한국미생물·생명공학회지 (Microbiology and Biotechnology Letters)

  • 제38권1호
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  • Pages.105-111
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  • 2010
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  • 1598-642X(pISSN)
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  • 2234-7305(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지

소목 추출물의 항균활성과 Brazilin의 구조분석

Antibacterial Activities of Caesalpinia sappan L. Extract and Structural Analysis of Its Related Brazilin

  • Kwon, Hyun-Jung (Department of Biology, College of Natural Science, Soonchunhyang University) ;
  • Kim, Yong-Hyun (Department of Biology, College of Natural Science, Soonchunhyang University) ;
  • Nam, Kung-Woo (Jung San Bio Technology Co.) ;
  • Kim, Sun-Ki (Sunbio Co.) ;
  • Bang, In-Soek (Department of Biology, College of Natural Science, Hoseo University) ;
  • Han, Man-Deuk (Department of Biology, College of Natural Science, Soonchunhyang University)
  • 투고 : 2010.01.13
  • 심사 : 2010.02.24
  • 발행 : 2010.03.28
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초록

소목은 전통적으로 타박상, 염증 등의 질병치료를 위해 민간의학에서 오래 동안 사용되어 온 한약재야다. 소목의 섬재로부터 항균효과를 조사하기 위하여 MeOH 추출에 의한 hexane, $CHCl_3$, EtOAc, n-BuOH, MeOH 및 $H_2O$ 분획을 얻었다. 분획 시료 중 3.94%의 가장 높은 수율의 EtOAc 분획이 조사된 인채 병원성 세균에 대해 가장 강한 항균활성 나타냈다. 또한 EtOAc 분획의 항균활성은 Gram (-) 균주 보다 Gram (+) 균주에 더 효과적이며, Gram (-) 균주에 대해서는 세균에 따른 항균활성의 차이를 보였다. EtOAc 분획은 silica gel adsorption column chromatography 및 Sephadex LH-20을 이용하여 분리하였으며 수득한 주요 성분은 채결정을 통하여 얻어진 단일물질을 HPLC를 이용하여 순도를 측정하였다. 300g의 C. sappan으로부터 3.94%의 EtOAc 분획을 얻었으며 EtOAc분획물은 분석한 결과 10 mg에는 1.67 mg의 brazilin을 포함하였다. 분리된 활성울질은 짧은 황색 결정의 단일 화합물로 MS에 의해 분자량이 285로 나타났으며, $^1H$-NMR, $^{13}C$-NMR에 의한 구조분석으로 brazilin($C_{16}H_{14}O_5$을 동정하였다. 이 같은 결과는 소목의 MeOH 추출에 의한 EtOAc 분획에 존재하는 brazilin이 S. aureus와 같은 인체 병원성 Gram (+) 균주에 대한 생약치료제로 활용이 가능할 것을 시사한다.

Caesalpinia sappan L. has long been commonly used in oriental folk medicines to treat diseases. To investigate the antibacterial effects from C. sappan L. heart wood, the MeOH soluble extract was successively fractionated by using hexane, $CHC1_3$, EtOAc, BuOH, MeOH, and $H_2O$. Among of these extracts, the EtOAc fraction which partitioned to 3.94% of the highest yields was to be the most active against all human pathogenic bacteria in this experiment. In addition, the antibacterial activities of the EtOAc fraction were more effective against Gram (+) bacteria compared to those against Gram (-) bacteria, which showed difference of the antibacterial activities against Gram (-) bacteria. To confirm the identity of the active substances, the EtOAc fraction was further separated by silica gel adsorption column, high performance liquid chromatography, and 98.48% purity of brazilin (1.67 mg)/EtOAc (10 mg) fraction was obtained from 300 g of C. sappan L. heart wood. The isolated active substance was a single compound of yellow crystalline, and was identified as brazilin ($C_{16}H_{14}O_5$) by MS, and $^lH$-NMR and $^{13}C$-NMR. These results suggest that the brazilin in the EtOAc fraction from MeOH extract of C. sappan L. has a potential as a natural therapeutic agent against human pathogenic Gram (+) bacteria such as Staphylococcus aureus.

