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Immune Stimulation and Anti-Metastasis of Crude Polysaccharide from Submerged Culture of Hericium erinaceum in the Medium Supplemented with Korean Ginseng Extracts

수삼추출물 첨가 배지에서 배양된 노루궁뎅이버섯 균사체 심부발효물 조다당획분의 면역 및 암전이 억제활성

  • Kim, Hoon (Division of Food and Biotechnology, Chungju National University) ;
  • Park, Chang-Kyu (Division of Food and Biotechnology, Chungju National University) ;
  • Jeong, Jae-Hyun (Division of Food and Biotechnology, Chungju National University) ;
  • Jeong, Heon-Sang (Dept. of Food Science and Technology, Chungbuk National University) ;
  • Lee, Hyeon-Yong (Dept. of Biomaterials Engineering, Kangwon National University) ;
  • Yu, Kwang-Won (Division of Food and Biotechnology, Chungju National University)
  • 김훈 (충주대학교 식품생명공학부) ;
  • 박창규 (충주대학교 식품생명공학부) ;
  • 정재현 (충주대학교 식품생명공학부) ;
  • 정헌상 (충북대학교 식품공학과) ;
  • 이현용 (강원대학교 생물소재공학전공) ;
  • 유광원 (충주대학교 식품생명공학부)
  • Published : 2009.11.30

Abstract

To find the new use of Korean ginseng and mushroom, crude polysaccharides were prepared from submerged cultures of Hericium erinaceum in the medium supplemented with Korean ginseng extracts. When we fractionated crude polysaccharides (HE-GE-CP-1, 3, and 5) from hot-water extracts of submerged cultures of H. erinaceum with ginseng extracts (1%, 3%, and 5% addition of total medium), the yields of HE-GE-CP-1, 3, and 5 were identified at 5.7, 5.1, and 4.8%, respectively. Among crude polysaccharide fractions, HE-GE-CP-5 was significantly higher (1.89-fold of the saline control) than those of HE-GE-CP-1 (1.64-fold) or HE-GE-CP-3 (1.76-fold) on mitogenic activity of splenocytes. HE-GE-CP-5 also had the more potent bone marrow cell proliferation (1.83-fold) rather than HE-CP or HE-GE-CP-1 or HE-GE-CP-3 (1.59- or 1.44- or 1.69-fold, respectively), and anti-metastatic activity as anti-cancer effect showed the highest prophylactic value (72.4% inhibition of tumor control) in 5% supplementation of ginseng extract. However, the lysosomal phosphatase of macrophage was significantly stimulated after HE-GE-CP-3 treatment (2.03-fold). In addition, the immunostimulating and anti-metastatic crude polysaccharide, HE-GE-CP-5, contained mainly neutral sugars (63.2%) with considerable amounts of uronic acid (19.3%) and a small amount of proteins (8.8%). HE-GE-CP-5 can stimulate immune system to inhibit tumor metastasis, and its anti-tumor metastasis may be associated with macrophages, splenocytes and Peyer's patch cells activation.

수삼과 버섯의 새로운 용도를 개발하기 위하여, 수삼추출물이 첨가된 액체배지에서 배양된 노루궁뎅이 균사체 심부발효물의 열수추출물로부터 조다당획분을 조제하였다. 수삼추출물이 액체배지 전체양의 1%, 3%와 5% 첨가된 노루궁뎅이버섯 균사체 심부발효물의 열수추출물로부터 조다당획분(HE-GE-CP-1, 3과 5)이 분획되었을 때, 수율은 각각 5.7, 5.1과 4.8%이었다. 이러한 조다당획분 중 HE-GE- CP-5(saline 대조군의 1.89배)는 HE-GE-CP-1(1.64배)과 HE-GE-CP-3(1.76배)보다 유의적으로 높은 비장세포의 마이토젠 활성을 보여주었다. 또한 HE-GE-CP-5(1.83배)는 소장의 국소 면역조직인 Peyer's patch를 경유한 골수세포 증식활성에서도 HE-CP(1.59배), HE-GE-CP-1(1.44배)과 HE-GE-CP-3(1.69배)보다 높은 활성을 나타내었으며, 항암효과로서의 암전이 억제활성 측정에서는 수삼추출물이 5% 첨가된 조다당획분에서 72.4%의 높은 암전이 억제활성도 보여주었다. 그러나 복강 내 마크로파지의 lysosomal phosphatase의 활성측정에서는 HE-GE-CP-3에서 2.03배의 가장 높은 마크로파지 자극활성을 확인할 수 있었다. 한편, 이러한 면역 및 암전이 억제활성을 갖는 조다당획분인 HE-GE-CP-5는 중성당(63.2%)과 함께 상당량의 산성당(19.3%)을 함유하였으며 단백질은 8.8%가 구성분으로 분석되었다. 이러한 결과로부터 HE-GE-CP-5는 암전이를 억제하기 위한 면역계를 자극할 수 있음이 확인되었으며, 이러한 암전이 억제의 항종양 활성은 마크로파지, 비장세포 및 Peyer's patch cell 등의 활성화 기작과 연계되어 있음을 추정할 수 있었다.

