• KSBB Journal
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• v.26 no.6
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• pp.517-522
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• 2011

In our previous study, we isolated and characterized a 1-deoxynojirimycin (DNJ)-producing bacterium, Bacillus subtilis MORI, from chungkookjang, a Korean traditional food. B. subtilis MORI was subjected to ${\gamma}$-irradiation and the resulting bacteria were screened for increased DNJ production. A mutant was identified that produced 7.6 times more DNJ and named B. subtilis MORI 3K-85. In this study, the protein profiles of both strains were compared by one-dimensional and two-dimensional gel electrophoresis (1-DE and 2-DE, respectively) under both native and denaturing conditions. The 1-DE native-PAGE and 1-DE SDS-PAGE analyses identified 5 and 7 bands, respectively, that were found at higher concentrations in B. subtilis MORI 3K-85 than in B. subtilis MORI. Similarly, 2-DE analyses identified 20 protein spots which were found at higher concentrations in B. subtilis MORI 3K-85. The peptide mass profiles of these 20 proteins were analyzed by MALDI-TOF and compared with peptide sequences of B. subtilis and B. amyloliquefaciens in the MASCOT database. This screening suggested that three dehydrogenases, an aldolase, a synthetase, an isomerase, a reductase, and a peroxidase are elevated in B. subtilis MORI 3K-85. Based on this data, one or more of the elevated 8 enzymes might be related to the DNJ biosynthetic pathway.

• Journal of Life Science
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• v.27 no.9
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• pp.1031-1039
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• 2017

The aim of this study was to investigate the potential effects of extracts from silkworm Bombyx mori L. fermented with Bacillus subtilis KACC 91157 at levels of 5%(v/w) and 10%(v/w) in Sprague-Dawley rats intoxicated with 1%(w/w) orotic acid (OA) for 10 days. The rats were divided into a normal group (N), a control group (C: OA), and treatment groups (SP10: OA + 10% extracts from B. mori L.; BSP5: OA + 5% extracts from B. mori L. fermented with B. subtilis KACC 91157; BSP10: OA + 10% extracts from B. mori L. fermented with B. subtilis KACC 91157). Serum activities of aspartate aminotransferase (AST), alanine transferase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) increased following OA feeding, but the rise was slightly reduced by administration of BSP10. The total lipid, free fatty acid, phospholipid, total cholesterol, and triglyceride contents in serum were significantly lower in the OA treatment groups than in the N group. However, the contents slightly increased following the administration of BSP10. Glutathione concentrations in liver and serum were reduced in the OA-induced fatty liver, but they increased following the administration of BSP10. Hepatocytes in the OA-induced fatty liver contained numerous large droplets. However, SP10, BSP5, and BSP10 feeding prevented OA-induced lipid droplet accumulation in hepatocytes. Accordingly, extracts from silkworm powder fermented with B. subtilis could be an ideal material as a dietary supplement in healthy functional foods to improve the effects of fatty liver.

• Microbiology and Biotechnology Letters
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• v.39 no.1
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• pp.49-55
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• 2011

Thirty Streptomyces sp. and 200 Bacillus sp. isolated from Korean soils and traditional foods were screened for their abilities to inhibit ${\alpha}$-glucosidase and produce 1-deoxynojirimycin (DNJ). This screening identified a Bacillus sp. bacterium that strongly inhibited ${\alpha}$-glucosidase and produced high levels of DNJ from Chungkookjang, a Korean traditional food. The bacterium was characterized in terms of its biochemical and molecular biological properties such as sugar utilization, cellular quinone composition, cell wall fatty acid composition, and 16S rDNA sequence. In addition, scanning electron microscopy was used to visualize the morphology of the bacterium. These analyses identified the bacterium as B. subtilis, a bacterium with Generally Recognized as Safe (GRAS) status. The selected strain was named B. subtilis MORI.

