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Antitumor-active Polysaccharides Isolated from the Fruiting Body of Hericium erinaceum, an Edible and Medicinal Mushroom Called yamabushitake or houtou a
a
ab
Takashi Mizuno , Tetsuya Wasa , Hitoshi Ito , Chiharu Suzuki
ac
ad
& Nobuo Ukai
a
Faculty of Agriculture, Shizuoka University, Ohya, Shizuoka 422, Japan
b
School of Medicine, Mie University, Tsu, Mie 514, Japan
c
Fuji Biocenter, Fuji Seito Co., Ltd., Shibakawa-cho, Shizuoka 419–03, Japan
d
Research Institute, Kagome Co., Ltd., Nishinasuno-cho, Tochigi 329–27, Japan Published online: 12 Jun 2014.
To cite this article: Takashi Mizuno, Tetsuya Wasa, Hitoshi Ito, Chiharu Suzuki & Nobuo Ukai (1992) Antitumor-active Polysaccharides Isolated from the Fruiting Body of Hericium erinaceum, an Edible and Medicinal Mushroom Called yamabushitake or houtou, Bioscience, Biotechnology, and Biochemistry, 56:2, 347-348, DOI: 10.1271/bbb.56.347 To link to this article: http://dx.doi.org/10.1271/bbb.56.347
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http:// www.tandfonline.com/page/terms-and-conditions
Biosci. Biotech. Biochem., 56 (2), 347-348, 1992
Note
Antitumor-active Polysaccharides Isolated from the Fruiting Body of Hericium erinaceum, an Edible and Medicinal Mushroom Called yamabushitake or houtout Takashi
MIZUNO,
Tetsuya
WASA,
Hitoshi
ITO, *
Chiharu
SUZUKI, **
and Nobuo
UKAI***
Downloaded by [New York University] at 13:22 05 June 2015
Faculty of Agriculture, Shizuoka University, Ohya, Shizuoka 422, Japan *School of Medicine, Mie University, Tsu, Mie 514, Japan ** Fuji Biocenter, Fuji Seito Co., Ltd., Shibakawa-cho, Shizuoka 419--03, Japan *** Research Institute, Kagome Co., Ltd., Nishinasuno-cho, Tochigi 329-27, Japan Received July 31, 1991
Yamabushitake, or Hericium erinaceum, is an edible mushroom fungus belonging to Aphyllophorales, Hydnaceae (Hericiaceae), Hericium. It grows widely in Japan and the whole of China. In recent years, Hericium erinaceum fungus has been cultivated by wood-log culture (Photo. 1) or culture-bed methods such as bottleculture and bag-culture. Several newly isolated compounds from this mushroom have shown interesting biological activities such as cytotoxic effects on HeLa cells,
promotion of the synthesis of neuro-growth factor, and inhibition of pollen tube germination, and moreover the fruiting body of Hericium erinaceum, or its liquid-cultured mycelia, or an alcohol-precipitated fraction of its culture filtrate showed antitumor activities based on immunopotentiation. 1- 5) Previoulsy, we have reported some antitumor polysaccharides obtained from several fungi belonging to Polyporaceae and Agaricaceae. 6 - 8) In this note, we describe the evaluation of antitumor activities of the polysaccharides fractionated from the fruiting bodies of yamabushitake mushroom. Table I. Antitumor Activities of Some Polysaccharides Isolated from the Fruiting Bodies of Hericium erinaceum Polysaccharide Control FIo-a FIo-a-cc Fl o-a-f1 FIo·b FII-l FIII-2b
Photo 1. Typical Fruiting Bodies ofYamabushitake, Hericium erinaceum cultivated in Wood-log Culture. Table II.
F1o-a FIo-a-cc FI o-a-f1 FIo-b FII-l FIII-2b
b
e d
e
0 40.3 65.9 73.0 63.8 67.9 75.9
Complete regression
Mortality
0/10 1/7 3/7 4/7 2/7 1/7 3/7
10/10 3/7 2/7 1/7 3/7 1/7 0/7
1.00 1.54 1.78 1.86 1.71 1.81 1.75
Note: In four-week old female ICR/Slc mice, 6 x 106 cells/mouse of 7-day-old Sarcoma 180 from an ascites tumor were implanted subcutaneously in the right groin and 10 mg/kg of fraction from Flo were injected i.p. once daily for 10 consecutive days starting 24 hours after tumor implantation. The tumor size was measured at 28 days after tumor implantation. The survival time and mortality were measured 61 days after tumor implantation.
