Characteristics of Crude Protein-bound Polysaccharide from Agaricus blasei Murill by Extraction and Precipitation Conditions and Its Antitumor Effect

추출 및 침전조건에 따른 아가리쿠스버섯에서 분리한 조단백다당류의 특성 및 암세포 증식억제 효과

  • Hong, Joo-Heon (Department of Food Science and Technology, Kyungpook National University) ;
  • Youn, Kwang-Sup (Faculty of Food Science and Industrial Technology, Catholic University of Daegu) ;
  • Choi, Yong-Hee (Department of Food Science and Technology, Kyungpook National University)
  • 홍주헌 (경북대학교 식품공학과) ;
  • 윤광섭 (대구가톨릭대학교 식품산업학부) ;
  • 최용희 (경북대학교 식품공학과)
  • Published : 2004.08.31

Abstract

Efficient extraction method of crude protein-bound polysaccharide (CPBP) from Agaricus blasei Murill was established. CPBP yields by ultrasonic and hot water extractions were 13.0 and 7.8%, respectively. Pressure extraction for 3 hr gave the highest ${\beta}-glucan$ content; no significant difference was observed between 2 and 3 hr extraction. Four volumes added ethanol gave the highest yields of CPBP and ${\beta}-glucan$ contents at 10.89 and 35.97%, respectively. Decomposition temperature of CPBP was $240-365^{\circ}C$, showing relatively good thermal stability. In SRB (sulforhodamine B) assay, CPBP treatment at $1,000\;{\mu}g/mL$ for 72 hr inhibited proliferations to A549, MCF-7, and AGS cancer cells by 43.9, 21.4, and 32.5%, respectively.

아가리쿠스버섯에 함유된 생리활성 물질인 조단백다당류의 효율적인 추출공정을 확립하기 위하여 다양한 추출조건에서의 화학적 특성을 조사하였으며, 추출 분리된 조단백다당류의 열분해 특성 및 SRB assay에 의한 인체 암세포 증식억제 효과를 조사하였다. 조단백다당류의 추출수율은 초음파추출이 13.0%로 가장 높았으며, 열수추출이 7.8%로 가장 낮았다. 추출된 조단백다당류의 당 조성은 약 80%의 glucose와 fucose, galactose 및 mannose로 구성되어 있었으며 가압조건에서 2시간과 3시간의 추출은 ${\beta}-glucan$ 함량이 각각 32.28%와 32.34%로 뚜렷한 차이가 발견되지 않아 가압추출 2시간이 생리활성물질의 추출조건으로 효과적이었다. 또한 침전조건에 따른 조단백다당류의 침전수율 및 ${\beta}-glucan$ 함량은 4배 첨가 시 각각 10.89%와 35.97%로 가장 많이 함유되어 있었다. 열분해 특성은 네 단계로 이루어져 있었으며, $240-365^{\circ}C$에서 67.8%의 무게 감소를 보여주어 높은 온도에서도 열적 안정성이 있었다. SRB assay에 의한 인체 암세포 증식억제 효과를 검토한 결과, A549의 경우 $1,000{\mu}g/mL$ 농도에서 72시간 처리 시에는 43.9%의 암세포 손상을 보여주어 MCF-7과 AGS에 비해 암세포 증식억제 효과가 높음을 확인할 수 있었다.

