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• W2013294405 startingPage "899" @default.
• W2013294405 abstract "OBJECTIVES: Immunogenic gluten proteins implicated in celiac disease (CD) largely resist degradation by human digestive enzymes. Here we pursued the isolation of gluten-degrading organisms from human feces, aiming at bacteria that would digest gluten under acidic conditions, as prevails in the stomach. METHODS: Bacteria with gluten-degrading activities were isolated using selective gluten agar plates at pH 4.0 and 7.0. Proteins in concentrated bacterial cell sonicates were separated by diethylaminoethanol chromatography. Enzyme activity was monitored with chromogenic substrates and gliadin zymography. Elimination of major immunogenic gluten epitopes was studied with R5 and G12 enzyme-linked immunosorbent assays. RESULTS: Gliadin-degrading enzyme activities were observed for 43 fecal isolates, displaying activities in the ˜150–200 and <50 kDa regions. The active strains were identified asPseudomonas aeruginosa. Gliadin degradation in gel was observed from pH 2.0 to 7.0. Liquid chromatography–electrospray ionization–tandem mass spectrometry analysis identified the enzyme as pseudolysin (lasB), a metalloprotease belonging to the thermolysin (M4) family proteases. Its electrophoretic mobility in SDS–polyacrylamide gel electrophoresis and gliadin zymogram gels was similar to that of a commercial lasB preparation, with tendency of oligomerization. Pseudolysin eliminated epitopes recognized by the R5 antibody, while those detected by the G12 antibody remained intact, despite destruction of the nearby major T-cell epitope QPQLPY. CONCLUSIONS: Pseudolysin was identified as an enzyme cleaving gluten effectively at extremely low as well as near-neutral pH values. The potential to degrade gluten during gastric transport opens possibilities for its application as a novel therapeutic agent for the treatment of CD." @default.
• W2013294405 created "2016-06-24" @default.
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• W2013294405 date "2015-06-01" @default.
• W2013294405 modified "2023-10-15" @default.
• W2013294405 title "Identification of Pseudolysin (lasB) as an Aciduric Gluten-Degrading Enzyme with High Therapeutic Potential for Celiac Disease" @default.
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• W2013294405 doi "https://doi.org/10.1038/ajg.2015.97" @default.
• W2013294405 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/4461489" @default.
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