• Journal of Microbiology and Biotechnology
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• 제7권4호
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• pp.262-266
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• 1997

To increase the high production of 1-deoxynojirimycin (DNJ) from Streptomyces sp. SID9135, the effect of various carbon sources, nitrogen sources, cationic metal ions, the initial pH of the medium, and agitation speed were investigated. The most effective carbon and nitrogen sources were found to be lactose 2.5% (w/v) and soybean meal 2.0% (w/v), respectively. None of the cationic metal ions examined had any detectable stimulating effect on DNJ production except $Fe^{+2}$ ion. The initial optimum pH for DNJ production ranged from 6-8 and agitation speed was most effective at 400 rpm. In the jar fermentor experiments under optimal culture conditions, the accumulation of DNJ reached about $640{\mu}g$/ml after 5 days of cultivation and the level remained the same for a further two days.

• 한국식품영양과학회지
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• 제37권11호
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• pp.1401-1407
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• 2008

본 연구는 $\alpha$-glucosidase의 강력한 효소저해제인 1-deoxynojirimycin(DNJ)을 고효율로 생산하는 균주의 선발 및 동정, 배양최적화 및 균 배양액의 혈당강하효과 등에 관하여 조사하였다. 먼저 토양으로부터 $\alpha$-glucosidase에 대해 강력한 저해능을 나타내는 9균주를 대상으로 이 중에서 DNJ 생산성이 가장 우수한 한 개 균주를 최종 선발하였고, 이 균주를 16S rDNA 염기서열분석으로 균주동정을 하여 Bacillus subtilis S10이라 명명하였다. 본 균주의 배양액을 이온교환수지법(Amberlyst 15 $H^+$ form, Dowex $1{\times}2$-100 $OH^-$ form, Amberlite CG-50 $NH_3^+$ form)에 의해 DNJ를 정제하고, 정제된 DNJ를 LC-MS로 분석한 결과 표준 DNJ와 동일한 물질임을 확인하였다. DNJ 대량생산을 위한 S10균주의 배양 최적탄소원 및 최적질소원과 이들의 최적농도를 조사한 결과 각각 1% galactose, 1.6% polypeptone임이 밝혀졌으며, 확립된 최적화 조건에서의 DNJ 생산량은 0.75 g/L이었다. 고혈당 유도 마우스를 대상으로 각종 혈당강하제의 효과를 알아보기 위해 혈당치를 비교한 결과 무처리구의 혈당치 $510{\pm}10.9\;mg/dL$에 비해 acarbose 처리구는 $139.4{\pm}33.1\;mg/dL$, 누에분말 처리구는 $209.1{\pm}19.6\;mg/dL$, 그리고 S10 균주 배양액 처리구는 $208.6{\pm}39.8\;mg/dL$로써 무처리구에 비해 각각 72.7%, 59.0%, 그리고 59.1%의 혈당 억제율을 나타냈다. 이상의 연구결과 S10균주 배양액은 누에분말과 같은 수준의 고혈당 억제 및 완화효과가 있음이 확인되었으며 향후 S10균주의 대량생산을 통해 식후혈당조절을 위한 건 강기능식품 개발이 기대된다 하겠다.

• International Journal of Industrial Entomology and Biomaterials
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• 제35권2호
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• pp.111-117
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• 2017

Mulberry leaves containing 1-deoxynojirimycin (DNJ) known to be a strong inhibitory effect for ${\alpha}-glucosidase$. Thus, DNJ has been recognized as a potentially important source for prevent or treat hyperglycemia. More effective method for the DNJ high-production is needed because DNJ content of natural mulberry leaf are as low as 0.1%. Many researchers have studied for the DNJ high-production in mulberry leaves such as the harvest season, fermentation using microorganisms, optimal culture conditions, and optimal extraction conditions. In order to provide for useful data that is anticipated at the level of industrial scale, we investigated ${\alpha}-glucosidase$ inhibitory activity, pH value and DNJ content in large-scale based on the optimal culture conditions for mulberry leaf fermentation of small-scale in our previous study. The ${\alpha}-glucosidase$ inhibitory activity, pH value, and DNJ content in this study were measured from the mulberry leaf fermentation broth for 7 days. During mulberry leaf fermentation, the ${\alpha}-glucosidase$ inhibitory activity and DNJ content was increased until 2 to 4 days, but after 4 day was decreased. The pH value showed a decreasing trend up to 2 day, and little changes in 2 to 4 days. However, the pH was started to increase after 4 days.

