Ju, Wan-Taek
(Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA)
,
Kim, Hyun-Bok
(Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA)
,
Kim, Kee-Young
(Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA)
,
Sung, Gyoo-Byung
(Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA)
,
Kim, Yong-Soon
(Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA)
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 l...
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.
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.
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제안 방법
To test the α-glucosidase inhibitory activity in the culture supernatants of the isolated strains, a TLC analysis was performed by spotting 10 μL of the supernatant of the culture broths onto Silica Gel 60 F254 TLC plates (Merck, Darmstadt Germany) and separating the samples in a TLC chamber containing a solvent mixture of butanol-propanol-acetic acid-water (5: 3: 3: 1.5, v/v/v/ v) at room temperature. As the standard control, 10 mM glucose and various disaccharides (10 mM fructose, maltose, sucrose) were spotted under the same conditions.
To measure the DNJ content of test materials, HPLC was used SHISEIDO SP3203 for fluorescence detection (EM 254, EX 322) and a C18 (100 x 4.6mm, ID 3 μm) column. The absorbance of the effluent was monitored at 254 nm and the flow rate was 1 mL/min.
To examine the use of silkworm powder (SP), silkworm feces powder (SFP), mulberry leaf powder (MLP) as nitrogen sources in the culture broth for the inoculated microorganisms, the α-glucosidase inhibitory activity of these powders was monitored during pre-fermentation (Fig. 1). When the sericultural products were fermented for 5 d, α-glucosidase inhibitory activity increased during pre-fermentation.
대상 데이터
Silkworm powder (SP), silkworm feces powder (SFP), Mulberry leaf powder (MLP) as nitrogen sources were collected from the Sericulture and Apiculture Division for Department of Agricultural Biology, RDA, Suwon, Republic of Korea. The samples were kept in plastic bags and stored at 4℃ until use.
samples. Approxymately 60 colonies were selected as candidates DNJ producers (Table 1). On LB agar plates, 3 strains (JS-1, OS21-2, and YS-5) exhibited a Bacillus-like morphological shape with a slimy and sticky surface, and were highly mucous forming, similar to B.
참고문헌 (31)
Asai A, Nakagawa K, Higuchi O, Kimura T, Kojima Y, and Kariya J (2011) Effect of mulberry leaf extract with enriched 1-deoxynojirimycin content on postprandial glycemic control in subjects with impaired glucose metabolism. J. Diabetes. Investig. 2, 318-323.
Asano N, Yamashita T, Yasuda K, Ikeda K, Kizu H, Kameda Y, Kato A, Nash RJ, Lee HS, and Ryu KS (2001) Polyhydroxylated alkaloids isolated from mulberry trees (Morus alba L.) and silkworms (Bombyx mori L.). J. Agric. Food Chem., 49, 4208-4213.
Ezure Y, Maruo S, Miyazaki K, and Kawamata M (1985) Moranoline (1-deoxynojirimycin) fermentation and its improvement. Agric. Biol. Chem., 49, 1119-1125.
Floris AV, Peter LL, Reinier PA, Eloy HV, Guy ER, and Chris VW (2005) $\alpha$ -Glucosidase inhibitors for patients with type 2 diabetes. Diabetes Care, 28, 154-162
Fujita H, Yamagami T, and Ohshima K (2001) Efficacy and safety of Touchi extract, an α-glucosidase inhibitor derived from fermented soybeans, in non-insulin-dependent diabetic mellitus. J. Nutr. Biochem. 12, 351-356.
Fujita H, Yamagami T, and Ohshima K (2003) Long-term ingestion of Touchi-extract, an α-glucosidase inhibitor, by borderline and mild type-2 diabetic subjects is safe and significantly reduces blood glucose levels. Nutr. Res. 23, 713-722.
Han J, Inoue S, and Isoda H (2007) Effects of silkworm powder on glucose absorption by human intestinal epithelial cell line Caco-2. J. Nat. Med., 61(4), 387-390.
Hardick DJ, Hutchinson DW, Trew SJ and Wellington EMH (1991) The biosynthesis of deoxynojirimycin and deoxymannonojirimycin in Streptomyces subrutilus. J. Chem. Soci. & Chem. Communications., 10, 729-730.
Kim JW, Kim SU, Lee HS, Kim IS, Anh MY, and Ryu KS (2003) Determination of 1-deoxynojirimycin in Morus alba L. leaves by derivatization with 9-fluorenylmethyl chloroformate followed by reversed-phase high-performance liquid chromatography. J. Chromatogr. A., 1002, 93-99.
