不同结构酪蛋白深度水解物的致敏性分析

Allergenicity analysis of casein deep hydrolysates with different structures

以酶凝酪蛋白和酸凝酪蛋白为研究对象,探究深度水解对不同结构酪蛋白致敏性的影响。分别采用高效液相色谱(HPLC)分析水解物的分子质量分布、酶联免疫吸附(ELISA)测定水解物的致敏性、液相色谱串联质谱(LC-MS/MS)解析过敏原肽段切割情况。结果表明:2种酪蛋白均符合深度水解的要求,其中酶凝酪蛋白中分子质量小于0.2 kDa约占27.43%,酸凝酪蛋白中分子质量小于0.2 kDa约占24.85%,表明酶凝酪蛋白的水解更完全。ELISA结果显示,酶凝酪蛋白的致敏性在水解后下降10%~20%,酸凝酪蛋白的致敏性在水解后还可能会出现增大的现象。LC-MS/MS结果显示,酶凝酪蛋白的切割效率为96.77%,酸凝酪蛋白的切割效率为69.35%。综上所述,酶凝酪蛋白水解物的致敏性低于酸凝酪蛋白水解物。

Using enzyme coagulated casein and acid coagulated casein as subjects,the effect of deep hy-drolysis on the allergenicity of casein with different structures was explored.The molecular mass distribu-tion of hydrolysate was analyzed by high performance liquid chromatography(HPLC),the allergenicity of hydrolysate was determined by enzyme-linked immunosorbent assay(ELISA),and the cleavage of aller-gen peptide was analyzed by liquid chromatography tandem mass spectrometry(LC-MS/MS).The results showed that the 2 kinds of casein met the requirements of deep hydrolysis,among which the enzyme co-agulated casein with molecular mass less than 0.2 kDa accounted for 27.43%,and the acid coagulated casein with molecular mass less than 0.2 kDa accounted for 24.85%,indicating that the hydrolysis of enzyme coagulated casein was more complete.The results of ELISA showed that the allergenicity of en-zyme coagulated casein was decreased by 10%-20%after hydrolysis,while that of acid coagulated ca-sein might increased after hydrolysis.The LC-MS/MS results showed that the cleavage efficiency of en-zyme coagulated casein was 96.77%,while that of acid coagulated casein was 69.35%.In summary,the allergenicity of enzymatic coagulated casein hydrolysates was lower than that of acid coagulated casein hydrolysates.