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Enzyme hydrolyses of wheat gluten for the gluten free application
Svobodová, Dominika ; Vránová, Dana (referee) ; Ing.Silvia Mošovská,Ph.D. (advisor)
Celiac disease is attributable to autoimmune diseases, where treatment is based on absolute respect for gluten-free diet. As the gluten used widely in all areas of food production, strict adherence to a gluten-free diet really can be difficult for people with celiac disease. Products suitable also for those suffering from the disease are usually expensive. Finally, patients that undergo exclusively on a gluten-free diet are robbing the important nutritional components. Therefore, interest in this type of products innovation enhance as to the nutritional, technological, as well as organoleptic properties. The aim of this final thesis was to draw up literature review of the celiac disease and gluten allergen, or his particular part of gliadin. In the experimental part, attention was focused on the first hydrolytic cleavage of starch, the substrate was sterilized wheat flour and water, where is incorporated spiral just gluten. The best ability of degrading the starch grains, the combination of enzymes and MT3K and GLUAMK concentration of 0, 1% over 4 hours of action, as indicated by a reduction in the starch content of the original amount of 72,65 % at a concentration of 29,00 %. The next item of work distribution used different proteases, to reduce the amount of gliadin to below 20 ppm per kilogram of final product. Low levels investigated allergen (10,79 ppm/kg) recorded by the enzyme combination exoprotease Flavourzyme (0,1 %) and endoprotease Neutral Protease (0,1 %) at 7 hours hydrolytic action. The conclusion of the thesis was prepared several alternatives gluten-free breads and through sensory evaluation assessed the characteristics of color, aroma and taste to predict the location of the products on the commercial market.
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Changes of gluten proteins during beer processing
Porubiaková, Otília ; Mikulíková, Renata (referee) ; Márová, Ivana (advisor)
The aim of thesis was monitoring of changes in the content of gluten proteins in the biotechnological process of beer production. During the production process of wheat and barley beer, the samples were collected and analysed using the electrophoresis and immunoassay method. The results of the analyses were compared with commercial Czech beers. The theoretical part contains description and composition of gluten proteins, malt and beer technology, the changes that occur in this process, and methods of gluten proteins analysis. The experimental part contains procedures for laboratory production of barley and wheat beer and analyses of gluten proteins. To identify the individual gluteal protein fraction acid and SDS electrophoresis methods were used. For quantification, enzyme immunoassay was used and evaluated spectrophotometrically. The identification of the gluten‘s fractions by electrophoretic methods has been shown to be less specific for samples with lower content of gluten proteins and for barley specimens. A decrease in the concentration of gliadins and glutenins in the beer production process was demonstrated. A significant change was found during wort production with 98% decrease of gluten content compared to the feedstock and during the fermentation, when the gluten concentration dropped below 10 mg/kg. This value is acceptable from the legislation for products labelled „gluten-free“.
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Screening of probiotics bacterial strains aimed at proof of gluten degradation
Korvasová, Lucie ; Obruča, Stanislav (referee) ; Ryšávka, Petr (advisor)
No other treatment for celiac disease and other gluten-related disorders has been found so far except for a lifelong diet, which is significantly restrictive for the patient. Moreover, it is not always possible to adhere strictly to this diet, mainly due to random gluten ingestion, which leads to health complications associated with these diseases. Currently, vaccination is being tested as an alternative treatment, and there is also increasing attention paid to enzymes that could break down the immunogenic part of gluten, specifically in its gliadin component. This thesis examined bacteria that could potentially break down this sequence in gliadin and thereby reduce its concentration during detection. A considerable proportion of the tested bacteria belong to probiotics to ensure their future use for humans is safe. To obtain data, the fact was utilized that as bacteria grow, the turbidity increases proportionally, which was subsequently detected to determine the amount of bacterial growth. The RIDASCREEN® Gliadin competitive kit was used for the gliadin detection test, which works on the principle of a competitive ELISA method with R5 antibodies. The result of the study was the finding that only the bacteria Bacillus subtilis was able to break down gliadin among the examined bacteria. It was also measured how much gliadin the bacteria could break down at different concentrations, and according to the data, it was evaluated that it is appropriate to adapt the culture to the presence of gliadin for a greater amount of broken gliadin.
