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首页> 美国卫生研究院文献>Nutrients >Prototype Gluten-Free Breads from Processed Durum Wheat: Use of Monovarietal Flours and Implications for Gluten Detoxification Strategies
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Prototype Gluten-Free Breads from Processed Durum Wheat: Use of Monovarietal Flours and Implications for Gluten Detoxification Strategies

机译：来自加工硬质小麦的原型无麸质面包：使用单血抗面粉和对麸质解毒策略的影响

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摘要

In this investigation, we reported the production of prototype breads from the processed flours of three specific Triticum turgidum wheat genotypes that were selected in our previous investigation for their potential low toxic/immunogenic activity for celiac disease (CD) patients. The flours were subjected to sourdough fermentation with a mixture of selected Lactobacillus strains, and in presence of fungal endoproteases. The breads were characterized by R5 competitive enzyme linked immunosorbent assay in order to quantify the residual gluten, and the differential efficacy in gluten degradation was assessed. In particular, two of them were classified as gluten-free (<20 ppm) and very low-gluten content (<100 ppm) breads, respectively, whereas the third monovarietal prototype retained a gluten content that was well above the safety threshold prescribed for direct consumption by CD patients. In order to investigate such a genotype-dependent efficiency of the detoxification method applied, an advanced proteomic characterization by high-resolution tandem mass spectrometry was performed. Notably, to the best of our knowledge, this is the first proteomic investigation which benefitted, for protein identification, from the full sequencing of the Triticum turgidum ssp. durum genome. The differences of the proteins’ primary structures affecting their susceptibility to hydrolysis were investigated. As a confirmation of the previous immunoassay-based results, two out of the three breads made with the processed flours presented an exhaustive degradation of the epitopic sequences that are relevant for CD immune stimulatory activity. The list of the detected epitopes was analyzed and critically discussed in light of their susceptibility to the detoxification strategy applied. Finally, in-vitro experiments of human gastroduodenal digestion were carried out in order to assess, in-silico, the toxicity risk of the prototype breads under investigation for direct consumption by CD patients. This approach allowed us to confirm the total degradation of the epitopic sequences upon gastro-duodenal digestion.
机译：在这次调查中，我们报告了在我们之前选择的三种特异性Triticum Turgidum小麦基因型的原型面包的生产，这些面粉在我们之前选择的腹泻（CD）患者的潜在低毒性/免疫原性活性。将面粉与选定的乳杆菌菌株的混合物进行酵母发酵，并在真菌内载酶存在下。通过R 5竞争酶连接的免疫吸附测定表征面包，以定量残留的麸质，评估麸质降解中的差异疗效。特别地，其中两种分别被分类为无麸质（<20ppm）和非常低谷蛋白含量（<100ppm）面包，而第三种单血管原型保留了谷蛋白含量，其远高于规定的安全阈值CD患者直接消费。为了研究所应用的解毒方法的这种基因型依赖性效率，通过高分辨率串联质谱法进行先进的蛋白质组学表征。值得注意的是，尽我们所知，这是从Turgidum SSP的全部测序中获得蛋白质鉴定的第一个受益的蛋白质组学调查。杜兰姆基因组。研究了影响它们对水解敏感性的蛋白质的主要结构的差异。作为先前的免疫测定结果的确认，用加工面粉制成的三种面包中的两个出来呈现了与CD免疫刺激活性相关的表位序列的详尽降解。分析了检测到的表位的清单，并根据易受应用的解毒策略的敏感性讨论。最后，进行了人胃胃胃病消化的体外实验，以便在硅中进行评估，在CD患者直接消费的调查下进行原型面包的毒性风险。这种方法使我们能够在胃十二指肠消化时确认表位序列的总劣化。

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