키워드

참고문헌

  1. Badami, S., S. Moorkoth, S. R. Rai, E. Kannan, and S. Bhojraj. 2003. Antioxidant activity of Caesalpinia sappan heart wood. Biol. Pharm. Bull. 26: 1534-1537. https://doi.org/10.1248/bpb.26.1534
  2. Baek, N. I., S. G. Jeon, E. M. Ahn, J. T. Hahn, J. H. Bahn, J. S. Jang, S.W. Cho, J. K. Park, and S. Y. Choi. 2000. Anticonvulsant compounds from the wood of Caesalpinia sappan L. Arch. Pharmacol. Res. 23: 344-348. https://doi.org/10.1007/BF02975445
  3. Beuchat, L. R. and D. A. Golden. 1989. Antimicrobials occurring naturally in foods. Food Technol. 43: 134-138.
  4. Chiang, L. C., W. Chiang, M. C. Liu, and C. C. Lin. 2003. In vitro antiviral activities of Caesalpinia pulcherrima and its related flavonoids. J. Antimicrob. Chemother. 52: 194-198. https://doi.org/10.1093/jac/dkg291
  5. Cho, J. Y., I. Choi, and E. K. Hwang. 2003. Antimicrobial activity of extracts from medicinal herbs against Escherichia coli. Korea J. Vet. Res. 43: 625-631.
  6. Choi, S. Y., K. M. Yang, S. D. Jeon, J. H. Kim, L. Y. Khil, T. S. Chang, and C. K. Moon. 1997. Brazilin modulates immune function mainly by augmenting T cell activity in halothane administered mice. Planta. Med. 63: 405-408. https://doi.org/10.1055/s-2006-957722
  7. de Oliveira, L. F. C., H. G. M. Edwards, E. S. Velozo, and M. Nesbitt. 2002. Vibrational spectroscopic study of brazilin and brazilein, the main constituents of brazilwood from Brazil. Vib. Spectrosc. 28: 243-249. https://doi.org/10.1016/S0924-2031(01)00138-2
  8. Hu, C. M., J. J. Kang, C. C. Lee, C. H. Li, J. W. Liao, and Y. W. Cheng. 2003. Induction of vasorelaxation through activation of nitric oxide synthase in endothelial cells by brazilin. Eur. J. Pharmacol. 468: 37-45. https://doi.org/10.1016/S0014-2999(03)01639-X
  9. Huang, K. C., 1993. The pharmacology of Chinese herbs. America CRC Press. 266.
  10. Hwang, G. S., J. Y. Kim, T. S. Chang, S. D. Jeon, D. S. So, and C. K. Moon. 1998. Effects of Brazilin on the phospholipase A2 activity and changes of intracellular free calcium concentration in rat platelets. Arch. Pharm. Res. 21: 774-778. https://doi.org/10.1007/BF02976775
  11. Kang, J. M., I. H. Cha, Y. K. Lee, and H. S. Ryu. 1997. Identification of volatile essential oil, and flavor characterization and antimicrobial effect of fractions from Houttuynia cordata thunb. J. Korean Soc. Food Sci. Nutr. 26: 209-213.
  12. Kim, D. S., N. I. Baek, S. R. Oh, K. Y. Jung, I. S. Lee, and H. K. Lee. 1997. NMR assignment of brazilein. Phytochemistry. 46: 177-178. https://doi.org/10.1016/S0031-9422(96)00874-6
  13. Kim, K. J., H. H. Yu, S. I. Jeong, J. D. Cha, S. M. Kim, and Y. O. You. 2004. Inhibitory effects of Caesalpinia sappan on growth and invasion of methicillin-resistant Staphylococcus aureus. J. Ethnopharmacol. 91: 81-87. https://doi.org/10.1016/j.jep.2003.11.017
  14. Kim, S. G., Y. M. Kim, L. Y., Khil, S. D. Jeon, D. S. So, C. H. Moon, and C. K. Moon. 1998. Brazilin inhibits activities of protein kinase C and insulin receptor serine kinase in rat liver. Arch. Pharm. Res. 21: 140-146. https://doi.org/10.1007/BF02974018
