Journal of the Korean Applied Science and Technology (한국응용과학기술학회지)

The Korean Society of Applied Science and Technology (한국응용과학기술학회)
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Fermentation of Momordica charantia Extract using Leuconostoc mesenteroidesies and Physiological Activity of Product

Leuconostoc mesenteroidesies 균주를 이용한 여주 추출물 발효 및 생산물의 생리활성 특성

  • 강정훈 (청주대학교 바이오메디컬학과)
  • Received : 2018.09.18
  • Accepted : 2018.12.25
  • Published : 2018.12.31
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Abstract

In this study, Momordica charantia (MC) fermented with Leuconostoc mesenteroides (MC-LM) were assessed for the antioxidant and the antidiabetic activities. Antioxidant activities of MC and MC-LM were evaluated using 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)(ABTS) radical. Although MC-treated groups showed little activity, 47% of activity was observed at $500{\mu}g/mL$ concentration for MC-LM and increased significantly(p<0.05) as MC-LM concentration increased. MC-LM more effectively inhibited the oxidative damage of DNA by peroxyl radical than MC and the inhibition of the strand breakage increased significantly as MC-LM concentration increased(p<0.05). Measuring the inhibition of ${\alpha}-glucosidase$ activity, which is closely related to the regulation of blood sugar, resulted in MC reduced the activity of ${\alpha}-glucosidase$ by 30% at 8 mg/mL and MC-LM at the same concentration by 60%. In addition, the effect of MC-LM on the cell viability of alloxan-treated RIN-m5F resulted in a significant increase in cell survival(p<0.05) in the group treated with MC-LM and a 20% increase in the concentration of $1000{\mu}g/mL$. As a result of insulin secretion by alloxan-treated RIN-m5F cell, the level of insulin secretion tended to increase in all group treated with MC-LM. At the concentration of $1000{\mu}g/mL$, the insulin secretion was increased by 15% in MC-LM group than in MC group. In conclusion, the results of this study suggest that fermented bitter gourd has antioxidant and antidiabetic effects.

여주(Momordica charantia) 열수 추출물을 젖산균(Leuconostoc mesenteroidesies)으로 발효하여 얻은 생산물의 항산화 및 항당뇨 효과를 관찰하였다. 여주 발효물(MC-LM)과 여주 추출물(MC)의 항산화 활성은 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)(ABTS) radical 소거 측정법을 사용하여 관찰하였다. MC 처리군은 radical 소거활성이 거의 나타나지 않았으나 MC-LM의 경우 $500{\mu}g/mL$ 농도에서 47%의 활성이 관찰되었으며 MC-LM의 농도가 증가함에 따라 활성이 유의하게(p<0.05) 증가하는 것을 확인 할 수 있었다. MC-LM은 MC에 비해 peroxyl radical에 의한 DNA의 산화적 손상을 더 효과적으로 억제하였으며 MC-LM의 농도가 증가함에 따라 strand breakage의 억제가 유의적으로 증가 하였다(p<0.05). 혈당량과 밀접한 관련이 있는 ${\alpha}-glucosidase$의 활성 억제정도를 측정한 결과 MC는 8 mg/mL 농도에서 ${\alpha}-glucosidase$의 활성을 30.1% 감소시켰고 같은 농도에서 MC-LM은 58.9% 감소 시켰다. 또한 alloxan에 의한 RIN-m5F 췌장세포 사멸에 발효물의 영향을 관찰한 결과 MC-LM을 농도별로 처리한 군에서 세포생존율이 유의적으로(p<0.05) 증가하였으며 $1000{\mu}g/mL$ 농도에서 MC군 보다 20% 높게 증가되었다. Alloxan에 의한 ${\beta}-cell$파괴를 유도하여 RIN-m5F cell이 분비한 인슐린의 측정결과 MC-LM을 농도별로 첨가한 모든 군에서 인슐린 분비능이 증가하는 경향을 보였으며, $1000{\mu}g/mL$ 농도에서 MC군 보다 15% 높게 증가되었다. 결론적으로 본 연구에서 관찰된 결과들을 통해 여주 발효물은 항산화 및 항당뇨 효과가 있는 것으로 확인되었다.