Some Properties of Antitumor-active Polysaccharides Isolated from the Fruiting Bodies of H. erinaceum
Polysaccharide
a
Inhibition Survival time (%) (T/C)
Molecular weight
Protein (%)b
Total sugar (%)C
118,000 89,000 175,000 565,000 155,000 30,000
34.2 51.5 4.2 22.1 5.7 20.2
53.4 40.5 89.7 71.4 91.4 75.7
Q
Components sugar" Gle
Xyl
Man
Gal
100
333 100 233 104 179 51
34 4 58 10
4
100 100 100 100
13 15 17 32
IR peak cm- le 880 868 889 893 893 899
Gel filtration by using a Toyopearl HW-65 column. Modified Lowry method. Phenol-sulfuric acid colorimetry. Each polysaccharide was hydrolyzed with 0.5 to 1.0 M sulfuric acid at 100°C for 3 to 6 hours and the hydrolyzate thus obtained was treated by ppc. Further, its alditol acetate derivative was identified and measured by GC method with the use of a Shimadzu GC-4CM -PF apparatus. KBr tablet method by using JIR-RFX-3001 apparatus of Nippon Bunko.
t Studies on the Host-mediated Antitumor Polysaccharides. Part XVII. For Part XVI, see ref. 6.
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et al.
MIZUNO
Table III. Solid State 13C-NMR and lH-NMR Spectral Chemical Shifts for Water-insoluble Autitumor Polysaccharides Isolated from Yamabushitake Mushroom 13C Shift (ppm) Polysaccharide
C-l
C-3 2
Downloaded by [New York University] at 13:22 05 June 2015
FI o-a-f3 FI o-b-f3 FII-l FIII-2b
103.3 103.1 108.1 104.2
91.0
4 85.8 85.6 81.3 87.5
78.3
Fruiting bodies of the fresh mushroom were extracted stepwisely with 85% ethanol (30°C, 24 hr), hot water (lOO°C, 3 hr), 1% ammonium oxalate (100°C, 5 hr), and 5% sodium hydroxide (30°C, 12-24 hr) as described previously.6) The hot water extract (FI), 1% ammonium oxalate extract (FII), and 5% sodium hydroxide extract (FIll), were further fractionated by DEAE-cellulose chromatography, Toyopearl HW65F gel filtration, and Con A-AF-Toyopear1650 affinity chromatography. Finally, five· antitumor-active polysacchride fractions were obtained. Table I shows the evaluation of the antitumor activities of the yamabushitake polysaccharides obtained during the fractionation and purification steps. Table II shows the physicochemical properties of the active polysaccharide fractions. FI o-a-f3 (an unadsorbed fraction from Con A-AF-Toyopearl 650 affinity column was a heteroglycan containing mainly xylose, glucose, and mannose and FIo-a-1X (an adsorbed fraction) was a xylan-protein complex, and both of them had f3-(l--t3) and (1--t6) glucan chains in those molecules based on the IR and NMR spectra (Table III). These f3-glucan parts of the molecules may be significant in the expression of antitumor activities. FII-I showed a good antitumor effect (Table I). FII-I was a glucoxylan having f3-(1--t3) and (l--t6) glucosidic linkages that gave characteristic spctra in lH_ and 13C-NMR (Table III). The spectrometric data were as follows: IR I' max (KBr): 893cm- 1. lH-NMR spectrum b: 4.88,4.74, 4.51, 4.25 ppm. 13C-NMR spectrum b: 108.1,91.0,81.3,78.3,74.9,74.0, 69.0, 65.8 ppm. FIII-2b was a galactoxyloglucan-protein complex and showed antitumor activity. This fraction was prepared from alkali-solubilized FIll by fractional precipitation with acetic acid followed by gel filtration on Toyopearl HW-65F. The structure of the FIII-2b was analyzed with the solid state 13C-NMR, which showed a characteristic spectrum of f3(l--t3) and (1--t6)-glucosidic linkages whose peaks (ppm) appeared at 4.50 and 4.25 (anomeric proton); 104.2 (C-l), 87.5 (C-3), 75.2 (C-5),
C-5
C-2
C-4
C-6
5
6
7
8
74.0 71.1
68.4 68.4 69.0 69.8
62.4 62.1 65.8 63.5
74.3 74.3 74.9 75.2
lH Shift (ppm) 2 4.51 4.24 4.51 4.50
3.11 3.11 4.25 4.25
71.1 (C-2), 69.8 (C-4), and 63.5 (C-6). Acknowledgments. The authors express appreciation to Mr. Tsutomu Takigawa of Otsuka Electronics Co., Ltd. for measuring solid state lH_ and 13C-NMR spectra, and to Assistant Professor Hirokazu Kawagishi of our University for measuring lH_ and 13C-NMR spectra. Further, the authors are grateful to Mr. Y oshikazu Mayuzumi of Myogi Kinoko-en, Gunma, Japan for providing us yamabushitake mushroom for study.
