollin and lectins dominating post-processing epitopes. Bioinformatics highlighted conserved allergenic motifs (e.g., VLVKPIQIR, LPQQADAE) in phaseollin and lectins, alongside cross-reactive sequences homologous. Paradoxically, autoclaving reduced intact allergen content but amplified immunoreactive diversity through conformational exposure of epitopes and hydrophobic residue enrichment. These findings underscore the necessity for epitope-specific risk assessment in legume processing, balancing benefits of thermal intervention against unintended allergenic consequences.</p>

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Autoclave processing modulates allergenic potential of red kidney beans (Phaseolus vulgaris L.): proteomic and bioinformatics insights after in vitro gastric digestion

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年8月6日

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  • Rong Zhang,
  • Kailin Feng,
  • Yongfei Wang,
  • Shudong He,
  • Sophia Xue,
  • Xinping Qian,
  • John Shi

摘要

Red kidney beans (Phaseolus vulgaris L.), a nutrient-dense legume, face significant limitations in food applications due to persistent allergenicity. The dual effects of industrial autoclaving (121 °C, 15 min) on allergenicity modulation were evaluated through in vitro gastrointestinal digestion models in this study, integrated with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), immunoblotting, and computational epitope prediction. Electrophoretic profiling demonstrated partial degradation of major allergens for 43 kDa phaseollin-like protein, lectin subunits (31/25 kDa). Legumin α-subunits exhibited pepsin resistance (74.8% stability), contrasting with rapid degradation of α-amylase inhibitor (α-AI) and group 3 late embryogenesis abundance (G3LEA) proteins. Chromatographic immunoreactivity screening revealed autoclaved samples generated 8 antigenic fractions, including novel hydrophobic peptides with enhanced immunoglobulin E (IgE)-binding potential. Mass spectrometry identified 849 and 1 516 unique peptides in raw and autoclaved digests, respectively, with phaseollin and lectins dominating post-processing epitopes. Bioinformatics highlighted conserved allergenic motifs (e.g., VLVKPIQIR, LPQQADAE) in phaseollin and lectins, alongside cross-reactive sequences homologous. Paradoxically, autoclaving reduced intact allergen content but amplified immunoreactive diversity through conformational exposure of epitopes and hydrophobic residue enrichment. These findings underscore the necessity for epitope-specific risk assessment in legume processing, balancing benefits of thermal intervention against unintended allergenic consequences.