体外模拟淡水鱼小清蛋白对胃肠消化的影响（英文）

发布时间：2018-08-18 10:40

【摘要】：大多数食物过敏原的理化性质决定了蛋白质在消化道中的稳定性。模拟胃液(simulated gastric fluid,SGF)的稳定性是评估食物过敏的一个重要参数。为了解一种主要的鱼类过敏原小清蛋白(parvalbumin,PV)与来自鲤鱼(Cyprinus carpio)和鲢鱼(Hypophthalmichthys molitrix)肌肉中的非致敏性蛋白在SGF和模拟肠液(simulated intestinal fluid,SIF)中的稳定性差异,本研究采用3种蛋白水解酶(胃蛋白酶、胰蛋白酶和来自猪的胰凝乳蛋白酶)来模拟人(Homo sapiens)体肠道的消化蛋白酶;结合两次三氯乙酸沉淀法和凝胶过滤层析法纯化PV,通过Tricine-十二烷基硫酸钠聚丙烯酰胺凝胶电泳(Tricine-sodium dodecyl sulfate polyacrylamide gel electrophoresis,Tricine-SDS-PAGE)和Western blot对3种蛋白在模拟肠胃液中的稳定性进行了评估。结果显示,鲤鱼和鲢鱼的PV都获得了分子量约10 k D的蛋白,同时在SGF中鲤鱼和鲢鱼的纯化PV也获得了相同的分子量蛋白。胃蛋白酶使原先的PV带几乎在60 min内完全降解,并观察到一些稳定的肽片段,而胰蛋白酶和胰凝乳蛋白酶在240 min后都无法有效降解PV。在SGF中的非致敏性鱼浆蛋白在很短的时间内迅速降解,而PV的消化时间延长。Western blot分析表明,抗鲢鱼PV多克隆抗体可以特异性地检测到PV及其降解产物。PV比非致敏性蛋白更能抵抗蛋白酶消化,相比胰蛋白酶和胰凝乳蛋白酶,胃蛋白酶处理能更有效地减少超敏反应。研究结果为未来低致敏性鱼产品的开发提供了理论参考。
[Abstract]:The physical and chemical properties of most food allergens determine the stability of proteins in the digestive tract. The stability of simulated gastric fluid (SGF) is an important parameter in assessing food allergies. Lmichthys molitrix) the stability of the non sensitizing proteins in muscle in SGF and simulated simulated intestinal fluid (SIF). 3 proteolytic enzymes (pepsin, trypsin and chymotrypsin from pig *) were used to simulate the digestive protease of human (Homo sapiens) body, combined with two times three chloroacetic acid precipitation. PV was purified by precipitation and gel filtration chromatography. The stability of the three proteins in simulated gastrointestinal juice was evaluated by Tricine-sodium dodecyl sulfate polyacrylamide gel electrophoresis (Tricine-SDS-PAGE) and Western blot. The purified PV of carp and silver carp in SGF also obtained the same molecular weight protein. Pepsin degraded the original PV band almost completely within 60 minutes, and some stable peptide fragments were observed. Trypsin and chymotrypsin could not degrade PV effectively after 240 minutes. Western blot analysis showed that polyclonal antibodies against silver carp PV could specifically detect PV and its degradation products. PV was more resistant to protease digestion than non-allergic proteins, and pepsin treatment was more effective than trypsin and chymotrypsin. The results provide a theoretical reference for the development of low-sensitivity fish products in the future.
【作者单位】：中国计量大学浙江省生物计量及检验检疫技术重点实验室;浙江省农业科学院食品科学研究所;
【基金】：国家自然科学基金(No.31571845) 浙江省自然科学基金(No.LY14C200012) 浙江省公共技术应用研究项目(No.2015C32023) 国家质量监督检验检疫总局(No.201410083)
【分类号】：R57

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