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Mulberry leaves have been used as the sole food for silkworms in sericulture, and also as a traditional medicine for diabetes prevention. Mulberry leaf components, for example 1-deoxynojirimycin (1-DNJ), inhibit the activity of -glucosidase and prevent increased blood glucose levels, and they are highly toxic to caterpillars other than silkworms. The -glucosidase inhibitory activity of mulberry leaves changes with the season, but it is unknown which environmental conditions influence the -glucosidase inhibitory activity. We investigated in this study the relationship between the -glucosidase inhibitory activity and environmental conditions of temperature and photoperiod. The results demonstrate that low temperatures induced decreasing -glucosidase inhibitory activity, while the induction of newly grown shoots by the scission of branches induced increasing -glucosidase inhibitory activity. These results suggest that the -glucosidase inhibitory activity was related to the defense mechanism of mulberry plants against insect herbivores. Key words:-glucosidase inhibitor; 1-deoxynojirimycine; mulberry; resistance to insect herbivore Mulberry leaves have been historically used as feed for silkworms and consumed as a beverage or health food by humans. Mulberry leaf components inhibit the activity of a kind of digestive enzyme for sugar absorption, -glucosidase, and promote slow absorption of glucose into the blood vessels. [1][2][3] The most important substance in mulberry leaves, 1-deoxynojirimycin (1-DNJ), is known to be a strong intestinal -glucosidase inhibitor. Mulberry leaves contain 1-DNJ, 4) so that mulberry leaf products have been commercialized as health foods.In contrast, mulberry leaves are highly toxic to caterpillars other than the silkworm, Bombyx mori, because they contain 1-DNJ and other -glucosidase inhibitors that affect common insects. 5,6) The silkworm has evolved adaptive enzymes to circumvent the toxic effects of -glucosidase inhibitors and made it able to feed on mulberry leaves. 6,7) 1-DNJ and the -glucosidase inhibitor from mulberry leaves are therefore used to both improve the symptoms of diabetes mellitus, and to avoid caterpillars by inhibiting growth.Mulberry plants are distributed throughout the world, the number of mulberry cultivars in Japan alone being over 1,000. These cultivars match the climatic characteristics in various areas. The typical cultivars of mulberry plants are maintained and administered in Japan at the National Institute of Agrobiological Sciences (Tsukuba, Ibaraki, Japan) and the Experimental Farm of Textile Science and Technology of Shinshu University (Ueda, Nagano, Japan). It has been reported that the -glucosidase inhibitory activity and 1-DNJ concentration of mulberry leaves differ among various cultivars and seasons, [8][9][10][11] but it is unknown which environmental conditions influence the -glucosidase inhibitory activity. We therefore investigated in this study the relationship between the environmental conditions and -glucosidase inhibitory activity. Materia...

1-Deoxynojirimycin (DNJ) is a potent α-glucosidase inhibitor and thus beneficial for prevention of diabetes. While we have succeeded in obtaining the culture supernatant extract (CSE) rich in DNJ from microorganism source, information regarding its anti-hyperglycemic effect and safety were still limited. Therefore, this study was aimed to evaluate the anti-hyperglycemic effect and safety of microorganism DNJ. Oral sucrose tolerance test was performed, and the result showed that CSE was able to significantly suppress the blood glucose elevation and suggested DNJ as the main active compound. To determine its safety, the absorption and excretion of microorganism DNJ were evaluated using 15N labeling method. Our findings investigated the recovery rate of 15N from DNJ reached 80% up to 48 hours after oral administration, suggesting its rapid excretion, suggesting the safety of DNJ. This study verified the functional properties and safety of DNJ from microorganisms, suggesting its potential use for functional purpose.

1-Deoxynojirimycin (DNJ)is an effective sugar analogue that suppresses the elevation of postprandial blood glucose. Therefore, food products containing DNJ are expected to be promising functional foods in preventing the onset of diabetes. To examine the absorption kinetics of DNJ, we prepared 15 N-labeled DNJ from the culture medium of a Bacillus amyloliquefaciens DSM7 strain. In the process, we found that lactose markedly increased the production of DNJ at a carbon/nitrogen ratio of 6.25:1, reaching 1140 mg/L with 5 days of cultivation. Comparisons between lactose and glucose on cell growth and DNJ production suggested that lactose made the DSM7 strain grow more slowly and maintained DNJ production in the long term. These findings may be useful for efficient microbial DNJ production or in the development of DNJ-containing fermented food products.

The medaka (Oryzias latipes) egg envelope (chorion) is composed of three major glycoproteins, Zona Interna (ZI)-1, -2, and -3, that originate in the spawning female liver as the precursor proteins Choriogenin (Chg.)H, Chg.Hm, and Chg.L, respectively. These ZI and Chg. proteins contain a structural ZP protein domain that is conserved among the egg envelope proteins of all animals. While ovarian expression of ZP proteins (e.g., ZPCs and ZPB) has been reported in medakas, the functions of these proteins remain unknown. Thus, the present study aimed to determine whether the ovary-expressed medaka ZP protein, mZPC5, is involved in forming the chorion matrix.The mZPC5 gene (mzpc5) was expressed in the ovaries but not the livers of mature female medakas, as shown by reverse transcription-polymerase chain reaction assays with mzpc5-specific primers. In situ hybridization analysis revealed that ovarian mzpc5 expression was restricted to the ooplasm of early (stage I–III) previtellogenic oocytes, and its expression signal weakened with oocyte growth. Following sodium-dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting analysis with anti-mZPC5 antibodies, two immunoreactive proteins were detected in the ovary and chorion extracts. These proteins were approximately 50 and 74 kDa in size, like ZI-3 and ZI-2, respectively.Immunohistochemical assays using anti-mZPC5 and anti-Chg.H antibodies localized the mZPC5 protein in the ooplasm of early previtellogenic oocytes. With oocyte growth, mZPC5 tended to accumulate in the chorion, co-localizing with Chg.H.We previously showed that ovary-expressed ZP proteins could not compensate for Chg.L function loss in gene knock-out (chg.l -/-) medakas. As in our previous study, the chg.l-/- females produced oocytes with thin chorions, resulting in infertile soft eggs. However, in the present study, mZPC5 and Chg.H were co-localized in the chg.l-/- chorions. These results suggested that in the medaka previtellogenic oocyte, 1) mZPC5 is secreted from the ooplasm and deposited on the outer surface of its plasma membrane, creating the thin chorion layer; and 2) following the accumulation of liver-derived Chgs., the 3D structure of the chorion matrix is formed cooperatively with mZPC5 and Chgs. during oogenesis. More research is needed to confirm the functions of mZPC5 in chorion structure and physiology.

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