Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
권호 표지
DOI QR코드

DOI QR Code

Changes of Lectin Activity of Kidney Beans by floating and Fermentation

강낭콩의 열처리 및 발효에 의한 렉틴의 활성변화

  • Published : 2002.02.01
Download PDF
⟨ Previous Next ⟩

Abstract

Synthesis of new active protein was investigated by heat-treatment and fermentation of kidney beans with Bacillus subtilis ATCC 51189. The amount of water-soluble protein in raw kidney bean (raw protein, RP) was greatly reduced by heating (heated protein, HP) and several new amino acids were synthesized by fermentation. The molecular weights of proteins determined by SDS-PAGE were 118 kDa for RP and a new band of 18.0 kDa For protein (fermented protein, FP) in kidney beans heated and fermented with B. subtilis ATCC 51189. Hemagglutinating activities of RP, HP and FP were 128 HU, 4 HU and 32 HU respectively. Both of RP and FP showed anticancer activity against stomach cancer cell line (SNU-1) at 50 $\mu\textrm{g}$g/mL and lymphocyte stimulating activity at 1 $\mu\textrm{g}$/mL, and stimulated PBMC to secrete IFN-${\gamma}$ and IL-12. However, HP did not show any kinds of activities. Taken together, these results suggested that lectin in kidney beans was destroyed by heating, hut new active lectin-like Protein was derived by fermentation with B. subtilis ATCC 51189.

강낭콩을 가열한 후 B. subtilis ATCC 51189로 발효시켜 새로운 활성 단백질의 생성여부를 확인하였다. 생강낭콩의 수용성 단백질은 열처리에 의하여 감소하였으며, 발효과정에서 단백질의 양은 변화하지 않았으나, 새로운 아미노산이 합성되었다. 생강낭콩의 수용성 단백질(raw protein, RP)은 분자량 118 kDa의 PHA(phytohemagglutinin)의 전형적인 모습을 보였으나, 열처리한 단백질(heated protein, HP)의 경우, RP의 band는 사라지고, 저분자 peptide로 변화되었으며, 발효과정에 의하여 분자량 18.0 kDa의 새로운 단백질(fermented protein, FP)이 생성된 것을 확인하였다. 또한, RP는 128 HU, HP는 4HU, FP는 32HU의 적혈구응집효과를 보였으며, 위암세포(SNU-1)에 대하여 RP 및 FP는 비교적 높은 농도($IC_{50}$/ $\mu\textrm{g}$/mL)에서 항암활성을 보였다. 그리고, RP 및 FP의 경우 1 $\mu\textrm{g}$/mL에서 림프구 증식효과를 보였고, IFN-${\gamma}$, IL-12의 분비를 촉진하였다. 그러나 HP는 항암효과, 림프구 증식효과, cytokine의 분비촉진효과를 보이지 않았다. 따라서, 강낭콩 lectin은 가열에 의하여 활성물질이 파괴되나, B. subtilis에 의한 발효과정에서 새로운 cytokine 분비촉진물질이 생성되는 것을 확인할 수 있었다.