• Microbiology and Biotechnology Letters
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• v.49 no.4
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• pp.566-575
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• 2021

Galactose and soybean meal were selected as the best carbon and nitrogen sources, repectively, for the efficient production of α-glucosidase inhibitor (AGI) and 1-deoxynojirimycin (DNJ) by a newly isolated Bacillus subtilis MORI. The optimal concentrations of the galactose and soybean meal for the production of AGI and DNJ were investigated by response surface methodology. For the production of AGI, the optimal galactose and soybean meal concentrations were 4.3% (w/v) and 3.2% (w/v), respectively, and for DNJ, 4.5% (w/v) and 3.0% (w/v), respectively. The nearly identical optimal concentrations of galactose and soybean meal for the production of both AGI and DNJ indicated a close correlation between the production of AGI and DNJ. The maximum production of AGI (50,880 GIU/ml) and DNJ (824 ㎍/ml) obtained from the statistically optimized medium in a jar fermenter was 2.33 and 2.38-fold, respectively, higher than those (21,798 GIU/ml and 346 ㎍/ml, respectively) of the pre-optimized medium. The production of both AGI and DNJ was greatly increased by jar fermentation (AGI of 38,524 GIU/ml and DNJ of 491 ㎍/ml) compared with flask fermentation.

• Journal of Life Science
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• v.28 no.6
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• pp.697-707
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• 2018

The aim of this study was to investigate the potential effects of extracts from silkworm (Bombyx mori L.) that was fermented with Bacillus subtilis KACC 91157 at the levels of 1%(v/w), 2.5%(v/w), and 5%(v/w) in alcohol-fed rats. Sprague-Dawley rats were divided into seven groups: normal group (N), alcohol treated group (C), silymarin treated group (PC, positive control), 5% silkworm powder treated group (SP5), 1% Bacillus subtilis-fermented silkworm powder treated group (BSP1), 2.5% BSP treated group (BSP2.5), and 5% BSP treated group (BSP5). The activities of AST, ALT, ALP, and LDH in the serum and the triglyceride concentrations in the liver and serum were increased by alcohol feeding but were reduced in the BSP5 group. In addition, the contents of total lipids, free fatty acids, and total cholesterol were increased in the alcohol-fed group but were reduced in the BSP5 group. The activities of ADH, ALDH and ADH, ALDH protein levels in the liver were increased in the BSP5 group. The TBARS contents in the liver, serum, liver mitochondria, and liver microsomes were slightly decreased in the BSP5 group compared to the C group. The hepatocytes in alcohol-fed rats contained numerous large droplets; however, BSP5 treatment prevented alcohol-induced lipid droplet accumulation in the hepatocytes. Based on these results, extracts from Bacillus subtilis-fermented silkworm (Bombyx mori L.) have significant potential for development into a functional health food that can improve alcoholic fatty liver conditions.

• Journal of Life Science
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• v.27 no.12
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• pp.1470-1478
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• 2017

The aim of this study was to investigate biological activity and biochemical properties of extracts from Bacillus subtilis-fermented silkworm (Bombyx mori L., SP) powder of different origin (Buan, Namwon, and Boeun). An additional aim was to determine the inhibition of cancer cell (B16-F10, HT-29, LNcaP, and MCF-7) proliferation and nitric oxide (NO) production from lipopolysaccharide (LPS)-induced RAW264.7 cells. Biological activities (${\alpha},{\alpha}^{\prime}$-diphenyl-${\beta}$-picrylhydrazyl [DPPH], free radical scavenging activity, fibrinolytic activity, antiproliferation activity, and anti-inflammatory activity) and biochemical properties (compositional amino acid contents, and mineral contents) were examined in water extracts from silkworm powder and B. subtilis-fermented silkworm powder. The highest amino acid contents were detected in Buan silkworm powder (BU). After fermented, the highest contents were found in B. subtilis-fermented Buan silkworm powder (BBO). The major minerals detected were K, Ca, and Mg. Rates of these minerals, especially those of Na increased after fermented. DPPH radical scavenging activity and fibrinolytic activity were stronger in the fermented group than non-fermented group. DPPH radical scavenging activity and fibrinolytic activity were highest in the extract from BBO. The inhibition activities of LNcaP and MCF-7 cells viability were significantly decreased in the BBO, and there was no inhibition activity in other cancer cells (B16-F10 and HT-29). An SRB assay of the cell viability of RAW 264.7 cells exposed to extracts of silkworm powder and B. subtilis-fermented silkworm powder revealed no toxicity in any of the groups. Compared with the LPS-treated group, the biggest reduction in NO production was detected in the BBO group. Based on these results, extracts from Boeun silkworm powder fermented with B. subtilis could be a candidate material as a dietary supplement for use in healthy functional foods.

