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Recent advances in Cordyceps sinensis

polysaccharides: Mycelial fermentation,
isolation, structure, and bioactivities: A review
ARTICLE in JOURNAL OF FUNCTIONAL FOODS JANUARY 2013
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Recent advances in Cordyceps sinensis

polysaccharides: Mycelial fermentation, isolation,
structure, and bioactivities: A review
Jing-Kun Yana,b,*, Wen-Qiang Wanga, Jian-Yong Wua,*
a
Department of Applied Biology & Chemical Technology, PolyU Shenzhen Research Institute, The Hong Kong Polytechnic University,
Hung Hom, Kowloon, Hong Kong
b
School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, Jiangsu, China

A R T I C L E I N F O

A B S T R A C T

Article history:

Cordyceps (Ophiocordyceps sinensis) sinensis, the Chinese caterpillar fungus, is a unique and

Received 7 July 2013

precious medicinal fungus in traditional Chinese medicine which has been used as a pres-

Received in revised form

tigious tonic and therapeutic herb in China for centuries. Polysaccharides are bioactive

23 November 2013

constituents of C. sinensis, exhibiting several activities such as immunomodulation, antitu-

Accepted 27 November 2013

mour, antioxidant and hypoglycaemic. As natural C. sinensis fruiting body-caterpillar com-

Available online 22 December 2013

plexes are very rare and expensive, the polysaccharides documented over the last 15
20 years from this fungal species were mostly extracted from cultivated fungal mycelia

Keywords:

(intracellular polysaccharides) or from mycelial fermentation broth (exopolysaccharides).

Cordyceps sinensis

Extraction and purification of the polysaccharides is a tedious process involving numerous

Polysaccharide

steps of liquid and solid phase separations. Nevertheless, a large number of polysaccharide

Production
Extraction
Structural elucidation
Bioactivity

structures have been purified and elucidated. However, relationships between the structures and activities of these polysaccharides are not well established. This review provides
a comprehensive summary of the most recent developments in various aspects (i.e., production, extraction, structure, and bioactivity) of the intracellular and exopolysaccharides
from mycelial fermentation of C. sinensis fungi. The contents and data will serve as useful
references for further investigation, production and application of these polysaccharides in
functional foods and therapeutic agents.
2014 Published by Elsevier Ltd.

Contents
1.
2.
3.
4.

Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Production of biomass and polysaccharides by mycelial fermentation
Extraction, isolation and purification of polysaccharides . . . . . . . . . . .
Physicochemical characterization . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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* Corresponding authors. Address: School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, Jiangsu, China. Tel.:
+86 15952819661 (J.-K. Yan). Tel.: +852 34008671 (J.-Y. Wu).
E-mail addresses: [email protected] (J.-K. Yan), [email protected] (J.-Y. Wu).
1756-4646/$ - see front matter 2014 Published by Elsevier Ltd.
http://dx.doi.org/10.1016/j.jff.2013.11.024

34

5.

6.

1.

JOURNAL OF FUNCTIONAL FOODS

6 ( 2 0 1 4 ) 3 3 4 7

4.1. Monosaccharide composition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

4.2. Average molecular weight . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
4.3. Chemical structures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
4.4. Conformational features . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Bioactivities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
5.1. Immunomodulatory activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
5.2. Antitumour activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
5.3. Antioxidant activity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
5.4. Hypoglycemic effect . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
5.5. Other bioactivities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Conclusions and future perspectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Introduction

Cordyceps (Ophiocordyceps) sinensis (Berk.) Sacc., the Chinese

caterpillar fungus or DongChongXiaCao (winter worm-summer grass) in Chinese or Tochukaso in Japanese, is a valuable
medicinal fungus in traditional Chinese medicine (TCM). C.
sinensis is a parasitic fungus to the moth larvae (Lepidoptera)
of Hepialus armoricanus (and Thitarodes). In late summer or
early autumn, the larvae are infected by the fungal spores
and gradually consumed by the fungal mycelia, and turned
into stiff worms in winter. In spring and early summer of
the following year, a stroma or fruiting body forms on the larva head, grows and emerges out of the ground like a grass
(Fig. 1) (Chen, Wang, Nie, & Marcone, 2013; Holliday, & Cleaver,
2008; Li, & Tsim, 2004; Lo, Hsieh, Lin, & Hsu, 2013; Winkler,
2010; Zhang, Li, Wang, Li, & Liu, 2012; Zhu, Halpern, & Jones,
1998). The natural C. sinensis fruiting body-caterpillar

38
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40
40
41
41
42
42
42
42
43
43

complexes are mainly distributed on the high plateaus of

35005000 m above sea level in Tibet, Qinghai, Sichuan, and
Yunnan Provinces in China (Li et al., 2011).
C. sinensis has been used in China for more than 700 years,
mainly as a tonic to invigorate the lungs and to nourish the
kidneys (Dong & Yao, 2008). Modern pharmacological studies
have shown its therapeutic effects on a wide range of diseases
and conditions, such as respiratory, renal, liver, nervous system, and cardiovascular diseases, as well as tumours, aging,
hyposexuality, and hyperlipidaemia (Ding et al., 2011; Ji
et al., 2009; Liu, Li, Zhao, Tang, & Guo, 2010; Lo et al., 2013;
Marchbank, Ojobo, Playford, & Playford, 2011; Song, Ming,
Peng, & Xia, 2010; Yue, Ye, Zhou, Sun, & Lin, 2013; Zhang,
Wang, Zhang, & Ye, 2011; Zhu et al., 1998). C. sinensis has been
listed as an herbal drug in the official Chinese Pharmacopoeia
by the Committee of Pharmacopoeia and Chinese Ministry of
Health since 1964. During the outbreak of the Severe Acute

Fig. 1 Cordyceps (Ophiocordyceps) sinensis fruiting body-caterpillar complexes: morphology and natural habitat (Paterson,
2008; Winkler, 2010).

