Highly Efficient Gluten Degradation by Lactobacilli and Fungal Proteases during Food Processing: New Perspectives for Celiac Disease

Department of Plant Protection and Applied Microbiology, University of Bari, Bari, Italy.
Applied and Environmental Microbiology (Impact Factor: 3.67). 08/2007; 73(14):4499-507. DOI: 10.1128/AEM.00260-07
Source: PubMed


Presently, the only effective treatment for celiac disease is a life-long gluten-free diet. In this work, we used a new mixture of selected sourdough lactobacilli and fungal proteases to eliminate the toxicity of wheat flour during long-time fermentation. Immunological (R5 antibody-based sandwich and competitive enzyme-linked immunosorbent assay [ELISA] and R5 antibody-based Western blot), two-dimensional electrophoresis, and mass spectrometry (matrix-assisted laser desorption ionization-time of flight, strong-cation-exchange-liquid chromatography/capillary liquid chromatography-electrospray ionization-quadrupole-time of flight [SCX-LC/CapLC-ESI-Q-TOF], and high-pressure liquid chromatography-electrospray ionization-ion trap mass spectrometry) analyses were used to determine the gluten concentration. Assays based on the proliferation of peripheral blood mononuclear cells (PBMCs) and gamma interferon production by PBMCs and intestinal T-cell lines (iTCLs) from 12 celiac disease patients were used to determine the protein toxicity of the pepsin-trypsin digests from fermented wheat dough (sourdough). As determined by R5-based sandwich and competitive ELISAs, the residual concentration of gluten in sourdough was 12 ppm. Albumins, globulins, and gliadins were completely hydrolyzed, while ca. 20% of glutenins persisted. Low-molecular-weight epitopes were not detectable by SCX-LC/CapLC-ESI-Q-TOF mass spectrometry and R5-based Western blot analyses. The kinetics of the hydrolysis of the 33-mer by lactobacilli were highly efficient. All proteins extracted from sourdough activated PBMCs and induced gamma interferon production at levels comparable to the negative control. None of the iTCLs demonstrated immunoreactivity towards pepsin-trypsin digests. Bread making was standardized to show the suitability of the detoxified wheat flour. Food processing by selected sourdough lactobacilli and fungal proteases may be considered an efficient approach to eliminate gluten toxicity.

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Available from: Carlo Giuseppe Rizzello, Sep 30, 2015
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    • "Di Cagno and others (2006) found that certain sourdough cultures increased bread volume and crumb firmness, eliminating the need for baking texture additives. Furthermore, the sensory qualities of foods made with hydrolyzed-gluten wheat flour are often superior to products made from nonwheat flours (Rizzello and others 2007). Unfortunately, there is often a tradeoff between degradation of reactive gluten and retention of gluten for basic baking properties. "
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    ABSTRACT: The role of wheat, and particularly of gluten protein, in our diet has recently been scrutinized. This article provides a summary of the main pathologies related to wheat in the human body, including celiac disease, wheat allergy, nonceliac wheat sensitivity, fructose malabsorption, and irritable bowel syndrome. Differences in reactivity are discussed for ancient, heritage, and modern wheats. Due to large variability among species and genotypes, it might be feasible to select wheat varieties with lower amounts and fewer types of reactive prolamins and fructans. Einkorn is promising for producing fewer immunotoxic effects in a number of celiac research studies. Additionally, the impact of wheat processing methods on wheat sensitivity is reviewed. Research indicates that germination and fermentation technologies can effectively alter certain immunoreactive components. For individuals with wheat sensitivity, less-reactive wheat products can slow down disease development and improve quality of life. While research has not proven causation in the increase in wheat sensitivity over the last decades, modern wheat processing may have increased exposure to immunoreactive compounds. More research is necessary to understand the influence of modern wheat cultivars on epidemiological change.
    Comprehensive Reviews in Food Science and Food Safety 02/2015; 14(3). DOI:10.1111/1541-4337.12129 · 4.18 Impact Factor
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    • "Other approaches may consider the use of gliadinases during food processing to eliminate the gluten toxicity before consumption. This strategy has been adopted through the use of selected lactic acid bacteria alone (Gerez, Dallagnol, Rollan & Font de Valdez, 2012) or through the cooperative action of lactobacilli and fungal proteases during sourdough fermentations (Rizzello et al., 2007; De Angelis et al., 2010). However, only a handful of gliadinases have been described so far while more than fifty gliadins exist that account for tens of immunogenic peptides (Camarca, del Mastro & Gianfrani, 2012). "
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    ABSTRACT: For patients with celiac disease, gliadin detoxification, via the use of gliadinases, may provide an alternative to a gluten-free diet. A culture medium, in which gliadins were the sole source of nitrogen, was developed for screening for microorganisms with gliadinase activity. The problem of gliadin insolubility was solved by mild acid treatment, which renders an acid-hydrolysed gliadin/peptide mixture (AHG). This medium provided a sensitive and reliable means of detecting proteases, compared to the classical spectrophotometric method involving azocasein. When a sample of fermented wheat (a source of bacteria) was plated on an AHG-based culture medium, strains with gliadinase activity were isolated. These strains’ gliadinase profiles were determined using an AHG-based substrate in zymographic analyses.
    Food Chemistry 02/2015; 168:439-444. DOI:10.1016/j.foodchem.2014.07.085 · 3.39 Impact Factor
    • "Sourdough fermentation of wheat flour has some limitations such as the long fermentation period (24–72 hours) required for reducing the toxic peptides of gluten (Fig. 8.1). This kind of fermentation abolishes the techno-functional properties of gluten and, when it is fully hydrolysed, it is unsuitable for making baked goods (Rizzello et al. 2007). A biotechnology protocol was therefore used by Rizzello et al. (2014) for preparing bread with an intermediate content of gluten. "
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    ABSTRACT: In this chapter, we present the advances of enzymatic modification of gluten proteins and starches for gluten-free and low-glycaemic-index cereal foods. We also describe general information on wheat constituents which induce or trigger some diseases, their pathogenesis and the possible effects of these modified ingredients or foods.
    Advances in Food Biotechnology, First Edition edited by Ravishankar Rai V, 01/2015: chapter Enzymatic Modification of Proteins and Starches for Gluten-Free and Low-Glycaemic-Index Foods for Special Dietary Uses: pages 133-144; John Wiley & Sons, Ltd.., ISBN: 978-1-118-86455-5
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