• Journal of Microbiology
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• 제45권5호
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• pp.431-440
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• 2007

A silkworm (Bombyx mori L.) extract known to contain naturally occurring iminosugars, including 1-deoxynojirimycin (1-DNJ) derived from the mulberry tree (Morus alba L.), was evaluated in surrogate HCV and HBV in vitro assays. Antiviral activity of the silkworm extract and one of its purified constituents, 1-DNJ, was demonstrated against bovine viral diarrhea virus (BVDV) and GB virus-B (GBV-B), both members of the Flaviviridae family, and against woodchuck hepatitis virus (WHV) and hepatitis B virus (HBV), both members of the Hepadnaviridae family of viruses. The silkworm extract exhibited a 1,300 fold greater antiviral effect against BVDV in comparison to purified 1-DNJ. Glycoprotein processing of BVDV envelope proteins was disrupted upon treatment with the naturally derived components. The glycosylation of the WHV envelope proteins was affected largely by treatment with the silkworm extract than with purified 1-DNJ as well. The mechanism of action for this therapy may lie in the generation of defective particles that are unable to initiate the next cycle of infection as demonstrated by inhibition of GBV-B in vitro. We postulate that the five constituent iminosugars present in the silkworm extract contribute, in a synergistic manner, toward the antiviral effects observed for the inhibition of intact maturation of hepatitis viral particles and may complement conventional therapies. These results indicate that pre-clinical testing of the natural silkworm extract with regards to the efficacy of treatment against viral hepatitis infections can be evaluated in the respective animal models, in preparation for clinical trials in humans.

• International Journal of Industrial Entomology and Biomaterials
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• 제34권2호
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• pp.38-44
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• 2017

Mulberry leaves containing 1-deoxynojirimycin (DNJ) have been recognized as a potentially important source for prevent or treat hyperglycemia. However, DNJ content of natural mulberry leaf are as low as 0.1%. Thus, the most effective method for increasing ${\alpha}$-glucosidase inhibitory activity with the DNJ high-production is needed. In this study, we investigated the influence of ${\alpha}$-glucosidase inhibitory activity according to different pH values (6-9) and inoculation amounts (0.1-0.5%) when Bacillus subtilis cultured on mulberry leaf powder media. We confirmed that ${\alpha}$-glucosidase inhibitory activity was difference according to culture conditions of different pH values, inoculation amounts, and fermentation times. The results of mulberry leaf fermentation according to pH values and inoculation amounts were shown that the optimal conditions for ${\alpha}$-glucosidase inhibitory activity were defined as pH 7 and 9, inoculation amount 0.4%, and incubation until 2 to 4 days. These results can be provided a basic data for the optimal culture conditions increasing ${\alpha}$-glucosidase inhibitory activity from mulberry leaf fermentation.

• Journal of Microbiology and Biotechnology
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• 제23권5호
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• pp.637-643
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• 2013

We have demonstrated that 1-deoxynojirimycin (DNJ) isolated from Bacillus subtilis MORI could enhance the levels of adiponectin and its receptors in differentiated 3T3-L1 adipocytes, which has been shown to be effective in lowering blood glucose levels and enhancing insulin sensitivity. DNJ was not toxic to differentiated 3T3-L1 adipocytes for up to a concentration of $5{\mu}M$. In terms of expression levels of adiponectin and its receptors (AdipoR1 and AdipoR2), DNJ in concentrations as low as $0.5{\mu}M$ elevated both mRNA and protein levels of adiponectin and transcript levels of AdipoR1 and AdipoR2. In addition, DNJ increased phosphorylation of 5' adenosine monophosphate-activated protein kinase (AMPK) in a statistically significant manner. Finally, treatment with DNJ resulted in increased mRNA expression of glucose transporter 4 (GLUT4), which encodes for a glucose transporter, along with a significant increase in glucose uptake into the adipocytes based on results of a 2-deoxy-D-[$^3H$] glucose uptake assay. Our findings indicate that DNJ may greatly facilitate glucose uptake into adipose tissues by increasing the action of adiponectin via its up-regulated expression as well as its receptor genes. In addition, the glucose-lowering effects of DNJ may be achieved by an increased abundance of GLUT4 protein in the plasma membrane, as a consequence of the increased transcript levels of the GLUT4 gene and the activation of AMPK.

