Professional Documents
Culture Documents
Abstract
The process for the removal of protein from Pleurotus ostreatus crude polysaccharide was carried out by Sevga treatment after
enzymatic hydrolysis. Then isolated by DEAE-cellulose-52 ion exchange column chromatography, POP-I, POP-II and POP-III
were obtained and purified by Sephadex G-100 column chromatography again. After that four homogenous pure polysaccharides
were got and named POP-I-1, POP-II-1, POP-III-1 and POP-III-2. Neither protein nor nuclear acid characteristic absorption peaks
were shown in the UV-visible spectra of POP-I-1and POP-II-1, and the result of IR, TLC and NMR showed they were mainly
composed of glucose and and glycosidic bond.
Keywords: Pleurotus Ostreatus Polysaccharide (POP); Isolation and Purification; UV-Visible Spectra; IR; TLC; NMR
*
1 1, 2 1 1
1. 225127
2. 211600
(Pleurotus ostreatus)
[1]
[2]
[3-4]
[5-7]
- 42 www.jma-journal.org
(IR)(TLC)(NMR)
1.1
CIL
1.2
HH-6
SHZ-RE-52D
Centrifuge 5804 Eppendorf UV-2550
BS2000S Alphal-2 LD Plus Christ AVANCE
600 BrukerCary 5000 -- Varian
1.3
1.3.1
40 1:40(gml)
90 3.5h5000r/min 10min 4
[8-11]
1.3.2
Sevag TCA
Sevga [12-16]
-Sevag 4
[17] DEAE-52
(1.6cm 50cm) 0.1mol/L-0.5mol/L NaCl
0.67mL/min 5mL--
24h 24h 6h [18-19]
Sephadex G-100 (2.6cm40cm)
[20-21]
1.3.3
10mg/mL 200-600nm
[22-23]
1.3.4
2g 1cm 1cm
85 10min[24]
1.3.5
2.1
POP 3.25%
-Sevag 82.3%POP
2.2 DEAE-52
DEAE-52 POP 5ml 1 1
NaCl POP
8 20 POP-I 0.1mol/L NaCl
39 60 POP-II 0.1-0.3mol/L
NaCl 67 89 POP-III
POP POP
1 (POP)DEAE-52
11 23 POP-III-1
23 40 POP-III-2
POP-IPOP-II POP-III Sephadex G-100 POP-I POP-II
POP-III POP-III-1 POP-III-2
POP-I-1 POP-II-1
2.4
5 POP-I-1 POP-II-1 POP-I-1 POP-II-1 260nm 280nm
5.000
5.000
4.000
4.000
Abs.
Abs.
2.000
2.000
POP-I-1
POP-II-1
0.000
0.000
-1.000
200.00
300.00
400.00
nm.
500.00
600.00
-1.000
200.00
300.00
5 POP-I-1 POP-II-1
6 POP-I-1(1) POP-II-1(2)
- 45 www.jma-journal.org
400.00
nm.
