Abstract: In this study, casein was hydrolyzed with seven food-grade proteases, including flavourzyme, trypsin, chymotrypsin, alkaline protease, papain, bromelain, and neutral protease and the allergenicity of the resultant hydrolysates was assessed using a serum pool of patients with milk allergy. Moreover, the molecular mass distribution, degree of hydrolysis and residual epitope information were evaluated in order to select the most suitable protease for the production of casein products with reduced allergenicity. The results showed that under the same conditions, the binding ability of casein to specific antibodies decreased to the greatest extent after being treated with chymotrypsin or flavourzyme, so they could be used as a candidate protease for the preparation of subsequent hypoallergenic casein hydrolysates. The results of T-cell epitope prediction and analysis of casein hydrolysates showed that only one T-cell epitope peptide consisting of more than nine amino acids LHSMKEGIHAQQK remained after chymotrypsin hydrolysis of casein.

Key words: milk allergy; protease; hydrolysis; casein; allergenicity assessment

摘要： 以酪蛋白为研究对象，选取复合风味蛋白酶、胰蛋白酶、胰凝乳蛋白酶、碱性蛋白酶、木瓜蛋白酶、菠萝 蛋白酶和中性蛋白酶共7 种食品级蛋白酶，对牛乳酪蛋白进行酶解。通过牛乳过敏患者血清池，评估酶解产物致敏 原性，同时结合水解产物的分子质量分布、水解度、残留表位信息等指标，筛选出最适合用于生产低致敏酪蛋白产 品的蛋白酶。结果表明：在相同条件下经胰凝乳蛋白酶、复合风味蛋白酶处理后酪蛋白与特异性抗体结合能力降低 程度最大，可用于后续低致敏酪蛋白制备的候选蛋白酶；结合T细胞表位预测结果，对酪蛋白水解产物进一步分析 发现，酪蛋白经过胰凝乳蛋白酶酶解后，仅剩余1 条大于9 个氨基酸的T细胞表位肽段LHSMKEGIHAQQK。

关键词: 牛乳过敏；蛋白酶；水解；酪蛋白；致敏性评估