JOURNAL OF FUNCTIONAL FOODS

Respiratory Syndrome (SARS) in China in 2003, there was a

notable increase in the use of C. sinensis. Over the last
10 years, the demand as well as the price for C. sinensis has increased dramatically in China, Japan, Korea and India (Au
et al., 2012; Jeffrey, 2012; Winkler, 2009). As natural C. sinensis
are very limited and cannot meet the increasing demand, fermentation technology has been widely exploited for largescale production of C. sinensis fungal mycelia and other useful
constituents. The fermentation-cultivated mycelia of some
fungal strains isolated from the natural C. sinensis have been
shown to produce the similar pharmacological efficacy to
the wild fungal materials and been widely applied to various
health food products (Zhu et al., 1998).
The multiple pharmacological effects of C. sinensis can be
attributed to its chemical ingredients, including polysaccharides, proteins, nucleotides, mannitol, ergosterol, aminophenol, fatty acids, and trace elements. Actually, several
reviews on these compounds and their properties and bioactivities of C. sinensis have been published in the last few years
(Chen, Wang et al., 2013; Paterson, 2008; Shashidhar, Giridhar,
Sankar, & Manohar, 2013; Zhao, Xie, Wang, & Li, 2013; Zhong
et al., 2009). In particular, polysaccharides represent one of
the most abundant components in the fungus and a major
group of bioactive constituents which have been extracted
and isolated from the fruiting bodies, cultured mycelium,
and fermentation broth, which are structurally diverse biomacromolecules with various physiochemical properties.
Polysaccharides have been the target for the development
and quality control of C. sinensis health products. More recently, some reviews on extraction, isolation, structure and
bioactivities of polysaccharides from C. sinensis have appeared (Lo et al., 2013; Nie, Cui, Xie, Phillips, & Phillips,
2013; Xiao, 2008; Zhong et al., 2009). However, these reviews
mainly focus on separation techniques, structural features
and bioactivities of intracellular polysaccharides (IPSs) from
the fruiting body of natural C. sinensis and mycelia of cultured
C. sinensis, but the mycelial fermentation, physicochemical
properties and pharmacological activities of extracellular
polysaccharides or exopolysaccharides (EPSs) isolated from
culture broth of C. sinensis are less involved. In addition, the
relationships are still not well established between the molecular structure and the bioactivity of C. sinensis derived polysaccharides. Herein, this review summarizes and compares
the recent studies on the production, extraction, and purification of IPSs and EPSs from C. sinensis as well as the characterization of their structural features, chain conformations, and
bioactivities.

2.
Production of biomass and polysaccharides
by mycelial fermentation
Wild or natural C. sinensis is becoming increasingly scarce because of reckless harvesting, geographical limitation, and
unfavourable weather conditions for its proliferation (Yao,
2004). Cultivation of fungal mycelia is a more reliable alternative for mass production of the fungal materials. Several species of fungi have been successfully isolated from the natural
C. sinensis such as Paecilomyces sinensis, Cephalosporium sinensis, Tolypocladium sinensis, and Hirsutella sinensis (Yin & Tang,

6 (2 0 14 ) 3 347

35

1995). Some of these species have been cultivated in large

quantities of mycelial biomass by fermentation technology.
The fungal mycelia have been reported to exert similar pharmacological effects to those of wild C. sinensis species (Dong &
Yao, 2008; Zhu et al., 1998).
To improve the efficiency and productivity of mycelial fermentation processes, many investigators have studied the effects of various process factors on the maximal production of
mycelial biomass and EPSs and to optimize the fermentation
conditions, such as medium composition, temperature, pH,
and culture vessel (Hsieh, Tsai, Hsu, Chang, & Lo, 2005; Kim
& Yun, 2005; Liu & Wu, 2012). Table 1 provides a summary
of the strains, culture conditions, mycelial biomass and EPS
yields of C. sinensis which have been reported in the literature.
The biomass and EPS yields varied in a wide range from 10 to
54 g/L, and <1.0 to >40 g/L with the fungal species and culture
conditions, respectively. In particular, our group has previously demonstrated the optimization of submerged culture
conditions for C. sinensis (Cs-HK1), such as temperature, initial pH, carbon and nitrogen levels, minerals, and surfactants
(Tween 80) (Leung, & Wu, 2007; Leung, Zhang, & Wu, 2006; Liu
& Wu, 2012). However, the production of bioactive EPS by liquid fermentation of edible or medicinal fungi (e.g. C. sinensis)
is still a new area of research without much industrial application. Thus, there is a need to enhance the EPS productivity
through effective strategies of process intensification in the
future.

3.
Extraction, isolation and purification of
polysaccharides
C. sinensis polysaccharides can be classified into two types
according to their locations in the fungal cells, intracellular
polysaccharides (IPSs) and extracellular polysaccharides (exopolysaccharides, EPSs). IPSs are extracted from the fruiting
body (or worm) and mycelium of C. sinensis with pure water,
aqueous acidic/alkaline solutions, aqueous buffers under
heating (Guan, Zhao, Feng, Hu, & Li, 2011; Kiho, Ookubo, Usui,
Ukai, & Hirano, 1999; Kiho, Tabata, Ukai, & Hara, 1986; Kiho,
Yamane, Hui, Usui, & Ukai, 1996; Wang, Wang et al., 2009;
Wu, Hu, Pan, Zhou, & Zhou, 2007; Wu, Ishurd, Sun, & Pan,
2005; Wu, Sun, & Pan, 2005; Wu, Sun, & Pan, 2006; Wu, Sun,
Qin, Pan, & Sun, 2006; Yan, Wang, Li, & Wu, 2011). Extraction
in hot or boiling water is the most common and convenient
method for extracting water-soluble mushroom polysaccharides. However, the major drawbacks of hot water extraction
are the high extraction temperature, long extraction time
and low extraction efficiency. Various methods have been
used to improve the extraction efficiency such as treatment
with enzymes, microwave and high power ultrasound (Wang,
Wang et al., 2011; Xie, Shan, & Zhang, 2009). The application
of high-power or high-intensity ultrasound or ultrasound-assisted extraction (UAE) has been widely studied for extracting
polysaccharides from different plant materials. The enhancement of extraction efficiency by UAE is mainly attributed to
the mechanical effects of ultrasound, particularly the shear
forces arising from acoustic cavitation (Velickovic, Milenovic,
Ristic, & Veljkovic, 2006). On the other hand, for extraction of
EPSs, the fermentation broth of C. sinensis was sequentially

