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Obesity is a growing health problem that many countries face, mostly due to the consumption of a Westernized diet. In this present study we observed the effects of a soybean extract fermented by Bacillus subtilis MORI (BTD-1) containing 1-deoxynojirimycin against high fat diet-induced obesity. The results obtained from this study indicated that BTD-1 reduced body weight, regulated hepatic lipid content and adipose tissue, and also affected liver antioxidant enzymes and glucose metabolism. These results suggest that administration of BTD-1 affects obesity by inhibiting hyperglycemia and free radical-mediated stress; it also reduces lipid accumulation. Therefore, BTD-1 may be potentially useful for the prevention of obesity and its related secondary complications.

Obesity is a serious health problem which may continuously increase the morbidity and mortality associated with a variety of acute and chronic diseases.This study aimed to examine the antiobesity effect of soy bean extract fermented by Bacillus subtilis MORI (BTD-1) and to elucidate the mechanisms underlying such effects using 3T3-L1 preadipocytes. Lipid accumulations were significantly inhibited by BTD-1 treatment, which were accompanied by the decreased expression of CCAAT element binding protein α (C/EBPα) and the increased phosphorylation of acetyl-CoA carboxylase (ACC) protein expression. In addition, BTD-1 treatment increased the expression of glucose transporter 4 (GLUT4) along with significantly increasing glucose uptake into the adipocytes based on results of a 2-deoxy-D-[3H] glucose uptake assay. Our findings suggest that BTD-1 may suppress the differentiation of 3T3-L1 adipocytes and greatly facilitate glucose uptake into the adipocytes. Therefore, BTD-1 has a potential as a functional food ingredient in improving obesity and related metabolic disorders.

Organic acid (OA) profiling analysis was performed in culture media from Lactobacillus pentosus K34 (L. pentosus K34) and Pediococcus lolli PL24 (P. lolli PL24) by gas chromatography-mass spectrometry (GC-MS) following methoxime/ tert-butyldimethylsilyl derivatives. 12 OAs were positively identified in culture media. Most of OA levels from L. pentosus K34 of hetero lactic fermentation were found to be higher when compared with those from P. lolli PL24 of homo lactic fermentation, which may explain different OA metabolism in each strain. In addition, the distorted dodecagonal star patterns were readily dis- tinguishable, and the characteristics of each strain were well represented. The present study demonstrates that the OA metabolic profiling method by GC-MS combined with star pattern recognition is useful for the monitoring study of characteristic OA metabolism in various microorganisms.

The purpose of this study was to investigate the anti-hyperlipidemic effect of soy bean extract solution fermented by Bacillus subtilis MORI (BTD-1E) in obese db/db mice. Eight-week-old male db/db mice were administered 33.3 mg/kg BTD-1E solution orally once a day for four weeks. The BTD-1E group showed significantly lower body weight compared with the db control group (P

Department of Life science, The University of Suwon, Hwaseong 445-743, Republic of Korea(Received 09 June 2011; Accepted 30 August 2011) 1-Deoxynojirimycin (DNJ) is an alkaloid that is foundat relatively high concentrations in mulberry leaf andtissues of the silkworm, Bombyx mori. DNJ is a wellknown inhibitor of α-glucosidase, an enzyme that isinvolved in the early stages of the N-linked glycopro-tein synthesis pathway. α-Glucosidase activity in thecell extract from B. mori-derived Bm5 cells showedapproximately 40-fold less sensitivity to DNJ than α-glucosidase activity in the cell extract from Spodopterafrugiperda-derived Sf9 cells. The replication of B. morinucleopolyhedrovirus (BmNPV) was not inhibitedwhen it was propagated in BmN cells that were grownin medium containing up to 10 mM DNJ. In contrast,the replication of Autographa californica multiple NPV(AcMNPV) was reduced by 67% when it was propa-gated in Sf9 cells that were grown in medium contain-ing 10 mM DNJ. The viability of Bm5 and Sf9 cellsthat were grown in medium containing up to 10 mMDNJ was not affected. Our results suggested that thereduced replication of AcMNPV was the result of thehigher sensitivity of α-glucosidase activity in Sf9 cellsto DNJ.Key words: 1-Deoxynojirimycin, Bombyx mori nucleo-polyhedrovirus, BmNPV, Baculovirus, α-Glucosidaseinhibitor