References 1)
H. Kawagishi, M. Ando, and'T. Mizuno, Tetrahedron Lett., 31, 373-376 (1990). 2) H. Kawagishi, M. Ando, T. Mizuno, H. Yokota, and S. Konishi, Agric. BioI. Chem., 54, 1329-1331 (1990). 3) H. Kawagishi, M. Ando, H. Sakamoto, T. Yoshida, and A. Furukawa, Nippon Nogeikagaku Kaishi, 65, 364 (1991); Kagaku to Seibutsu, 29, 640--646 (1991). 4) Y. Kimura, M. Nishibe, A. Shimada, H. Nakajima, T. Hamazaki, and A. Tsuneda, Nippon Nogeikagaku Kaishi, 65,309 (1991); Agric. Bioi. Chem., 55, 2673-2674 (1991). 5) N. Sato, T. Iimori, M. Machida, M. Yadai, Z. Magai, M. Yamazaki, and K. Nagai, Nippon Nogeikagaku Kaishi, 65, 364 (199l). 6) T. Mizuno, T. Hagiwara, T. Nakamura, H. Ito, K. Shimura, and A. Asakura, Agric. Bioi. Chem:, 54, 2889-2896, 2897-2905 (1990): ibid., 50, 1679-1688 (1986); ibid., 55, 2701-2710 (1991); Biosci. Biotech. Biochem., 56, 34---41 (1992). 7) H. Ito and H. Hidaka, Cancer Lett., 19, 215-221 (1983); Japan J. Pharmacol., 33,403--408 (1983). 8) T. Mizuno, S. Ohtahara, and J. Li, Bull. Fac. Agr. Shizuoka Univ., 38, 37-46 (1988).
NII-Electronic Library Service
Bioscience, Biotechnology, and Biochemistry Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/tbbb20
Antitumor-active Polysaccharides Isolated from the Fruiting Body of Hericium erinaceum, an Edible and Medicinal Mushroom Called yamabushitake or houtou a
a
ab
Takashi Mizuno , Tetsuya Wasa , Hitoshi Ito , Chiharu Suzuki
ac
ad
& Nobuo Ukai
a
Faculty of Agriculture, Shizuoka University, Ohya, Shizuoka 422, Japan
b
School of Medicine, Mie University, Tsu, Mie 514, Japan
c
Fuji Biocenter, Fuji Seito Co., Ltd., Shibakawa-cho, Shizuoka 419–03, Japan
d
Research Institute, Kagome Co., Ltd., Nishinasuno-cho, Tochigi 329–27, Japan Published online: 12 Jun 2014.
To cite this article: Takashi Mizuno, Tetsuya Wasa, Hitoshi Ito, Chiharu Suzuki & Nobuo Ukai (1992) Antitumor-active Polysaccharides Isolated from the Fruiting Body of Hericium erinaceum, an Edible and Medicinal Mushroom Called yamabushitake or houtou, Bioscience, Biotechnology, and Biochemistry, 56:2, 347-348, DOI: 10.1271/bbb.56.347 To link to this article: http://dx.doi.org/10.1271/bbb.56.347
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http:// www.tandfonline.com/page/terms-and-conditions
Biosci. Biotech. Biochem., 56 (2), 347-348, 1992
Note
Antitumor-active Polysaccharides Isolated from the Fruiting Body of Hericium erinaceum, an Edible and Medicinal Mushroom Called yamabushitake or houtout Takashi
MIZUNO,
Tetsuya
WASA,
Hitoshi
ITO, *
Chiharu
SUZUKI, **
and Nobuo
UKAI***
Downloaded by [New York University] at 13:22 05 June 2015
Faculty of Agriculture, Shizuoka University, Ohya, Shizuoka 422, Japan *School of Medicine, Mie University, Tsu, Mie 514, Japan ** Fuji Biocenter, Fuji Seito Co., Ltd., Shibakawa-cho, Shizuoka 419--03, Japan *** Research Institute, Kagome Co., Ltd., Nishinasuno-cho, Tochigi 329-27, Japan Received July 31, 1991
