水产学报2017，Vol.41Issue(6)：870-876,7.DOI:10.11964/jfc.20160710498
甘露糖—小清蛋白美拉德反应产物的免疫活性
Study on the immunoreactivity of mannose-parvalbumin conjugate
摘要
Abstract
In order to determine the influence of Maillard reaction on the structural and immunological properties of parvalbumin, the major allergen of silver carp (Hypophthalmichthys molitrix), recombinant silver carp PV (rPV) was used and incubated with mannose at 100 ℃ for 100 min, and glycated PV (M-rPV) was then obtained after dialysis. Tricine-SDS-PAGE and Dot-blotting was performed to elucidate the change of mobility and immunoreactivity of rPV after glycation. Simulated gastrointestinal digestion experiment was carried out to compare the digestibility of rPV and M-rPV. Circular dichroism spectroscopy (CD) and scanning electron microscopy (SEM) were used to determine the change of the secondary structure and the microstructure of rPV after Maillard reaction. The results showed that M-rPV had significantly high amounts of protein-bound mannose of approximately 150 μg per microgram M-rPV as measured by the anthrone sulfuric acid method. Dimers and polymers formed in the glycated sample, which might be attributed to the covalent bonds formed during Maillard reaction as analyzed by reducing Tricine-SDS-PAGE. Formation of compact aggregates after Maillard reaction was further confirmed by SEM, which might block the access of immunoglobulins to the epitopes, and therefore decrease the immunoreactivity of M-rPV, as demonstrated by dot-blotting using mouse anti-silver carp parvalbumin monoclonal antibody. M-rPV exhibited less resistance against gastrointestinal digestion in vitro as revealed by Tricine-SDS-PAGE and dot-blotting. Maillard reaction also showed a minor effect on the secondary structures of rPV with a decrease of α-helix and an increase of β-strand content. The aggregation formation and change in the secondary structure of M-rPV might explain the decrease of rPV immunoreactivity and digestibility after Maillard reaction. Our work might be useful for understanding the effects of Maillard reaction on the decrease of immunoreactivity of food allergens in general.关键词
鲢/小清蛋白/甘露糖/美拉德反应/免疫活性Key words
Hypophthalmichthys molitrix/parvalbumin/mannose/Maillard reaction/immunoreactivity分类
轻工纺织引用本文复制引用
杨汝晴,赵勇娟,张凌晶,谢泽伟,刘光明,曹敏杰..甘露糖—小清蛋白美拉德反应产物的免疫活性[J].水产学报,2017,41(6):870-876,7.基金项目
国家自然科学基金(31301440) （31301440）
福建省自然科学基金(2016J01709) （2016J01709）
福建省教育厅专项(JA14170) National Natural Scientific Foundation of China (31301440) （JA14170）
National Scientific Foundations of Fujian Province (2016J01709) （2016J01709）
Fujian Education Department Fund (JA14170) （JA14170）