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The role of innate immunity cells in the pathogenesis of celiac disease
Dáňová, Klára ; Palová Jelínková, Lenka (advisor) ; Černý, Jan (referee)
Celiac disease is an autoimmune disease which occurs in susceptible individuals after ingestion of food containing gluten. Gluten and its monomeric fraction gliadin induce inflammatory damage of the small intestine by activating the immune cells that react strongly to gluten peptides. Gluten peptides have the ability to activate cells of adaptive as well as innate immune system. This work is focused on the production of interleukin (IL)-1 in antigen presenting cells stimulated with peptic gliadin digest. We found that monocytes and peripheral blood mononuclear cells (PBMC) isolated from blood of celiac patients secrete significantly more IL-1α and IL-1β than cells of healthy donors after stimulation with gliadin digest. The gliadin-induced IL-1β expression is controlled by a signaling cascade that includes MAPK kinase family molecules and transcription factor NF-κB. Moreover, we found that the adaptor proteins MyD88 and TRIF as well as Toll-like receptor (TLR) 2 and 4 play a role in the signaling cascade underlying gliadin-induced IL-1β expression by using murine bone marrow derived dendritic cells (BMDC). The precursor form of IL-1β in gliadin- stimulated PBMC and murine BMDC is maturated by caspase-1. In celiac PBMC the gliadin- induced maturation and secretion of IL-1β depends on the potassium...
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The possibilities to influence the response of innate immune cells to gliadin
Drašarová, Hana ; Tučková, Ludmila (advisor) ; Bezouška, Karel (referee)
Gluten sensitive entheropathy - celiac disease is a lifelong, genetically predisposed, immunologically mediated susceptibility to dietary wheat gluten, most frequently demonstrated by small-bowel damage and malabsorption syndrome. Strict adherence to gluten-free diet is the sole rational therapy of the disease. One of the possible therapeutic strategy for the treating of celiac disease is to utilize the synthetic polymer P(HEMA-co-SS). This polymer is capable specifically bound gliadin in gastrointestinal tract and by this way to neutralize the damaging effect of this alimentary protein on mucosa of small intestine in celiac patients. The in vitro study on human PBMC and specimens of small intestinal biopsies of celiac patients in our laboratory demonstrated that putative therapeutic ability of P(HEMA-co-SS) is substantially influenced by degree of proteolytic processing of gliadin and P(HEMA- co-SS) and also by different timing of per os administration of both components in organism. Another putative adjuvant therapy of celiac disease is employing of the beneficial probiotic bacterial strains. Our experiments were based on the findings of Prof. Y Sánz and her group demonstrating the significant differences in the composition of bacterial microflora in patients with active form of celiac disease,...
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Quality assessment of gluten-free foods with a focus on pastries
ZIMMELOVÁ, Helena
The topic of this thesis is the production of gluten-free bread from naturally gluten-free flour, which will be enriched with suitable additives that will increase antioxidant activity and naturally effect on the structure of the pastry, so it will meet the sensory requirements of final consumers. The literary part define, which proteins can be found in cereals, what does the gluten means, its function in the pastry and which diseases can cause. In connection with these diseases, a gluten-free diet for improving the patient's health and suitable foodstuff are briefly described. The literary part is concluded with the theory of sensory evaluation. The practical part deals with the production of naturally gluten-free breads enriched with ingredients from kitchen onions (Allium cepa L.) and subsequent sensory evaluation of this bread. Selected statistical procedures were used to obtain, processed and evaluated data. Antioxidant activity was also monitored and evaluated. By using texture-meter was find out, what effect has additives on aging of gluten-free bread.
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Changes of gluten proteins during beer processing
Porubiaková, Otília ; Mikulíková, Renata (referee) ; Márová, Ivana (advisor)
The aim of thesis was monitoring of changes in the content of gluten proteins in the biotechnological process of beer production. During the production process of wheat and barley beer, the samples were collected and analysed using the electrophoresis and immunoassay method. The results of the analyses were compared with commercial Czech beers. The theoretical part contains description and composition of gluten proteins, malt and beer technology, the changes that occur in this process, and methods of gluten proteins analysis. The experimental part contains procedures for laboratory production of barley and wheat beer and analyses of gluten proteins. To identify the individual gluteal protein fraction acid and SDS electrophoresis methods were used. For quantification, enzyme immunoassay was used and evaluated spectrophotometrically. The identification of the gluten‘s fractions by electrophoretic methods has been shown to be less specific for samples with lower content of gluten proteins and for barley specimens. A decrease in the concentration of gliadins and glutenins in the beer production process was demonstrated. A significant change was found during wort production with 98% decrease of gluten content compared to the feedstock and during the fermentation, when the gluten concentration dropped below 10 mg/kg. This value is acceptable from the legislation for products labelled „gluten-free“.
