大氣電漿處理對花生中過敏原含量及結構之影響

Effect of atmospheric plasma treatment on the content and structure of peanut allergen

黃暄貽(Huang Shiuan-Yi)

指導教授 ： 余旭勝

國立屏東科技大學/農學院/食品科學系所/碩士(2021年)

https://doi.org/10.6346/NPUST202100313

摘要

花生 (Arachis hypogaea) 為歐美地區公認最重要的過敏食物之一，其過敏症狀包含蕁麻疹、腹痛或腹瀉，嚴重可能會有過敏性休克之情況發生。Ara h 2是花生中主要的過敏原之一，其耐熱性佳不易受到熱破壞。大氣電漿技術為一種新興非熱加工技術，依據相關文獻表明此技術具誘導蛋白質結構變化能力，同時保留花生中的營養成分，但不清楚對Ara h 2致過敏性的影響，其他文獻的研究也缺乏有關大氣電漿對Ara h 2的抗原性之處理條件說明，因此本實驗將探討大氣電漿不同處理時間減少Ara h 2過敏原的結構變化之相關性。首先利用大氣電漿產生濃度約400 ± 10 ppm臭氧與花生分別進行反應0、5、15、30、45及60分鐘，並各別萃取其花生蛋白，以十二烷基硫酸鈉聚丙烯醯胺凝膠電泳 (Sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE) 評估蛋白條帶差異，再以酵素連結免疫吸附分析法 (Enzyme-linked immunosorbent assay, ELISA) 檢測花生中Ara h 2的濃度，另外利用傅立葉轉換紅外光譜儀 (Fourier-transform infrared spectroscopy, FTIR) 檢測花生經大氣電漿處理後Ara h 2二級結構的變化。結果顯示，花生中Ara h 2經大氣過電漿處理後從SDS-PAGE中可知蛋白質條帶濃度有產生減弱的現象；ELISA之結果顯示，與對照組相比下，Ara h 2的濃度在電漿處理5~30分鐘皆無明顯變化，但在反應45分鐘及60分鐘後則分別減少0.67 ± 0.16 mg/g及0.60 ± 0.13 mg/g，另外在結構方面，從光譜圖中的Amide I區 (1700-1600 cm–1) 波峰範圍處觀察到α-helix, β-sheet, β-turn, and Random coil之二級結構的變化，隨著電漿處理時間越長α-helix跟β-sheet的結構面積皆呈現減少，β-turn和Random coil的結構面積則增加，與未經電漿處理的Ara h 2相比較，大氣電漿處理45分鐘的α-helix的面積下降48.99%、β-sheet則下降15.74 %、β-turn及Random coil的面積各增加28.49%及36.24%，表示二級結構有明顯改變。綜合上述結果，由大氣電漿可破壞花生中Ara h 2，是有效減少花生過敏原Ara h 2的非熱加工技術。

關鍵字

Ara h 2 ； 食物過敏 ； 大氣電漿 ； 傅立葉轉換紅外光譜 ； 二級結構

並列摘要

Peanut (Arachis hypogaea) is recognized as one of the most important allergic foods in Europe and America. Its symptoms can cause hives, stomach, or diarrhea. In severe cases, it can cause anaphylactic shock. The main allergen in peanuts is Ara h 2, which has a heat resistance well. However, atmospheric plasma is an emerging non-thermal processing technology. According to relevant literature, this technology has the ability to induce protein conformation changes, while reducing the allergenicity of Ara h 2 and retaining the nutrients in peanuts. It lacks relevant optimal atmospheric plasma conditions for reducing the antigenicity of Ara h 2 are explained. Therefore, this study will explore the correlation between different treatment times of atmospheric plasma to reduce the sensitization of the structural of Ara h 2 changes. This study utilizes atmospheric plasma to generate ozone at a concentration of about 400 ± 10 ppm and reaction time for 0, 5, 15, 30, 45, and 60 minutes. Extract peanut protein components individually, and use Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to evaluate the difference in protein bands. Enzyme-linked immunosorbent assay (ELISA) was used to detect the concentration of Ara h 2 in peanuts. In addition, Fourier transform infrared spectroscopy (FTIR) was used to detect the changes in the secondary structure of Ara h 2 after atmospheric plasma treatment of peanuts. The results showed that after atmospheric plasma treatment of Ara h 2, the protein bands in SDS-PAGE were weakened; In part of ELISA showed that compared with the control group, the concentration of Ara h 2 did not change significantly after plasma treatment for 5 to 30 minutes. After 45 and 60 minute treat, the concentration of Ara h 2 were reduced by 0.67 ± 0.16 mg/g and 0.60 ± 0.13 mg/g, respectively. Other results showed that the structure, which the conformational changes of the secondary structure of α-helix, β-sheet, β-turn, and Random coil were observed in the Amide I (1700-1600 cm–1) in the spectrum. The longer treatment time, the percentages of α-helix and β-sheets decreased and the percentages of β-turns and random coils increased. Compared with Ara h 2 without atmospheric plasma treatment, the Ara h 2 treated for 45 minutes that α-helix decreased 48.99%, β-sheet decreased 15.74%, β-turn and random coil increased 28.49% and 36.24%, indicating that the secondary structure has significantly changed. From the above results, atmospheric plasma can destroy the Ara h2 protein in peanuts, which is an effective method for reducing peanut allergens in the current non-thermal technology.

並列關鍵字

Ara h 2 ； Food allergy ； Atmospheric plasma ； Fourier-transform infrared spectroscopy ； Secondary structure

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