500.00
600.00
2.5
POP-I-1 POP-II-1 6 POP-I-1 1 POP-II-1 2
D-D-D-D-D-L- DPOP-I-1 Rf 0.455,0.477,0.420,0.500,0.592,0.477,0.699,0.443POP-II-1
Rf 0.446,0.441,0.379,0.497,0.610,0.492,0.718,0.452POP-I-1 POP-II-1
Rf POP-I-1 POP-II-1
2.6
POP-I-1 POP-II-1 7 3200-3600cm-1 O-H
2800-3000 cm-1 C-H
1650 cm-1 C=O 1000-1200 cm-1
POP-I-1 872.7 cm-1 C
POP-II-1 913.9 cm-1 D- C-O-C
100
100
1
2097.2
90
2
2125.2
90
872.7
1246.7
80
80
70
70
913.9
2934.1
50
1352.7
1417.3
1250.9
60
800.1
1650.8
40
10
1077.5
2929.2
1647.7
1418.6
602.7 537.5
30
1149.2
20
801.1
50
40
603.3 577.5
30
60
1046.2
1149.4
20
3390.6
10
3389.9
1076.9
1045.6
0
0
3800 3600 3400 3200 3000 2800 2600 2400 2200 2000 1800 1600 1400 1200 1000
800
600
400
200
3800
3000 2800
1400 1200
1000
800
600
400
200
7 POP-I-1 POP-II-1
2.7
POP-I-1 POP-II-1 1HNMR 8 5.0ppm
5.0ppmPOP-I-1 3.0-5.5ppm POP-I-1
POP-II-1 3.0-5.0ppm 5.09ppm POPII-1
POP-I-1
POP--1
REFERENCES
[1]
Lize Wang. Mushroom cultivation [M]. Hefei: Anhui Science and Technology Press, 1993: 82-97
[2]
Honghua Wu, Yi Wang, Lei Qian, et al. Extract Technical Study on Pleurotus Ostreatus [J]. Journal of Zhejiang University of
Traditional Chinese Medicine, 2010, 34 (3): 424-426
[3]
Xueyan Xu, Peng Zhou, Shan Jiang. Application Progress of Modern Separation and Purification Technology of Polysaccharides,
Strait Pharmaceutical Journal, 2011, 23 (4): 10-13
[4]
Huiyang Zhang, Junze Qin. A Study of the Macroporous Resins Absorption and Purification of the Phellinus Linteus
Polysaccharide [J]. Journal of ShanXi University of Science & Technology, 2011, 29 (1): 73-76
[5]
Jing Xu, Ting Zhao, Wei Liu, et al. Carboxymethylation of a Polysaccharide Extracted from Ganoderma Lucidum Enhances its
Antioxidant Activities in Vitro [J]. Carbohydrate Polymers, 2009 (78):227-234
[6]
Grigory M. Lipkinda, L Exander S.S Hashkovy, Uriya. Knlre, et al. A Computer-assisted Structural Analysis of Regular
Polysaccharides on the Basis of 13C-N.M.R. Data [J]. Carbohydrate Research, 1988 (175): 59-75
[7]
Peng Zhou, Mingyong Xie, Shaoping Nie, et al. Structure Characterization of of Tea Polysaccharides TGC [J]. Science in China
Ser. C Life Sciences, 2004,34 (2): 178-185
[8]
Tsai Shu-Yao, Huang Shih-Jeng, Mau Jeng-Leun. Antioxidant Properties of Hot Water Extracts from Agrocybe cylindracea [J].
Food Chemistry, 2006 (98): 670-677
[9]
Jinxi Wang, Zhenwei Bao, Xiao Liu, et al. Optimization of Polysaccharides Extraction Process from Pleurotus ostreatus by
Response Surface Methodology [J]. China Brewing, 2013, 32 (4): 108-112
[10] Jinxi Wang, Lin Gu, Fanwei Kong, et al. Antioxidant Activity in vitro of Crude Polysaccharides from Coprinus comatus [J]. Food
Science, 2012, 33 (13): 79-82
[11] Min Zhang, Ruizhan Chen, Ruiping Chen, et al. Comparison among Extraction Technologies of Polysaccharide from Pleurotus
ostreatus [J]. Journal of Jilin University ( Science Edition), 2011, 49 (6): 1141-1149
[12] Yingchun Gao, Jun Chen. Study on the comparison of different methods for removing of protein from polysaccharide of Chinese
yam [J]. Food Science and Technology, 2009, 34 (6): 71-74
[13] Qiaojuan Yan, Lujia Han, Zhengqiang Han.Removal of protein from the crude astragalus polysaccharides by proteases [J]. Food
Science and Technology, 2004, 6: 23-26
[14] Shanguang Wu, Taijun Lai, Jianhua Sun, et al.A Comparative Investigation into Deproteinization Methods for Silkworm Pupa
Polysaccharides [J]. Food Science, 2011, 32 (14): 21-24
[15] Meicheng Liu, Yin Wan, Zongcai Tu, et al. A Research on Deproteinization Methods for Lily-Polysaccharide [J]. Food Science,
2002, 23 (1): 89-90
[16] Kun Zhang, Hao Wu, Lingchong Wang, et al. Comparative Study on Deproteinization Methods for Polysaccharide from Mactra
veneriformis [J]. Food Science, 2011, 32 (8): 50-53
[17] Hongli Zhou, Xiaohong Yang. Removal of Proteins from Corn Silk Polysaccharide by a Combined Enzymatic-Sevag Method [J].