36

Table 1 Mycelial biomass and EPS production by mycelial fermentation of Cordyceps sinensis.
Fungi source

Fermentation conditions
Medium composition

pH Culture vessel

Mycelial EPS References

biomass yield
Period
(g/L)
(g/L)
(days)

25

4.4

20

4.0

18
2225

5-L Jar fermentor: agitation speed,

300 rpm
5-L Stirred-tank fermenter: aeration
rate,2 vvm; agitation speed,150 rpm
Rotary shaker at 150 rpm
Shaking incubator at 150 rpm

25

Rotary shaker at 150 rpm

3.2

Hsu, Shiao, Hsieh, and Chang

(2002), Hsieh et al. (2005)
Kim and Yun (2005)

16

20.9

4.1

40
7

22.1
23.2

3.4

54.0

28.4

Dong, and Yao (2005)

Leung et al. 2006; Leung and
Wu (2007)
Cha et al., 2007

6.5

Rotary shaker at 120 rpm

5.9

Quan, Wang, Du, and Liu (2007)

24

7.0

Rotary shaker at 140 rpm

3.9

26

7.0

24.5

25

5.5

48.9

Choi et al (2010)

24

5.5

1000-mL shake flask: aeration rate 1 L/

min, agitation speed 130 rpm
5-L Jar fermenter: aeration rate 1.0 vvm,
agitation speed, 150 rpm
Rotary shaker at 180 rpm

Lang, Qi, Hou, Zhao, and Jiang

(2009)
Wu, Chen, and Hao (2009)

2.2

Li, Jiang, and Guan (2010)

27

6.5

250-mL shake flask at 160 rpm

0.4

Wang, Liu, Zhu, & Kuang, 2011

25

Shaking incubator at 150 rpm

14.7

7.2

Liu and Wu (2012)

25.0

2.8

Yin, Qiao, Qin, Tang, and Jia

(2013)

250-mL shake flask

12.3

56
4

10.0

6 ( 2 0 1 4 ) 3 3 4 7

22

JOURNAL OF FUNCTIONAL FOODS

C. sinensis CCRC36421 Sucrose 6.17%, corn steep powder 0.5%,

(NH4)2HPO4 0.5%, KH2PO4 0.15% (w/v)
C. sinensis
Sucrose 20, corn steep powder 25, CaCl2 0.78,
MgSO47H2O 1.73 (g/L)
C. sinensis 762
Sucrose 50, peptone 10, yeast extract 3 (g/L)
C. sinensis Cs-HK1
Glucose 40, yeast extract 5, peptone 5, KH2PO4
1, MgSO47H2O 0.5 (g/L);NH4Cl 10 mmol/L
C. sinensis 16
Sucrose 2%, yeast extract 0.9%, K2HPO4
0.3%,CaCl2 0.4% (w/v)
C. sinensis 1
Sucrose 3%, corn steep powder 5%, bean cake
4%, KH2PO4 0.1, MgSO47H2O 0.05%, vitamin
B1 0.01%
C. sinensis 383
Glucose 30, bean cake 20, MgSO4 2.0, KH2PO4
4.0 (g/L)
C. sinensis
Sucrose 20, yeast extract 2.0, KH2PO4 1.0,
MgSO47H2O 0.6 (g/L)
C. sinensis CCRC36421 Rice bran 1.5%, molasses 0.5%, CSL 3%,
KH2PO4 0.1%, MgSO4 0.05%
Hirsutella sinensis
Potato extract 20%, sucrose 2.5%, peptone
0.5%, K2HPO4 0.2%, MgSO4 0.05% (w/v)
C. sinensis CS001
Glucose 30, yeast extract 3, peptone 2, KH2PO4
0.6, MgSO47H2O 0.4, vitamin B1 0.01, palmitic
acid 1.0 (g/L)
C. sinensis Cs-HK1
Glucose 40, yeast extract 15, peptone 5,
KH2PO4 1, MgSO47H2O 0.5 (g/L);NH4Cl
10 mmol/L; Tween 80 1.5% (w/v)
C. sinensis
Glucose 30, peptone 15, KH2PO4 3.0,
MgSO47H2O 1.5, potato 200 (g/L)

Temperature
(C)

JOURNAL OF FUNCTIONAL FOODS

centrifuged and concentrated, and the resultant material was

precipitated by using ethanol, and then centrifuged to harvest
the crude EPSs. Fig. 2 summarizes the isolation procedures of
IPSs and EPSs from C. sinensis.
After extracting crude polysaccharides from C. sinensis, the
obtained polysaccharide precipitate was partially purified by
deproteination and decoloration, and then further purified

6 (2 0 14 ) 3 347

37

through column chromatography, such as ion-exchange chromatography, gel filtration chromatograph and affinity chromatography. Elution was conducted with an appropriate
running buffer, followed by collection, concentration, dialysis,
and lyophilization (Li, Su, Dong, & Tsim, 2002; Li et al., 2003;
Wang, Peng et al., 2011; Wang, Wang et al., 2009; Wu et al.,
2005; Wu et al., 2006; Wu et al., 2007; Yan, Li, Wang, & Wu,

Fig. 2 Schematic diagram for the extraction of intracellular and extracellular polysaccharides from Cordyceps sinensis.

38

JOURNAL OF FUNCTIONAL FOODS

2010; Yan et al., 2011). In addition, based on the different solubility of polysaccharides in ethanol, isopropanol, and other
solvents, polysaccharides were simply and effectively fractionated. Huang, Siu, Wang, Cheung, and Wu (2013) recently
isolated EPS fractions from a fermentation medium of C. sinensis by gradient ethanol precipitation. Their results suggest
that the method is simple and workable for the initial fractionation of polysaccharides, proteins, and their complexes
with different molecular sizes and for further identification
of bioactive components.