Keywords

References

  1. Stillmard, H. : Cell agglutinating and sugar specific proteins. Science, 117, 949-950 (1972)
  2. Peumans, W.J. and Van Damme, E.J.M. : Recent advances in the purification and characterization of plant lectins. In COST 98: Effects of Antinutrients on the Nutritional value of Legume Diets, Bardocz, S., Gelencser, E. and Pusztai, A. (eds.), European Commission, Brussels, Belgium, Vol. 1, p.1-7 (1998)
  3. Goldstein, I.J. and Poretz, R.D. : Isolation, physicochemical characterization and carbohydrate-binding specificity of lectins. In The Lectins: Properties, Functions, and Applications in Biology and Medicine, 1st ed., Liener, I.E., Saron, N. and Goldstein, I.J. (eds.), Academic Press, London, UK, p. 233-237 (1986)
  4. Kovacs, E. : Serum levels of IL-12 and the production of IFNgamma, IL-2 and IL-4 by peripheral blood mononuclear cells (PBMC) in cancer patients treated with Viscum album extract. Biomed. Pharmacother., 54, 305-310 (2000) https://doi.org/10.1016/S0753-3322(00)80052-9
  5. Pusztai, A. : Lectins and their specificity. In Plant lectins, Pusztai, A. (ed.), Cambridge University Press, Cambridge, UK, p.3-31 (1991)
  6. Pryme, I.F., Pusztai, A.J., Grant, G. and Bardocz, S. : Dietary phytohemagglutinin slows down the proliferation of a mouse plasmacytoma (MPC-11) tumor in Balb/c mice. Cancer Lett., 103, 151-155 (1996) https://doi.org/10.1016/0304-3835(96)04206-1
  7. Pryme, I.F., Grant, G. and Bardocz, S. : Switching between control and phytohemagglutinin-containing diets affects growth of Krebs II ascites cells and produces differences in the levels of putrescine, spermidine and spermine. Cancer Lett., 93, 233-237 (1995) https://doi.org/10.1016/0304-3835(95)03815-E
  8. Pryme, I.F., Pusztai, A.J., Grant, G. and Bardocz, S. : Dietary phytohemagglutinin slows down the proliferation of a mouse plasmacytoma (MPC-11) tumour in Balb/c mice. Cancer Lett., 103, 151-155 (1996) https://doi.org/10.1016/0304-3835(96)04206-1
  9. Pryme, I.F., Pusztai, A.J., Bardocz, S. and Ewen, S.W. : The induction of gut hyperplasia by phytohaemagglutinin in the diet and limitation of tumour growth. Histol. Histopathol., 13, 575-583 (1998)
  10. Pryme, I.F., Pusztai, A.J., Bardocz, S. and Ewen, S.W. : The growth of an established mourine non-Hodgkin lymphoma tumour is limited by switching to a phytohaemagglutinincontaining diet. Cancer Lett., 146, 87-91 (1999) https://doi.org/10.1016/S0304-3835(99)00242-6
  11. Ogawara, M., Utsugi, M., Yamazaki, M. and Sone, S. : Induction of human monocyte-mediated tumor cell killing by a plant lectin, wheat germ agglutinin. Japn. J. Cancer Res. (Gann), 76, 1107-1144 (1985)
  12. Ogawara, M., Sone, S. and Ogura, T. : Human aveolar macrophages: Wheat germ agglutinin-dependent tumor cell killing. Japn. J. Cancer Res. (Gann), 78, 288-295 (1987)
  13. Lafont, J., Rouanet, J.M., Gabrion J., Assouad, J.L., Zambonino, I.J.L. and Resancon, P. : Duodenal toxicity of dietary Phaseolus vulgaris lectins in the rat: An integrative assay. Digestion, 41, 83-93 (1988) https://doi.org/10.1159/000199736
  14. Rodhouse, J.C., Haugh, C.A., Roberts, D. and Gilbert, R.J. : Red kidney bean poisoning in the UK : an analysis of 50 suspected incidents between 1976 and 1989. Epidermiol. Infect., 105, 485-491 (1990) https://doi.org/10.1017/S095026880004810X
  15. Grant, G., Dorward, P.M., Buchan, W.C., Armour, J.C. and Pusztai, A. : Consumption of diets containing raw soya beans, kidney beans, cowpeas or lupin seeds by rats for up to 700 days: effects on body composition and organ weights. Br.J. Nutr., 73, 17-29 (1995) https://doi.org/10.1079/BJN19950005
  16. Kordas, K., Burghardt, B., Kisfalvi, K., Bardocz, S., Pusztai, A. and Varga, G. : Diverse effects of phytohaemagglutinin on gastrointestinal secretion in rats. J. Physiol. Paris, 94, 31-36 (2000) https://doi.org/10.1016/S0928-4257(99)00106-0
  17. Pusztai, A., Clarke, E.M.W., Grant, G. and King, T.P. : The toxicity of Phaseolus vulgaris lectins; nitrogen balance and immunochemical studies. J. Sci. Food and Agric., 32, 1037-1046 (1981) https://doi.org/10.1002/jsfa.2740321014
  18. Lee, S.K., Heo, S., Bae, D.H. and Choi, K.H. : Medium optimization for fibrinolytic enzyme production by Bacillus subtilis KCK-7 isolated from Korean traditional chungkookjang. Kor. J. Appl. Microbiol. Biotechnol., 26, 226-231 (1998)
  19. Lim, S.I., Kim, H.K. and Yoo, J.Y. : Characteristics of protease produced by Bacillus subtilis PCA 20-3 isolated from Korean traditional meju. Korean J. Food. Sci. Technol., 32, 154-160 (2000)
  20. SAS Institute : SAS/STAT User Guide. release 6.30 edition (1988)

Cited by

  1. Enhancement of Cyclo-His-Pro (CHP) Content from Soybean Fermented with Bacillus amyloliquefaciens CHP-12 and Its Anti-diabetic Effect vol.26, pp.1, 2011, https://doi.org/10.7841/ksbbj.2011.26.1.041
  2. Physicochemical characteristics of acorn tea by processing methods vol.23, pp.3, 2016, https://doi.org/10.11002/kjfp.2016.23.3.335