An isolate of actinomycetes was obtained from farmland soil and designated as Streptomyces sp. A1022 based on cultural characteristics, 16S rDNA, DNA gyrase subunit B (gyrB), and fatty acid analyses. When strain A1022 was grown on chrome azurol S agar plate, it did not produce siderophore. However, strain A1022 had a strong antagonism and broad spectrum against six out of nine tested phytopathogens including Alternaria logipes, Cercospora canescece, Colletotrichum gloeosporioides, Diaporthe citri, Magnaporthe grisea, and Sclerotinia soleroforum. These results suggest that Streptomyces sp. A1022 could be employed as a potential biocontrol agent for the development of biopesticide having broad spectrum.

Dept. of Life Science, The University of Suwon, Gyeonggido 445-743, KoreaAbstractThis study was conducted to investigate the effect of administration of calcium supplementary food containing fermented product of Bacillus subtilis SE4 highly producing poly-γ-glutamic acid on the growth- parameters of adolescent male rats. Four-week old male Sprague-Dawley (SD) rats were fed for 4 weeks and assigned to the following 4 groups: two groups administered orally with new calcium supplementary food (such as 150 mg/kg and 300 mg/kg) containing fermented product of B. subtilis SE4, one group administered with conventional calcium supplementary food product (150 mg/kg) and one saline group as control. Daily weight gain and daily food intake in the two new food product groups were higher than those of conventional food product group and control group. Especially, the content of serum IGF-Ⅰ in the two new food product groups were significantly higher than those in conventional food product group and in control group (p

Dept of Life Science, The University of Suwon, Gyeonggido 445-743, Korea Abstract1-Deoxynojirimycin (DNJ) is a strong α-glucosidase inhibitor which inhibits hyperglycemia in animals. To select the Bacillus strains highly producing DNJ, 4,000 strains were isolated from soil and grain samples. By the inhibitory activity against α-glucosidase, nine Bacillus strains were selected and then identified by 16S rDNA sequencing. B. subtilis S10 was finally selected as the best strain for the production of DNJ. Various carbon sources and nitrogen sources in culture medium were evaluated for the highest production of DNJ. As the results, the optimized concentration of carbon source and nitrogen source was 1.0% galactose and 1.6% polypeptone and the concentration of DNJ produced was 0.75 g/L. The effect of culture supernatant of B. subtilis S10 on lowering blood glucose level was investigated in streptozotocin (STZ)-induced diabetic mice model. Mice were randomly assigned to control group (saline) and three test groups such as acarbose group, silkworm powder group and B. subtilis S10 group. After eight-week oral feeding, blood glucose levels of the B. subtilis S10 and silkworm powder groups were respectively 209.1±19.6 mg/dL (59.1%) and 208.6±39.8 mg/dL (59.0%) lower than 510±10 mg/dL of the control group. These results indicated that the culture supernatant of B. subtilis S10 was able to reduce the blood glucose level in STZ-induced diabetic mice.Key words: Bacillus subtilis S10, 1-deoxynojirimycin (DNJ), polyhydroxylated alkaloids, α-glucosidase inhibitor, hyperglycemic mice