Keywords

References

  1. Chang S.T. 1993. Mushrooms and mushroom biology. In Genetics and Breeding of Edible Mushrooms. Chang ST, Buswell JA, Miles PG, eds. Gordon & Breach Science Publisher, Philadelphia, USA. p 1-13
  2. Soccol C.R, Dalla Santa H.S, Rubel R, Vitola F.M, Leifa F, Pandey A. 2008. Mushrooms-A promising source to produce nutraceuticals and pharmaceutical byproducts. In Current Topics on Bioprocesses in Food Industry. Koutinas AA, Pandey A, Larroche C, Larroche A, eds. Asiatech Publishers Inc., New Delhi, India. p 439-448
  3. Kidd P.M. 2000. The use of mushroom glucans and proteoglycans in cancer mushroom. Alter Med Rev 5: 4-27
  4. Zhong J.J, Tang Y.J. 2004. Submerged cultivation of medicinal mushrooms for production of valuable bioactive metabolites. Adv Biochem Eng Biotechnol 87: 25-59
  5. Kim S.W, Hwang H.J, Park J.P, Cho Y.J, Song C.H, Yun J.W. 2002. Mycelial growth and exo-biopolymer production by submerged culture of various edible mushroom under different media. Lett Appl Microbiol 34: 56-61 https://doi.org/10.1046/j.1472-765x.2002.01041.x
  6. Sugihara T.F, Humfeld H. 1954. Submerged culture of the mycelium of various species of mushroom. Appl Microbiol 2: 170-172
  7. Yang X.M. 1988. The cultivation of edible mushrooms in China. Agriculture Press, Beijing, China
  8. Liu C.Y. 1981. Technique of cultivation of monkey head mushroom. Edible Fungi 4: 33-34
  9. Arnone A, Cardillo R, Nasini G, de Pava O.V. 1994. Secondary mold metabolites: Part 46. Hericenes A-C and erinapyrone C, new metabolites produced by the fungus Hericium erinaceum. J Nat Prod 57: 602-606 https://doi.org/10.1021/np50107a006
  10. Baek G.Y. 2001. Studies on the growth characteristics and functional screening of mycelia and fruiting body of Hericium erinaceum. MS Thesis. Chungju National University, Chungbuk, Korea
  11. Ha T.M, Ji J.H, Jeong H.K. 1996. A liquid cultivation of Hericium erinaceum hyphae using by-product of food, and health drink compositions containing hyphae extract thereof. Korean Patent 10-0187892
  12. Malinowska E, Krzyczkowski W, Herold F, Lapienis G, Slusarczyk J, Suchocki P, Kuras Turło J. 2009. Biosynthesis of selenium-containing polysaccharides with antioxidant activity in liquid culture of Hericium erinaceum. Enz Micro Technol 44: 334-343 https://doi.org/10.1016/j.enzmictec.2008.12.003
  13. Ko H.G, Park H.G, Park S.H, Choi C.W, Kim S.H, Park W.M. 2005. Comparative study of mycelial growth and basidiomata formation in seven different species of the edible mushroom genus Hericium. Bioresour Technol 96: 1439- 1444 https://doi.org/10.1016/j.biortech.2004.12.009
  14. Chung J.Y, Kim C.S. 2008. Antioxidant activities of domestic garlic (Allium sativum L.) stems from different areas. J Korean Soc Food Sci Nutr 37: 972-978 https://doi.org/10.3746/jkfn.2008.37.8.972
  15. Dubois M, Gilles K.A, Hamilton J.K, Rebers P.A, Smith F. 1956. Colorimetric method for determination of sugars and related substances. Anal Chem 28: 350-356 https://doi.org/10.1021/ac60111a017
  16. Blumenkrantz N, Asboe-Hansen G. 1973. New method for quantitative determination of uronic acid. Anal Biochem 54: 484-489 https://doi.org/10.1016/0003-2697(73)90377-1
  17. Bradford M.M. 1976. A rapid and sensitive method for the quantitation 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
  18. Sugawara I, Kimoto M, Fujimoto M, Ishizaka S, Tsuji T, Nishiyama T. 1984. MTT assay, rapid colorimetric assay applicable to cellular proliferation and cytotoxicity assay. Igakuno Ayumi 123: 733-735
  19. Yu K.W, Kim Y.S, Shin K.S, Kim J.M, Suh H.J. 2005. Macrophage-stimulating activity of exo-biopolymer from cultured rice bran with Monascus pilosus. Appl Biochem Biotechnol 126: 35-48 https://doi.org/10.1007/s12010-005-0004-6
