In this study, we determined whether solid-state fermentation could degrade major allergens and reduce potential allergenicity of soybean meal (SBM). Solid-state fermentation was realized through a mixture of Lactobacillus casei, yeast, and Bacillus subtilis. High-performance liquid chromatography, size exclusion-high-performance liquid chromatography, and capillary liquid chromatography/tandem mass spectrometry coupled with electrospray ionization were used to examine the total amino acids and molecular weight distribution of the fermented soybean meal (FSBM). In addition, the potential allergenicity of FSBM was assessed by conducting in vitro competitive inhibition ELISA and oral sensitization and challenge of a BALB/c mice model. The results indicated that the total amino acid content increased and soy protein was degraded into polypeptides with low molecular weights that were derived from the hydrolysis of the allergen sequences N232-D383, G253-I265, E169-S215, G68-G98, A365-I375, and V153-A167. Moreover, the FSBM group exhibited a lower in vitro immunoglobulin E (IgE)-binding capacity than the SBM group. The BALB/c model indicated that the FSBM group manifested milder damage to the intestine, lower mMCP-1 and IgE levels, and higher IFN-γ levels as compared to the SBM group. These findings suggested that the potential allergenicity of SBM was reduced by the solid-state fermentation induced by the mixture of Lactobacillus casei, yeast, and Bacillus subtilis.

Microbial transglutaminase (MTGase) has been developed as a new tofu coagulant in recent years due to its good hydrophilicity, high catalytic activity, and strong thermal stability. This study aimed to investigate the effect of MTGase on the physicochemical properties and immunoreactivity of tofu relative to conventional coagulants [brine and glucono-δ-lactone (GDL)]. Structural changes of the MTGase cross-linked soymilk protein were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), circular dichroism (CD) spectroscopy, ultraviolet (UV) absorption spectroscopy, and fluorescence spectroscopy. The IgE-binding capacity of MTGase cross-linked proteins was tested by enzyme-linked immunosorbent assay (ELISA). The physicochemical properties, quality characteristics, and surface microstructures of five different types of tofu were determined by the Kjeldahl nitrogen method, texture analysis, and scanning electron microscopy (SEM). The digestibility of tofu was evaluated in vitro by simulated gastrointestinal (GIS) digestion. A cell sensitization experiment was performed in vitro to evaluate the capability of tofu digestion products to induce the release of bioactive mediators from human basophil leukemia (KU812) cells. Results indicated that MTGase significantly changed the advanced structure of the soymilk protein. Compared with tofu without MTGase, the composite coagulant tofu containing MTGase exhibited better quality. MTGase improved the water-holding capacity (WHC) of the internal mesh structure and increased the yield of tofu. The digestion products of the composite coagulant tofu, especially the GDL plus MTGase tofu, induced KU812 cells to release fewer bioactive mediators compared with those of MTGase-free tofu. MTGase can not only improve the quality of conventional coagulant tofu but also reduce the potential allergenicity of tofu to a certain extent.

Given that roasted peanut (Ro) products are commonly used in daily life, peanut allergenicity is a foremost concern. Analyzing the changes in the structure and potential allergenicity of individual allergens can promote the exploration of the structural basis of the alterations in the potential allergenicity of Ro. This work focused on four major allergens in raw peanut (Ra) and Ro. Structural changes were analyzed on the basis of circular dichroism, ultraviolet and fluorescence spectroscopy, and molecular dynamic simulation. The IgE recognition capability of allergens was assessed via western blot analysis. The IgE binding capacity of allergens was detected by conducting enzyme-linked immunosorbent assay. The potential allergenicity of allergens was evaluated using the KU812 cell degranulation model. The results showed that roasting induced different changes in the overall structures of allergens and altered the structures and electrostatic potential of IgE epitopes, especially Ara h 1 and Ara h 6. These alterations affected the potential allergenicity of allergens. Ara h 1 and Ara h 6 in Ro showed significantly enhanced IgE binding capacities and abilities to elicit KU812 cell degranulation, while Ara h 2 and Ara h 3 did not change significantly. For total protein, the roasted peanut protein showed decreased abilities to elicit KU812 cell degranulation. The results indicated that different allergens in Ro showed different changes of structures and potential allergenicity and that the conformational structure plays a crucial role in potential allergenicity of allergens.