Antifungal Activity of Methylene Chloride Fraction of Pimpinella brachycarpa Against Aspergillus niger

참나물 Methylene Chloride 분획의 Aspergillus niger에 대한 항진균 활성

  • Received : 2011.05.12
  • Accepted : 2011.05.30
  • Published : 2011.06.28

Abstract

In order to develop safe and economic novel antifungal agents, we prepared 73 methanol extracts from medicinal and edible herbs and examined their 365 solvent fractions using n-hexane, methylene chloride, ethylacetate, butanol and water residue based on the sequential organic solvent fraction method. When using the various fractions in the screening step for antifungal activity, we discovered ethylacetate fraction of Morus alba L., methylene chloride fraction of Pimpinella brachycarpa (MCPB), and n-hexane fraction of Salvia miltiorrhiza Bunge, which all have activities in methanol extracts, as potential sources of antifungal agents. Amongst these, the antifungal activity of P. brachycarpa has not to date been reported on. In addition, the mycelial growth inhibition and spore germination inhibition activities of MCPB against A. niger were confirmed by disc-diffusion assay in a 10 day culture. The MIC and MFC of MCPB were determined as 0.25 and 0.5 mg/ml, respectively. The MCPB has no hemolytic activity against human RBC at 0.5 mg/ml and glycoside-flavonoids are theorized to be active constituents. These results suggest that MCPB has a prominent antifungal activity and that the application of sequential organic solvent fractions, instead of simple natural product extracts, is useful in the screening process of novel bioactive substances.

본 연구에서는 안전하고 효율적인 항진균제 개발 연구의 일환으로 73종의 약용 및 식용식물의 메탄올 추출물을 조제하고 이들의 hexane, methylene chloride, ethylacetate, butanol 분획물 및 water 잔류물 365종을 각각 조제하여 Aspergillus niger에 대한 항진균력을 조사하였다. 활성 선별단계에서 비교적 경제적이면서도 손 쉬운 순차적 유기용매 분획물을 제조하여 항진균 천연물 탐색을 시도함으로서 기존에 천연물 추출물에서 검색되지 않은 참나물, 뽕나무, 단삼의 항진균 활성을 확인하였다. 특히 항진균 활성이 보고된 바 없는 참나물의 경우 methylene chloride 분획에서 A. niger에 대한 MIC, MFC 가 각각 0.25 및 0.5 mg/mL임을 확인하여 강력한 항진균 활성을 확인하였으며, 0.25 mg/mL 농도에서 포자발아 억제활성도 나타냄을 확인하였다. 또한 활성분획은 0.5 mg/mL농도까지 인간 적혈구에 대해 용혈활성을 나타내지 않았으며, 활성물질은 당이 결합된 flavonoid로 추측되었다. 본 연구결과는 참나물 methylene chloride 분획의 우수한 항진균 활성을 제시하며, 비교적 간단한 유기 용매 분획법을 이용한 천연물 시료제조 및 이를 이용한 활성탐색이 새로운 활성물질 선별에 효과적임을 제시하고 있다.

