Comparison of Extraction Methods and Separation and the Study on Purification of Crude Polysaccharides from Grifola frondosa

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1 13 1 Vol. 13 No Journal of Fungal Research Mar * 1 1 1** CO % CO % 69. 9% 61. 9% CO % 50. 1% CO 2 R DOI /j. jfr A J Comparison of Extraction Methods and Separation and the Study on Purification of Crude Polysaccharides from Grifola frondosa JIN Ling-yun 1 HUA Zheng-gen 1 LI Ye 1** YAO Wei-xi 2 1. Fujian Xianzhilou Biological Science and Technology Co. Ltd. Research Center of Medicinal Mushroom Cultivation and Deep Processing of Local and National United Engineering Fuzhou China 2. Research Center for Eco-Environmental Sciences Chinese Academy of Sciences Beijing China Abstract The supercritical CO 2 extraction process was compared with several conventional extraction methods in extracting the crude polysaccharide from Grifola frondosa in the meantime the methods of deproteinization and decolorization crude polysaccharide were studied. The results showed that the yield of crude polysaccharide by hot water method was the highest 28. 1% while the purity of crude polysaccharide 39. 3% was the highest when the supercritical CO 2 extraction method was used. Polyamide performed well in the remove of protein and pigment of Grifola frondosa crude polysaccharide and the removal rate of protein and pigment were 69. 9% and 61. 9% respectively with the low adsorption rate of polysaccharide. Under the optimal dosage of polyamide the yield and purity of Grifola frondosa polysaccharide by supercritical CO 2 method were 79. 3% and 50. 1% respectively. So polyamide method is a convenient efficient way for the isolation of Grifola frondosa crude polysaccharide. Key words Grifola frondosa polysaccharide supercritical CO 2 isolation and purification! S2011C !! lee@ xianzhilou. com

2 Grifola frondosa Dicks. Fr. S. F. Gray Basidiomycotina Hymenomycetes Holobasidiomyceidae Aphyllophorales Polyporaceae Grigola Grifolan β mm g2 500 ml W 9 30 min g2 500 ml 200 W 6 min 12 CO g2 500 ml 6 h CO kg 9 L 50% CO 2 2 / MPa h 2 h 6 MPa 40 CO L /h CO L /32 MPa CO V V = SIGMA Sevag 20 ml 1. 25% G Sevag V V = min r /min 10 min AR1140 XW - 80A ml 1. 25% TDL - 40B g UV h

3 G ml g 30 2 h 1 ml G ml 10 min595 nm = OD - OD / OD 100% = - / 100% / 100% 2. 4 = - / 100% = ml = / 5 g min 100% r /min 30 min Table 1. Comparison of Grifola frondosa crude polysaccharide yield and purity in different extracting methods Extraction methods /% Yield of polysaccharide /% Purity of crude polysaccharide Hot water extraction method ± 0. 4 aa ± 0. 7 bb Microwave extraction method ± 0. 5 aa ± 0. 4 cc Ultrasonic extraction method ± 0. 7 bb ± 0. 5 cc CO 2 Supercritical CO 2 extraction method 4. 5 ± 0. 2 cc ± 0. 9 aa Duncan Note The significant differences were showed by Duncan multiple comparisons % CO % CO % CO 2 CO % 39. 3% 1. 8 CO % ~ 21. 8% CO 2 CO 2

4 Sevag % % 2 Sevag Table 2. Effect of Sevag treatment number on the deproteinization of Grifola frondosa crude polysaccharide % Treatment number Removal rate of protein Adsorption rate of polysaccharide ± ± ± ± ± ± ± ± % 2. 5 g 69. 9% 16. 5% Sevag 1 Fig. 2. Effect of active charcoal dosage on the de- colorization of crude polysaccharide Fig. 1. Effect of polyamide dosage on the deproteinization of crude polysaccharide 3. 3 CO nm 297 nm 2 2 ~ 3 5 g 57. 9% % Fig. 3. Effect of polyamide dosage on the decolorization of crude polysaccharide 5 g < 38. 3% g 61. 9%

5 ml 20 g 79. 3% 50. 1% 4 21 DA201 CO 2 78% 2 85% 2 CO 2 D CO 2 DA201-C % 87. 8% 85% 10 CO 2 42% 4. 2 CO % 28. 1% 27. 2% 69. 9% 61. 9% 22. 6% 39. 3% 1. 8 Sevag J J Fang J P Wang Y Lv X F Shen X K Ni X Y Ding K. Structure of a β-glucan from Grifola frondosa and its antitumor effect C O NH 18 by activating Dectin-1 /Syk /NF-κB signaling J. Glycoconjugate Journal / Soares R Meireles M Rocha A Pirraco A Obiol D Alonso E Sevag Joos G Balogh G. Maitake D fraction mushroom extract induces apoptosis in breast cancer cells by BAK-1 gene activation J. Journal of Medicinal Food Gu C Q Li J W Chao F H. Inhibition of hepatitis B virus by D- fraction from Grifola frondosa Synergistic effect of combination % Sevag - Sevag 20 6 Yang B K Gu Y A Jeong Y T Jeong H 80% tion process J. International Journal of Biological Macromole- cules with interferon-α in HepG J. Antiviral Research Song C H. Chemical characteristics and immuno-modulating activities of exo-biopolymers produced by Grifola frondosa during submerged fermenta

6 Kubo K Aoki H Nanba H. Anti-diabetic activity present in the fruit body of Grifola frondosa Maitake I J. Biological and Pharmaceutical Bulletin J J J CO J J. J J J D J J J J J J J 櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧櫧 CABI CA ZR AGRIS CSA 20 A http xuebao. jlau. edu. cn jlndxb@ vip. sina. com