Yamabushitake, or Hericium erinaceum, is an edible mushroom fungus belonging to Aphyllophorales, Hydnaceae (Hericiaceae), Hericium. It grows widely in Japan and the whole of China. In recent years, Hericium erinaceum fungus has been cultivated by wood-log culture (Photo. 1) or culture-bed methods such as bottleculture and bag-culture. Several newly isolated compounds from this mushroom have shown interesting biological activities such as cytotoxic effects on HeLa cells,
promotion of the synthesis of neuro-growth factor, and inhibition of pollen tube germination, and moreover the fruiting body of Hericium erinaceum, or its liquid-cultured mycelia, or an alcohol-precipitated fraction of its culture filtrate showed antitumor activities based on immunopotentiation. 1- 5) Previoulsy, we have reported some antitumor polysaccharides obtained from several fungi belonging to Polyporaceae and Agaricaceae. 6 - 8) In this note, we describe the evaluation of antitumor activities of the polysaccharides fractionated from the fruiting bodies of yamabushitake mushroom. Table I. Antitumor Activities of Some Polysaccharides Isolated from the Fruiting Bodies of Hericium erinaceum Polysaccharide Control FIo-a FIo-a-cc Fl o-a-f1 FIo·b FII-l FIII-2b
Photo 1. Typical Fruiting Bodies ofYamabushitake, Hericium erinaceum cultivated in Wood-log Culture. Table II.
F1o-a FIo-a-cc FI o-a-f1 FIo-b FII-l FIII-2b
b
e d
e
0 40.3 65.9 73.0 63.8 67.9 75.9
Complete regression
Mortality
0/10 1/7 3/7 4/7 2/7 1/7 3/7
10/10 3/7 2/7 1/7 3/7 1/7 0/7
1.00 1.54 1.78 1.86 1.71 1.81 1.75
Note: In four-week old female ICR/Slc mice, 6 x 106 cells/mouse of 7-day-old Sarcoma 180 from an ascites tumor were implanted subcutaneously in the right groin and 10 mg/kg of fraction from Flo were injected i.p. once daily for 10 consecutive days starting 24 hours after tumor implantation. The tumor size was measured at 28 days after tumor implantation. The survival time and mortality were measured 61 days after tumor implantation.
Some Properties of Antitumor-active Polysaccharides Isolated from the Fruiting Bodies of H. erinaceum
Polysaccharide
a
Inhibition Survival time (%) (T/C)
Molecular weight
Protein (%)b
Total sugar (%)C
118,000 89,000 175,000 565,000 155,000 30,000
34.2 51.5 4.2 22.1 5.7 20.2
53.4 40.5 89.7 71.4 91.4 75.7
Q
Components sugar" Gle
Xyl
Man
Gal
100
333 100 233 104 179 51
34 4 58 10
4
100 100 100 100
13 15 17 32
IR peak cm- le 880 868 889 893 893 899
Gel filtration by using a Toyopearl HW-65 column. Modified Lowry method. Phenol-sulfuric acid colorimetry. Each polysaccharide was hydrolyzed with 0.5 to 1.0 M sulfuric acid at 100°C for 3 to 6 hours and the hydrolyzate thus obtained was treated by ppc. Further, its alditol acetate derivative was identified and measured by GC method with the use of a Shimadzu GC-4CM -PF apparatus. KBr tablet method by using JIR-RFX-3001 apparatus of Nippon Bunko.
t Studies on the Host-mediated Antitumor Polysaccharides. Part XVII. For Part XVI, see ref. 6.
NII-Electronic Library Service
348
T.
et al.
