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Immunochemical evaluation of proteolysis of cereal proteins causing celiac disease by microbial proteases

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Celiac disease is one of the most prevalent food intolerances worldwide, with a gluten-free diet as the only effective therapy. Therefore, searching for new alternative approaches is a priority, and one of them is the idea of using proteinases for degradation of gluten proteins. The aim of the study was a trial of enzymatic cleavage of prolamins extracted from wheat, barley, rye, triticale and oat. The proteolysis was performed by microbial proteases from Bacillus stearothermophilus, Bacillus licheniformis, Bacillus thermoproteolyticus and Streptomyces griseus. The reaction was performed up to 60 min, stopped by addition of appropriate synthetic inhibitor and products of limited proteolysis were analyzed by SDS-PAGE. Most of the prolamins were susceptible to proteolysis by the examined microbial enzymes and as the result of hydrolysis the molecules of lower molecular weight and peptides occurred (<30,000 Da). The immunoreactivity of proteolysis products was evaluated using the reaction with polyclonal and monoclonal antibodies by Western blot and ELISA analysis, respectively. Immunochemical analyses confirmed that celiac-active sequences of amino acids in prolamins were at least partially destroyed and have lost their “toxicity”, so application of microbial proteases seems to be an alternative method for decreasing of celiac activity of prolamins.
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Keywords: Prolamins; celiac disease; immunoreactivity; proteolysis

Document Type: Research Article

Affiliations: 1: Małopolska Centre of Food Monitoring, Faculty of Food Technology, University of Agriculture in Kraków, Poland 2: Research Centre AgroBioTech, Slovak University of Agriculture in Nitra, Nitra, Slovakia 3: Department of Biochemistry and Biotechnology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Nitra, Slovakia

Publication date: November 1, 2016

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