PyroGlu-Leu is present in certain food protein hydrolysates and traditional Japanese fermented foods. Our previous study demonstrated that the oral administration of pyroGlu-Leu (0.1 mg/kg body weight) attenuates dysbiosis in mice with experimental colitis. The objective of this study was to elucidate why such a low dose of pyroGlu-Leu attenuates dysbiosis in different animal models. High fat diet extensively increased the ratio of Firmicutes/Bacteroidetes in feces of rats compared to control diet. Oral administration of pyroGlu-Leu (1 mg/kg body weight) significantly attenuated high fat diet-induced dysbiosis. By focusing on the production of intestinal antimicrobial peptides, we found that pyroGlu-Leu significantly increased the level of 4962 Da peptides, which identified as the propeptide of rattusin or defensin alpha 9, in ileum. We also observed increased tryptic fragment peptides from rattusin in the lumen. Here, we report that orally administered pyroGlu-Leu attenuates dysbiosis by increasing in the host antimicrobial peptide, rattusin.

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Proteome analysis of membrane proteins by two-dimentional electrophoresis (2-DE) is still insufficient due to the problem of membrane proteins solubilization. Additionally, nucleic acids and lipids are known to interfere with the profile on 2-DE, and removal of these substances is often required. In this study, three reagents from Bio-X Inc. were used to remove these interfering substances from protein. Prior to loading, samples were treated with the reagents for 30-60 min at 30°C. Pretreatment with the reagents led to high-resolution separations of proteins. Here, we describe how sample pretreatment using the reagents improved proteomic maps of proteins extracted from Escherichia coli and Saccharomyces cerevisiae. To remove lipids enhanced the solubility and separation of membrane proteins from cells.

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