  15. Lee, B. W. and D. H. Shin. 1991. Screening of natural antimicrobial plant extract on food spoilage microorganism. Kor. J. Food Sci. Technol. 23: 200-204.
  16. Lee, K. T. and J. H. Kim. 1997. Brazilin as a new sunless tanning agent. J. Soc. Cosmet. Sci. Korea 23: 82-85.
  17. Lee, S. G. 2003. Antimicrobial activity of Caesalpinia sappan against animal husbandry disease. Kor. J. Microbiol. Biotechnol. 31: 242-249.
  18. Lee, S. H., W. S. Moon, and K. N. Park. 2000. Antimicrobial activity of Caesalpinia sappan L. extracts and its effect on preservation of ground meats. J. Kor. Soc. Food sci. Nutr. 29: 888-892.
  19. Lim, M. Y., J. H. Jeon, E. Y. Jeong, C. H. Lee, and H. S. Lee. 2007. Antimicrobial activity of 5-hydroxy-1,4-naphtho-quinone isolated from Caesalpinia sappan toward intestinal bacteria. Food Chem. 100: 1254-1258. https://doi.org/10.1016/j.foodchem.2005.12.009
  20. Mok, M. S., S. D. Jeon, K. M. Yang, D. S. So, and C. K. Moon. 1998. Effects of brazilin on induction of immunological tolerance by sheep red blood cells in C57BL/6 female mice. Arch. Pharmacol. Res. 21: 769-773. https://doi.org/10.1007/BF02976774
  21. Moon, C. K., K. S. Park, S. G. Kim, H. S. Won, and J. H. Chung. 1992. Brazilin protects cultured rat hepatocytes from $BrCCl_3-induced$ toxicity. Drug Chem. Toxicol. 15: 81-91. https://doi.org/10.3109/01480549209035174
  22. Namikoshi, M., H. Nakata, M. Nuno, T. Ozawa, and T. Saitoh. 1987. Homoisoflavonoids and related compounds. III. Phenolic constituents of Caesalpinia japonica SIEB. Et ZUCC. Chem. Pharm. Bull. 35: 3568-3575. https://doi.org/10.1248/cpb.35.3568
  23. Nguyen, M. T., S. Awale, Y. Tezuka., J. Y. Ueda, Q. Tran, and S. Kadota. 2006. Xanthine oxidase inhibitors from the flowers of Chrysanthemum sinense. Planta. Med. 72: 46-51. https://doi.org/10.1055/s-2005-873181
  24. Oh, S. R., D. S. Kim, I. S. Lee, K. Y. Jung, J. J. Lee, and H. K. Lee. 1998. Anticomplementary activity of constituents from the heartwood of Caesalpinia sappan. Planta. Med. 64: 456-458. https://doi.org/10.1055/s-2006-957481
  25. Otvos, L. Jr. 2000. Antibacterial peptides isolated from insects. J. Pept. Sci. 6: 497–511.
  26. Parameshwar, S., K. K. Srinivasan, and C. M. Rao. 2002. Oral antidiabetic activities of different extracts of Caesalpinia bonducella seed kernels. Pharm. Biol. 40: 590-595. https://doi.org/10.1076/phbi.40.8.590.14656
  27. Park, K. J., S. Yang, Y. A. Eun, S. Y. Kim, H. H. Lee, and H. Kang. 2002. Cytotoxic effects of Korean medicinal herbs determined with hepatocellular carcinoma cell lines. Pharm. Biol. 40: 189-195. https://doi.org/10.1076/phbi.40.3.189.5824
  28. Park, U. Y., D. S. Chang, and H. R. Cho. 1992. Screening of antimicrobial activity for medicinal herbs extracts. J. Kor. Soc. Food Nutr. 21: 91-96.
  29. Puchtler, H., S. N. Meloan, and F. B. Waldrop. 1986. Application of current concepts to metal-hematein and - brazilein stains. Histochemistry. 85: 353-364. https://doi.org/10.1007/BF00982665