Most of the available drugs for the treatment of diabetes mellitus (DM) produce detrimental side effects, which has prompted an ongoing search for plant with the antidiabetic potential. The present study investigated the effect of soybean extracts fermented with Bacillus subtilis MORI, fermented soybean extracts (BTD-1) was investigated in streptozotocin (STZ)-induced diabetic rats. The possible effects of BTD-1 against hyperglycemia and free radical-mediated oxidative stress was investigated by assaying the plasma glucose level and the activity of enzymatic antioxidants, such as superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), and malondialdehyde (MDA). A significant increase in the levels of both plasma glucose and reactive oxygen species (ROS) was observed in the diabetic rats when compared to normal control group. After administration of BTD-1 (500 and 1000 mg/kg/day), the elevated plasma glucose level was significantly reduced while the plasma insulin level and the activities of SOD, GSH-Px, CAT and MDA were significantly increased. The results suggest that administration of BTD-1 can inhibit hyperglycemia and free radical-mediated oxidative stress. The administration of BTD-1 also inhibited the contractile response by norepinephrine (10(-10)-10(-5) M) in the presence of endothelium, and caused significant relaxation by carbachol (10(-8)-10(-5) M) in rat aorta. These findings indicate that BTD-1 improves vascular functions on STZ-induced diabetic rats. Therefore, subchronic administration of BTD-1 could prevent the functional changes in vascular reactivity in STZ-induced diabetic rats. The collective findings support that administration of BTD-1 may prevent some diabetes-related changes in vascular reactivity directly and/or indirectly due to its hypoglycaemic effect and inhibition of production of ROS.

1-Deoxynojirimycin (DNJ), a D-glucose analogue with a nitrogen atom substituting for the ring oxygen, is a strong inhibitor of intestinal α-glucosidase. DNJ has several promising biological activities, including its antidiabetic, antitumor, and antiviral activities. Nevertheless, only limited amounts of DNJ are available because it can only be extracted from some higher plants, including the mulberry tree, or purified from the culture broth of several types of soil bacteria, such as Streptomyces sp. and Bacillus sp. In our previous study, a DNJ-producing bacterium, Bacillus subtilis MORI, was isolated from the traditional Korean fermented food Chungkookjang. In the present study, we report the identification of the DNJ biosynthetic genes in B. subtilis MORI 3K-85 strain, a DNJ-overproducing derivate of the B. subtilis MORI strain generated by γ-irradiation, xhe genomic DNA library of B. subtilis MORI 3K-85 was constructed in Escherichia coli, and clones showing α-glucosidase inhibition activity were selected. After DNA sequencing and a series of subcloning, we were able to identify a putative Operon which consists of gabT1, yktc1, and gutB1 genes predicted to encode putative transaminase, phosphatase, and oxidoreductase, respectively. When a recombinant plasmid containing this Operon sequence was transformed into an E. coli strain, the resulting transformant was able to produce DNJ into the culture medium. Our results indicate that the gabT1, yktc1, and gutB1 genes are involved in the DNJ biosynthetic pathway in B. subtilis MORI, suggesting the possibility of employing these genes to establish a large-scale microbial DNJ overproduction system through genetic engineering and process optimization.

Discovery of alpha-glucosidase inhibitors has been actively pursued with the aim to develop therapeutics for the treatment of diabetes and the other carbohydrate-mediated diseases. We have identified four novel alpha-glucosidase inhibitors by means of a drug design protocol involving the structure-based virtual screening under consideration of the effects of ligand solvation in the scoring function and in vitro enzyme assay. Because the newly identified inhibitors reveal in vivo antidiabetic activity as well as a significant potency with more than 70% inhibition of the catalytic activity of alpha-glucosidase at 50 microM, all of them seem to deserve further development to discover new drugs for diabetes. Structural features relevant to the interactions of the newly identified inhibitors with the active site residues of alpha-glucosidase are discussed in detail.

Discovery of alpha-glucosidase inhibitors has been actively pursued with the aim to develop therapeutics for the treatment of diabetes and the other carbohydrate mediated diseases. We have been able to identify 13 novel alpha-glucosidase inhibitors by means of a computer-aided drug design protocol involving homology modeling of the target protein and the virtual screening with docking simulations under consideration of the effects of ligand solvation in the binding free energy function. Because the newly discovered inhibitors are structurally diverse and reveal a significant potency with IC(50) values lower than 50 microM, all of them can be considered for further development by structure-activity relationship studies or de novo design methods. Structural features relevant to the interactions of the newly identified inhibitors with the active site residues of alpha-glucosidase are discussed in detail.