4.

Physicochemical characterization

The physicochemical and structural features of a polysaccharide mainly include monosaccharide composition, molecular
weight (MW), configuration of glycosidic linkages, type of glycosidic linkage, position of glycosidic linkage, sequence of
monosaccharide, number and location of appended non-carbohydrate groups, and molecular chain conformation (Cui,
2005; Nie, & Xie, 2011; Zhang, Cui, Cheung, & Wang, 2007).
Polysaccharides with different monosaccharide constituents
and chemical structures have been isolated from wild or cultured C. sinensis. Many research groups have elucidated the
chemical structures of purified IPSs and EPSs using infrared
spectroscopy, liquid-state nuclear magnetic resonance
(NMR) (one and two dimensions), solid-state NMR, gas chromatography (GC), GC-mass spectroscopy (GCMS), high-performance liquid chromatography (HPLC), acid hydrolysis,
methylation analysis, periodate- oxidation, and Smith degradation (Akaki et al., 2009; Kiho, Hui, Yamane, & Ukai, 1993;
Kiho et al., 1996; Kiho et al., 1999; Nie et al., 2011; Wang,
Yin, Chen, & Wang, 2009; Wang, Wang et al., 2009; Wang, Peng
et al., 2011; Wu et al., 2005; , 2006; Wu et al., 2007; Yan et al.,
2010; Yan et al., 2011; Wu et al., 2005). A wide range of bioactive polysaccharides of different structural characteristics
from C. sinensis have been investigated based on differences
in source materials, isolation protocols, and fractionation
protocols. The sources, molecular properties, chemical structures, and bioactivities are summarized in Table 2.

4.1.

Monosaccharide composition

Monosaccharide composition analysis usually involves cleavage of glycosidic linkages by acid hydrolysis, derivatization,
and detection and quantification by GC. In addition, high-performance anion-exchange chromatography with pulsed
amperometric detection has been gradually developed to supplement traditional methods as it doesnt require derivatization
of
monosaccharide
with
high
resolution
(Panagiotopoulos, Sempere, Lafont, & Kerherve, 2001). Recently, a 1-phenyl-3-methy-5-pyrazolone pre-column derivation method has been used to determine monosaccharide
composition (Chen, Siu, Wang, Liu, & Wu, 2013; Wang, Yin
et al., 2010; Wang, Wang et al., 2009).
Although many different IPSs and EPSs have been obtained, the monosaccharide composition is usually glucose
(Glu), mannose (Man), and galactose (Gal) in various mole ratios (Cha et al., 2007; Gong et al., 1990; Kiho et al., 1999; Li
et al., 2003; Miyazaki, Oikawa, & Yamada, 1977; Nie et al.,

6 ( 2 0 1 4 ) 3 3 4 7

2011; Wang, Wang et al., 2009; Wu et al., 2006; Yan et al.,

2010). However, the IPSs are also found to only contain
D-Glu to be composed of different polyglucans (Akaki et al.,
2009; Wu et al., 2005; Wu et al., 2005; Wu et al., 2006; Yan
et al., 2011), but so far, only one study has been reported that
EPS isolated from culture broth of C. sinensis was a b-glucan
(Yamada et al., 1984). In addition, IPSs and EPSs may also contain uronic acid, proteins and inorganic elements (Kiho et al.,
1986; Wang, Wang et al., 2011; Wang, Peng et al., 2011). These
polysaccharide conjugates isolated from natural or
cultured C. sinensis also represent a major class of bioactive
compounds and may exert more important pharmacological
effects than neutral polysaccharides.

4.2.

Average molecular weight

Various techniques such as viscometry, osmometry, sedimentation, and HPLC have been used to determine the average
polymer molecular weight (MW) and polydispersity index.
Among them, high-performance gel permeation chromatography (HPGPC) is a common method for determining the
MW of polysaccharides and has also been used by many
researchers for MW of IPSs and EPSs. Size-exclusion chromatography with multi-angle laser light scatter detection is also
an efficient method for the evaluation of the absolute MW of
polysaccharides and provides greater resolution than traditional gel permeation chromatography (Boukari et al., 2009;
Hilliou et al., 2009). Different MWs ranging from 103 to
106 Da have been found in various source materials of C. sinensis and experimental conditions (Nie et al., 2013; Yan et al.,
2011; Zhao et al., 2013; Zhong et al., 2009; Zhou, Gong, Su, Lin,
& Tang, 2009).

4.3.

Chemical structures

IPSs from natural and cultured C. sinensis usually consist of

glucose, mannose, and galactose with 1 ! 4(6)-glucopyranosyl (Glcp), 1 ! 6-mannopyranosyl (Manp), and 1 ! 4(6)-galactopyranosyl (Galp) (Guan et al., 2011; Nie et al., 2013; Zhong
et al., 2009; Zhou et al., 2009). The earliest reports on IPSs by
Miyazaki et al. (1977) and Kiho et al. (1986) included a galactomannan designated CS-I from a hot-water extract and a
water-soluble, protein-containing galactomannan (CT-4N)
isolated from a 5% sodium carbonate extract of C. sinensis,
respectively. Both IPSs contained (1 ! 2)-linked D-mannopyranosyl main-chains and (1 ! 5)-linked D-galactofuranosyl
side-chains. However, significant differences between CS-I
and CT-4N were observed, i.e., CS-I contains (1 ! 2,3) linkages
of D-mannopyranose residues, as well as (1 ! 3) and (1 ! 6)
linkages of galactofuranose, but CT-4 N has (1 ! 4,6) linkages
of mannopyranose and (1 ! 6) linkages of galactopyranose.
Thus, galactomannans from C. sinensis have a core of D-mannopyranosyl residues and D-galactofuranosyl side chains, and
some differences in the linkage mode are observed. In addition, a galactoglucomannan named CS-F10 (Kiho et al., 1999)
isolated from C. sinensis mycelium had a comb-type structure
composed of non-reducing terminal a-D-glucopyranosyl residues, (1 ! 5 and/or 6)-linked b-D-galactofuranosyl residues,
as well as (1 ! 2)-linked and branched a-D-mannopyranosyl
residues. Recently, some water-soluble IPSs isolated from

