Reference Number: 2
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 ionizationquadrupole- 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.
SIGNIFICANCE OF THIS STUDY
Celiac disease (CD) is an inflammatory disorder of the small intestine that affects genetically predisposed individuals when they ingest gluten from any Triticum species and similar proteins of barley and rye and their crossbred varieties. Following a gluten free diet (GFD) remains the only effective therapy for CD. Nevertheless, a GFD also has some drawbacks. It is expensive, gluten-free products have poor sensory and shelf life properties, and the diet is hard to follow and needs continuous monitoring by dieticians mainly due to occasional malnutrition problems. On the other hand, the development of wheat varieties that are free of toxic polypeptide sequences is considered unlikely because of the high degree of sequence homology existing among members of the cereal protein family and because cereals like wheat are hexaploid. The current paper explores the novel option of using sourdough lactobacilli to help eliminate the toxic effects of gluten. As a result, the flour contained <20ppm of gluten which can be considered gluten free according to the Codex Alimentarius Commission.
Although these results showed promising results in using sourdough lactobacilli in gluten degradation, these are highly controlled laboratory experiments and we do not encourage coeliac individuals to consume sourdough bread (store brought or home-made) without consulting a doctor.