• 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.

• 한국식품과학회지
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• 제52권2호
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• pp.191-199
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• 2020

본 연구에서는 발효식품으로부터 항당뇨 효능을 지닌 유산균을 분리하여 뽕잎 추출물을 발효하고, 제조된 유산균 발효 뽕잎 추출물의 항당뇨 효능을 평가하였다. 갓김치에서 분리된 Lactobacillus plantarum SG-053은 α-glucosidase 저해 활성이 96.8%로 가장 우수하고, 뽕잎을 발효하여 뽕잎에 존재하는 항당뇨 지표물질인 1-deoxynojirimycin (DNJ)의 함량을 2.2배 증가시키며 우수한 생육 활성을 나타내어 본 연구에서 뽕잎 추출액의 발효에 사용하였다. L. plantarum SG-053으로 발효한 뽕잎 추출물은 L6 근관세포에 대한 세포독성은 없었고, IRS-1, PI3K p85α, GLUT-4 유전자 발현을 각각 1.4, 2.2, 1.4배로 증가시켰으며, 세포의 2-deoxyglucose 흡수를 1 μM의 인슐린보다 높은 수준인 40.7% 증가시켜 인슐린 신호전달 경로를 활성화시킴으로써 세포의 인슐린 저항성을 개선하는 것으로 확인되었다. 경구 당 부하 검사를 통해 유산균 발효 뽕잎 추출물이 포도당 섭취에 의해 증가한 혈당을 빠르게 감소시키고, α-glucosidase의 저해를 통해 maltose의 분해에 의한 혈당 증가를 억제하였다. 유산균 발효 뽕잎 추출물은 SD rat의 허벅지 골격근 조직의 PI3K p85α와 GLUT-4 유전자 발현을 각각 6.4, 2.1배 증가시켜 L6 근관세포에서와 유사한 결과를 나타냈으며, in vivo 근육 조직에서도 인슐린 신호전달경로의 활성화에 영향을 준다는 것을 확인하였다. 본 연구 결과로 L. plantarum SG-053이 뽕잎을 효과적으로 발효하여 DNJ의 함량을 증진시킨다는 것을 확인하였으며, L. plantarum SG-053으로 발효한 뽕잎 추출물은 우수한 항당뇨 효능을 지니는 것을 세포 및 동물실험 수준에서 확인하였다.

• 한국잠사학회:학술대회논문집
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• 한국잠사학회 2001년도 제44회 춘계 학술연구 발표자료
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• pp.32-32
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• 2001

• International Journal of Industrial Entomology and Biomaterials
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• 제7권2호
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• pp.161-164
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• 2003

Several polyhydroxylated alkaloids were isolated from the extracts of freeze-dried silkworm powder, and purified by ion exchange chromatographic analysis. Through the HPLC analysis, we could identify 1-deoxynojirimycin (DNJ) and a kind of calystegin $B_2$ (HS-58) as well as a noble compound (HS-74) from the purified polyhydroxylated alkaloids. In order to know the characteristics of these isolated alkaloids as enzyme inhibitors, glycosidase inhibition activities of these identified alkaloids including other two non-purified alkaloids (SWP 3-1 and SWP 3-2) were investigated.

• 한국식품저장유통학회지
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• 제18권2호
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• pp.244-249
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• 2011

꾸지뽕잎의 물 및 70% 에탄올 추출물에 대한 세포의 독성 및 지방생성에 미치는 영향과 DNJ 함량과 rutin 함량을 측정하였다. Cell viability test에서 꾸지뽕잎 추출물 0.5-5 mg/mL의 농도일 때 물 및 에탄올 추출물 모두 세포의 생장에 영향을 미치지 않는 것으로 나타나 세포에 대한 독성이 없다고 판단되었다. 동일한 농도로 3T3-L1 지방세포의 지방축적에 미치는 영향은 물 및 에탄올 추출물 모두 농도 의존적으로 지방축적을 저해하는 것으로 나타났으며 추출물 농도가 5mg/mL 일 때, 물 추출물은 86%, 에탄올 추출물 50%의 지방축적을 보였다. DNJ 함량은 꾸지뽕잎의 에탄올 추출물에서 $3921.3{\mu}g/g$로 높았고 rutin의 함량은 꾸지뽕잎의 물 추출물에서 $4592.9{\mu}g/g$로 높게 나타냈다. 따라서 꾸지뽕잎 추출물에는 기능성 성분인 DNJ 함량과 rutin 함량이 다량 함유되어 있고 지방의 축적을 저해하는 효과가 있어 비만과 대사관련 질환개선을 위한 기능성 소재로 활용될 수 있다고 생각한다.

