Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
권호 표지

Isolation of Polyphenol Compounds from the Leaves of Korean Persimmon (Diospyrus kaki L. Folium)

한국산 감잎로부터 Polyphenol계 생리활성물질 분리

  • An, Bong-Jeun (Faculty of Life Resources and Engineering, Kyungsan University) ;
  • Bae, Man-Jong (Faculty of Life Resources and Engineering, Kyungsan University) ;
  • Choi, Hee-Jin (Department of Food Science & Technology Yeungnam University) ;
  • Zhang, Yun-Bin (Department of Food Science & Technology Yeungnam University) ;
  • Sung, Tae-Soo (Korea, Department of Food & Nutrition, Changwon Junior College) ;
  • Choi, Cheong (Department of Food Science & Technology Yeungnam University)
  • 안봉전 (경산대학교 생명자원공학부) ;
  • 배만종 (경산대학교 생명자원공학부) ;
  • 최희진 (영남대학교 생물산업공학부) ;
  • 장운빈 (영남대학교 생물산업공학부) ;
  • 성태수 (창원전문대학 식품영양과) ;
  • 최청 (영남대학교 생물산업공학부)
  • Published : 2002.11.30
Download PDF
⟨ Previous Next ⟩

Abstract

We purified polyphenols from persimmon leaf and tested their biological activity. The 60% acetone extract was lyophilized and applied to test enzyme inhibition of glucosyltransferase and tyrosinase. GTase was 82.4% inhibited at $1.8{\times}10^{-1}$ mg/ml and tyrosinase 21.7% inhibited at 0.8 mg/ml. The acetone extract was fractionated into F-1, 2, 3, 4, 5 by Sephadex Q-50 gel filtration and the fraction-1 and 2 showed higher enzyme inhibition activity than the other fractions. To the Proteinase K treatment and autoclaving of the two fractions had no effect on the enzyme activity, but these results suggested that active fraction was not protein but phenol ring completed compounds. By Sephadex LH-20, MCI-gel and Bondapak $C_{18}$ column chromatographies, compouds 1, 2, 3 and 4 from F-1 fraction, compounds 5 and 6 from F-2 fraction and compounds 7 , 8 from F-3 fraction were purified and re-crystallized. The purified compounds was assumed to be condensed tannins of frame flavan-3-ol frame on the basis of color reagent reaction and to be a mixture of monomer, dimer and trimer according to TLC analysis.

옛부터 민간 약제로 사용했던 감나무 잎을 식품, 화장품 기능 생물 신소재로 활용하기 위해 감나무잎의 주성분으로 알려진 폴리페놀 화합물을 분리하고 생리활성 기능을 조사하였다. 감나무 잎을 60%의 아세톤으로 추출한 용액을 동결 건조시켜 농도별로 조제하고 glucosyltransferase, tyrosinase 효소저해를 관찰한 결과 glucoslytransferase는 $1.8{\times}10^{-1}$ mg/ml에서 82.84% 저해를, tyrosinase는 0.8 mg/ml에서 21,65% 저해 활성이 확인되었다. Sephadex G-50으로 gel filtration한 결과 fraction 1, 2, 3, 4, 5로 분획되었고 이중 fraction 1, 2에서 효소 저해 활성을 나타내었다. 활성 분획물의 물질군을 확인하기 위해 proteinase K 처리와 멸균처리한 결과 효소저해에 영향은 없어 활성물질군은 단백질군이 아니며, UV spectrum에서 자외선 영역에 최대 흡광치를 보여주므로 페놀링을 갖는 화합물로 추정하게 되었다. 추정화합물을 Sephadex LH-20, MCI-gel 및 Bondapak $C_{18}$ column에서 분리한 결과 F1 fraction에서 compound $1{\sim}4$가지 4종류의 화합물, F2 fraction에서는 compounds 5, 6, F3 fraction에서 compound 7, 8, 9를 순수 정제하여 재결정하였다. 분리된 화합물은 TLC 발색반응에서 flavan-3-ol 골격을 기본으로 하는 축합탄닌 반응을 보였고 전개정도에서 monomer가 2종류, dimer가 3종류, trimer가 1종류로 확인되었다.