Keywords

HGOHBI_2018_v35n4_1250_f0001.png 이미지

Fig. 1. ABTS radical scavenging activity of fermented Momordica charantia (MC) with Leuconostoc mesenteroidesies (MC-LM). The reaction mixture contained 7 mM ABTS and 2.5 mM potassium persulfate in the presence or absence of various concentrations of MC (black bar), MC-LM (grey bar) at 37℃ for 10 min. The values represent the mean±S.D. for triplicate experiments. Significantly different from the ascorbic acid treated group.

HGOHBI_2018_v35n4_1250_f0002.png 이미지

Fig. 2. Protective effects of fermented Momordica charantia (MC) with Leuconostoc mesenteroidesies (MC-LM) on DNA strand breakage induced by peroxyl radical. pUC 19 DNA was incubated with 10 mM AAPH in the presence of various concentrations of MC-LM at 37℃ for 2 h. (A) Lane 1, control DNA; lane 2, lane 1 + AAPH; lane 3, lane 2 + 0.1 mg/mL MC; lane 4, lane 2 + 1 mg/mL MC; lane 5, lane 2 + 10 mg/mL MC; lane 6, lane 2 + 20 mg/mL MC. (B) Lane 1, control DNA; lane 2, lane 1 + AAPH; lane 3, lane 2 + 0.1 mg/mL MC-LM; lane 4, lane 2 + 1 mg/mL MC-LM; lane 5, lane 2 + 10 mg/mL MC-LM; lane 6, lane 2 + 20 mg/mL MC-LM. (C) Relative staining intensity of agarose gel was analyzed by densitometric scanning.

HGOHBI_2018_v35n4_1250_f0003.png 이미지

Fig. 3. Protective effects of fermented Momordica charantia (MC) with Leuconostoc mesenteroidesies (MC-LM) on the inhibition of α- glucosidase activity in the presence of various concentrations of MC (black bar) or MC-LM (grey bar).

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Fig. 4. Protective effects of fermented Momordica charantia (MC) with Leuconostoc mesenteroidesies (MC-LM) on cell death induced by alloxan. RIN-m5F cells were treated with 4 mM alloxan the presence or absence of varying concentrations of MC (black bar) or MC-LM (grey bar) and cell viabilities were estimated by with a colormetric assay using MTT. The values represent the mean±S.D. for triplicate experiments. Significantly different from the control.

HGOHBI_2018_v35n4_1250_f0005.png 이미지

Fig. 5. Protective effects of fermented Momordica charantia (MC) with Leuconostoc mesenteroidesies (MC-LM) on the secretion of insulin from the cell induced by alloxan. RIN-m5F cells were treated with 4 mM alloxan in the presence or absence of varying concentrations of MC (black bar) or MC-LM (grey bar) and the amounts of insulin were estimated by using Rat/Mouse Insulin ELISA kit (Merk Millipore). The values represent the mean±S.D. for triplicate experiments. Significantly different from the control.

