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Journal of Modern Agriculture

July 2014, Volume 3, Issue 3, PP.42-48

A Preliminary Study on Extraction, Isolation,


Purification and Structure Identification of
Pleurotus Ostreatus Polysaccharides
Xiao Liu 1, Jinxi Wang 1, 2, Lin Gu 1, #, Hai Yu 1
1. College of Food Science and Engineering/Yangzhou University, Yangzhou 225127, China
2. Jiangsu Fufeng Biotechnology Co., Ltd, Jinhu 211600, China
#Email: gulin@yzu.edu.cn

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

(POP) Sevga DEAE-52

POP-IPOP-II POP-III Sephadex G-100 POP-I-1POP-II-1POP-III-1 POP-III-2


POP-I-1 POP-II-1

(Pleurotus ostreatus)
[1]
[2]

[3-4]
[5-7]

Sevag DEAE-52 Sephadex G-100


*
2012BAD36B05

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(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

20mg 6mL 1mol/L 100 8hBaCO3


4000r/min 10min D-D-D-
L-D-D- D- 1mg/mL
=13:3:3:1 10mL 85%100mL 2mL
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2g 1cm 1cm
85 10min[24]
1.3.5

1mg KBr 4004000cm-1 10mg 0.5mL


[25-30]

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

2 POP-I Sephadex G-100

2.3 Sephadex G-100


(POP) DEAE-52 POP-IPOP-II POPIIIPOP-II POP-III Sephadex G-100
234
2 POP-I Sephadex G-100 POP-I
15 40 POP-I-1
3 POP-II Sephadex G-100 11 40
29 11 29
POP-II-1
4 POP-III Sephadex G-100 POP-III
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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

3 POP-II Sephadex G-100

4 POP-III Sephadex G-100

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)
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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

3600 3400 3200

3000 2800

2600 2400 2200

2000 1800 1600

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

8 POP-I-1 POP-II-1 1HNMR


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DEAE-52 POP-IPOP-II POP-III

Sephadex G-100 POP-I-1POP-II-1POP-III-1


POP-III-2 POP-I-1 POP-II-1

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1991-

1987-

Email: monkeyet@126.com

Email: 867068809@qq.com

1956-

Email: gulin@yzu.edu.cn
4

1973-

Email: yuhai@yzu.edu.cn

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