Keywords

References

  1. Misuno T. Antitumor activity and some properties of water soluble polysaccharides from fruiting body of Agaricus blasei Murill. Agric. Biol. Chem. 54: 2889-2896 (1990) https://doi.org/10.1271/bbb1961.54.2889
  2. Hirokazu K, Ryuichi I, Teturo K, Takashi M. Fractionation and antitumor activity of the water-in-soluble residue of Agricus blasei fruiting bodies. Carbohydr. Res. 186: 267-273 (1989) https://doi.org/10.1016/0008-6215(89)84040-6
  3. Sasaki T, Takasuka N. Further studies of the structure of lentinan, An antitumor polysaccharides from Lentinus edodes. Carbohydr. Res. 47: 99-106 (1976) https://doi.org/10.1016/S0008-6215(00)83552-1
  4. Komatsu N, Okubo S, Kikumoto S, Kimura K, Saito G, Sasaki S. Host mediated antitumor action of Schizophyllum commune. Gann. Jpn. J. Cancer Res. 60: 557-563 (1971)
  5. Tsugagoshi S, Ohash F. Protein-bound polysaccharides preparation, PS-K, effective against sarcoma 180 and rat asites hepatoma AH-13 by use. Gann. Jpn. J. Cancer Res. 65: 557-565 (1974)
  6. Regina CR, Notoya M, Mario SM. Antimutagenic effects of the mushroom Agaricus blasei Murill extracts on V79 cells. Mutation Res. 496: 5-13 (2001) https://doi.org/10.1016/S1383-5718(01)00227-3
  7. Hirokazu K, Aya N, Takayuki Y, Takashi M. Isolation and prorerties of a lectin from the fruiting bodies of Agaricus blasei. Carbohydr. Res. 183: 150-154 (1988) https://doi.org/10.1016/0008-6215(88)80057-0
  8. Nakajima A, Ishida T, Koga M, Takeuchi M. Effect of hot water extract from Agaricus blasei Murill on antibody-producing cells in mice. Int. Immunopharmacol. 2: 1205-1211 (2002) https://doi.org/10.1016/S1567-5769(02)00056-5
  9. Hui LC, Guei RC, Chin CC, Jeng LM. Non-volatile taste components of Agaricus blasei, Antrodia camphorata and Cordyceps militaris mycelia. Food Chem. 74: 203-207 (2001) https://doi.org/10.1016/S0308-8146(01)00127-3
  10. Mizuno M, Morimoto M, Minate K, Tsucjida H. Polysaccharides from Agaricus blasei stimulate lymphocyte T-cell subsets in mice. Biosci. Biotechnol. Biochem. 62: 434-437 (1998) https://doi.org/10.1271/bbb.62.434
  11. Qun D, Jian Y, Xiao Y, Jinian F. Structural characterization of a water-soluble $\beta-D-glucan$ from fruiting bodies of Agaricus blasei Murr. Carbohydr. Res. 337: 1417-1421 (2002) https://doi.org/10.1016/S0008-6215(02)00166-0
  12. Mizuno T, Inagaki R, Kanto T, Hagiwara T, Nakamura T, Ito H, Shimura K, Sumiya T, Asakura A. Antitumor activity and some properties of water-insoluble polysaccharides from Himematsutake, the fruiting body of Agaricus blasei Murill. Agric. Biol. Chem. 54: 2897-2905 (1990) https://doi.org/10.1271/bbb1961.54.2897
  13. Maziero R, Bononi VL. Science and Cultivation of Edible Fungi. Balkema, Rotterdam, Netherlands. pp. 887-892 (1995)
  14. Cavazzoni V, Adami A. Exopolysaccharides produced by mycelial edible mushrooms. Ital. J. Food Sci. 1: 9-15 (1992)
  15. Hyun CK, Park KH, Kim YB, Yoon IH. Differential scanning calorimeter of rice starch. Korean J. Food Sci. Technol. 20: 331-337 (2000)
  16. Masayuki K, John WD. Calorimetric determination of the amylose content of starches based on formation and melting of the amylose-lysolecithin complex. J. Food Sci. 46: 765-771 (1981) https://doi.org/10.1111/j.1365-2621.1981.tb15344.x
  17. Papazisis KT, Geromichalos GD, Dimitriadis KA, Kortsaris AH. Optimization of the sulforhodamine B colorimetric assay. J. Immunol. Meth. 208: 151-158 (1997) https://doi.org/10.1016/S0022-1759(97)00137-3