  30. Rivero-Cruz, J. F. 2008. Antimicrobial compounds isolated from Haematoxylon brasiletto. J. Ethnopharmacol. 119: 99-103. https://doi.org/10.1016/j.jep.2008.06.005
  31. Rodriguez-Lopez V., L. Salazar, and S. Estrada. 2003. Spasmolytic activity of several extracts obtained from some Mexican medicinal plants. Fitoterapia. 74: 725-728. https://doi.org/10.1016/S0367-326X(03)00187-4
  32. Safitri, R., P. Tarigan, H. J. Freisleben, R. J. Rumampuk, and A. Murakami. 2003. Antioxidant activity in vitro of two aromatic compounds from Caesalpinia sappan L. Biofactors. 19: 71-77. https://doi.org/10.1002/biof.5520190109
  33. Saitoh, T., S. Sakashita, H. Nakata, T. shimokawa, J. Kinjo, J. Yamahara, M. Yamasaki, and T. Nohara. 1986. 3-Benzylchroman derivatives related to brazilin from Sappan lignum. Chem. Pharm. Bull. 34: 2506-2511. https://doi.org/10.1248/cpb.34.2506
  34. Shin, D. H., M. S. Kim, and J. S. Han. 1997. Antimicrobial effect of ethanol extracts from some medicinal herbs and their fractionates against food-borne bacteria. Kor. J. Food Sci. Technol. 29: 808-816.
  35. Shin, D. W. 2003. Screening and using of antioxidative effect and antimicrobial activity from plant. Food Science industry. 36: 81-89.
  36. Sitaram, N. and R. Nagaraj. 2002. Host-defense antimicrobial peptides: Importance of structure for activity. Curr. Pharm. Des. 8: 727-742. https://doi.org/10.2174/1381612023395358
  37. Son, E. J., J. H. Kim, H. A. Kim, S. H. Baek, Y. H. Kho, M. R. Kim, and C. H. Lee. 2003. A Caspase inducing inhibitor isolated from Caesalpinia sappan. Kor. J. Food. Sci. Technol. 35: 680-683.
  38. Ueda, J. Y., Y. Tezuka, A. H. Banskota, Q. L. Tran, Q. K. Tran, Y. Harimaya, I. Saiki, and S. Kadota. 2002. Antiproliferative activity of Vietnamese medicinal plants. Biol. Pharm. Bull. 25: 753-760. https://doi.org/10.1248/bpb.25.753
  39. Woldemichael, G. M., M. P. Singh, W. M. Maiese, and B. N. Timmermann. 2003. Constituents of antibacterial extract of Caesalpinia paraguariensis Burk. Z. Naturforsch. 58: 70-75.
  40. Wu, M., E. Maier, R. Benz, and R. E. Hancock. 1999. Mechanism of interaction of different classes of cationic antimicrobial peptides with planar bilayers and with the cytoplasmic membrane of Escherichia coli. Biochemistry. 38: 7235-7242. https://doi.org/10.1021/bi9826299
  41. Xie, Y. W., D. S. Ming, H. X. Xu, H. Dong, and P. P. But. 2000. Vasorelaxing effects of Caesalpinia sappan Involvement of endogenous nitric oxide. Life Sci. 67: 1913-1918. https://doi.org/10.1016/S0024-3205(00)00772-4
  42. You, E. J., L. Y. Khil, W. J. Kwak, H. S. Won, S. H. Chae, B. H. Lee, and C. K. Moon. 2005. Effects of brazilin on the production of fructose-2,6-bisphosphate in rat hepatocytes. J. Ethnopharmacol. 102: 53-57. https://doi.org/10.1016/j.jep.2005.05.020
  43. Zhao, H., H. Bai, Y. Wang, W. Li, and K. A. Koike. 2008. New homoisoflavan from Caesalpinia sappan. Nat. Med. 62: 325-327. https://doi.org/10.1007/s11418-008-0231-6