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가지 열매 lectin의 생화학적 성질

Biochemical Properties of Eggplant Fruit Lectin.

  • 발행 : 2008.03.31

초록

가지 열매로부터 neutral saline에 의한 추출, $(NH_4)_2SO_4$ 침전 및 Sephadex-G100을 이용한 affinity chromatography에 의해 분리한 lectin의 생화학적 특성을 연구하였다. Trypsin을 처리하지 않은 사람, 쥐와 토끼의 적혈구에서는 혈구 응집반응이 일어나지 않았으며, Trypsin을 처리한 사람, 쥐와 토끼의 적혈구 중에서는 쥐의 적혈구에서만 혈구 응집반응이 일어났다. SDS-PAGE에 의해 가지 lectin의 분자량을 확인한 결과, 19.3 kDa의 단일 band 임이 확인되었다. D-glucose를 포함하는 7개의 탄수화물은 100 mM 이하의 농도에서 lectin과 적혈구의 응집을 저해시키지 않았으므로, 탄수화물에 대한 특이성이 나타나지 않았다. 최적 반응온도 범위는 $10-20^{\circ}C$로서, $20-70^{\circ}C$에서 열에 대해 안정하였으며, 안정된 pH 범위는 pH 6.2-7.2로 확인되었다. $Ca^{2+},\;Co^{2+},\;Cu^{2+},\;Fe^{2+},\;Mg^{2+}$$Mn^{2+}$ 20 mM 이하의 농도에서는 lectin과 적혈구의 응집을 저해시키지 않았으므로, 금속이온에 대한 특이성이 나타나지 않았다

Biochemical characterization including hemagglutination of erythrocytes, molecular weight, optimum temperature, thermal stability, optimum pH, carbohydrate specificity, and inhibitory effect of metal ion were studied in lectin of eggplant (Solanum melongena L.) fruit prepared by ammonium sulfate fractionation and affinity chromatography. This lectin was agglutinated by trypsin-treated rat blood erythrocyte. The molecular weight of this lectin by SDS-PAGE was estimated to be approximately 19.3 kDa of a single band. This lectin has no activity by 7 carbohydrates containing D-glucose. The optimum range of temperature and pH were $10-20^{\circ}C$ and pH 6.2-7.2, respectively. This lectin was relatively stable at $20-70^{\circ}C$. And the activity of this lectin was not inhibited by $Ca^{2+},\;Co^{2+},\;Cu^{2+},\;Fe^{2+},\;Mg^{2+}$, and $Mn^{2+}$.