• 한국잠사곤충학회지
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• 제48권1호
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• pp.25-27
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• 2006

누에장려품종 중 혈당강하제 개발에 적합한 품종 결정을 위하여 DNJ 함량을 정량한 결과, 봄에 사육한 금옥잠이 5.45 mg/gDW으로 DNJ 함량이 가장 높았다. 또한 누에 유전자원 원종 66품종의 DNJ 함량을 결정한 결과, 품종간 DNJ 함량 차이가 매우 큰 것으로 밝혀져, 혈당 강하제용 누에 육종시 기초자료가 될 수 있으며, 또한 DNJ 함량이 낮은 품종은 DNJ 축적 기작 연구에 많은 도움이 될 것으로 사료된다.

• International Journal of Industrial Entomology and Biomaterials
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• 제27권2호
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• pp.277-281
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• 2013

Silkworm powder, which contains 1-deoxynojirimycin (DNJ), is a promising complementary and alternative medicine (CAM) in Korea. Silkworm powder was produced from Yeonnokjam pupae at d 3 of the 5th instar at the National Academy of Agricultural Science. The powder was derivatized with 9-fluorenylmethyl chloroformate (FMOC-Cl), and the DNJ-FMOC content was measured by HPLC. We investigated the content of 1-DNJ in the silkworm powder and its glucose-lowering effect when it was treated at different temperatures. The content of 1-DNJ was the lowest at $150^{\circ}C$, while it was constant at other temperatures. The silkworm extract powder was orally administered to diabetic mice (20 mg/kg/d) for 4 wk. Water intake did not significantly change when compared with the control group (T0). The blood glucose levels significantly decreased when mice where administered silkworm powder treated at $60^{\circ}C$ (T60) compared to the control group, but no difference was observed between the groups T100 and T150. Moreover, the blood levels of TG significantly decreased compared with the control group. Based on these results, we surmise that the properties of the silkworm extract powder were stable upon heating at $100^{\circ}C$ but not at $150^{\circ}C$.

• Nutrition Research and Practice
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• 제12권6호
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• pp.469-478
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• 2018

BACKGROUND/OBJECTIVES: Mulberry leaf (ML) has been shown to have an inhibitory effect on ${\alpha}$-glucosidase, and suppresses postprandial hyperglycemia, which may be related to its deoxynojirimycin (DNJ) content. This study was conducted to investigate the hypoglycemic and dyslipidemic effects of rice coated with ML rich in DNJ in a type 2 diabetes mouse model. MATERIALS/METHODS: The mice were divided into four groups (n = 8 each): non-diabetic normal control (NC); diabetic control (DM-C), fed with 10% polished rice powder (DM-R); and fed with 10% polished rice powder coated with DNJ-rich ML (DM-DNJR). RESULTS: Supplementation with DNJR for six weeks decreased levels of fasting blood glucose, plasma insulin, triglyceride, total cholesterol, and blood glycosylated hemoglobin; conversely, levels of glucagon-like peptide-1 and high-density lipoprotein-cholesterol showed an increase in the same treatment. In addition, weights of mesenteric, epididymal, and total adipose tissues decreased with DNJR supplementation, when compared with diabetic control db/db mice, while maltase, lactase, and sucrase activity in the small intestine were inhibited. The anti-diabetic effects were marginally greater in the DM-DNJR group than in the DM-R group. CONCLUSIONS: These results suggest that rice coated with ML rich in DNJ can reduce hyperglycemia and hyperlipidemia in db/db mice, and may prove useful for individuals with diabetes.

• KSBB Journal
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• 제26권6호
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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.