• Journal of Life Science
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• v.23 no.6
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• pp.789-795
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• 2013

Current study was to investigate the potential effects of silkworm, Bombyx mori L. fermented with Bacillus subtilis (BFSP) and Aspergillus kawachii (AFSP) at the 5% (w/w) levels in Sprague-Dawley rats, which was intoxicated with 1% (w/w) orotic acid (OA) for 10 days. The rat group administered silkworm powder showed improvements in fatty liver condition. Consumption of fermented silkworm powder reduced triglyceride concentrations in the liver tissues and serum and increased the serum lipid concentrations to normal levels, thereby aiding in improving fatty liver conditions. These effects were more pronounced in the BFSP than that in SP or AFSP in orotic acid-induced hepatotoxicity and oxidative stress. Based on these results, fermented silkworms are considered to be a material with significant potential for development into a functional health food that can improve fatty liver conditions.

• Applied Biological Chemistry
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• v.35 no.5
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• pp.361-366
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• 1992

The antagonistic bacteria, showing distinguished effect against Fusarium oxysporum and Rhizoctonia solani were isolated from the rhizosphares of horticultural plants and identified as Bacillus subtilis. The strains were studied for their chracteristics of biochemistry, physiology, antagonistic effect against plant pathogenic fungi, and growth promoting effect on horticultural plants. The Bacillus thuringiensis(BT) HD-1 toxin gene was introduced into these B. subtilis. The BT toxin genes on chromosome of the bacteria were identified by southern blotting, but its proteins were not detected by SDS-PAGE. These transformed bacteria showed growth promoting effect and showed also insecticidal and antagonistic effects against Bombix mori and fungi F. oxysporum and R. solani but not against nematode Bursaphelenchus xylophilus.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.4
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• pp.452-458
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• 2008

The effect of fermented soybean culture of Bacillus subtilis MORI mixed with mulberry leaves on the lowering the levels of blood glucose and cholesterol was examined using neonatal streptozotocin-induced diabetic (n-STZ) rats. B. subtilis MORI produces 1-deoxynojirimycin (DNJ), an $\alpha$-glucosidase inhibitor. The content of DNJ of soybean fermented culture mixed with mulberry was higher (4.1$\pm$0.0 mg/g dry base) than that (1.5$\pm$0.0 mg/g) of mulberry. The concentration of 50% inhibition (IC50) against rat intestinal $\alpha$-glucosidase of soybean fermented culture mixed with mulberry was $5.6{\pm}0.1{\mu}g$/mL and that of mulberry was $17.0{\pm}0.5{\mu}g$/mL. Experimental groups of diabetic rats were randomly assigned to normal control group (NC group), diabetic control group (DC group) and three diabetic groups fed with DNJ food product. One of the three diabetic groups was M group (60 mg DNJ food product/kg) (DNJ food product containing 30% mulberry) and the other two were MM-60 group (60 mg/kg) and MM-120 group (120 mg/kg) (DNJ-fortified food product containing 30% fermented soybean culture mixed with mulberry leaves). The glucose in serum was significantly decreased in the MM-60 and MM-120 groups fed with DNJ-fortified food product for 4 weeks, compared with DC group. Total cholesterol and triglyceride in serum were also lower in MM-60 and MM-120 groups than the DC group. These results support that the fermented soybean culture of B. subtilis MORI mixed with mulberry leaves improved the metabolism of blood glucose and lipid in the n-STZ rat model.

• Korean Journal of Food Science and Technology
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• v.50 no.3
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• pp.297-307
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• 2018

The aim of this study was to develop a seasoning paste fermented by Aspergillus oryzae and Bacillus subtilis using three edible insects: Tenebrio molitor larvae, Gryllus bimaculatus, and Bombyx mori pupa. No significant changes in pH and titratable acidity were observed between the insect pastes and soybean control during a 5-week fermentation process. The amino nitrogen was 0.35-0.50% (w/w) in the early stage of the fermentation and increased to 0.72-0.78%. The total nitrogen was 2.36-3.62% (w/w) in the early stage and was preserved during the fermentation period. Free amino acids of the insect pastes were similar to those in soybean paste. In general, the fermented insect pastes showed a lower flavor and color than soybean paste but B. mori pupa-fermented paste did not show any significant difference in preference from soybean. Glutamate, aspartate, lysine, phenylalanine, arginine, and serine were found to affect taste preference. It was possible to ferment edible insect protein to produce a fermented seasoning paste like Korean doenjang.