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风味酶和碱性蛋白酶处理对花生过敏原Ara h 1结构及致敏性的影响

Effects of Flavourzyme and Alkaline Protease Treatment on Structure and Allergenicity of Peanut Allergen Ara h 1.

作者信息

Shu Erlian, Wang Shuo, Kong Xiangxiang, Sun Xiaodong, Yang Qiaoling, Chen Qin, Niu Bing

机构信息

School of Life Sciences, Shanghai University, No. 99, Shangda Road, Baoshan District, 200444 Shanghai, PR China.

School of Medicine, Shanghai University, No. 99, Shangda Road, Baoshan District, 200444 Shanghai, PR China.

出版信息

Food Technol Biotechnol. 2024 Mar;62(1):4-14. doi: 10.17113/ftb.62.01.24.8064.

Abstract

RESEARCH BACKGROUND

Peanut allergy poses a significant threat to human health due to the increased risk of long-term morbidity at low doses. Modifying protein structure to affect sensitization is a popular topic.

EXPERIMENTAL APPROACH

In this study, the purified peanut allergen Ara h 1 was enzymatically hydrolysed using Flavourzyme, alkaline protease or a combination of both. The binding ability of Ara h 1 to antibodies, gene expression and secretion levels of the proinflammatory factors interleukin-5 and interleukin-6 in Caco-2 cells was measured. Changes in the secondary and tertiary structures before and after treatment with Ara h 1 were analysed by circular dichroism and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).

RESULTS AND CONCLUSIONS

The results indicated a decrease of the allergenicity and proinflammatory ability of Ara h 1. The evaluation showed that the Flavourzyme and alkaline protease treatments caused particle shortening and aggregation. The fluorescence emission peak increased by 3.4-fold after the combined treatment with both proteases. Additionally, the secondary structure underwent changes and the hydrophobicity also increased 8.95-fold after the combined treatment.

NOVELTY AND SCIENTIFIC CONTRIBUTION

These findings partially uncover the mechanism of peanut sensitization and provide an effective theoretical basis for the development of a new method of peanut desensitization.

摘要

研究背景

由于低剂量时长期发病风险增加,花生过敏对人类健康构成重大威胁。改变蛋白质结构以影响致敏是一个热门话题。

实验方法

在本研究中,使用风味酶、碱性蛋白酶或两者的组合对纯化的花生过敏原Ara h 1进行酶解。测定了Ara h 1与抗体的结合能力、Caco-2细胞中促炎因子白细胞介素-5和白细胞介素-6的基因表达及分泌水平。通过圆二色性和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析了Ara h 1处理前后二级和三级结构的变化。

结果与结论

结果表明Ara h 1的致敏性和促炎能力降低。评估显示风味酶和碱性蛋白酶处理导致颗粒缩短和聚集。两种蛋白酶联合处理后荧光发射峰增加了3.4倍。此外,联合处理后二级结构发生变化,疏水性也增加了8.95倍。

新颖性与科学贡献

这些发现部分揭示了花生致敏机制,为开发新的花生脱敏方法提供了有效的理论依据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a3/11002447/d79c2a604653/FTB-62-4-f1.jpg

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