Gell Gyöngyvér, Bugyi Zsuzsanna, Florides Christakis George, Birinyi Zsófia, Réder Dalma, Szegő Zsuzsanna, Mucsi Edina, Schall Eszter, Ács Katalin, Langó Bernadett, Purgel Szandra, Simon Katalin, Varga Balázs, Vida Gyula, Veisz Ottó, Tömösközi Sándor, Békés Ferenc
Department of Biological Resources, Agricultural Institute, Centre for Agricultural Research, EötvösLoránd Research Network, Martonvásár, Hungary.
Department of Applied Biotechnology and Food Science, Research Group of Cereal Science and Food Quality, Budapest University of Technology and Economics, Budapest, Hungary.
Front Nutr. 2021 Sep 29;8:702352. doi: 10.3389/fnut.2021.702352. eCollection 2021.
The use of pure oats (oats cultivated with special care to avoid gluten contamination from wheat, rye, and barley) in the gluten-free diet (GFD) represents important nutritional benefits for the celiac consumer. However, emerging evidence suggests that some oat cultivars may contain wheat gliadin analog polypeptides. Consequently, it is necessary to screen oats in terms of protein and epitope composition to be able to select safe varieties for gluten-free applications. The overall aim of our study is to investigate the variability of oat protein composition directly related to health-related and techno-functional properties. Elements of an oat sample population representing 162 cultivated varieties from 20 countries and the protein composition of resulting samples have been characterized. Size distribution of the total protein extracts has been analyzed by size exclusion-high performance liquid chromatography (SE-HPLC) while the 70% ethanol-extracted proteins were analyzed by RP-HPLC. Protein extracts separated into three main groups of fractions on the SE-HPLC column: polymeric proteins, avenins (both containing three subgroups based on their size), and soluble proteins, representing respectively 68.79-86.60, 8.86-27.72, and 2.89-11.85% of the total protein content. The ratio of polymeric to monomeric proteins varied between 1.37 and 3.73. Seventy-six reversed phase-HPLC-separated peaks have been differentiated from the ethanol extractable proteins of the entire population. Their distribution among the cultivars varied significantly, 6-23 peaks per cultivar. The number of appearances of peaks also showed large variation: one peak has been found in 107 samples, while 15 peaks have been identified, which appeared in less than five cultivars. An estimation method for ranking the avenin-epitope content of the samples has been developed by using MS spectrometric data of collected RP-HPLC peaks and bioinformatics methods. Using ELISA methodology with the R5 antibody, a high number of the investigated samples were found to be contaminated with wheat, barley, or rye.
在无麸质饮食（GFD）中使用纯燕麦（经过特殊培育以避免受到小麦、黑麦和大麦的麸质污染的燕麦）对乳糜泻患者具有重要的营养益处。然而，新出现的证据表明，一些燕麦品种可能含有小麦麦醇溶蛋白类似多肽。因此，有必要根据蛋白质和表位组成对燕麦进行筛选，以便能够选择用于无麸质应用的安全品种。我们研究的总体目标是研究与健康相关和技术功能特性直接相关的燕麦蛋白质组成的变异性。对代表来自20个国家的162个栽培品种的燕麦样本群体的元素以及所得样本的蛋白质组成进行了表征。通过尺寸排阻高效液相色谱（SE-HPLC）分析了总蛋白提取物的尺寸分布，而70%乙醇提取的蛋白质则通过反相高效液相色谱（RP-HPLC）进行分析。蛋白提取物在SE-HPLC柱上分离为三个主要的组分：聚合蛋白、燕麦蛋白（两者均根据其大小包含三个亚组）和可溶性蛋白，分别占总蛋白含量的68.79-86.60%、8.86-27.72%和2.89-11.85%。聚合蛋白与单体蛋白的比例在1.37至3.73之间变化。从整个群体的乙醇可提取蛋白中区分出了76个反相高效液相色谱分离峰。它们在不同品种间的分布差异显著，每个品种有6-23个峰。峰的出现次数也显示出很大差异：在107个样本中发现了一个峰，而有15个峰只在不到五个品种中出现。通过使用收集的RP-HPLC峰的质谱数据和生物信息学方法，开发了一种对样本燕麦蛋白表位含量进行排名的估计方法。使用R5抗体的酶联免疫吸附测定（ELISA）方法，发现大量被研究的样本受到小麦、大麦或黑麦的污染。
