†Istituto di Scienze dell'Alimentazione, Consiglio Nazionale delle Ricerche, Via Roma 64, I-83100 Avellino, Italy.
‡Istituto di Chimica Biomolecolare, Consiglio Nazionale delle Ricerche, Via Campi Flegrei 34, I-80078 Pozzuoli (Napoli), Italy.
J Agric Food Chem. 2015 Apr 8;63(13):3579-86. doi: 10.1021/acs.jafc.5b00631. Epub 2015 Mar 30.
Wheat beer is a traditional light-colored top-fermenting beer brewed with at least 50% malted (e.g., German Weissbier) or unmalted (e.g., Belgian Witbier) wheat (Triticum aestivum) as an adjunct to barley (Hordeum vulgare) malt. For the first time, we explored the proteome of three Weissbier samples, using both 2D electrophoresis (2DE)-based and 2DE-free strategies. Overall, 58 different gene products arising from barley, wheat, and yeast (Saccharomyces spp.) were identified in the protein fraction of a representative Weissbier sample analyzed in detail. Analogous to all-barley-malt beers (BMB), barley and wheat Z-type serpins and nonspecific lipid transfer proteins dominated the proteome of Weissbier. Several α-amylase/trypsin inhibitors also survived the harsh brewing conditions. During brewing, hundreds of peptides are released into beer. By liquid chromatography-electrospray tandem mass spectrometry (LC-ESI MS/MS) analysis, we characterized 167 peptides belonging to 44 proteins, including gliadins, hordeins, and high- and low-molecular-weight glutenin subunits. Because of the interference from the overabundant yeast-derived peptides, we identified only a limited number of epitopes potentially triggering celiac disease. However, Weissbier samples contained 374, 372, and 382 ppm gliadin-equivalent peptides, as determined with the competitive G12 ELISA, which is roughly 10-fold higher than a lager BMB (41 ppm), thereby confirming that Weissbier is unsuited for celiacs. Western blot analysis demonstrated that Weissbier also contained large-sized prolamins immunoresponsive to antigliadin IgA antibodies from the pooled sera of celiac patients (n = 4).
小麦啤酒是一种传统的浅色顶部发酵啤酒,用至少 50%的麦芽(例如德国小麦白啤酒)或未麦芽化的(例如比利时 Witbier)小麦(Triticum aestivum)作为大麦(Hordeum vulgare)麦芽的辅料酿造。我们首次使用基于 2D 电泳(2DE)和无 2DE 策略探索了三种小麦白啤酒样品的蛋白质组学。总体而言,在详细分析的代表性小麦白啤酒样品的蛋白质部分中鉴定出了来自大麦、小麦和酵母(Saccharomyces spp.)的 58 种不同的基因产物。类似于全麦芽啤酒(BMB),大麦和小麦 Z 型丝氨酸蛋白酶抑制剂和非特异性脂质转移蛋白在小麦白啤酒的蛋白质组中占主导地位。几种α-淀粉酶/胰蛋白酶抑制剂也在恶劣的酿造条件下存活下来。在酿造过程中,有数百种肽释放到啤酒中。通过液相色谱-电喷雾串联质谱(LC-ESI MS/MS)分析,我们鉴定了 167 种肽,这些肽属于 44 种蛋白质,包括麦醇溶蛋白、麦谷蛋白和高分子量和低分子量谷蛋白亚基。由于酵母衍生肽的大量存在的干扰,我们只鉴定出了少数可能引发乳糜泻的表位。然而,小麦白啤酒样品含有 374、372 和 382ppm 的麦醇溶蛋白等价肽,通过竞争 G12 ELISA 测定,这大致是拉格 BMB(41ppm)的 10 倍,从而证实小麦白啤酒不适合乳糜泻患者。Western blot 分析表明,小麦白啤酒还含有大分子量醇溶蛋白,对乳糜泻患者血清池中的抗麦醇溶蛋白 IgA 抗体有反应(n=4)。