• Title/Summary/Keyword: 1-Deoxynojirimycin (1-DNJ)

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Analysis by GC of 1-Deoxynojirimycin in Leaves and Teas of Cudrania tricuspidata Bureau (꾸지뽕잎 및 꾸지뽕 잎차에 함유된 1-deoxynojirimycin의 GC 분석)

  • Lee, Jin-Shik;Lee, Sung-En
    • Journal of the Korean Applied Science and Technology
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    • v.27 no.3
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    • pp.337-343
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    • 2010
  • 1-Deoxynojirimycin(1-DNJ) was hardly detected by general UV detector. Therefore, in this study, we analyzed 1-DNJ, a effective functional material in which was contained the leaves of Cudrania tricuspidata and its tea by gas-chromatography. Synthesized a TMS derivative by attaching trimethylsilyl group to 1-DNJ, analyzed this by GC, and could detect a good peak. The leaves of Cudrania tricuspidata contains $1154.83{\pm}56.67$ ug/g.d.w of 1-DNJ and tea of Cudrania tricuspidata leaves does $8.01{\pm}0.61$ ug/g.d.w. This means the contents of 1-DNJ was nearly identical to each other. The larger size of the leaves had the more contents of 1-DNJ and the middle region of collection was the highest than any other collective regions of leaves.

Quantitative Analysis of 1-Deoxynojirimycin Content Using Silkworm Genetic Resources

  • Ju, Wan-Taek;Kim, Kee-Young;Sung, Gyoo-Byung;Kim, Yong-Soon
    • International Journal of Industrial Entomology and Biomaterials
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    • v.29 no.2
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    • pp.162-168
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    • 2014
  • 1-Deoxynojirimycin(1-DNJ or DNJ), a component in silkworm powder, prevents glucose from being absorbed into the bloodstream from the small intestine by inhibiting ${\alpha}$-glucosidase activity. This study compared the functional components of 1-DNJ from various silkworm species using a gene database with those of 1-DNJ produced by silkworms bred through cross-combinations. We utilized comparisons of geographical origins and species of silkworms using a gene database and discovered that 1-DNJ activity was ranked in the following order by species, Japanese (SK-1) > Chinese (C48) > European (Rock191). 1-DNJ constituted varying percentages of silkworm organs in the following order, blood > epithelial tissue > body fat > silk glands. With regard to sex, 1-DNJ, levels were higher in males than females. However, 1-DNJ levels with respect to various genetic traits (blood color, silk color, and egg color) were consistent. In order to study 1-DNJ changes that occurred during cross breeding of the silkworm gene, we bred cross-combinations utilizing SK-1 and C48 silkworms. In conclusion, in order to provide information about the constituents of functional materials contained in silkworm powder, it is imperative that silkworm cross breeding occurs so that the database of functional materials extracted from silkworms will expand.

Screening of 1-deoxynojirimycin (DNJ) producing bacteria using mulberry leaf

  • Ju, Wan-Taek;Kim, Hyun-Bok;Kim, Kee-Young;Sung, Gyoo-Byung;Kim, Yong-Soon
    • International Journal of Industrial Entomology and Biomaterials
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    • v.31 no.2
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    • pp.48-55
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    • 2015
  • 1-Deoxynojirimycin (DNJ) has been extensively investigated for its applications as an a-glucosidase inhibitor in postprandial hyperglycemia, and has been applied to nutraceuticals and medicines to prevent or delay the progression of type 2 diabetes. However, the amount of DNJ in mulberry leaves is low (approximately 0.1%), therefore, a more effective extraction method is needed. In this study, microbial DNJ production was developed as an alternative to chemical methods. We identified fermented sericultural products and bacteria that produce DNJ in large quantities using high performance liquid chromatography and thin layer chromatography. The inhibition of a-glucosidase activity was examined with respect to DNJ production or non-production. Crude DNJ from the isolated strains exhibited greater than 70% a-glucosidase activity. An investigation of the effect of mulberry leaf powder concentration (1~5%), using high DNJ producing bacteria, provided evidence for microbial mass production of DNJ.