References

  1. J. Lee, H. Park, "Relationships between diabetic knowledge, self-care behaviors and HbA1c in diabetic patients using public hospitals", J. Korean Acad. Fundam. Nurs, Vol.21, No.3 pp. 243-252, (2014). https://doi.org/10.7739/jkafn.2014.21.3.243
  2. A. K. Meena, P. Bansal, S. Kumar, "Plants-herbal wealth as a potential source of ayurvedic drugs", Asian J. Traditional, Vol.4, No.4 pp. 152-170, (2009).
  3. S. Y. Lee, S. L. Park, Y. D. Nam, S. H. Yi, and S. I. Lim, "Anti-diabetic effects of fermented green tea in KK-Ay diabetic mice", Korean J. Food Sci. Technol, Vol.45, No.4 pp. 488-494, (2013). https://doi.org/10.9721/KJFST.2013.45.4.488
  4. J. O. Kim and G. D. Lee, "Hypoglycemic effects of a medicinal herb mixture prepared through the traditional antidiabetic prescription", Korean J. Food Preserv, Vol.18, No.6 pp. 923-929, (2011). https://doi.org/10.11002/kjfp.2011.18.6.923
  5. W. K. Yun, H. J. Bae, Y. J. Kim, J. O. Kwon, M. H. Im, H. D. Cho, T. Kim, "Effect of the supplementation of Coconopsis lanceolata extract on lipid metabolism amelioration in type 2 diabetes mouse model induced by high fat diet", Korean J. Food Preserv, Vol.21, No.1 pp. 107-113, (2014). https://doi.org/10.11002/kjfp.2014.21.1.107
  6. H. K. Son, H. J. Han, and J. J. Lee, "Anti-diabetic effect of the mixture of mulberry leaf and green teapowder in rats with streptozotocin-induced diabetes", Korean J. Food Preserv, Vol.21, No.4 pp. 549-559, (2014). https://doi.org/10.11002/kjfp.2014.21.4.549
  7. H. S. Lee, H. J. Kong, E. H. Lee, S. J. Hwang, H. A. Jung, M. L. Kim, E. M. Choi, J. H. Jang, K. M. Yang, "Hypoglycemic effects of boiled rice made from unpolished rice, job' tear, and extract from medicinal herbs mixture on diabetic rat", Korean J. Herbology, Vol.29, No.3 pp. 59-70, (2014). https://doi.org/10.6116/kjh.2014.29.3.59
  8. H. J. Lee, J. H. Moon, W. M. Lee, S. G. Lee, A. K. Kim, Y. H. Woo, D. K. Park, "Charantin contents and fruit characteristics of bitter gourd (Momordica charantia L.) accessions", J Bio-enviroment control, Vol.21, No.4 pp. 379-84, (2012). https://doi.org/10.12791/KSBEC.2012.21.4.379
  9. Y. Park, H. O. Boo, Y. L. Park, D. H. Cho, H. H. Lee "Antioxidant activity of Momordica charantia L. extracts" Korean J. Medicinal Crop. SAci. Vol.15, No.1 pp 56-61, (2007).
  10. E. Y. Kim, I. H. Baik, J. H. Kim, S. R. Kim, M. R. Rhyu, "Screening of the antioxidant activity of some medicinal plants" Korean J. Food Sci Thechnol. Vol.36, No2 pp. 333-338, (2004).
  11. H. S. Yang, Y. J. Choi, H. H. Oh, J. S. Moon, H. K. Jung, K. J. Kim et al, "Antioxidative activity of mushroom water extracts fermented by lactic acid bacteria", J. Korean Soc. Food Sc.i Nutr, Vol.43, No.1 pp. 80-85, (2014). https://doi.org/10.3746/jkfn.2014.43.1.080
  12. H. S. Park, W. K. Kim, H. P. Kim, Y. G. Yoon, "The efficacy of lowering blood glucose levels using the extracts of fermented bitter melon in the diabetic mice" J. Appl. Biol. BChem. Vol.58, No.3 pp. 259-265, (2015). https://doi.org/10.3839/jabc.2015.041
  13. W. Brand-Williams, M. E. Cuvelier, C. Berset, "Use of a free radical method to evaluate antioxidant activity", Food Sci. Technol, Vol.28, No.1 pp. 25, (1995).
  14. J. H. Kang, S. M. Kim, "DNA cleavage by hydroxyl radicals generated in the Cu,Zn-Superoxide dismutase and hydrogen peroxide system", Mol. Cells, Vol.7, No.6 pp. 777-782, (1997).
  15. M. Choe, D. J. Kim, H. J, Lee, J. K. You, D. J. Seo, J. H. Lee, M. J. Chung, "A study on the glucose regulating enzymes and antioxidant activities of water extracts from medicinal herbs", J. Korean Soc. Food Sci. Nutr, Vol.37, No.5 pp. 542-547, (2008). https://doi.org/10.3746/jkfn.2008.37.5.542
  16. J. L. Kim, C. R. Bae, Y. S. Cha, "Helianthus tuberosus extracts has anti-diabetes effects in HIT-T15 cells" J. Korean Soc. Food. Sci. Nutr. Vol.39, No.1 pp. 31-35 (2010). https://doi.org/10.3746/jkfn.2010.39.1.031