• 한국잠사학회:학술대회논문집
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• 한국잠사학회 2003년도 제46회 춘계 학술연구 발표회
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• pp.53-53
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• 2003

Alpha-glucosidase inhibitor, 1-deoxynojirimycin, has been shown to eliminate the production of ER-budding viruses in the ER of host cell. This glucose-derived iminosugar derivative, 1-DNJ, has been largely purified from midgut of silkworm, Bombyx mori, Here, we have screened the antiviral activity in the GBV-B infected marmoset hepatocyte cell line. (omitted)

• International Journal of Industrial Entomology and Biomaterials
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• 제27권2호
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• pp.237-242
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• 2013

We investigated the 1-deoxynojirimycin (1-DNJ) content of extracts from silkworm larvae at each developmental stage within three silkworm varieties. We also compared the content of the following polyhydroxylated alkaloids in the silkworm extracts: 1-DNJ, fagomine, and 1,4-dideoxy-1,4-imino-d-arabinitol (DAB). In addition, we evaluated the glucose-lowering effects of silkworm extract powder in db/db mice. The 1-DNJ content was the highest in Yeonnokjam $5^{th}$ instar $3^{rd}$ d larvae and Hansaengjam $5^{th}$ instar $3^{rd}$ d larvae, which contained 18.4 mg/100 g dry weight and 18.3 mg/100 g dry weight, respectively. The $5^{th}$ instar $3^{rd}$ d larvae exhibited a higher content of 1-DNJ than that of $5^{th}$ instar $5^{th}$ d larvae among all varieties. The glucose-lowering effects of silkworm extracts and Yeonnokjam powder were tested on db/db mice, and the blood glucose levels were found to decrease significantly in the YR70 group. Silkworm extracts (180 mg/kg, 90 mg/kg, 45 mg/kg, and 22.5 mg/kg) and acarbose (50 mg/kg) were administered orally for 4 wk. Changes in water intake were not statistically significant between control and silkworm extract-treated groups. Compared to the control group, blood glucose levels in the silkworm extract powder-treated group decreased in the 22.5 mg/kg/d group after being administered for 4 wk. This decrease was statistically significant. Furthermore, biochemical changes in the AST(Aspartate aminotransferase), ALT(Alanine aminotransferase), TCHO(Total Cholesterol), TG(Triglyceride), LDL(Low density lipoprotein), and HDL(High density lipoprotein) levels in blood were not observed. However, statistically significant decreases in blood GLU in the 22.5 mg/kg/d group compared to that of the control group occurred. In addition, the epididymal fat weight of the silkworm extract powder-treated group decreased significantly in both the 22.5 mg/kg/d group and 180 mg/kg/d group compared to that of the control group, but there were no statistically significant changes in perirenal fat weight. These results demonstrate that silkworm extracts inhibit changes in blood glucose levels in model diabetic mice.

