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2025年4月5日 星期六
  2016, Vol. 24 Issue (12): 1908-1914    
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
体外模拟淡水鱼小清蛋白对胃肠消化的影响
陶淼1,梁提松2,肖朝耿3,刘明启4,关荣发1
1. 中国计量学院
2. 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
3.
4. 浙江省杭州市下沙高教园区中国计量学院生命学院食品系
Effect of Parvalbumins on Gastrointestinal Digestion in Freshwater Fish with In vitro Simulation
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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 kD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。
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陶淼
梁提松
肖朝耿
刘明启
关荣发
关键词 淡水鱼小清蛋白纯化过敏原鉴定    
Abstract:The physicochemical properties of most food allergens confer stability to the proteolytic in the digestive tract. Stability of simulated gastric fluid (SGF) is an important parameter for the estimation of food allergenicity. To study the digestive stabilities of a major fish allergen — parvalbumin (PV) and non-allergenic proteins from the muscle of carp (Cyprinus carpio) and silver carp (Hypophthalmichthys molitrix) in SGF and simulated intestinal fluid (SIF), three kinds of proteinases including pepsin, trypsin, and chymotrypsin from porcine (Sus scrofa) were used to simulate digestive proteinases from humans (Homo sapiens). Two runs of trichloroacetic acid precipitation and gel filtration chromatography were used to purify the fish PV. Results were evaluated via Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Tricine-SDS-PAGE) and Western blot. The results showed that PVs of carp and silver carp with molecular weight of approximate 10 kD was obtained. Similar results were obtained in SGF assays of purified PVs of carp and silver carp. The original PV band with pepsin was almost completely degraded within 60 min, and some stable peptide fragments were observed. Both trypsin and chymotrypsin could not effectively degrade PV in 240 min. In the SGF digestion on non-allergenic fish sarcoplasmic proteins were rapidly degraded within a short period of time, whereas the digestion time of PV was prolonged. Western blot analysis indicated that the polyclonal antibody against silver carp PV could specifically detect the PV and its degraded products. In conclusion, results indicated that fish PV was more resistant to proteinase digestion than non-allergenic proteins, and pepsin treatment was more effective than trypsin and chymotrypsin in reducing hypersensitivity. This study provides a theoretical reference for the development of hypoallergenic fish products in the future.
Key wordsFreshwater fish    Parvalbumin    Purification    Allergen    Identification
收稿日期: 2016-04-08      出版日期: 2016-11-07
通讯作者: 关荣发     E-mail: rfguan@163.com;rongfaguan@163.com
引用本文:   
陶淼 梁提松 肖朝耿 刘明启 关荣发. 体外模拟淡水鱼小清蛋白对胃肠消化的影响[J]. , 2016, 24(12): 1908-1914.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2016/V24/I12/1908
 
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