  17. R. L. Prior, X. Wu, K. Schaich, "Standardized method for the determination of antioxidant capacity and phenolics in foods and dietary supplement", J. Agric. Food Chem, Vol.53, No.10, pp. 4290, (2005). https://doi.org/10.1021/jf0502698
  18. M. Sato, N. Ramarathnam, Y. Suzuki, T. Ohkubo, M. Takeuchi, H, Ochi, "Varietal differences in the phenolic content and superoxide radical scavenging potential of wines from different sources", J. Agric. Food Chem, Vol.44, No.1 pp. 37-41, (1996). https://doi.org/10.1021/jf950190a
  19. D. F. Fitzpatrick, S. L. Hirschfield, R. G. Coffey, "Endothelium-dependent vasorelaxing activity of wine and other grape products", Am. J. Physiol, Vol.265, pp. H774-H778, (1993).
  20. S. Yoon, H. K. Kwak, Y. K. Kim, H. K. Kim, M. S. Park, K. J. Yeum, H. S. Oh, M. J. Lee, J. H. Lee, G. E. Ji, "Functional foods. Life Science Publishing Co", Seoul, Korea, pp. 222-231, (2006).
  21. Z. Jia, M. Tang, J. Wu, "The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals", Food Chem, Vol.64, No.4 pp. 555-559, (1999). https://doi.org/10.1016/S0308-8146(98)00102-2
  22. M. G. L. Hertog, P. C. H. Hollman, M. B. Katan, "Content of potentially anticarcinogenic flavonoids of 28 vegetables and 9 fruits commonly consumed in the Netherlands." J. Agric. Food Chem, Vol.40, No.12 pp. 2379-2383, (1992). https://doi.org/10.1021/jf00024a011
  23. R. L. Lee, "Nutritional Components and Antioxidant Activity of Dry Bitter Melon (Momordica charantia L.) J Korean Soc", Food Sci. Nutr, Vol.45, No.4 pp. 518-523, (2016). https://doi.org/10.3746/jkfn.2016.45.4.518
  24. E. Niki, "Antioxidants in relation to lipid peroxidation", Chem. Phys. Lipids, Vol.44, No.2-4 pp. 227 (1987). https://doi.org/10.1016/0009-3084(87)90052-1
  25. K. Hiramoto, H. Hohkoh, K. Sako, K. Kikugawa, "DNA breaking activity of the carbon-centered radical generated from 2,2'-azobis(2-amidinopropane) hydrochloride(AAPH)", Free Radic. Res. Commun, Vol.19, No.5 pp. 323-332, (1993). https://doi.org/10.3109/10715769309056521
  26. B. N. Ames, "Dietary carcinogens and anticarcinogens. Oxygen radicals and degenerative diseases", Science, Vol.221, No.4617 pp. 1256-1264, (1983). https://doi.org/10.1126/science.6351251
  27. P. A. Cerutti, "Prooxidant states and tumor promotion", Science, Vol.227, No.4685 pp. 375-381, (1985). https://doi.org/10.1126/science.2981433
  28. J. L. Sagripanti, K. H. Kraemer, "Site-specific oxidative DNA damage at polyguanosines produced by copper plus hydrogen peroxide", J. Biol Chem., Vol.264, No.3 pp. 1729-1734, (1989).
  29. T. Finkel, N. J. Holbrook, "Oxidants, oxidative stress and the biology of ageing", Nature, Vol.408, No.6809 pp. 239-247, (2000). https://doi.org/10.1038/35041687
  30. S. Kawanishi, Y. Hiraku, S. Oikawa, "Mechanism of guanine-specific DNA damage by oxidative stress and its role in carcinogenesis and aging", Mutat Res, Vol.488, No.1 pp. 65-76, (2001). https://doi.org/10.1016/S1383-5742(00)00059-4
  31. J. E. Kim, S. I. Joo, J. H. Seo, S. P. Lee, "Antioxidant and ${\alpha}$-glucosidase inhibitory effect of tartary backwheat extract obtained by the treatment of different solvents and enzymes", J. Korean Soc. Food Sci. Nut. Vol.38 pp. 989-995, (2009). https://doi.org/10.3746/jkfn.2009.38.8.989
  32. J. H. Park, M. R. Baek, B. H. Lee, G. H. Yon, S. Y. Ryu, Y. S. Kim, "${\alpha}$-Glucosidase and ${\alpha}$-amylase inhibitory activity of compounds from roots extract of Pueraria thunbergiana" Korean J. Medicinal Crop Sci Vol.17, No.5 pp. 357-362. (2009).
  33. C. H. Lee, Y. R. Lee, "Antioxidant and antidiabetic activities of methanol extracts from different parts of Jerusalem artichke(Helianthus tuberosus L)" Korean J. Food Nutr. Vol.29, No.1 pp. 128-133. (2016). https://doi.org/10.9799/ksfan.2016.29.1.128
  34. D. C. Weaver, M. L. McDaniel, S. P. Naber, C. D. Barry, P. E. Lacy, "Alloxan stimulation and inhibition of insulin release from isolated rat islets of Langerhans" Diabetes Vol.27, No.12 pp. 1205-1214. (1978). https://doi.org/10.2337/diab.27.12.1205