Table 2 Polysaccharides originated from Cordyceps sinensis fungi: source, chemical structures and bioactivities.
Components

Molecular
weight

Linkages and types

Bioactivities

References

Paecilomyces sinensis(Cs-4)
Paecilomyces sinensis(Cs-4)

Mycelium
Mycelium

Man:Gal = 1:1
Man:Gal = 3:5

23 kDa

CS-I Galactomannan
CT-4 N Galactomannan

Miyazaki et al. (1977)

Kiho et al., 1986

Paecilomyces sinensis(Cs-4)

Mycelium

Hot water
5% Sodium
carbonate
Hot water

Gal:Glc:Man = 62:28:10

45 kDa

CS-F30

Hypoglycemic activity

Paecilomyces sinensis(Cs-4)
Cephalosporium sinensis Chen
sp. nov.
Paecilomyces sinensis(Cs-4)

Mycelium
Mycelium

Hot water
Hot water

Gal:Glu:Man = 43:33:24
Glu:Man:Gal = 1:0.6:0.75

15 kDa
210 kDa

CS-F10 Galactoglucomann
CSP-1

Mycelium

Hot water

b-Glu

13.6 kDa

Wu et al. (2005)

Paecilomyces sinensis(Cs-4)

Mycelium

Hot water

b-Glu

12.9 kDa

Antitumour activity

Wu et al. (2005)

Paecilomyces sinensis(Cs-4)

Mycelium

a-Glu

184 kDa

Wu et al. (2005)

Paecilomyces sinensis(Cs-4)

Mycelium

Glu:Man = 9:1

7.7 kDa

Mannoglucan

Antitumour activity

Wu et al. (2007)

Paecilomyces sinensis(Cs-4)

Mycelium

0.05 M
phosphate
buffer
0.05 M
acetate
buffer
Hot water

Neutral (1 ! 3),(1 ! 4)-b-D- glucan

(1 ! 3)-b-D-glucan
Cordyglucans
SCI-I

Hypoglycemic activity
Antioxidant activity;
Hypoglycemic activity

Kiho et al. (1993), Kiho et al.

(1996)
Kiho et al., 1999
Li et al. (2003), Li et al. (2006)

Glc:Man:Gal = 21:2:1

Immunomodulating

Akaki et al. (2009)

Paecilomyces sinensis(Cs-4)

Mycelium

Hot water

Glc:Man:Gal = 2:1:1

460 kDa

Mycelium
Mycelium
Mycelium

Hot water
Hot water
Hot water

Glc:Man:Gal = 2.8:2.9:1
8.1 kDa
Man, Glu, Gal, Uronic acid 27 kDa
Man:Glu: Gal = 4:11:1
43.9 kDa

Tolypocladium sinensis

Mycelium

Hot water

a-Glu

1180 kDa

Tolypocladium sinensis

Mycelium

a-Glu

1150 kDa

AIPS a-D-(1 ! 4)-glucan

(86%), (1 ! 6) -a-D-glucose
(14%)

Tolypocladium sinensis
Paecilomyces sinensis(Cs-4)

Mycelium
Mycelium

1.25 M
NaOH /
0.04%
NaBH4
Hot water
Hot water

Cholesterol esterase
inhibitory activity
Antioxidant activity
Immunomodulating
Protection of chronic
renal failure
Antitumour and
Immuno-stimulating
effects
Antitumour and
Immuno-stimulating
effects

Kim (2010)

Cephalosporium sinense Chen

Cephalosporium sinense Chen
Cephalosporium sinense Chen

CS-Pp 1,3-b-D-glucan with

1,6-branched chain
! 3-a-D-Glcp-1 ! 3-b-DGlcp-1 ! 3-b-D-Galp-1 !
CPS1 Glucomannogalactan
CAPS
Galactoglucomanno- -glycan
(CPS-2)
WIPS a-D-(1 ! 4)-glucan

Man:Glu:Gal = 3.3:2.3:1
Glu(95%), Man, Gal

260 kDa

APS
CBHP a-1,4-linked-Glcp

Antioxidant activity
Antifibrotic effect

Cordyceps ophioglossoides
Paecilomyces sinensis(Cs-4)
Tolypocladium sinensis

Culture filtrate
Culture broth
Culture broth

Ethanol
Ethanol
95% Ethanol

b-Glu
Man:Gal:Glc = 10.3:3.6:1
Man:Glu: Gal = 23:1:2.6

632 kDa
43 kDa
104 kDa

CO-1
CS-81002
EPS

Paecilomyces sinensis
Tolypocladium sinensis
Tolypocladium sinensis
Tolypocladium sinensis

Culture
Culture
Culture
Culture

Ethanol
95% Ethanol
Ethanol
Ethanol

Glu:Man:Gal = 2.4:2:1
Glu:Man:Gal = 15.2:3.6:1
Glcp:GlcUp = 8:1
ManNH2:GalNH2:
Gal= 1.0:1.1:0.3

82 kDa
40 kDa
36 kDa
6 kDa

Cordysinocan
EPS-1A
AEPS-1
Poly-N-acetylhexosamine

Antitumour activity
Immunomodulating
Immunomodulatory and
antitumour
Immunomodulating

Immunomodulating
Antioxidant activity

broth
broth
broth
broth

Wang et al. (2009)

Wang et al. (2009)
Wang et al. (2010)
Yan et al. (2011)

Yan et al. (2011)

6 (2 0 14 ) 3 347

Polysaccharides Extraction
source
medium

JOURNAL OF FUNCTIONAL FOODS

Living strains

Shen et al. (2011)