Keywords

References

  1. Ahn, S. M., D. S. Lee, M. S. Kim, S. J. Choi, C. S. Choi, J. B. Lee, H. S. Jang, and H. Y. Sohn. 2009. Bioactivity of the extract of Coptis chinensis: In-vitro antifungal activity against Phytophthora capsici and growth promoting effect in red-pepper. Kor. J. Microbiol. Biotechnol. 37: 280-286.
  2. Ahn, S. M., H. Y. Ryu, D. K. Kang, I. C. Jung, and H. Y. Sohn. 2009. Antimicrobial and antioxidant activity of the fruit of Prunus avium L. Kor. J. Microbiol. Biotechnol. 37: 371-376.
  3. Chang, K. M. 2007. A study cookery utilization of Pimpinella brachycarpa N for developing as functional foods. Kor. J. Food Culture 22: 274-282.
  4. Chang, K. M., M. S. Chung, M. K. Kim, and G. H. Kim. 2007. Analysis of mineral and volatile compounds in Pimpinella brachycarpa N. by ICP-AES and SDE, HSSPME- GC-MS. Kor. J. Food Culture 22: 246-253.
  5. Choi, G. J., J. C. Kim, K. S. Jang, H. K. Lim, I. K. Park, S. C. Shin, and K. Y. Cho. 2006. In vivo antifungal activities of 67 plant fruit extracts against six plant pathogenic fungi. J. Microbiol. Biotechnol. 16: 491-495.
  6. Choi, M. H. and G. H. Kim. 2002. A study on quality characteristics of Pimpinella brachycarpa kimchi during storage at different temperatures. J. Kor. Soc. Food Sci. Nutr. 31: 45-49. https://doi.org/10.3746/jkfn.2002.31.1.045
  7. Choi, N. S., S. S. Oh, and J. M. Lee. 2001. Change of biologically functional compounds of Pimpinella brachycarpa (Chamnamul) by blanching conditions. Kor. J. Dietary Culture 16: 388-397.
  8. Choo, M. H., J. J. Lee, and M. Y. Lee. 2007. Effect of Pimpinella brachycarpa ethanol extract on chronically ethanol-induced liver damage in rats. J. Life sci. 17: 1406- 1413. https://doi.org/10.5352/JLS.2007.17.10.1406
  9. Grayer, R. J. and J. B. Harbone. 1994. A survey of antifungal compounds from higher plants. Phytochemistry 37: 19-24. https://doi.org/10.1016/0031-9422(94)85005-4
  10. Kim, B. S., H. S. Jang, C. S. Choi, J. S. Kim, G. S. Kwon, I. S. Kwun, K. H. Son, and H. Y. Sohn. 2008. Antifungal activity of Zanthaxylium schinifolium against Fusarium graminearum. J. Life Sci. 18: 974-979. https://doi.org/10.5352/JLS.2008.18.7.974
  11. Kim, H. G., Y. E. Kim, J. R. Do, Y. C. Lee, and B. Y. Lee. 1995. Antioxidative activity and physiological activity of some Korean medicinal plants. Kor. J. Food Sci. Technol. 27: 80-85.
  12. Kim J. I., H. S. Jang, J. S. Kim, and H. Y. Sohn. 2009. Evaluation of antimicrobial, antithrombin, and antioxidant activity of Dioscorea batatas Decne. Kor. J. Microbiol. Biotechnol. 37: 133-139.
  13. Kum, E. J., S. J. Park, B. H. Lee, J. S. Kim, K. H. Son, and H. Y. Sohn. 2006. Antifungal activity of phenanthrene derivatives from aerial bulbils of Dioscorea batatas Decne. 2006. J. Life Sci. 16: 647-652. https://doi.org/10.5352/JLS.2006.16.4.647
  14. Kwon, C. S., Y. S. Kwon, Y. S. Kim, G. S. Kwon, I. Jin, G. C. Ryu, and H. Y. Sohn. 2004. Inhibitory activities of edible and medicinal herbs against human thrombin. J. Life. Sci. 14: 509-513. https://doi.org/10.5352/JLS.2004.14.3.509
  15. Lee, C. B. 2003. Illustrated flora of Korea. Hwang Moon Sa, Seoul.