Keywords

References

  1. Chung, S. H., Moon, K. D., Kim, J. K., Seong, J. H. and Sohn, T. H. (1994) Change of chemical components in persimmon leaves dunng growth for processing persimmon leaves tea. Korean J. Food Sci. Technol. 26, 141-146
  2. Choi, H. J., Son, J. H., Woo, H. S., An, B. J., Bae, M. J. and Choi, C. (1998) Changes of composition in the species of persimmon leaves during growth. Korean J. Food Sci. Technol. 30, 529-534
  3. Choi, S. H. (1990) The aroma components of duchung tea and persimmon leaf tea. Korean J. Food Sci. Technol. 22, 405-410
  4. Kim, M. J. and Oh, S. L. (1999) Effect of pre-treatment methods on the quality improvement of persimmon leaf tea. Korean J. Postharvest Sci. Tectmol. 6, 435-441
  5. Choi, S. J., Jun, W. J., Yu, K. W., Shin, D. H., Hong, B. S., Cho, H. Y. and Yang, H. C. (2000) PuriScadon and characterization of angiotensin I converdng enzyme inhibitor from porphura yezoensis. J. Korean Soc. Food Sci. Nutr. 29, 719-725
  6. Park, J. Y, Park, E. M., Lee, M. K., Jang, J. Y, Kim, M. J. and Cho, S. Y. (2000) Effect of persimmon leaves extract on serum and liver lipid concentradons in hypercholesterolemic rats. J. Korean Soc. Food Sci. Nutr. 29, 537-542
  7. Kim, 0. K. (2001) Protective effects of extracts of Diospyrus kaki Folium aganist hepatotoxicity in carbon tetra chlodde intoxicated rats. J. Korean Soc. Food Sci. Nutr. 30, 97-101
  8. Song, H. S., Lee, H. K. and Kang, M. H. (2000) Antimutagenic effects of persimmon leaf tea extract (PLTE) in mice using micronucleus & induction (MN) test. J. Korean Soc. Food Sci. Nutr. 29, 881-887
  9. Uchida, S., Ohta, H., Niwa, M., Mon, A., Nonaka, G., Nishioka, I. and Ozaki, M. (1990) Prolongadon of life span of stroke-prone spontaneously hypertensive rats (SHRSP) ingesting persimmon tannin. Chem. Pharm. Bull. 38, 1049-1054 https://doi.org/10.1248/cpb.38.1049
  10. Okonogi, T., Hatton, Z., Ogiso, A. and Mitsui, S. (1970) DetoxiScation by persimmon tannin of snake venoms and bacteiial toxins. Toxicon. 17, 524-529 https://doi.org/10.1016/0041-0101(79)90287-3
  11. Nose, K. (1984) Inhibidon by flavonoids of RNA synthesis in permeable WI-38 cells and of transcription by RNA polymerase II. Biochem. Pharm. 33, 3823-3828 https://doi.org/10.1016/0006-2952(84)90046-7
  12. Kim, B. G., Rhew, T. H., Choe, E. S., Chung, H. Y, Park, K. Y. and Rhee, S. H. (1933) Effect of selected persimmon leaf components against sarcoma 180 induced tumor in mice. J. Korean soc. Food Nutr. 22, 334-340
  13. Kameda, K., Takaku, T, Okuda, H. and Kimura, Y. (1978) Inhibitory effects of various flavonoids isolated from leaves of persimmon on angiotensin-converting enzyme activity. J.Naturat Pmducts 50, 680-686
  14. Moon, S. H., Kim, J. 0., Rhee, S. H,, Park, K. Y, Kim, K. H. and Rhew, T. H. (1993) Antimutagenic effects and compounds identified from hexane fraction of persimmon leaves. J. Korean Soc. Food Nutr. 22, 307-402
  15. Moon, S. H., Kim, J. 0. and Rhee, S. H. (1993) Antimutagenic effects and compounds identified from hexane fraction of persimmon leaves. J. Korean Soc. Food Nutr. 22, 307-312
  16. Kim, B. G., Rhew, T. H., Choe, E. S., Chung, H. Y, Park, K. Y. and Rhee, S. H. (1990) Effect of selected persimmon leaf components against spontaneosly hypertensive rats (SHRSP) ingesting persimmon tannin. Chem. Pharm. Bull. 38, 1049-1054 https://doi.org/10.1248/cpb.38.1049
  17. Funayama, S. and Hikino, H. (1979) Hypotensive principles of Diospyros kaki leaves. Chem. Pharm. Bull. 27, 2865-2870 https://doi.org/10.1248/cpb.27.2865
  18. Osawa, T. and Namiki, M. (1981) A novel type of antioxidant isolated from leaf wax of Eucalyptus leaves. Aeric. Biol. Chem. 45, 735-740
  19. Uchida, S., Edamatsu, R., Hiramatsu, M., Moii, A., Nonaka, G.Y, Nishioka, I., Niwa, M. and Ozaki, M. (1987) Condensed tannins scavenge active oxygen free radicals. Med. Sci. Res. 15, 831-836
  20. Hashimoto, F, Nonaka, G. and Nishioka, I. (1989) Novel Chalcan-flavan Dimers, Assamicans A, B and C and a New Flavan-3-ol and Proanthocyanidins from the Fresh Leaves of Camollia sinensis L. Chem. Pharm. Bull. 37, 77-8 https://doi.org/10.1248/cpb.37.77
  21. Endo, T., Tanaka, 0. and Shibata, S. (1971) Studies on glucosyltranferase inhibitor from Aspergillus terrus. Syotakagaku Zasshi. 25, 28-32
  22. Yagi, K. (1987) Lipid peroxides and human disease. Chem. Phys. Lipids. 45, 337-341 https://doi.org/10.1016/0009-3084(87)90071-5
  23. Hirose, M., Hoshiya, T, Takahashi, S., Hara, Y. and Ito, N. (1991) Inhivition of carcinogenesis by green tea catechin in rats. Proceedings of the Intemational Symposium on Tea Science (Japan). 210-215
  24. Ikeda, I., Imasato, Y, Sasaki, S., Nakyama, M., Nagano, H., Takeo, T., Yayave, F. and Sugano, M. (1995) Tea catechins decrease micellar solubility and intestinal absorption of cholesterol in rats. Proeceedings of the Intemational Symposium on Tea Science (Japan). 215-221
  25. Muramatsu, K., Sugiyama, K., Amano, S., Nakashima, J. and Saeki, S. (1995) Effect of green tea on cholesterol metabolism in rats. Proceedings of the Intemational Symposium on Tea Science (Japan). 220-226
  26. Choi, J. S., Park, S. H. and Choi, J. H. (1989) Nithte scavenging effect by flavonoids and its structure effect relationship. Arch. Pharm. Res. 12, 26-31 https://doi.org/10.1007/BF02855742
  27. Matsuo, T. and Ito, S. (1978) The chemical structure of kaki-tannin from tmmature fruit of the persimmon (Diospyros kaki L.). Agric. Biol. Chem. 42, 1637-3642