Nie et al. (2011), Zhang, Li et al.
(2012), Zhang, Cheung et al.
(2012)
Yamada et al., 1984
Gong et al., 1990
Yang et al. (2005), Chen et al.
(2006), Sheng et al. (2011)
Cheung et al., 2009
Yan et al. (2010)
Wang, Liu, Zhu, & Kuang, 2011
Chen, Siu et al. (2013)

39

40

JOURNAL OF FUNCTIONAL FOODS

the cultured C. sinensis have been identified as glucomannogalactans, whose backbone were mainly composed of
(1 ! 2) and (1 ! 4)-linkage of mannose, (1 ! 3)-linkage of galactose, (1 ! ), and (1 ! 3,6)-linkage of glucose (Nie et al., 2011;
Wang, Wang et al., 2009). Furthermore, a novel neutral
mannoglucan isolated from C. sinensis mycelium has a backbone of predominantly (1 ! 4)-linked a-D-Glcp (61.3%) together with a proportion of (1 ! 3)-linked a-D-Glcp residues
(28.0%), with single a-D-Manp units (10.7%) as the side chains
attached to C-6 of (1 ! 3)-linked D-Glcp residue (Wu et al.,
2007). Similarly, Wang, Yin et al. (2010) reported the chemical
structure of a water-soluble polysaccharide (CPS-2) isolated
from cultured C. sinensis, which was composed mostly of a
a-(1 ! 4)-D-glucose and a a-(1 ! 3)-D-mannose branched with
a-(1 ! 4,6)-D-glucose every twelve residues on average. Based
on the above reports, it can be concluded that heteropolysaccharides are the most common bioactive polysaccharides in
the fruiting bodies and mycelia of C. sinensis.
Some researchers have also reported that IPSs are polyglucans with different structural characteristics. For instance,
Wu et al. (2005) found that cordyglucans obtained from C. sinensis mycelia is mainly composed of a (1 ! 3)-b-D-glucan
linked backbone with short (1 ! 6)-b-D-glucan linked
branches. Wu et al. (2006) also reported that the structure of
a polysaccharide (SCP-I) isolated from C. sinensis mycelium,
i.e., SCP-I, is a D-glucan containing an a-(1 ! 4)-linked backbone and a branched short a-(1 ! 6)-linkage. Furthermore,
Akaki et al. (2009) reported that an insoluble polysaccharide
(CS-Pp) purified from the cultured mycelium of C. sinensis
was a 1,3-b-D-glucan with some 1,6-branched chains. Recently, Yan et al. (2011) isolated the two polysaccharides,
WIPS and AIPS, from hot water and dilute alkaline extracts,
respectively, of the mycelial biomass of a C. sinensis fungus
Cs-HK1, which were characterized as a-D-glucans with a backbone of (1 ! 4) linked a-D-glucopyranosyl (Glcp) (>60%). WIPS
is found to have a short branch of (1 ! 6)-linked a-D-Glcp
(14%), whereas AIPS is a highly linear glucan.
In addition to the IPSs extracted from the mycelium or
fruiting bodies of C. sinensis, EPSs isolated from the culture
broth of C. sinensis have been reported. Gong et al. (1990) reported an EPS called CS-81002 isolated from the fermentation
medium of C. sinensis and characterized as a branched heteropolysaccharide. Recently, Yan et al. (2010) reported the isolation and structure of EPS-1A from a fermentation broth of C.
sinensis Cs-HK1. EPS-1A was found to be a slightly branched
polysaccharide with its backbone being composed of (1 ! 6)a-D-glucose residues (77%) and (1 ! 6)-a-D-mannose residues (23%). Branching occurred at the O-3 position of
(1 ! 6)-a-D-mannose residues of the backbone with (1 ! 6)a-D-mannose residues and (1 ! 6)-a-D-glucose residues and
terminated with b-D-galactose residues. Meanwhile, Wang,
Peng et al. (2011) reported that on the acidic polysaccharide
AEPS-1, which had a linear backbone of (1 ! 3)-linked
a-D-Glcp residues with two branches, namely, a-D-Glcp and
a-D-pyrano-glucuronic acid (GlcUp), attached to the main
chain by (1 ! 6) glycosidic bonds at every seventh a-D-Glcp
unit. In addition, a novel poly-N-acetylhexosamine (polyhexNAc) of about 6 kDa was isolated from the low-MW fraction
of EPS produced from the liquid fermentation of C. sinensis

6 ( 2 0 1 4 ) 3 3 4 7

Cs-HK1. The molecular structure was elucidated as a [-4-b-DManNAc-(1 ! 3)-b-D-GalNAc-(1 ! ] disaccharide repeating
unit in the main chain with a Gal branch randomly occurring
at the 3-position of ManNAc (Chen, Siu et al., 2013).

4.4.