Food Science, 2011, 32 (8): 129-132
[18] Yongxu Sun, Jicheng Liu. Purification, Structure and Immunobiological Activity of a Water-soluble Polysaccharide from the
Fruiting Body of Pleurotus ostreatus [J]. Bioresource Technology, 2009 (100): 983-986
- 47 www.jma-journal.org
[19] Lin Gu, Jun Jiang. StudyonConstitutes, SeparationandPurificationTechnologyofYamPolysoccharides [J]. Food Science, 2007, 28
(9): 158-161
[20] Xiangyang Wu, Guanghua Mao, Ting Zhao, et al. Isolation, Purification and in Vitro Anti-tumor Activity of Polysaccharide from
Ginkgo Biloba Sarcotesta [J]. Carbohydrate Polymers, 2011 (86): 1073-1076
[21] Song Pan, ChangjiangLiu, Shuang Liang. Isolation, Purification and MolecularWeight Determination of Polysaccharides from
Actinidia arguta Sieb. et Zucc. Fruits [J]. Food Science, 2012, 33 (15): 66-70
[22] Liyan Zhao, Yanhong Dong, Guitang Chen, et al. Extraction, Purification, Characterization and Antitumor Activity of
Polysaccharides from Ganoderma lucidum [J]. Carbohydrate Polymers, 2010 (80): 783-789
[23] Guo Qi, Ruiguang Sun, Guoyun Guo, et al. Isolation and Purification, its Structure Spectral Analysis, and Microscopy Observation
of Lycium barbarum polysaccharide [J]. Chinese Traditional and Herbal Drugs, 2012, 43 (4): 645-648
[24] Yuping Zhao. Effections of Seven Chromatographic Solutions on Components Analysis of Fungi polysaccharides [J]. Science and
Technology of Food Industry, 2006, 27 (11): 109-110, 113
[25] Guanghua Huo, Laisheng Li, Yinyu Gao. Spectroscopy in Structural Analysis of Polysaccharides [J]. Chemistry of Life, 2002, 22
(2): 194-196
[26] Xiuju Du, Jingsong Zhang, Yingjie Pan. Application of the NMR Techniques in Structural Analysis of Polysaccharide from Edible
Fungi [J]. Edible Fungi of China, 2010, 29 (1): 3-6, 19
[27] Chengmin Hou, Wenning Chen, Yufang Chen, et al. Characteristics of Spectral Analysis for Carbohydrates Structures [J]. Nat
Prod Res Dev, 2012, 24: 556-561
[28] Lin Gu, Jun Jiang, Jing Sun. Isolation, Purification and Structure Determination of the Yam [J]. Food Science and Technology,
2007 (5): 109-112
[29] Omaira Gutierrez de G, Maritza Martnez, Lilian Sanabria, et al. 1D- and 2D-NMR spectroscopy studies of the polysaccharide
gum from Spondias purpurea var. Lutea [J]. Food Hydrocolloids, 2005 (19): 37-43
[30] Qingyang Liu , Ting Zhao, Hong Wei, et al. Carboxymethylation of polysaccharides from Auricularia auricula and their
antioxidant activities in vitro [J]. International Journal of Biological Macromolecules, 2011 (49): 1124-1130
1991-
1987-
Email: monkeyet@126.com
Email: 867068809@qq.com
1956-
Email: gulin@yzu.edu.cn
4
1973-
Email: yuhai@yzu.edu.cn
- 48 www.jma-journal.org