  18. Mosman T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J. Immunol. Meth. 65: 55-63 (1983) https://doi.org/10.1016/0022-1759(83)90303-4
  19. Saha SK, Brewer CF. Determination of the concentrations of oligosaccharides, complex type carbohydrates, and glyco-proteins using the phenol-sulfuric acid method. Carbohydr. Res. 254: 157-167 (1994) https://doi.org/10.1016/0008-6215(94)84249-3
  20. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin-phenol reagents. J. Biol. Chem. 193: 265-275 (1954)
  21. Pamela M, Laura P. Beta-glucans in edible mushrooms. Food Chem. 68: 315-318 (2000) https://doi.org/10.1016/S0308-8146(99)00197-1
  22. Nelly B, Gustav AH. New method for quantitave determination od uronic acids. Anal. Biochem. 54: 484-489 (1973) https://doi.org/10.1016/0003-2697(73)90377-1
  23. SAS Institute, Inc. SAS User's guide. Statistical Analysis systems Institute, Cary, NC, USA (1993)
  24. Choi SH. Extraction and purification of physiologically active materials from Agaricus blasei fruiting bodies. MS thesis, Sogang Univ., Seoul, Korea (2000)
  25. Mason TJ, Paniwnyk L, Lorimer JP. The use of ultrasound in food technology. Ultrason. Sonochem. 3: 253-260 (1996) https://doi.org/10.1016/S1350-4177(96)00034-X
  26. Lee JH, Cho SM, Song KS, Han SB, Kim HM, Hong ND, Yoo ID. Immunostimulating activity and characterization of polysaccharides from Mycelium of Phellinus linteus. J. Microbiol. Biotechnol. 6: 213-218 (1996)
  27. Jong SC, Birmingham JM. Medicinal and therapeutic value of the shiitake mushroom. Adv. Appl. Microbiol. 39: 152-184 (1993)
  28. Kim GY, Park HS, Nam BH, Lee SJ, Lee JD. Purification and characterization of acidic proteo-heteroglycan from the fruiting body of phellinus linteus Teng. Biores. Technol. 89: 81-87 (2003) https://doi.org/10.1016/S0960-8524(02)00273-0
  29. Ballance GM, Manners DJ. Structural analysis and enzymic solubilization of barley endosperm cell walls. Carbohydr. Res. 61: 107-113 (1978) https://doi.org/10.1016/S0008-6215(00)84471-7
  30. Cho SM, Lee JH, Han SB, Kim HM. Immuno-stimulating polysaccharides from the fruiting bodies of Formitella fraxinea (II). Korean J. Mycol. 23: 340-347 (1995)
  31. Park KS, Lee JY, Lee SJ, Kim SH, Lee JS. Extraction and separation of protein-bound polysaccharide produced by Coriolus versicolor (Fr) Quel. Korean J. Mycol. 20: 72-76 (1992)
  32. Chen HK. Studies on the characteristics of taste-active components in mushroom concentrate and its powderization. MS thesis. National Chung-Hsing Univ. Taichung, Taiwan (1986)
  33. Choi JM, Koo SJ. Effect of $\beta$-glucan from Agaricus blasei Murill on blood glucose and lipid composition in db/db mice. Korean J. Food Sci. Technol. 32: 1418-1425 (2000)
  34. Kim HH, Park SK, Kim HS. Preparation of activated coffee char by KOH activation. Appl. Chem. 2: 1048-1051 (1998)
  35. Abdellatif AM, Patricia RD. The effect of mixing and wheat protein/ gluten on the gelatinization of wheat starch. Food Chem. 81: 533-545 (2003) https://doi.org/10.1016/S0308-8146(02)00487-9
  36. Sharma HS. Analysis of the components of lignocellulose degraded by Agaricus bisporus and Pleurotus ostreatus. Thermochim. Acta. 173: 241-252 (1990) https://doi.org/10.1016/0040-6031(90)80610-B
  37. Chun HS, Choi EH, Kim HJ, Choi CW, Hwang SJ. In vitro and in vivo antitumor activities of water extracts from Agaricus blasei Murill. Food Sci. Biotechnol. 10: 335-340 (2001)