食品科学 ›› 2018, Vol. 39 ›› Issue (20): 20-26.doi: 10.7506/spkx1002-6630-201820004

• 食品化学 • 上一篇    下一篇

磷酸化对鲢小清蛋白抗原性降低的作用

杨汝晴1,林灵海1,万楚君1,陈玉磊1,张凌晶1,2,曹敏杰1,2,*   

  1. (1.集美大学食品与生物工程学院,福建?厦门 361021;2.福建省海洋生物资源开发利用协同创新中心,福建?厦门 361102)
  • 出版日期:2018-10-25 发布日期:2018-10-24
  • 基金资助:
    国家自然科学基金面上项目(31471640);福建省科技计划高校产学合作项目(2017N5011)

Effect of Phosphorylation on Reducing the Allergenicity of Silver Carp Parvalbumin

YANG Ruqing1, LIN Linghai1, WAN Chujun1, CHEN Yulei1, ZHANG Lingjing1,2, CAO Minjie1,2,*   

  1. (1. College of Food and Biological Engineering, Jimei University, Xiamen 361021, China; 2. Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources, Xiamen 361102, China)
  • Online:2018-10-25 Published:2018-10-24

摘要: 通过建立鱼类主要过敏原小清蛋白(parvalbumin,PV)磷酸化反应的条件,研究磷酸化对PV的抗原性及结构影响。将PV和葡萄糖-6-磷酸二钠盐(D-glucose-6-phosphate disodium salt hydrate,G6P-Na2)混合,在不同质量比、反应时间以及反应温度条件下进行干法磷酸化反应。采用Tricine-十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和Dot-blotting检验其聚合类型以及抗原性变化。通过扫描电镜对聚合物的微观结构进行分析,利用圆二色谱仪和ANS探针法分析磷酸化产物的二级结构和疏水性变化。结果显示,在PV与G6P-Na2质量比为1∶4、反应温度80?℃、反应时间80?min条件下生成的磷酸化产物的抗原性最低。磷酸化反应后,产物呈现聚集状态,其二级结构变化较为明显,疏水性明显提高。磷酸化反应对蛋白结构的改变可能是影响PV抗原性降低的主要原因。

关键词: 小清蛋白, 葡萄糖-6-磷酸二钠盐, 磷酸化反应, 抗原性, 二级结构

Abstract: The aims of this work were to establish and optimize the phosphorylation reaction conditions of parvalbumin (PV), the major allergen of silver carp (Hypophthalmichthys molitrix), and to determine the influence of phosphorylation reaction on the structural and immunological properties of PV. Phosphorylation of PV was carried out by dry heating of its mixture with D-glucose-6-phosphate disodium salt hydrate (G6P-Na2) under varying conditions of mass ratio between reactants, reaction time and temperature. The type of polymerization was determined by tricine-SDS-PAGE, and the antigenicity was detected by dot blotting. The microstructure of the obtained polymer was examined by scanning electron microscope (SEM), and its secondary structure and surface hydrophobicity were measured by circular dichroism (CD) spectroscopy and the hydrophobic fluorescence probe 8-anilino-1-naphthalenesulfonate (ANS), respectively. The results indicated that the optimal conditions for phosphorylation reaction of PV were as follows: reaction temperature, 80 ℃; reaction time, 80 min; and ratio of PV to G6P-Na2, 1:4. The aggregate formation and the changes in the secondary structures of G6P-Na2-PV might explain the decrease of PV antigenicity after the phosphorylation reaction. Our present work strongly suggested an effect of phosphorylation on reducing the antigenicity of food allergens.

Key words: parvalbumin, D-glucose-6-phosphate disodium salt hydrate, phosphorylation reaction, antigenicity, secondary structure

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