Di Cagno Raffaella, De Angelis Maria, Auricchio Salvatore, Greco Luigi, Clarke Charmaine, De Vincenzi Massimo, Giovannini Claudio, D'Archivio Massimo, Landolfo Francesca, Parrilli Giampaolo, Minervini Fabio, Arendt Elke, Gobbetti Marco
Department of Plant Protection and Applied Microbiology, University of Bari, 70126 Bari, Italy.
Appl Environ Microbiol. 2004 Feb;70(2):1088-96. doi: 10.1128/AEM.70.2.1088-1096.2004.
This work was aimed at producing a sourdough bread that is tolerated by celiac sprue (CS) patients. Selected sourdough lactobacilli had specialized peptidases capable of hydrolyzing Pro-rich peptides, including the 33-mer peptide, the most potent inducer of gut-derived human T-cell lines in CS patients. This epitope, the most important in CS, was hydrolyzed completely after treatment with cells and their cytoplasmic extracts (CE). A sourdough made from a mixture of wheat (30%) and nontoxic oat, millet, and buckwheat flours was started with lactobacilli. After 24 h of fermentation, wheat gliadins and low-molecular-mass, alcohol-soluble polypeptides were hydrolyzed almost totally. Proteins were extracted from sourdough and used to produce a peptic-tryptic digest for in vitro agglutination tests on K 562(S) subclone cells of human origin. The minimal agglutinating activity was ca. 250 times higher than that of doughs chemically acidified or started with baker's yeast. Two types of bread, containing ca. 2 g of gluten, were produced with baker's yeast or lactobacilli and CE and used for an in vivo double-blind acute challenge of CS patients. Thirteen of the 17 patients showed a marked alteration of intestinal permeability after ingestion of baker's yeast bread. When fed the sourdough bread, the same 13 patients had values for excreted rhamnose and lactulose that did not differ significantly from the baseline values. The other 4 of the 17 CS patients did not respond to gluten after ingesting the baker's yeast or sourdough bread. These results showed that a bread biotechnology that uses selected lactobacilli, nontoxic flours, and a long fermentation time is a novel tool for decreasing the level of gluten intolerance in humans.
这项研究旨在制作一种乳糜泻(CS)患者能够耐受的酸面团面包。筛选出的酸面团乳酸菌具有特殊的肽酶,能够水解富含脯氨酸的肽,包括33肽,这是CS患者肠道来源的人T细胞系最有效的诱导剂。这个在CS中最重要的表位,在用细胞及其细胞质提取物(CE)处理后被完全水解。用乳酸菌启动由小麦(30%)与无毒燕麦、小米和荞麦粉混合制成的酸面团。发酵24小时后,小麦醇溶蛋白和低分子量醇溶性多肽几乎被完全水解。从酸面团中提取蛋白质,并用于制备胃蛋白酶-胰蛋白酶消化物,以对人源K 562(S)亚克隆细胞进行体外凝集试验。最小凝集活性比化学酸化面团或用面包酵母启动的面团高约250倍。用面包酵母或乳酸菌及CE制作了两种含约2克麸质的面包,并用于CS患者的体内双盲急性激发试验。17名患者中有13名在摄入面包酵母面包后肠道通透性出现明显改变。当食用酸面团面包时,同样的13名患者排出的鼠李糖和乳果糖值与基线值无显著差异。17名CS患者中的另外4名在摄入面包酵母或酸面团面包后对麸质无反应。这些结果表明,一种使用筛选出的乳酸菌、无毒面粉和长时间发酵的面包生物技术是降低人类麸质不耐受水平的一种新工具。