Kimura T, Nakagawa K, Saito Y, Yamagishi K, Suzuki M, Yamaki K, Shinmoto H, and Miyazawa T (2004) Determination of 1-deoxynojirimycin in mulberry leaves using hydrophilic interaction chromatography with evaporative light scattering detection. J. Agric. Food Chem., 52, 1415-1418.
Kimura T, Nakagawa K, Kubota H, Kojima Y, Goto Y, and Yamagishi K (2007) Food-grade mulberry powder enriched with 1-deoxynojirimycin suppresses the elevation of postprandial blood glucose in human. J. Agric. Food Chem., 55, 5869-5874.
Konno K, Ono H, Nakamura M, Tateishi K, Hirayama C, Tamura Y, Hattori M, Koyama A, and Kohno K (2006) Mulberry latex rich in antidiabetic sugar-mimic alkaloids forces dieting on caterpillars. Proceedings of National Academy of Sciences in USA, 103, 1337-1341.
McCue P, Kwon YI, and Shetty K (2005) Anti-diabetic and antihypertensive potential of sprouted and solid-state bioprocessed soybean. Asian Pacific Journal of Clinic Nutrition, 14(2), 145-152.
Murata K, Yatsunami K, Fukuda E, Onodera S, Mizukami O, Suzuki N, and Kamei T (2005) Effect of propolis and mulberry leaf extract (Quapolis Tm) on type 2 diabetes. Food Func., 1(1), 26-30.
Paek NS Kang DJ, Lee HS, Kim TH, and Kim KW (1997) Production of 1-deoxynojirimycin by Streptomyces sp. SID9135. J. Microbiol. Biotechnol., 7, 262-266.
Romaniouk AV, Silva A, Feng J, and Vijay IK (2004) Synthesis of a novel photoaffinity derivative of 1-deoxynojirimycin for active sitedirected labeling of glucosidase I. Glycobiology 2004, 14, 301-310.
Ryu KS, Lee HS, Chung SH, and Kang PD (1997) An activity of lowering blood-glucose levels according to preparative conditions of silkworm powder. Korean J. Seric. Sci. 39, 79-85.
Ryu KS, Lee HS, Kim KY, Kim MJ, Sung GB, Ji SD, and Kang PD (2013) 1-Deoxynojirimycin content and blood glucose-lowering effect of silkworm (Bombyx mori) extract powder. Int. J. Indust. Entomol., 27(2), 237-242.
Shibano M, Fujimoto Y, Kushino K, Kusano G, and Baba K (2004) Biosynthesis of 1-deoxynojirimycin in Commelina communis: A difference between the microorganisms and plants. Phytochem., 65, 2661-2665.
Stein DC, Kopec LK, Yasbin RE, and Young FE (1984) Characterization of Bacillus subtilis DSM704 and its production of 1-deoxynojirimycin. Appl. Environmental. Microbiol., 48, 280-284.
Vichasilp C, Nakagawa K, Sookwong P, Higuchi O, Kimura F, and Miyazawa T (2012) A novel gelatin crosslinking method retards release of mulberry 1-deoxynojirimycin providing a prolonged hypoglycaemic effect. Food Chem., 134, 1823-1830.
Yagi M, Kouno T, Aoyagi Y, and Murai H (1976) The structure of moranoline, a piperidine alkaloid from Morus species. Nippon Nogeikagaku Kaishi, 50, 571-572.
Yamaki K and Mori Y (2006) Evaluation of alpha-glucosidase inhibitory activity in colored foods: A trial using slope factors of regression curves. Nippon Shokuhin Kagaku Kaishi 53, 229-231.
Yatsunami K, Murata K and Kamei T (2011) 1-Deoxynojirimycin content and alfa-Glucosidase inhibitory activity and heat stability of 1-deoxynojirimycin in silkworm powder. Food. Nutri. Sci., 2, 87-89.
Yin H, SHI XQ, SUN B, Ye JJ, Duan ZA, Zhau XL, Cui WZ and Wu XF (2010) Accumulation of 1-deoxynojirimycin in silkworm, Bombyx mori L. Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology)., 11(4), 286-291.
Zheng YG, Xue YP, and Shen YC (2006) Production of valienamine by a newly isolated strain: Stenotrophomonas maltrophilia. Enzyme and Microbial Technology, 39, 1060-1065.
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