Ara h 1 is the most abundant sensitizing protein in peanuts; it has high thermal stability and is difficult to degrade. The peanut sprout is a high-quality, natural food that has various beneficial effects and lower allergenicity than peanut seeds. In this study, ultrasonication (US) of peanut sprouts was used to alter their Ara h 1 content. We determined that the optimal parameters for the US process were 35 °C temperature, 30 min duration, 240 W power, and 100 kHz frequency. After 5 days of germination, the protease activity of the control (blank) group increased to 262.39 ± 0.10 U, whereas that of the US group increased to 290.1 ± 0.25 U. We also investigated the effects of US on Ara h 1 protein composition, structure, and related gene expression during germination. ELISA results showed that after 5 days of germination, Ara h 1 content in the blank group decreased from 20.63 ± 0.31 ppm to 3.35 ± 0.42 ppm, whereas in the US group, they decreased to below the detection limit. SDS-PAGE bands between 50 and 70 kDa from peanut sprout extracts gradually became lighter in both groups. The band almost disappeared at day 5 of germination in the US group, indicating that US reduced the Ara h 1 content of peanut sprouts, consistent with the ELISA results. The expression of the Ara h 1 gene in peanut seeds was 173.92 ± 26.37. In the BK control group, it decreased to 0.49 ± 0.17 on the fourth day and increased slightly to 0.75 ± 0.09 on the fifth day. In the US group, it decreased to 1.37 ± 0.28 on the first day, dropped sharply to 0.00 on the third day, and increased slightly to 0.04 ± 0.01 on the fourth and fifth days. Protein structure results showed that the α-helix structure of Ara h 1 decreased after US, whereas the content of β-fold structures increased. The surface hydrophobicity decreased, and the secondary and tertiary structures of Ara h 1 were loose.
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