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Diagnostics of the enzymatic defects in patients with coeliac disease
Bartoňová, Simona ; Herink, Josef (advisor) ; Semecký, Vladimír (referee)
Celiac sprue is a chronic, lifelong disease characterized by permanent intolerance to gluten. Small intestinal mucosa of these patients shows typical inflammatory changes. These changes lead to reduced absorption of nutrients, minerals and vitamins that may result malabsorption syndrome. Diagnosis of celiac disease must be a comprehensive evaluation of the results of immunological testing of autoantibodies and microscopic evaluation of the intestinal mucosa state. The bachelor thesis deals with the histochemical examination of intestinal biopsies, which allows better distinguish malabsorption syndromes of different origin. The aim of the study was to determine incidence of celiac sprue and evaluate the completeness of remission in patients on a gluten - free diet in patients aged over 50 years in intestinal biopsies samples taken in 2013. Strict gluten - free diet leads to reparation of mucosa and brush border enzyme activity. Biopsies were snap frozen immediately after collection, to preserve the morphology of tissue and the brush border enzyme activity. Reactions, demonstrating the enzymatic activity of the lactase, trehalase and dipeptidylpeptidase IV, were exercided on sections by previously described histochemical methods. Alteration of relief was assessed by Marsh score. In 2013 was recieved...
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The role of innate immunity cells in the pathogenesis of celiac disease
Dáňová, Klára ; Palová Jelínková, Lenka (advisor) ; Černý, Jan (referee)
Celiac disease is an autoimmune disease which occurs in susceptible individuals after ingestion of food containing gluten. Gluten and its monomeric fraction gliadin induce inflammatory damage of the small intestine by activating the immune cells that react strongly to gluten peptides. Gluten peptides have the ability to activate cells of adaptive as well as innate immune system. This work is focused on the production of interleukin (IL)-1 in antigen presenting cells stimulated with peptic gliadin digest. We found that monocytes and peripheral blood mononuclear cells (PBMC) isolated from blood of celiac patients secrete significantly more IL-1α and IL-1β than cells of healthy donors after stimulation with gliadin digest. The gliadin-induced IL-1β expression is controlled by a signaling cascade that includes MAPK kinase family molecules and transcription factor NF-κB. Moreover, we found that the adaptor proteins MyD88 and TRIF as well as Toll-like receptor (TLR) 2 and 4 play a role in the signaling cascade underlying gliadin-induced IL-1β expression by using murine bone marrow derived dendritic cells (BMDC). The precursor form of IL-1β in gliadin- stimulated PBMC and murine BMDC is maturated by caspase-1. In celiac PBMC the gliadin- induced maturation and secretion of IL-1β depends on the potassium...
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The possibilities to influence the response of innate immune cells to gliadin
Drašarová, Hana ; Tučková, Ludmila (advisor) ; Bezouška, Karel (referee)
Gluten sensitive entheropathy - celiac disease is a lifelong, genetically predisposed, immunologically mediated susceptibility to dietary wheat gluten, most frequently demonstrated by small-bowel damage and malabsorption syndrome. Strict adherence to gluten-free diet is the sole rational therapy of the disease. One of the possible therapeutic strategy for the treating of celiac disease is to utilize the synthetic polymer P(HEMA-co-SS). This polymer is capable specifically bound gliadin in gastrointestinal tract and by this way to neutralize the damaging effect of this alimentary protein on mucosa of small intestine in celiac patients. The in vitro study on human PBMC and specimens of small intestinal biopsies of celiac patients in our laboratory demonstrated that putative therapeutic ability of P(HEMA-co-SS) is substantially influenced by degree of proteolytic processing of gliadin and P(HEMA- co-SS) and also by different timing of per os administration of both components in organism. Another putative adjuvant therapy of celiac disease is employing of the beneficial probiotic bacterial strains. Our experiments were based on the findings of Prof. Y Sánz and her group demonstrating the significant differences in the composition of bacterial microflora in patients with active form of celiac disease,...
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