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Identification of a Hepatoprotective Peptide in Wheat Gluten Hydrolysate against d-Galactosamine-Induced Acute Hepatitis in Rats

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Division of Applied Life Sciences, Kyoto Prefectural University, Shimogamo, Kyoto 606 8522, Japan
Graduate School of Horticulture, Chiba University, 648 Matsudo, Matsudo-Shi, Chiba 271 8510, Japan
§ Graduate School of Science and Engineering, University of Toyama, 3190 Gofuku, Toyama 930 8555, Japan
Faculty of Education and Welfare Science, Oita University, 700 Dannohara, Oita 870 1192, Japan
Health Care Research Center, R&D Division, Nisshin Pharma Inc., 5-3-1 Tsurugaoka, Fujimino-city, Saitama 356 8511, Japan
Graduate School of Health and Sports Science, Juntendo University, 1-1 Hiraga-Gakuenndai, Inzai, Chiba 276 1695, Japan
*Tel/Fax: 81 75 723 3503. E-mail: [email protected]
Cite this: J. Agric. Food Chem. 2013, 61, 26, 6304–6310
Publication Date (Web):June 7, 2013
https://doi.org/10.1021/jf400914e
Copyright © 2013 American Chemical Society

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    Abstract

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    A hepatoprotective peptide, pyroglutamyl leucine (pyroGlu-Leu), was identified in wheat gluten hydrolysate through an in vivo activity-guided fractionation approach based on d-galactosamine-induced acute hepatitis in rats and fractionation of peptides with large-scale preparative ampholine-free isoelectric focusing. The active acidic fraction predominantly consisted of pyroglutamyl peptides and free pyroglutamic acid. Pyroglutamyl peptides were derivatized with phenyl isothiocyanate after removal of a pyroglutamyl residue by pyroglutamate aminopeptidase. The derivatives were purified by reversed-phase HPLC and subjected to sequence analysis. The active fraction contained pyroGlu-Ile, pyroGlu-Leu, pyroGlu-Gln, pyroGlu-Gln-Gln, and free pyroGlu. Ingestion of pyroGlu-Leu at 20 mg/kg body weight significantly decreased serum aspartate and alanine aminotransferases to approximately 30% and 20% of those values of the vehicle group, respectively, which were near the normal levels. Thirty minutes after ingestion of pyroGlu-Leu at 20 mg/kg, the concentration of pyroGlu-Leu in portal blood plasma increased to approximately 2 μM.

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