摘要
目的分离获得太平洋牡蛎致敏蛋白,考察酶解法对牡蛎致敏蛋白抗原性的影响。方法利用硫酸铵沉淀和葡聚糖凝胶G-75分离牡蛎致敏蛋白,聚丙烯酰胺电泳分析获得蛋白的分子量大小,使用木瓜蛋白酶、中性蛋白酶和动物蛋白水解酶(3 000 U/g)酶解牡蛎致敏蛋白,OPA法测定水解度,间接竞争ELISA法测定酶解产物的抗原性。结果分离到分子量约为40和98 kD的牡蛎致敏蛋白,经木瓜蛋白酶酶解的致敏蛋白特征条带消失,酶解产物水解度为(31.87±0.309)%,动物蛋白酶和中性蛋白酶对牡蛎致敏蛋白水解度分别为(24.13±0.153)%和(12.43±0.115)%,但致敏蛋白特征条带未完全消失;间接竞争ELISA检测表明3种蛋白酶均能不同程度地降低牡蛎致敏蛋白的抗原性或使致敏性基本消除。结论蛋白酶酶解法可有效水解牡蛎致敏蛋白,从而降低其抗原性,其中木瓜蛋白酶效果明显。
Objective Allergenic proteins of pacific oyster （ Crassostrea gigas） were separated and the change of protein molecular weight and antigenicity after proteases treatment were studied. Methods Oyster allergenic protein was isolated by ammonium sulfate precipitation and Sephadex G-75. The oyster allergenic protein was hydrolyzed by animal protein hydrolysis enzyme, papain, and neutrase protease, respectively. The degrees of hydrolysis were assessed by OPA and SDS- PAGE methods. The change of allergenic protein antigenicity were detected by indirect competitive ELISA. Results Oyster allergenic protein of 40 and 98 kD was separated and the 40 kD protein was used for further studies. The highest hydrolysis degree of （31.87 ± 0. 309）% was achieved by papain and the oyster allergy protein band was disappeared in SDS-PAGE analysis. While by animal protein hydrolysis enzyme and neutral protease, the proteolytic degree of oyster allergies were （24. 13 ± 0. 153 ）% and （12.43 ± 0. 115 ）% , respectively. The indirect competitive ELISA analysis also showed that the antigenicity of protein were obviously decreased by hydrolysis with these three proteases. Conclusion Antigenicity of oyster allergenic protein could be effectively reduced by papain hydrolysis.
出处
《中国食品卫生杂志》
北大核心
2014年第4期307-311,共5页
Chinese Journal of Food Hygiene
基金
"十二五"国家科技支撑计划项目子课题(2012BAD33B10-3)
海洋公益性行业科研专项子课题(2013418018)
国家青年科学基金项目(31101271)
省部产学研结合项目(2012B091000025)
广东省海洋经济创新发展区域示范专项项目(SZHY2012-B01-004)
广东省海洋经济创新发展区域专项项目(GD2013-B03-001)
关键词
牡蛎致敏蛋白
分离
酶解
抗原性
食品安全
致敏
食物致敏
水产品
Oyster allergenic protein
separation
enzyme hydrolysis
antigenicity
food safety
allergy
food allergy
aquatic product