Conformational features

The activities of polysaccharides depend on their MWs,

chemical structures, and chain conformations. In general,
polysaccharides in aqueous solutions exhibit different
forms of chain conformations, such as random coil (Senti
et al., 1955), various helical forms, single helix, double helix,
triple helix (Kashiwagi, Norisuye, & Fujita, 1981; Sato, Norisuye, & Fujita, 1984; Zhang, & Yang, 1995), and aggregate
(Ding, Jiang, Zhang, & Wu, 1998). A few reports are available
on the solution properties and chain conformations of C.
sinensis polysaccharides. For example, Cai, Li, and Lu
(1999) firstly developed a method to study the morphology
of a IPS by atomic force microscopy (AFM). Their results
show that this IPS has a multi-branched structure and various linkages between adjacent monosaccharides, which
make up small rings and helical structures. Very recently,
the morphological characteristics and chain conformation
of EPS isolated from a mycelial culture of C. sinensis CsHK1 have been analyzed by AFM together with the Congo
red test, optical rotation, and dynamic light scattering. Results suggest that this EPS forms large interwoven networks
in aqueous solution and is primarily connected with triplehelical conformation and occasionally single-helical conformation in solution. However, IPSs obtained from Cs-HK1
mycelium exhibit random coils in aqueous alkaline solutions (Wang, Cheung, Leung, & Wu, 2010; Wang, Peng
et al., 2011; Yan et al., 2011). In addition, the random coils
or aggregated networks of EPS-1 formed in aqueous solution
were transformed into the single helices after sulfation
(Yan, Wang, Ma, & Wu, 2013).
The relationships among solution properties, chain conformations of polysaccharides, and their biological activities
are difficult to elucidate. The detailed chain conformations
of C. sinensis polysaccharides in aqueous solutions require
further investigations using other technological means,
such as static and dynamic light scattering, viscosity analysis based on the theory of dilute polymer solutions, circular
dichroism analysis, transmission electron microscope, scanning electron microscopy, AFM, AFM-based single-molecule
force spectroscopy, fluorescence correlation spectroscopy,
and NMR spectroscopy (Yang, & Zhang, 2009). Diluted solution theory, molecular modeling, and computer-assisted energy minimization methods have also been used to analyze
the chain conformation of polysaccharides (Brant, 1981; PolFachin, Fernandes, & Verli, 2009; Perez, Kouwijzer, Mazeau,
& Engelsen, 1996; Strlegel, Plattner, & Willett, 1999).

5.

Bioactivities

Based on TCM theories, the major effects of C. sinensis are to

enrich the lung yin and yang. Its use includes treatment of
chronic lower back pain, sensitivity to cold, overabundance
of mucus and tears, chronic cough and wheezing, and blood

JOURNAL OF FUNCTIONAL FOODS

in phlegm due to consumption as a result of kidney yang

(shenyangqu). C. sinensis also has antibacterial activity, reduces asthma, lowers blood pressure, and strengthens heartbeat according to Western medicine (Zhu et al., 1998).
Polysaccharides represent a major class of bioactive constituents of C. sinensis, contributing to its health effects and pharmacological activities according to a large number of animal
and clinical studies. The multiple bioactivities and health
benefits of IPSs and EPSs are summarized and compared in
detail below.

5.1.

Immunomodulatory activity

Immunomodulation is the most notable biological function of

natural polysaccharides, which is associated with their putative role as biological response modifiers (Moradali, Mostafavi, Ghods, & Hedjaroude, 2007). The immunostimulating
and immunosuppressive properties of IPSs and EPSs have
been assessed on natural killer cells, T-cells, B-cells, and macrophage-dependent immune system responses (Koh, Yu, Suh,
Choi, & Ahn, 2002; Paterson, 2008; Zhong et al., 2009; Zhou
et al., 2009). Phagocyte release is an early step in the response
of macrophages to pathogen invasion of the human body.
Macrophages can also defend against pathogen invasion by
secreting proinflammatory cytokines [e.g., tumour necrosis
factor (TNF)-a and interleukin (IL)-1] and releasing cytotoxic
and inflammatory molecules [e.g., nitric oxide (NO) and reactive oxygen species] (Medzhitov, & Janeway, 2000).
The majority of studies on IPSs and EPSs immunomodulation have been evaluated by activating macrophages. EPSs
prepared from cultured C. sinensis induce the production of
TNF-a, IL-6, and IL-10 dose-dependently and elevate phagocytes in monocytes and PMN, but IPSs only moderately induce
TNF-a release, CD11b expression, and phagocytes at the same
concentration (Cheung et al., 2009; Kuo, Chang, Cheng, & Wu,
2007). This finding indicates that the immunomodulatory
components of cultured C. sinensis mainly reside in the culture filtrate and are similar to previously reported ones (Gong
et al., 1990). Recent reports have shown that polysaccharides
isolated from various natural or cultured C. sinensis have the
same immunomodulatory activity of stimulation of the release of several major cytokines in the mouse macrophage
cell line RAW264.7 and in mouse splenocyte cells by activating
the IjB-NF-jB pathway (Akaki et al., 2009; Chen, Zhang, Shen,
& Wang, 2010; Wang, Peng et al., 2011). More recently, Chen,
Yuan, Wang, Song, and Zhang (2012) reported that an acid
polysaccharide fraction (APSF) from C. sinensis fungus could
increase the expressions of TNF-a, IL-12 and iNOS, and reduce
the expression of IL-10 of Ana-1 cells, convert M2 macrophages to M1 phenotype by activating NF-jB pathway.

5.2.

Antitumour activity

Since the first report on the antitumour activity of mushroom

polysaccharides in the 1960s (Chihara, 1969), researchers have
isolated structurally diverse polysaccharides with strong antitumour activity from plants, animals, and fungi. Mushroom
polysaccharides exert inhibitory effects toward many kinds
of tumours, such as Sarcoma 180 solid tumour, Ehrlich solid
tumour, Sarcoma 37, Yoshida sarcoma, and Lewis lung

