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In this study, the 11S globulin allergen Zan b 2 was identified as the major allergen and characterized through structural and immunological analyses. Integrated immunoinformatics, molecular docking, and molecular dynamics simulations were used to predict B-cell epitopes and screen proteases capable of specifically cleaving these regions. Prediction results indicate that proteinase K targets the following epitopes: Glu13, Thr100, Tyr149, Glu164, Glu229, Glu231, Leu239, Glu241, Thr245, Val328, Glu368, Ala381, and thermolysin targets Arg182, Lys247, Ser273, Asn294, Gln332, and Arg380. Experimental validation using enzymatic hydrolysis, SDS-PAGE, indirect ELISA, LC\u2013MS\/MS, and proteomics showed that combined treatment with proteinase K and thermolysin markedly reduced the IgE-binding activity. These findings demonstrate that simulation-assisted targeted enzymatic hydrolysis is an effective strategy for reducing Zan b 2 immunoreactivity and supports the development of hypoallergenic Sichuan pepper products.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c08943","type":"journal-article","created":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T16:01:51Z","timestamp":1788883311000},"page":"29550-29563","source":"Crossref","is-referenced-by-count":0,"title":["Epitope-Guided\nTwo-Step Enzymatic Hydrolysis for Targeted\nControl of the Immunoreactivity of the Major Sichuan Pepper Allergen\nZan b 2"],"prefix":"10.1021","volume":"74","author":[{"given":"Tingyu","family":"Zhang","sequence":"first","affiliation":[{"name":"Ocean University of China , , 1299 Sansha Road , ,","place":["Qingdao, China, 266003"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xitong","family":"Yu","sequence":"additional","affiliation":[{"name":"Ocean University of China , , 1299 Sansha Road , ,","place":["Qingdao, China, 266003"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yumiao","family":"Bian","sequence":"additional","affiliation":[{"name":"Ocean University of China , , 1299 Sansha Road , ,","place":["Qingdao, China, 266003"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuai","family":"Ma","sequence":"additional","affiliation":[{"name":"Ocean University of China , , 1299 Sansha Road , ,","place":["Qingdao, China, 266003"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liwei","family":"Wang","sequence":"additional","affiliation":[{"name":"Ocean University of China , , 1299 Sansha Road , ,","place":["Qingdao, China, 266003"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Siddanakoppalu Narayana","family":"Pramod","sequence":"additional","affiliation":[{"name":"Davangere University , , Shivagangotri , , ,","place":["Davangere, Karnataka, India, 577007"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1016-3976","authenticated-orcid":true,"given":"Hao","family":"Wang","sequence":"additional","affiliation":[{"name":"Ocean University of China , , 1299 Sansha Road , ,","place":["Qingdao, China, 266003"]},{"name":"Ocean University of China , , 1299 Sansha Road , ,","place":["Qingdao, China, 266003"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7866-681X","authenticated-orcid":true,"given":"Zhenxing","family":"Li","sequence":"additional","affiliation":[{"name":"Ocean University of China , , 1299 Sansha Road , ,","place":["Qingdao, China, 266003"]},{"name":"Ocean University of China , , 1299 Sansha Road , ,","place":["Qingdao, China, 266003"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,8]]},"reference":[{"issue":"10","key":"2026092310085923300_cit1","doi-asserted-by":"publisher","first-page":"1934578X231209153","DOI":"10.1177\/1934578x231209153","article-title":"Zanthoxylum bungeanum Maxim: A review of its phytochemistry,\npharmacology, and pharmacokinetics","volume":"18","author":"Yan","year":"2023","journal-title":"Nat. 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