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참고문헌

  1. Bradford, M. M. 1976. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248-254. https://doi.org/10.1016/0003-2697(76)90527-3
  2. Buczkowska, H. 2000. Eggplant cultivation under cover- Climatic requirements. Owoce. 39, 12-19.
  3. Burger, M. M. 1967. Assays for agglutination with lectins. Proc. Natl. Acad. Sci. USA. 57, 359-367. https://doi.org/10.1073/pnas.57.2.359
  4. Chang, C. S., M. J. Oh and K. S. Roh. 1999. Purification and biochemical characterization of lectin from Viscum album. Kor. J. Biotechnol. Bioeng. 14, 578-584.
  5. Chung, S. R., K. H. Jeune, S. Y. Park and S. J. Jang. 1993. Toxicity and lectins constituents from the seed of Corus officinalis. Kor. J. Pharmacogn. 24, 177-182.
  6. Esteban, R. M., E. M. Molla, L. M. Robredo, F. J. Lopez-Andreu. 1992. Changes in chemical composition of eggplant fruits during development and ripening. J. Agr. Food Chem. 40, 998-1000. https://doi.org/10.1021/jf00018a017
  7. Gleddie, S., W. Keller, G. Setterfield. 1986. Eggplant, pp. 500-511, In Evans, D. A. and W. R. Sharp (eds.), Handbook of Plant Cell Culture, Vol. 3, Techniques for Propagation and Breeding. MacMillan, New York.
  8. Golestein, I. J., R. C. Hughes. M. Monsigny. T. Osawa, and N. Sharon. 1980. What should be called a lectin? Nature 285, 66-70.
  9. Gupta, N. and P. S. Srivastava. 1998. Purification and characterization of a lectin from seeds and cotyledonary callus of Zizyphus mauritiana, Plant Cell Reports, 17, 552-556. https://doi.org/10.1007/s002990050440
  10. Higuchi, M. and K. Iwai. 1985. Purification and some properties of the basic lectin from winged bean seeds. Agr. Biol. Chem. 49, 391-398. https://doi.org/10.1271/bbb1961.49.391
  11. Hinata, H. 1986. Eggplant (Solanum melongena L.), pp. 363-370, In Bajaj, Y. P. S. (ed.), Biotechnology in Agriculture and Forestry, Vol. 2, Springer, Berlin.
  12. Hirano, K., T. Teraoka, H. Yamanaka, A. Harashima, A. Kunisaki, H. Tahahashi and D. Hosokawa. 2000. Novel mannose-binding rice lectin composed of some isolectins and its relation to a stress-inducible salT gene. Plant Cell Physiol. 41, 258-267. https://doi.org/10.1093/pcp/41.3.258
  13. Kalloo, G. 1993. Eggplant (Solanum melongena), pp. 587- 604, In Kalloo, G. (ed.), Genetic Improvement of Vegetable Crops. Pergamon Press, Oxford.
  14. Kalsi, G., H. R. Das, C. R. Babu and R. H. Das. 1992.Isolation and characterization of a lectin from peanut roots. Biochem. Biophys. Acta. 1117, 114-119. https://doi.org/10.1016/0304-4165(92)90067-5
  15. Khan, R. 1979. Solanum melongena and its ancestral forms. pp. 629-636, In Hawkes, J., R. N. Lester and A. D. Skelding (eds.), The Biology and Taxonomy of Solanaceae. Academic Press, London.
  16. Kilpatrick, D. C. 1980. Purification and some properties of a lectin from the fruit juice of the tomato. Biochem. J. 185, 269-272. https://doi.org/10.1042/bj1850269
  17. Kim, H. S., S. Y. Son, S. Y. Hwang and B. S. Hong. 1999. Purification and chatacterization of the lectins from mushroom Flammulina velutipes. J. Kor. Soc. Agric. Chem. Biotechnol. 42, 304-309.
  18. Kuku, A. and O. B. Eretan. 2004. Purification and partial characterization of a lectin from the fresh leaves of Kalanchoe crenata (Andr.) Haw. J. Biochem. Mol. Biology. 37, 229-233. https://doi.org/10.5483/BMBRep.2004.37.2.229
  19. Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of head of bacteriophage T4. Nature, 227, 680-685. https://doi.org/10.1038/227680a0
  20. Lis, H. and N. Sharon. 1986. Lectins as molecules and as tools. Ann. Rev. Biochem. 55, 35-67. https://doi.org/10.1146/annurev.bi.55.070186.000343
  21. Maghil, E. P., D. P. Mercy, B. N. Renuka and S. S. Jeya. 2003. Purification and characterization of a sialic acid specific lectin from the hemolymph of the freshwater crab Paratelphusa jacquemontii. Eur. J. Biochem. 270, 4348-4355. https://doi.org/10.1046/j.1432-1033.2003.03828.x
  22. Moon, I. J., S. R. Chung and K. H. Jeune. 1995. Mitotic stimulation and cancer cell agglutination of the lectin from Lentinus edodes. Yakhak Hoeji. 39, 260-267.
  23. Nachbar, M. S., J. D. Oppebheim and J. O. Thomas. 1980. Isolation and characterization of a lectin from the tomato (Lycopersicon esculentum). J. Biol. Chem. 255, 2056-2061.
  24. Olsnes, S., F. Stirpe,, K. Sandvig and A. Phil. 1982. Isolation and characterization of viscumin, a toxic lectin from Viscum album L. (Mistletoe). J. Biol. Chem. 257, 13263-13270.
  25. Park, K. B., K. S. Lee, J. R. Kim and Y. J. Kim. 1981. Purification and characterization of a lectin obtained from banha (Pinellia ternata) roots. Kor. Biochem. 14, 137-147.
  26. Park, N. Y., S. P. Lee and K. S. Roh. 2007. Biochemical characterization of lectin isolated from cherry tomato fruit. J. Life Science. 17, 254-259. https://doi.org/10.5352/JLS.2007.17.2.254
  27. Park, W. B and J. Y. Park. 1999. Isolation and characterization of lectin from Aloe vera. Kor. J. Food. Sci. Technol. 31, 899-905.
  28. Roh, K. S. 2007. Biochemical characterization of lectin purified from kidney bean seedling. Kor. J. Biotechnol. Bioeng. 22, 53-57.
  29. Roh, K. S. and D. J. Lee. 2002. Purification and some properties of lectin from Canavalia ensiformis L. Kor. J. Biotechnol. Bioeng. 17, 484-489.
  30. Roh, K. S. and N. Y. Park. 2005. Characterization of the lectin purified from Canavalia ensiformis shoots. Biotechnol. Bioproc. Engineer. 10, 334-340. https://doi.org/10.1007/BF02931851
  31. Sharon, N. and H. Lis. 1972. Lectins: Cell-agglutinating and sugar-specific proteins. Science 177, 949-959. https://doi.org/10.1126/science.177.4053.949
  32. Sharon, N. 1977. Lectins. Sci. American 236, 108-119. https://doi.org/10.1038/scientificamerican0677-108
  33. Sharon, N. and H. Lis. 1987. Century of lectin research (1888-1988). Trends Biochem. Sci. 12, 488-491. https://doi.org/10.1016/0968-0004(87)90236-2
  34. Takatsy, G. 1967. The use of a microtitrator in serological procedures. In International Symposium on Immunological Method of Biological Study. 4, 275-280.
  35. Utarabhand, P. and P. Akkayanont. 1995. Purification of a lectin from Parkia javanica beans. Phytochem. 38, 281-285. https://doi.org/10.1016/0031-9422(94)00550-D
  36. Wagner, H., E. Jordan, and B. Feil. 1986. Studies on the atandardization of mistletoe prepartions. Oncology 43, 16-22. https://doi.org/10.1159/000226416
  37. Walkins, A. H. 1972. Carbohydrates in cell recognition. Sci. American 22, 74-81.
  38. Weber, K. and M. Osborn. 1969. The reliability of molecular weight determination by dodecyl-sulfate polyacrylamide gel electrophoresis. J. Biol. Chem. 244, 4406-4412.
  39. Zhang, W., W. J. Peumans, A. Barre, C. H. Astoul, P. Rovira, P. Rouge, P. Proost, P. Truffa-Bachi, A. A. H. Jalali and E. J. M. Van Damme. 2000. Isolation and characterization of a jacalin-related mannose-binding lectin from salt-stressed rice (Oriza sativa) plants. Planta 210, 970-978. https://doi.org/10.1007/s004250050705
  40. Ziska, P., H. Franz and K. Kindt. 1978. The lectin from Viscum album L. purification by biospecific affinity chromatography. Experientia 34, 123-124. https://doi.org/10.1007/BF01921941

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