  20. Hong T, Matsumoto T, Kiyohara H, Yamada H. 1998. Enhanced production of hematopoietic growth factors through T cell activation in Peyer's patches by oral administration of Kampo (Japanese herbal) medicine, "Juzen- Taiho-To". Phytomedicine 5: 353-360 https://doi.org/10.1016/S0944-7113(98)80017-2
  21. Page B, Page M, Noel C. 1993. A new fluorometric assay for cytotoxicity measurements in vitro. Int J Oncol 3: 473- 476
  22. Conrad R.E. 1981. Induction and collection of peritoneal exudates macrophages. In Manual of Macrophage Methodology. Herscowitz B.H, Holden H.T, Bellanti J.A, Ghaffar A, eds. Marcel Dekker Incorporation, New York, NY. p 5-11
  23. Suzuki I, Tanaka H, Kinoshita A, Oikawa S, Osawa M, Yadomae T. 1990. Effect of orally administered-glucan on macrophage function in mice. Int J Immunopharmacol 12: 675-684 https://doi.org/10.1016/0192-0561(90)90105-V
  24. Yoon T.J, Yoo Y.C, Kang T.B, Baek Y.U, Huh C.S, Song S.K, Lee K.H, Azuma I, Kim J.B. 1998. Prophylactic effect of Korean mistletoe (Viscum album coloratum) extract on tumor metastasis is mediated by enhancement of NK cell activity. Int J Immunopharmacol 20: 163-172 https://doi.org/10.1016/S0192-0561(98)00024-1
  25. Yang H.S, Yu K.W, Choi Y.M. 2004. Isolation of polysaccharides modulating mouse's intestinal immune system from peels of Citrus unshiu. J Korean Soc Food Sci Nutr 33: 1476-1485 https://doi.org/10.3746/jkfn.2004.33.9.1476
  26. Xavier-Santos S, Carvalho C.C, Bonfa M, Silva R, Capelari M, Gomes E. 2004. Screening for pectinolytic activity of wood-rotting basidiomycetes and characterization of the enzymes. Folia Microbiol (Praha) 49: 46-52 https://doi.org/10.1007/BF02931645
  27. Han J. 2003. Solid-state fermentation of cornmeal with the basidiomycete Hericium erinaceum for degrading starch and upgrading nutritional value. Int J Food Microbiol 80: 61-66 https://doi.org/10.1016/S0168-1605(02)00122-8
  28. Mowat A.M, Viney J.L. 1997. The anatomical basis of intestinal immunity. Immunol Rev 156: 145-166 https://doi.org/10.1111/j.1600-065X.1997.tb00966.x
  29. Trier J.S. 1991. Structure and function of intestinal M cells. Gastroenterol Clin North Am 20: 531-547
  30. Kadhim S, Penney C, Lagraoui M, Heibein J, Attardo G, Zacharie B, Connolly T, Gagnon L. 2000. Synergistic anti- tumor activity of a novel immunomodulator, BCH-1393, in combination with cyclophosphamide. Int J Immunopharmacol 22: 659-671 https://doi.org/10.1016/S0192-0561(00)00028-X
  31. Kudo C, Saito M, Yoshida T. 1995. Curative treatments of murine colon 26 solid tumors by immunochemotherapy with G-CSF and OK-432. Immunopharm 29: 235-243 https://doi.org/10.1016/0162-3109(95)00060-7
  32. Yoo Y.C, Saiki I, Sato K, Azuma I. 1994. MDP-Lys (L18), a lipophilic derivative of muramyl dipeptide, inhibits the metastasis of haematogenous and non-haematogenous tumors in mice. Vaccine 12: 175-180 https://doi.org/10.1016/0264-410X(94)90057-4
  33. Ha E.S, Hwang S.H, Shin K.W, Yu K.W, Lee K.H, Choi J.S, Park W.M, Yoon T.J. 2004. Anti-metastatic activity of glycoprotein fractionated from Acanthopanax senticosus, involvement of NK-cell and macrophage activation. Arch Pharm Res 27: 217-224 https://doi.org/10.1007/BF02980109
  34. Yoon T.J, Sung J.Y, Yu K.W, Lee H, Lee K.H. 2007. Induction of enhancement of anti-tumor immunity by polysaccharides fractionated from Acanthopanax senticosus. Kor J Pharmacogn 38: 117-122
  35. Yoon T.J, Yu K.W, Shin K.S, Suh H.J. 2008. Innate immune stimulation of exo-polymers prepared from Cordyceps sinensis by submerged culture. Appl Microbiol Biotechnol 80: 1087-1093 https://doi.org/10.1007/s00253-008-1607-y
  36. Fidler I.J. 1985. Macrophage and metastasis-a biological approach to cancer therapy. Cancer Res 45: 4714-4726

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