• 한국식품영양과학회지
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• 제32권1호
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• pp.15-21
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• 2003

본 연구는 뽕잎을 이용한 고부가가치의 기능성식품의 제조에 적합한 뽕잎 품종을 선별하고자 품종별(YK-209, Cheong il, Yongchon and Kaeryang), 산지별(영천 및 상주), 부위별(상위, 중위 및 하위엽) 및 생육시기별(5월, 6월 및 8월)에 따른 뽕잎의 기능성 지표물질인 DNJ, GABA및 flavonoid의 함량을 HPLC로 정량하였다. 4가지 뽕잎 중 YK-209 뽕잎의 기 능성 물질 (DNJ, GABA 및 flavonoid) 함량이 가장 높았다. 그리고 YK-209 뽕잎으로부터 분리된 4가지 flavonoid 중 rutin 함량이 가장 높았으며, 그 다음으로 isoquercitrin>kaempferol-3-Ο-rutinoside＞astragalin 순으로 나타났으나 그 외 3가지 뽕잎에서는isoquercitrin의 함량이 가장 높았고 그 다음으로 rutin이 차지 하였으나 품종별, 산지별, 부위별 및 성장시기별에 따라 flavonoid구성성분의 함량이 다소 차이가 있었다. 영천지역에서 수확한 YK-209뽕잎의 3가지 기능성물질의 함량이 상주에서 수확한 뽕잎보다 그 함량이 높았다. 한편, YK-209 뽕잎의 부위 중 상위엽이 중위 및 하위엽보다 3가지 기능성물질의 함량이 높았다. 끝으로YK-209뽕잎의 성장시기 중 5월에 수확한 것이 6월 및 8월에 수확한 뽕잎보다 기능성물질의 함량이 높았다. 이러한 연구결과를 미루어 볼 때 3가지 기능성 성분함량이 높은 YK-209 뽕잎(용천뽕과 개량뽕을 교배하여 얻은 것)의 5월산 상위엽을 향후 뽕잎의 이용한 기능성식품의 제조용 원료로 활용할 수 있을 것으로 기대된다.

• International Journal of Industrial Entomology and Biomaterials
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• 제33권1호
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• pp.31-35
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• 2016

The influence of commonly used steeping times and temperatures about mulberry tea on a-glucosidase inhibitory activity were studied. The effects of product and preparation variables on the in-cup chemical composition of mulberry tea extracts is of interest because the appearance and taste characteristics and the possible health effects of a tea liquor arise from the chemical components extracted from the leaf during mulberry tea preparation. A comprehensive study was therefore undertaken to determine the contributions of product and provide a basic data for development of high quality mulberry tea products against diabetes. Specific eight mulberry tea were collected from each region of Korea (A-I) and when it was extracted on 85℃ for 2min, a-glucosidase activity was best for 98-102%. According to various temperature and time extraction methods, this study was carried out to optimize teamaking conditions for maximal DNJ extraction from Korean mulberry tea and and it would be of particular interest for people who drink mulberry tea to control blood glucose levels.

• International Journal of Industrial Entomology and Biomaterials
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• 제21권2호
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• pp.163-167
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• 2010

[ $\alpha$ ]Glucosidase (EC 3.2.1.20) is a glycosidase that hydrolyzes disaccharides, oligosaccharides, and polysaccharides resulting in the release of α-D-glucose. In this study, $\alpha$-glucosidase activity in the hemolymph and midgut of the mulberry silkworm Bombyx mori and Japanese oak silkmoth Antheraea yamamai was measured using maltose, sucrose, trehalose, and p-nitrophenyl $\alpha$-D-glucopyranoside as substrates. In general, hemolymph $\alpha$-glucosidase activity was higher in B. mori than in A. yamamai. In contrast, midgut $\alpha$-glucosidase activity was higher in A. yamamai than in B. mori for all of the substrates tested. $\alpha$-Glucosidase activity in the midgut of both B. mori and A. yamamai showed similar responses to changes in pH and temperature for all of the substrates tested. Native (7.5%) PAGE of hemolymph and midgut proteins from B. mori and A. yamamai followed by staining with 4-methylumbelliferyl $\alpha$-D-glucoside (MUG) indicated that the $\alpha$-glucosidases of these related lepidopterans are functionally similar but structurally different. In comparison to $\alpha$-glucosidase activity from A. yamamai, $\alpha$-glucosidase activity from B. mori was generally less sensitive to the $\alpha$-glucosidase inhibitors, 1-deoxynojirimycin (DNJ), acarbose, and voglibose when the activity was determined using maltose, sucrose, and trehalose.