• Title/Summary/Keyword: DNJ

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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.

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.

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.

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.

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.

Characterization of DNJ production for large-scale fermentation of mulberry leaf

  • Kwon, O-Chul;Ju, Wan-Taek;Kim, Hyun-Bok;Sung, Gyoo-Byung;Kim, Yong-Soon
    • International Journal of Industrial Entomology and Biomaterials
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    • v.35 no.2
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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.

Accumulating Pattern of ${\alpha}-glycosidase$ Inhibitor in Various Silkworm Varities (누에품종별 혈당강하물질 축적양상 구명)

  • Kang, Pil-Don;Kim, Jin-Won;Sohn, Bong-Hee;Kim, Kee-Young;Jung, I-Yoen;Kim, Mi-Ja;Ryu, Kang-Sun
    • Journal of Sericultural and Entomological Science
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    • v.48 no.1
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    • pp.25-27
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    • 2006
  • [ ${\alpha}-Glycosidase$ ] inhibitors slows the velocity of the uptake of monosaccharides in the small intestine by retarding the speed of degradation of disaccharides to monosaccharides, which made it possible to develop ${\alpha}-glycosidase$ inhibitors as the antihyperglycemic ('antihyperglycemic' means 'blood-glucose-level-lowering') reagent for the diabetic patients such as acarbose and miglitol. Twenty kinds of ${\alpha}-glycosidase$ inhibitors have been reported to exist in mulberry, Morns alba, and some of them are also found in the silkworm, Bombyx mari, as the result of its daily feeding of mulberry leaves as the sole diet. 1-Deoxynojirimycin (DNJ), one of the most potent ${\alpha}-glycosidase$ inhibitor, is the most abundant among polyhydroxylated alkaloids with ${\alpha}-glycosidase$ inhibiting activity in both M alba and B. mari, therefore considered the antihyperglycemic criterion of the mulberry- or silkworm-based neutroceutical products. DNJ is thought to be accumulated in the body of silkworm because the its concentration in the silkworm body is two to three times as much as that in the mulberry leaves. Eighteen silkworm F1 varieties have been recommended for industrial rearing in Korea by some standards such as pathological strength. DNJ concentration in 18 recommendedwere measured at 3rd day in the 5th instar after lyophilization to determine the varieties that accumulates DNJ in its body most. GeumOk-Jam was the highest in the DNJ concentration of 5.45 mg/gDW among the recommended F1 varieties.

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.

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.