加工工序对苦杏仁球蛋白免疫反应性及消化稳定性的影响

doi:10.3864/j.issn.0578-1752.2023.24.011

摘要/Abstract

摘要：

【背景】近年来，过敏已经成为全球关注的健康问题，过敏人群数量持续上升。坚果是常见的过敏诱因之一，苦杏仁为常见的坚果，因其含有致敏的苦杏仁球蛋白（amandin），成为最易引起过敏的坚果之一，因此，脱敏已成为其研究热点。苦杏仁加工一般要经过去皮、脱苦和干制等工序，而在这些加工过程中，其致敏性是否会受到影响尚未见到相关报道。【目的】探究加工工序对苦杏仁致敏性的影响，并以苦杏仁致敏性、品质和营养特性为评价指标，优化加工工序使苦杏仁产品致敏性最低，为低致敏性苦杏仁产品加工提供理论依据和技术支撑。【方法】采用Western blotting和ELISA试验研究不同去皮、脱苦和干制方法对苦杏仁中amandin免疫反应性的影响；采用圆二色光谱、外源荧光光谱、表面疏水性和zeta电位测定以研究各加工工序对amandin结构和表面性质的影响，分析致敏蛋白免疫反应性变化的机理；最后采用体外模拟消化试验探究加工前后苦杏仁中amandin的消化稳定性，并对消化产物进行Western blotting分析，进一步探究苦杏仁潜在致敏性的变化情况。【结果】就致敏性而言，苦杏仁经饱和热空气去皮和热烫去皮后的amandin免疫反应性分别降低了8.41%和13.15%，再经超声快速脱苦后amandin的免疫反应性又降低6.79%，热水脱苦对其免疫反应性无显著影响；脱苦杏仁经自然干燥和热风干燥后amandin的免疫反应性分别显著上升4.58%和2.81%（P<0.05）。结合加工工序对苦杏仁品质和营养特性的影响，最终优化得到低致敏性苦杏仁的加工工序为饱和热空气去皮、超声快速脱苦和热风干制，经此工序加工后苦杏仁免疫反应性降低15.03%。就致敏蛋白结构而言，致敏蛋白的二级结构组成、三级结构、表面疏水性以及zeta电位在加工过程中都发生了一定程度变化，其中超声快速脱苦显著改变了amandin的三级结构并使其表面疏水性显著增强（P<0.05），从而使其免疫反应性降低最显著。就消化稳定性而言，加工后的amandin消化稳定性明显降低，致敏蛋白中与特异性抗原抗体反应相关结构降解速度加快，导致苦杏仁的潜在致敏性进一步降低。【结论】不同加工工序处理可通过改变amandin的结构而对苦杏仁致敏性产生影响，生产中可以通过合理的加工方式来降低苦杏仁的致敏性。

关键词: 苦杏仁, 苦杏仁球蛋白, 加工, 免疫反应性, 消化稳定性

Abstract:

【Background】In recent years, allergies have become a global health concern, and the number of allergic individuals continues to rise. Nut is one of the common origins of allergies, and apricot kernels, as a common nut containing the allergenic protein amandin, have become one of the most susceptible nuts to allergies. Therefore, allergy removal of nuts has become a research hotspot. The processing of apricot kernels generally involves procedures, such as peeling, debitterizing and drying, and there are no relevant reports on whether their allergenicity will be affected during these processing. 【Objective】The aim of this study was to explore the impact of processing on the allergenicity with the allergenicity, quality and nutritional characteristics of apricot kernels as the evaluation indicators, and to optimize the processing for reducing the allergenicity of apricot kernels, thus providing the theoretical basis and technical support for the processing of low allergenic nuts products of apricot kernels.【Method】Firstly, the methods of Western blotting and ELISA were used to investigate the effects of different peeling, debitterizing and drying methods on the amandin immunoreactivity in apricot kernels. Then, the circular dichroism spectroscopy, extrinsic fluorescence spectroscopy, surface hydrophobicity and zeta potential measurements were used to study the effects of various processing methods on the structure and surface properties of amandin, and to analyze the mechanism of the immune-reactivity changes of amandin. Finally, the vitro simulation digestion experiments were conducted to investigate the digestive stability of amandin in apricot kernels before and after processing, and Western blotting analysis was conducted on the digestion products to further explore the changes in potential allergenicity of apricot kernels.【Result】In terms of allergenicity, the amandin’s immunoreactivity after being peeled by the saturated hot air and blanched decreased by 8.41% and 13.15%, respectively. After being quickly debitterized by ultrasound, the amandin’s immunoreactivity decreased by 6.79%. Blanching debitterizing had no significant effects on its immunoreactivity. After natural drying and hot air drying, the immune reactivity of the amandin significantly increased by 4.58% and 2.81%, respectively (P<0.05). Based on the impact of processing on the quality and nutritional characteristics of apricot kernels, those suitable processing methods to decrease the allergenicity of apricot kernels were optimized like the saturated hot air peeling, ultrasonic rapid debitterizing and hot air drying, and the immune reactivity of apricot kernels decreased by 15.03% under the optimal conditions. In terms of the structure of amandin, the secondary structure composition, tertiary structure, surface hydrophobicity and zeta potential have undergone certain changes during the processing. Among them, the ultrasound rapid debitterizing significantly changed the tertiary structure of amandin and enhanced its surface hydrophobicity (P<0.05), resulting in the most significant decrease in its immune reactivity. The digestive stability of the amandin after processing was significantly reduced, and the accelerated degradation rate of structures was related to the specific antigen antibody reactions in allergenic proteins, leading to a further decrease in the potential allergenicity of apricot kernels.【Conclusion】Different processing steps could affect the allergenicity of apricot kernels by changing the structure of amandin, i.e. the reasonable processing methods could be used to reduce the allergenicity of apricot kernels.

Key words: apricot kernel, amandin, processing, immunoreactivity, digestive stability

龙斐斐, 张清安, 张志华. 加工工序对苦杏仁球蛋白免疫反应性及消化稳定性的影响[J]. 中国农业科学, 2023, 56(24): 4930-4943.

LONG FeiFei, ZHANG QingAn, ZHANG ZhiHua. Effects of Processing Technology on the Amandin Immunoreactivity and Digestive Stability of Apricot Kernel[J]. Scientia Agricultura Sinica, 2023, 56(24): 4930-4943.

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