title: Effect of barley endoprotease EP-B2 on gluten digestion in the intact rat.
abstract: Celiac Sprue is a multifactorial disease characterized by an intestinal inflammatory response to ingested gluten. Proteolytically resistant gluten peptides from wheat, rye, and barley persist in the intestinal lumen and elicit an immune response in genetically susceptible individuals. Here, we demonstrate the in vivo ability of a gluten-digesting protease ("glutenase") to accelerate the breakdown of a gluten-rich solid meal. The proenzyme form of endoprotease B, isoform 2 from Hordeum vulgare (EP-B2), was orally administered to adult rats with a solid meal containing 1 g of gluten. Gluten digestion in the stomach and small intestine was monitored as a function of enzyme dose and time by high-performance liquid chromatography and mass spectrometry. In the absence of supplementary EP-B2, gluten was solubilized and proteolyzed to a limited extent in the stomach and was hydrolyzed and assimilated mostly in the small intestine. In contrast, EP-B2 was remarkably effective at digesting gluten in the rat stomach in a dose- and time-dependent fashion. At a 1:25 EP-B2/gluten dose, the gastric concentration of the highly immunogenic 33-mer gliadin peptide was reduced by more than 50-fold within 90 min with no overt signs of toxicity. Evaluation of EP-B2 as an adjunct to diet control is therefore warranted in celiac patients.
authors.affiliation: Celiac Spruce Research Foundation, Palo Alto, California, USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Gass
authors.forename: Harmit
authors.initials: H
authors.lastname: Vora
authors.forename: Michael T
authors.initials: MT
authors.lastname: Bethune
authors.forename: Gary M
authors.initials: GM
authors.lastname: Gray
authors.forename: Chaitan
authors.initials: C
authors.lastname: Khosla
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003546:Cysteine Endopeptidases / Q000494:pharmacology*; D004063:Digestion; D004305:Dose-Response Relationship, Drug; D041981:Gastrointestinal Tract / Q000378:metabolism; D005983:Glutens / Q000378:metabolism*; D001467:Hordeum / Q000201:enzymology*; D008297:Male; D008969:Molecular Sequence Data; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results
chemical_list: D005983:Glutens; D003546:Cysteine Endopeptidases