Liang Xiaona, Yang Hui, Sun Jing, Cheng Jiao, Luo Xue, Wang Zongzhou, Yang Mei, Bing Tao Dong, Yue Xiqing, Zheng Yan
College of Food Science Shenyang Agricultural University ShenYang China.
Food Sci Nutr. 2020 Dec 30;9(2):985-993. doi: 10.1002/fsn3.2066. eCollection 2021 Feb.
Cow milk (CM) allergy is one of the most common food allergies worldwide; the most abundant CM proteins, such as casein (CN), β-lactoglobulin (β-LG), and ɑ-lactalbumin (ɑ-LA), are all potentially allergenic. Reducing the antigenicity of CM continues to be a major challenge. However, previous studies have focused on the antigenicity of individual allergic CM proteins. Thus, in the present study, we aimed to evaluate the effects of different food-grade enzymes on the antigenicity of CN, β-LG, ɑ-LA in natural CM. The degree of hydrolysis (DH) and molecular mass (MW) distribution of CM hydrolysates were assessed. Additionally, the residual antigenicity of CM hydrolysates was evaluated through enzyme-linked immunosorbent assay and Western blotting with anti-CN, anti-β-LG, and anti-ɑ-LA rabbit polyclonal antibodies. The results showed that Alcalase- and Protamex-mediated hydrolysis could efficiently reduce the antigenicity of CN, β-LG, and ɑ-LA, inducing a higher DH, the loss of density of CM proteins, and the increasing levels of low MW (<3 kDa) peptides in CM hydrolysates. Further, Protamex and Alcalase could more efficiently hydrolyze the major allergenic components of CM than the other enzymes, which could represent an advantage for the development of hypoallergenic CM. These findings add further knowledge about the study and development of hypoallergenic CM.
牛奶（CM）过敏是全球最常见的食物过敏之一；CM中最丰富的蛋白质，如酪蛋白（CN）、β-乳球蛋白（β-LG）和α-乳白蛋白（α-LA），都具有潜在的致敏性。降低CM的抗原性仍然是一项重大挑战。然而，以往的研究主要集中在单个致敏CM蛋白的抗原性上。因此，在本研究中，我们旨在评估不同食品级酶对天然CM中CN、β-LG、α-LA抗原性的影响。评估了CM水解产物的水解度（DH）和分子量（MW）分布。此外，通过酶联免疫吸附测定以及使用抗CN、抗β-LG和抗α-LA兔多克隆抗体进行蛋白质印迹法，评估了CM水解产物的残留抗原性。结果表明，碱性蛋白酶和中性蛋白酶介导的水解可以有效降低CN、β-LG和α-LA的抗原性，导致更高的DH、CM蛋白密度的降低以及CM水解产物中低分子量（<3 kDa）肽水平的增加。此外，与其他酶相比，中性蛋白酶和碱性蛋白酶能够更有效地水解CM的主要致敏成分，这可能是开发低敏CM的一个优势。这些发现为低敏CM的研究和开发增添了更多知识。
