Fei Dan-Xia, Liu Qing-Mei, Chen Feng, Yang Yang, Chen Zhong-Wei, Cao Min-Jie, Liu Guang-Ming
College of Food and Biological Engineering, Fujian Provincial Engineering Technology Research Center of Marine Functional Food, Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources, Jimei University, Xiamen, Fujian, China.
Department of Food, Nutrition, and Packaging Sciences, Clemson University, Clemson, SC, USA.
Mol Nutr Food Res. 2016 Jul;60(7):1707-18. doi: 10.1002/mnfr.201500936. Epub 2016 Apr 1.
The enzymatic cross-linking of an allergen by food processing may alter its sensitization potential. In this study, the IgE-binding activity and allergenicity of cross-linked thermal polymerized arginine kinase (CL-pAK) were investigated.
The IgE-binding activity and stability of CL-pAK were analyzed by immunological and proteomics methods. The sensitization and potency to induce oral tolerance of CL-pAK were tested using in vivo assays and a cell model. According to the results of inhibition of ELISA, the half inhibitory concentration of AK after cross-linking changed from 1.13 to 228.36 μg/mL. The results of in vitro digestion demonstrated that CL-pAK showed more resistance to gastrointestinal digestion than native AK. Low allergenicity and capacity to induce oral tolerance in mice were shown by the sera levels of AK-specific antibodies and T-cell cytokine production. Exposure of RBL-2H3 cells to CL-pAK compared with AK, resulted in lower levels of mast degranulation and histamine.
Enzymatic cross-linking with thermal polymerization of AK by tyrosinase and caffeic acid had high potential in mitigating IgE-binding activity and allergenicity, which were influenced by altering the molecular and immunological features of the shellfish protein.
食品加工过程中过敏原的酶促交联可能会改变其致敏潜力。在本研究中，对交联热聚合精氨酸激酶（CL-pAK）的IgE结合活性和致敏性进行了研究。
采用免疫学和蛋白质组学方法分析CL-pAK的IgE结合活性和稳定性。使用体内试验和细胞模型测试CL-pAK诱导口服耐受性的致敏性和效力。根据ELISA抑制结果，交联后AK的半数抑制浓度从1.13 μg/mL变为228.36 μg/mL。体外消化结果表明，CL-pAK比天然AK对胃肠道消化具有更强的抗性。AK特异性抗体的血清水平和T细胞细胞因子产生表明，CL-pAK在小鼠中具有低致敏性和诱导口服耐受性的能力。与AK相比，RBL-2H3细胞暴露于CL-pAK后，肥大细胞脱颗粒和组胺水平较低。
酪氨酸酶和咖啡酸对AK进行热聚合的酶促交联在减轻IgE结合活性和致敏性方面具有很大潜力，这是通过改变贝类蛋白的分子和免疫学特征来实现的。