Development and Validation of a Unique HPLC-ELSD Method for Analysis of 1-Deoxynojirimycin Derived from Silkworms (누에에 함유된 1-Deoxynojirimycin의 분석을 위한 HPLC-ELSD 분석법 밸리데이션)

  • Hyejin Cho;Sullim Lee;Myoung-Sook Shin;Joohwan Lee;Sanghyun Lee
    • Korean Journal of Pharmacognosy
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    • v.54 no.1
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    • pp.38-43
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    • 2023
  • A simple and accurate assay was developed for the quantitative analysis of 1-deoxynojirimycin (1-DNJ) derived from the silkworm (Bombyx mori). Normal-phase high-performance liquid chromatography coupled with an evaporative light scattering detector (HPLC-ELSD) and a hydrophilic interaction liquid chromatography column was used. Various parameters were applied to optimize the analysis method. The limits of detection and quantification of 1-DNJ were 2.97 × 10-3 and 9.00 × 10-3 mg/mL, respectively. The calibration curve showed good linearity results. The concentration range and the r2 value were 0.0625-1.0 mg/mL and 0.9997, respectively. The accuracy test demonstrated a significantly high recovery rate (89.95-103.22%). The relative standard deviation was ≤ 1.00%. Thus, a method for the accurate identification and quantitative analysis of 1-DNJ in silkworms was developed. Moreover, in this procedure, the process of derivatization of 1-DNJ, which was required in previous experiments, could be eliminated. This technique may be actively utilized for the development of pharmaceuticals and health functional foods using 1-DNJ.

Production of α-Glucosidase Inhibitor and 1-Deoxynojirimycin by Bacillus subtilis MORI

  • Park, Young Shik;Lee, Jae Yeon;Hwang, Kyo Yeol;Kim, Keun
    • 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.

Optimization of culture conditions of Bacillus subtilis with α-glucosidase inhibitory activity

  • Kim, Yong-Soon;Ju, Wan-Taek;Kim, Hyun-Bok;Sung, Gyoo-Byung
    • International Journal of Industrial Entomology and Biomaterials
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    • v.33 no.1
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    • pp.24-30
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    • 2016
  • 1-Deoxynojirimycin (DNJ) have been extensively investigated for their α-glucosidase inhibitor on postprandial hyperglycemia, and applied in nutraceuticals and medicine for preventing or delaying progression of type 2 diabetes. However, the amount of DNJ in mulberry leaves is low (about 0.1%), therefore, more effective extraction method is needed. This study was performed to develop microbial DNJ for biological methods of DNJ as an alternative to the chemical methods. In this study, we obtained evidence for Bacillus subtilis that produce DNJ in large quantities by high performance liquid chromatography. Inhibition of α-glucosidase activity was determined to DNJ production or non-production. Investigation of the effect of mulberry leaves powder concentration (1~5%), using the DNJ high-production bacteria, provided evidence for microbial mass production of DNJ. When the 4% mulberry leaf powder for 9 days was used, the α-glucosidase inhibitory activity was over the 85%. Also, the results presented in this study confirm DNJ yield's increasement in microbes using the various of nutrients and provide insight of ways to improve DNJ yields in microorganisms.

Regional and Varietal Variation of 1-Deoxynojirimycin (DNJ) Content in the Mulberry Leaves

  • Kim, Iksoo;Kim, Jin-Won;Lee, Heui-Sam;Ha, Nam-Kyu;Ryu, Kang-Sun
    • International Journal of Industrial Entomology and Biomaterials
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    • v.2 no.2
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    • pp.141-147
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    • 2001
  • This study was carried out to find out whether there exists any content variation in 1-deoxynojirimycin (DNJ) among regional and varietal mulberry leaves. HPLC analysis was performed for 22 samples collected from several localities in Korea and Tsushima Islands Japan. The highest content was observed in the Cheongilppong sample collected at Youngchun City, Kyungsangbuk Province (1,882.7 $\mu\textrm{g}$/g DW), whereas lowest content was observed in the Cheongilppong sample collected at Jiniu City, Kyungsangnam Province (395.65 $\mu\textrm{g}$/g DW). In the comparison of regional samples of Kaeryangppong variety, DNJ content was highest in the order of Woniu City (1,460.1) > Boungun (999.85) > Hweongsung-gun (939.12) > Cheongju City (688.78). In case of Cheongilppong, DNJ content was highest in the order of Youngchun City (1.882.8) > Sanchung-gun (1,125.3) > Sunchon City (649.17) > Byunsan-myon (441.54) > Jinju City (395.65). Average content of regional samples was higher in Kaeryangppong (1,021.7 $\mu\textrm{g}$/g DW) than that of Cheongilppong (898.87 $\mu\textrm{g}$/g DW). Average DNJ content of the wild mulberry loaves collected from Cheju Island and Tsushima Island (1,012.9$\mu\textrm{g}$/g DW) was high enough to reach to that of the regional Kaeryangppong samples. In the comparison among varietal samples collected in Suwon, the highest DNJ content was observed in YK209 (768.16 $\mu\textrm{g}$/g DW), amongst overall low DNJ content of the varietal samples. Considering a few pieces of information from our data, DNJ content appears not to be influenced solely by annual temperature of the planted areas.