  16. Lee, J. J., M. H. Choo, and M. Y. Lee. 2006. Effect of Pimpinella brachycarpa. extract on lipid metabolism in rats fed high cholesterol diet. J. Korean Soc. Food Sci. Nutr. 35: 1151-1158. https://doi.org/10.3746/jkfn.2006.35.9.1151
  17. Lee, J. J., M. H. Choo, and M. Y. Lee. 2007. Physicochemical compositions of Pimpinella brachycarpa. J. Korean Soc. Food Sci. Nutr. 36: 327-331. https://doi.org/10.3746/jkfn.2007.36.3.327
  18. Lee, M. K., S. H. Kim, S. S. Ham, S. Y. Lee, C. K. Chung, I. J. Kang, and D. H. Oh. 2000. The effect of far infrared rayvacuum drying on the quality changes of Pimpinella brachycarpa. J. Kor. Soc. Food Sci. Nutr. 29: 561-567.
  19. Lee, Y. M., J. J. Lee, and M. Y. Lee. 2008. Antioxidative effect of Pimpinella brachycarpa ethanol extract. 2008. J. Life Sci. 18: 467-473. https://doi.org/10.5352/JLS.2008.18.4.467
  20. Ryu, H. Y., S. M. Ahn, Y. K. Shin, and H. Y. Sohn. 2010. Antimicrobial and hemolytic activity of oriental medicinal herbs. Kor. J. Microbiol. Biotechnol. 38: 190-1976.
  21. Ryu, H. Y., S. M. Ahn, J. S. Kim. I. C. Jung, and H. Y. Sohn. 2010. Antimicrobial activity of fruit of Crataegus pinnatifida Bunge against multidrug resistant pathogenic Pseudomonas aeruginosa and Candida sp. Kor. J. Microbiol. Biotechnol. 38: 77-83.
  22. Sohn, H. Y., C. S. Kwon, and K. H. Son. 2010. Fungicidal effect of penylated flavonol, papyriflavonol A, isolated from Broussonetia papyrifera (L.) Vent, against Candida albicans. J. Microbiol. Biotechnol. 20: 1397-1402. https://doi.org/10.4014/jmb.1007.07026
  23. Sohn, H. Y., E. J. Kum, H. Y. Ryu, S. J. Jeon, N. S. Kim, and K. H. Spn. 2006. Antifungal activity of fistulosides, steroidal saponins, from Allium fistulosum L. J. Life Sci. 16: 310-314. https://doi.org/10.5352/JLS.2006.16.2.310
  24. Sohn, H. Y., E. J. Kum, Y. S. Kwon, G. S. Kwon, I. Jin, H.Y. Kwon, C. S. Kwon, and K. H. Son. 2003. Screening of anticandidiosis agent from medicinal and wild plants. Kor. J. Life Sci. 13: 604-617. https://doi.org/10.5352/JLS.2003.13.5.604
  25. Sohn, H. Y., K. H. Son, C. S. Kwon, G. S. Kwon, and S. S. Kang. 2004. Antimicrobial and cytotoxic activity of 18 prenylated flavonoids isolated from medicinal plants: Morus alba L., Morus mongolica Schneider, Broussnetia papyrifera (L.) Vent, Sophora flavescens Ait and Echinosophora koreensis Nakai. Phytomedicine 11: 666-672.. https://doi.org/10.1016/j.phymed.2003.09.005
  26. Son, S. I. and J. C. Kim. 1999. Cloning and characterization of homeodomain-zip gene, Phc5 in embryogenic callus derived from Pimpinella brachycarpa suspension cultured cells. Kor. J. Plant Tissue Culture 26: 121-126.
  27. Song, H. S., H. S. Choi, and M. S. Lee. 1997. Usefulness of Pimpinella brachycarpa as natural spice by sensory analysis. Kor. J. Food Sci. 13: 669-673
  28. Wong, K. S. and W. K. Tsang. 2009. In vitro antifungal activity f the aqueous extract of Scutellaria baicalensis Georgi root against Candida albicans. Int. J. Antimicrob. Agents 34: 284-285. https://doi.org/10.1016/j.ijantimicag.2009.03.007
  29. Yang, D., H. Hu, S. Huang, J. P. Chaumont, and J. Millet. 2000. Study on the inhibitory activity, in vitro, of baicalein and baicalin against skin fungi and bacteria. Zhong Yao Cai.23: 272-274.