MPIase is the first known glycolipid that is essential for membrane protein integration in the inner membrane of E. coli. Since the amount of natural MPIase available for analysis is limited and it contains structural heterogeneity, precisely designed synthetic derivatives are promising tools for further elucidation of its membrane protein integration mechanism. Thus, we synthesized the minimal unit of MPIase, a trisaccharyl pyrophospholipid termed mini-MPIase-3, and its derivatives. Integration assays revealed that the chemically synthesized trisaccharyl pyrophospholipid possesses significant activity, indicating that it includes the essential structure for membrane integration. Structure-activity relationship studies demonstrated that the number of trisaccharide units and the 6- O-acetyl group on N-acetylglucosamine contribute to efficient integration. Furthermore, anchoring in the membrane by a lipid moiety was essential for the integration. However, the addition of phosphorylated glycans devoid of the lipid moiety in the assay solution modulated the integration activity of MPIase embedded in liposomes, suggesting an interaction between phosphorylated glycans and substrate proteins in aqueous solutions. The prevention of protein aggregation required the 6- O-acetyl group on N-acetylglucosamine, a phosphate group at the reducing end of the glycan, and a long glycan chain. Taken together, we verified the mechanism of the initial step of the translocon-independent pathway in which a membrane protein is captured by a glycan of MPIase, which maintains its structure to be competent for integration, and then MPIase integrates it into the membrane by hydrophobic interactions with membrane lipids.

The e#ects on blood coagulation of dipicolinic acid (DPA, ,,0-pyridinedicarboxylic acid), an antibacterial substance known to be produced by Bacillus subtilis natto and contained in natto, a traditional Japanese fermented soybean food, were studied. It was found that addition of DPA with a final concentration of /῍+* ῌ-M results in substantial inhibition of platelet aggregation. DPA inhibition was found to be far stronger than that resulting from addition of aspirin. Furthermore, the clotting reaction of thrombinfibrinogen was also found to be inhibited by DPA. It was confirmed by examination of thromboelastogram patterns that the coagulation of whole rat blood was completely inhibited by addition of /῍+* ῌ-M DPA. From the point of view of the blood coagulation system, these results show that DPA contained in natto may be e#ective in the prevention of thrombosis.

Therapy with both statins improved lipid profiles and reduced proinflammatory responses; however, 2.5 mg of ROS have a potent LDL-C-lowering and hsCRP-lowering effect compared with 2 mg of PIT in patients with diabetes.

d -amino acids produced by Lactobacillus are thought to contribute to the taste quality and health functions; however, no studies have comprehensively evaluated the concentrations of the D- and L-forms of amino acids separately in individual Lactobacillus strains. To gain insight into amino acid concentrations in Lactobacillus , we evaluated amino acid concentrations in culture broth of Lactobacillus separately for the D- and L-forms. Lactobacillus strains were cultured in culture broth, and the amino acid concentrations in supernatant were assessed. The amino acid concentrations obtained by liquid chromatography-tandem mass spectrometry (LC-MS/MS) were subjected to cluster analysis based on Bray-Curtis distance with Ward's minimum variance method. In the analysis of amino acid concentrations under culture with different monosaccharides, the distances among strains cultured with the same monosaccharide were significantly greater than those among cultures of the same strain under different monosaccharides (p < 0.01). The cluster analysis of amino acid concentrations under culture with the same monosaccharide suggested that strains belonging to the same phylogenetic group of Lactobacillus exhibited similar concentrations of amino acids. Data analyses of 70 strains belonging to 17 Lactobacillus taxa indicated that the concentrations of amino acids were highly dependent on the phylogenetic group of Lactobacillus and that the group differences in amino acid concentration were strongly driven by differences in l -serine and d -alanine concentrations. Our results indicate that it is important to evaluate D- and l -amino acids separately when evaluating variations in amino acid concentrations. Because d -alanine has the potential to affect taste quality, the results of this study may provide insight into the taste quality of fermented food produced by Lactobacillus .

Abstracts: It was found that when dipicolinic acid is added to the culture solution of Bacillus subtilis natto, the area of fibrin dissolved by nattokinase (standard fibrin plate) and the amidase activity of nattokinase against Suc-Ala-Ala-Pro-PhepNA increased. For example, when manufacturing natto using steamed soybeans, the addition of 10-64 mM of dipicolinic acid increases amidase activity by more than 10 times.The concentration of vitamin K 2 (menaquinone-7) also increased by about 4 times with the addition of 10 mM of dipicolinic acid. No other food contains such a high concentration of vitamin K 2 . The results were the same for the shaking culture as well as for the stationary culture. If the concentration of dipicolinic acid is appropriately controlled, a product with excellent levels for both nattokinase activity and vitamin K 2 concentration could be manufactured.