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Quantitative Analysis of 1-deoxynojirimycin in Mulberry Leaves (뽕나무 유전자원의 1-deoxynojirimycin 함량 변이)

  • Kim, Hyun-Bok;Seo, Sang-Deok;Koo, Hui-Yeon;Seok, Young-Seek;Kim, Sun-Lim;Sung, Gyoo-Byung
    • Journal of Sericultural and Entomological Science
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    • v.51 no.1
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    • pp.20-29
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    • 2013
  • Countries of the world are competing for the collection and utilization of genetic resources, which is a resource of value-added industrialization. We are building database of silkworm and mulberry resources with morphological, genetic characteristics data etc. Moreover, recent bioactive compound has been receiving increasing attention. So we analyzed 1-deoxynojirimycin(1-DNJ) content as a this item in addition to the basic information using 363 strains(varieties) of mulberry genetic resources. They were grown under the same environment and conditions. Mulberry leaves were collected and then freeze-dried and powdered for 1-DNJ test. As a result, 1-DNJ mean content of 363 mulberry strains was $0.176{\pm}0.077%$, and the coefficient of variation (CV) was 43.5%. The variation between the strains was greatly severe. Among the tested strains, 'C1D89/29' was showed the highest content of 0.47%, whereas 'Pumbo 24' and 'Turkey E' were showed the lowest content of 0.05% respectively. The content of 1-DNJ of 16 mulberry varieties for silkworm rearing were compared. Average concentration was $0.17{\pm}0.04%$, and the coefficient of variation (CV) was 22.8%. Variation among cultivars was not significant. Finally we selected 1-DNJ high-containing 11 strains. They are as follows. 'C1D89/29', 'Hiihak', 'Jeokasibmunja', 'Gweonchil', 'Botongsibmunja', 'Jeokchuk', 'bulguksang', 'Geunsookgojo', 'Busanggeum', 'Guksang 20' and 'Taekwang' which are more than twice of the overall average content.

Isolation and Identification of a Bacillus sp. producing ${\alpha}$-glucosidase Inhibitor 1-deoxynojirimycin (알파글루코시다아제 저해제 1-deoxynojirimycin을 생산하는 Bacillus 균주의 분리 및 동정)

  • Kim, Hyun-Su;Lee, Jae-Yeon;Hwang, Kyo-Yeol;Cho, Yong-Seok;Park, Young-Shik;Kang, Kyung-Don;Seong, Su-Il
    • 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.

Effect of 1-deoxynojirimycin on the Replication of Baculoviruses, Bombyx Mori Nucleopolyhedrovirus and Autographa Californica Multiple Nucleopolyhedrovirus

  • Kang, Kyung-Don;Park, Joo-Sung;Cho, Yong-Seok;Park, Young-Shik;Lee, Jae-Yeon;Hwang, Kyo-Yeol;Yuk, Won-Jeong;Kamita, Shizuo George;Suzuki, Koichi;Seong, Su-Il
    • International Journal of Industrial Entomology and Biomaterials
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    • v.23 no.1
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    • pp.123-128
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    • 2011
  • 1-Deoxynojirimycin (DNJ) is an alkaloid that is found at relatively high concentrations in mulberry leaf and tissues of the silkworm, $Bombyx$ $mori$. DNJ is a well known inhibitor of ${\alpha}$-glucosidase, an enzyme that is involved in the early stages of the $N$-linked glycoprotein synthesis pathway. ${\alpha}$-Glucosidase activity in the cell extract from $B.$ $mori$-derived Bm5 cells showed approximately 40-fold less sensitivity to DNJ than ${\alpha}$-glucosidase activity in the cell extract from $Spodoptera$ $frugiperda$-derived Sf9 cells. The replication of $B.$ $mori$ nucleopolyhedrovirus (BmNPV) was not inhibited when it was propagated in BmN cells that were grown in medium containing up to 10 mM DNJ. In contrast, the replication of $Autographa$ $californica$ multiple NPV (AcMNPV) was reduced by 67% when it was propagated in Sf9 cells that were grown in medium containing 10 mM DNJ. The viability of Bm5 and Sf9 cells that were grown in medium containing up to 10 mM DNJ was not affected. Our results suggested that the reduced replication of AcMNPV was the result of the higher sensitivity of ${\alpha}$-glucosidase activity in Sf9 cells to DNJ.