美拉德反应对鲢鱼小清蛋白免疫活性的影响
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- 英文论文题名：Effectof Maillard Reaction on Immunoreactivity of Recombinant Silver Carp Parvalbumin
- 论文作者：杨汝晴 ; 赵勇娟 ; 曹敏杰 ; 刘光明 ; 谢泽伟 ; 蔡秋凤
- 英文论文作者：Yang Ruqing ; Zhao Yongjuan ; Cao Minjie ; Liu Guangming ; Xie Zewei ; Cai Qiufeng ; Academy of science ; Liaoning University ; National & Local Joint Engineering Research Center of Processing Technology for Aquatic Products
- 年：2016
- 作者机构：集美大学食品与生物工程学院;水产品深加工技术国家地方联合工程研究中心;
- 论文关键词：小清蛋白 ; 甘露糖 ; 美拉德反应 ; 免疫活性
- 英文论文关键词：parvalbumin ; mannose ; maillard reaction ; immunoreactivity
- 会议召开时间：2016-11-09
- 会议录名称：中国食品科学技术学会第十三届年会论文摘要集
- 英文会议录名称：Abstracts of the 13th Annual Meeting of CIFST
- 语种：中文
- 分类号：TS201.2
- 学会代码：ZGSP
- 会议名称：中国食品科学技术学会第十三届年会
- 会议地点：中国北京
- 主办单位：中国食品科学技术学会
- 学会名称：中国食品科学技术学会
- 页数：2
- 文件大小：111k
- 原文格式：O
- 会议级别：全国
摘要
本研究的目的是探讨以甘露糖为还原糖进行美拉德反应对小清蛋白(parrvalbumin,PV)免疫活性的影响。通过将鲢鱼重组小清蛋白(recormbinant PV,rPV)和甘露糖混合物在干热条件下(100℃)反应100 min制得糖化的rPV。采用Tricine-SDS-PAGE和Dot-blotting分析糖化前后rPV的聚合特性、免疫活性及消化稳定性的变化;采用圆二色谱仪和扫描电子显微镜分析糖化对rPV的二级结构及微观结构的影响。结果显示,rPV糖化后形成大分子量的片状聚集体;甘露糖糖化修饰的rPV的免疫活性及消化稳定性均显著下降;美拉德反应导致rPV二级结构中的α-螺旋、无规则卷曲的含量有一定程度的减少,而β-折叠的含量显著增加。本研究的结果表明,以甘露糖为还原糖进行美拉德反应可有效降低rPV的免疫活性及消化稳定性,这可能与糖化后rPV聚集体的形成及二级结构的改变有关。
The aim of this study was 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) was used to determine the change of the secondary structure and the microstructure of rPV after Maillard reaction.The results showed that M-rPV had sienificantlv high amounts of protein-bound mannose of approximately 150 μg per microgram M-rPV measuring 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 analyzing by reducing Tricine-SDS-PAGE.Formation of compact aggregates after Maillard reaction was further comfirmed by SEM,which might block the access of immunoglobulins to the epitopes,and therefore decrease the immunoreactivity of M-rPV,as demostrated by dot-blotting using mouse anti-silver carp parvalbumin monoclonal antibody.M-rPV exhibited less resistance against gastrointestinal digestion in vitro which revealed by Tricine-SDS-PAGE and dot-blotting.Maillard reaction also shown 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.
The aim of this study was 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) was used to determine the change of the secondary structure and the microstructure of rPV after Maillard reaction.The results showed that M-rPV had sienificantlv high amounts of protein-bound mannose of approximately 150 μg per microgram M-rPV measuring 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 analyzing by reducing Tricine-SDS-PAGE.Formation of compact aggregates after Maillard reaction was further comfirmed by SEM,which might block the access of immunoglobulins to the epitopes,and therefore decrease the immunoreactivity of M-rPV,as demostrated by dot-blotting using mouse anti-silver carp parvalbumin monoclonal antibody.M-rPV exhibited less resistance against gastrointestinal digestion in vitro which revealed by Tricine-SDS-PAGE and dot-blotting.Maillard reaction also shown 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.
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