MIZUNO
Table III. Solid State 13C-NMR and lH-NMR Spectral Chemical Shifts for Water-insoluble Autitumor Polysaccharides Isolated from Yamabushitake Mushroom 13C Shift (ppm) Polysaccharide
C-l
C-3 2
Downloaded by [New York University] at 13:22 05 June 2015
FI o-a-f3 FI o-b-f3 FII-l FIII-2b
103.3 103.1 108.1 104.2
91.0
4 85.8 85.6 81.3 87.5
78.3
Fruiting bodies of the fresh mushroom were extracted stepwisely with 85% ethanol (30°C, 24 hr), hot water (lOO°C, 3 hr), 1% ammonium oxalate (100°C, 5 hr), and 5% sodium hydroxide (30°C, 12-24 hr) as described previously.6) The hot water extract (FI), 1% ammonium oxalate extract (FII), and 5% sodium hydroxide extract (FIll), were further fractionated by DEAE-cellulose chromatography, Toyopearl HW65F gel filtration, and Con A-AF-Toyopear1650 affinity chromatography. Finally, five· antitumor-active polysacchride fractions were obtained. Table I shows the evaluation of the antitumor activities of the yamabushitake polysaccharides obtained during the fractionation and purification steps. Table II shows the physicochemical properties of the active polysaccharide fractions. FI o-a-f3 (an unadsorbed fraction from Con A-AF-Toyopearl 650 affinity column was a heteroglycan containing mainly xylose, glucose, and mannose and FIo-a-1X (an adsorbed fraction) was a xylan-protein complex, and both of them had f3-(l--t3) and (1--t6) glucan chains in those molecules based on the IR and NMR spectra (Table III). These f3-glucan parts of the molecules may be significant in the expression of antitumor activities. FII-I showed a good antitumor effect (Table I). FII-I was a glucoxylan having f3-(1--t3) and (l--t6) glucosidic linkages that gave characteristic spctra in lH_ and 13C-NMR (Table III). The spectrometric data were as follows: IR I' max (KBr): 893cm- 1. lH-NMR spectrum b: 4.88,4.74, 4.51, 4.25 ppm. 13C-NMR spectrum b: 108.1,91.0,81.3,78.3,74.9,74.0, 69.0, 65.8 ppm. FIII-2b was a galactoxyloglucan-protein complex and showed antitumor activity. This fraction was prepared from alkali-solubilized FIll by fractional precipitation with acetic acid followed by gel filtration on Toyopearl HW-65F. The structure of the FIII-2b was analyzed with the solid state 13C-NMR, which showed a characteristic spectrum of f3(l--t3) and (1--t6)-glucosidic linkages whose peaks (ppm) appeared at 4.50 and 4.25 (anomeric proton); 104.2 (C-l), 87.5 (C-3), 75.2 (C-5),
C-5
C-2
C-4
C-6
5
6
7
8
74.0 71.1
68.4 68.4 69.0 69.8
62.4 62.1 65.8 63.5
74.3 74.3 74.9 75.2
lH Shift (ppm) 2 4.51 4.24 4.51 4.50
3.11 3.11 4.25 4.25
71.1 (C-2), 69.8 (C-4), and 63.5 (C-6). Acknowledgments. The authors express appreciation to Mr. Tsutomu Takigawa of Otsuka Electronics Co., Ltd. for measuring solid state lH_ and 13C-NMR spectra, and to Assistant Professor Hirokazu Kawagishi of our University for measuring lH_ and 13C-NMR spectra. Further, the authors are grateful to Mr. Y oshikazu Mayuzumi of Myogi Kinoko-en, Gunma, Japan for providing us yamabushitake mushroom for study.
References 1)
H. Kawagishi, M. Ando, and'T. Mizuno, Tetrahedron Lett., 31, 373-376 (1990). 2) H. Kawagishi, M. Ando, T. Mizuno, H. Yokota, and S. Konishi, Agric. BioI. Chem., 54, 1329-1331 (1990). 3) H. Kawagishi, M. Ando, H. Sakamoto, T. Yoshida, and A. Furukawa, Nippon Nogeikagaku Kaishi, 65, 364 (1991); Kagaku to Seibutsu, 29, 640--646 (1991). 4) Y. Kimura, M. Nishibe, A. Shimada, H. Nakajima, T. Hamazaki, and A. Tsuneda, Nippon Nogeikagaku Kaishi, 65,309 (1991); Agric. Bioi. Chem., 55, 2673-2674 (1991). 5) N. Sato, T. Iimori, M. Machida, M. Yadai, Z. Magai, M. Yamazaki, and K. Nagai, Nippon Nogeikagaku Kaishi, 65, 364 (199l). 6) T. Mizuno, T. Hagiwara, T. Nakamura, H. Ito, K. Shimura, and A. Asakura, Agric. Bioi. Chem:, 54, 2889-2896, 2897-2905 (1990): ibid., 50, 1679-1688 (1986); ibid., 55, 2701-2710 (1991); Biosci. Biotech. Biochem., 56, 34---41 (1992). 7) H. Ito and H. Hidaka, Cancer Lett., 19, 215-221 (1983); Japan J. Pharmacol., 33,403--408 (1983). 8) T. Mizuno, S. Ohtahara, and J. Li, Bull. Fac. Agr. Shizuoka Univ., 38, 37-46 (1988).
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