6 (2 0 14 ) 3 347

41

carcinoma (Wasser, & Weis, 1999). The currently accepted

mechanism by which mushroom polysaccharides exert antitumour effects can be summarized as follows: (1) prevention
of oncogenesis by oral administration of polysaccharides
isolated from medicinal mushrooms, (2) enhancement of
immunity against bearing tumours, (3) direct antitumour
activity to induce the apoptosis of tumour cells, and (4) prevention of the spread or migration of tumour cells in the body
(Moradali et al., 2007; Wasser, 2002; Zhang et al., 2007).
Many studies have demonstrated that both of IPSs and
EPSs have strong antitumour activity through the above proposed mechanisms. As a simple method, the prevention of
the onset of tumour by oral administration is used to evaluate
the antitumour activity of polysaccharides in vivo. IPSs obtained from the mycelia of C. sinensis are effective against sarcoma 180, with almost 90% inhibition in ICR/JCL mice (Wu
et al., 2005; Zhang et al., 2007). In addition, EPSs obtained from
a cultured broth of C. sinensis significantly lowers c-Myc, cFos, and vascular endothelial growth factor expression in
B16 melanoma-bearing mice. Thus, EPSs can inhibit tumour
growth in the lungs and liver of mice and can be a potential
adjuvant in cancer therapy (Yang et al., 2005).
EPSs exert their antitumour effect mainly through the
enhancement and activation of the immune response of the
host organism. An EPS isolated from one of the anamorph
strains of C. sinensis belonging to Tolypocladium spp. is found
to significantly enhance the neutral red uptake capacity of
peritoneal macrophages and spleen lymphocyte proliferation
in B16-bearing mouse. The metastasis of B16 melanoma cells
in lungs and liver is also significantly inhibited. Moreover, this
EPS can markedly prevent H22 tumour growth and elevate
immunocyte activity in H22 tumour-bearing mice, indicating
that EPSs inhibit tumour cells mainly by activating the hosts
immune system (Zhang, Li, Qiu, Chen, & Zheng, 2008; Zhang,
Yang, Chen, Hou, & Han, 2005). To confirm this finding, Sheng,
Chen, Li, and Zhang (2011) investigated the effects of EPSs on
immunocytes in vitro, and results indicate that EPS treatment
significantly promotes the mRNA and protein levels of TNF-a
and IFN-c. This phenomenon supports the assumption that
antitumour activity is related to the promoted cytokine
expression of immunocytes. EPSs can also stimulate the maturation and activation of murine and human dendritic cells
by inhibiting STAT3 activation (Huang, Song, Yang, Yin, &
Zhang, 2011; Song, Lin, Yuan, & Zhang, 2011). Furthermore,
EPS may induce dendritic cell sarcoma (DCS) cells to exhibit
mature characteristics, and the mechanism involved is probably related to the inhibition of the JAK2/STAT3 signal pathway and promotion of the NF-jB signal pathway (Song
et al., 2013). On the other hand, IPSs can also activate many
immune cells to modulate the release of cell signal messengers such as cytokines, and increased cytokine production
in immune cells has been studied in mice and humans (Chen,
Shiao, Lee, & Wang, 1997; Yoon, Yu, Shin, & Suh, 2008).
Finally, the ability to induce apoptosis has been identified
and utilized in successful cancer chemotherapeutics (Chen
et al., 1997; Yoon et al., 2008), and studies have suggested that
C. sinensis can induce apoptosis (Buenz, Bauer, Osmundson,
& Motley, 2005). A 410 kDa polysaccharide fraction (IPS)
isolated from C. sinensis is found to inhibit cell proliferation,
promote the apoptosis of IL-1- and platelet-derived growth

42

JOURNAL OF FUNCTIONAL FOODS

6 ( 2 0 1 4 ) 3 3 4 7

factor-BB-activated human mesangial cells in vitro, and prevent the tyrosine phosphorylation of human mesangial proteins. Bcl-2 and Bcl-XL are probably among these proteins as
revealed by results of immunoprecipitation and immunoblotting (Yang, Chen, Kuo, & Lin, 1999; Yang, Huang, Hsieh, & Lin,
2003). Similarly, crude polysaccharides obtained from mycelium of C. sinensis also affect the induction of apoptosis and
expression of p53 gene in SP2/0 cells in vitro (Liu et al.,
2009). Shen, Shao, Ni, Xu, and Tong (2009) reported the effects
of the polysaccharide fraction of C. sinensis (PSCS) on triptolide (TPL)-induced apoptosis in HL-60 cells and the molecular
events and signaling pathway, enhancing TPL-induced apoptosis and inhibiting the expression of NF-jB and caspases 3, 6,
7, and 9. However, up to now, no studies revealed that EPSs
isolated from culture broth of C. sinensis could induce
apoptosis.

in tumour-bearing mice, and by reducing the MDA level in

the liver and brain of tumour-bearing mice (Chen et al., 2006).
A few studies have reported on the antioxidative activities
of EPSs isolated from culture broths of C. sinensis. Wu et al. recently studied the antioxidant action of an EPS (named CsHK1) isolated from the mycelial liquid culture of C. sinensis
fungus by ethanol precipitation in vitro. Results demonstrate
that Cs-HK1 shows moderate antioxidative activities, and
acidic degradation can improve its antioxidative activities
and radical-scavenging capacities (Leung, Zhao, Ho, & Wu,
2009; Yan et al., 2009). Very recently, EPS fractions were isolated from the fermentation broth of Cs-HK1 by gradient precipitation with ethanol at different volume ratios to the liquid
medium exhibited moderate antioxidant activities and their
activities showed a significant dependence on the protein
content (Huang et al., 2013).

5.3.

5.4.

Antioxidant activity

Oxidation phenomena have been implicated in many illnesses, such as diabetes mellitus, arteriosclerosis, nephritis,
Alzheimers disease, and cancer (Negre-Salvayre, Coatrieux,
Ingueneau, & Salvayre, 2008). Therefore, natural antioxidants
isolated from plants, fungi, and marine algae represent most
useful nutraceuticals and functional foods for health protection and disease prevention (Gutteridge, & Halliwell, 1994).
Antioxidant activity has become one of the focuses of studies
on mechanisms of the nutraceutical and therapeutical effects
of TCM using various assay methods and activity indices
(Dong & Yao, 2008; Schlesier, Harwat, Bohm, & Bitsch, 2002).
Li, Li, Dong, and Tsim (2001) studied the antioxidative
activity of water extracts of natural C. sinensis and mycelial
biomass of the Cordyceps anamorph fungus Cephalosporium
sinensis of various sources using xanthine oxidase, haemolysis induction, and lipid peroxidation. Their results show that
Cordyceps generally possesses strong antioxidative activity in
all assays, and cultured Cordyceps mycelia have antioxidative
activity as strong as that of natural C. sinensis. Further testing
suggests that several forms of IPSs prepared by ethanol
precipitation and DEAE column chromatography have great
antioxidative activities. Li et al. (2003) used column chromatography to purify a polysaccharide-enriched fraction isolated
from cultured C. sinensis that possesses strong oxidative activity. A polysaccharide named CSP-1 is then obtained using
activity-guided fractionation. CSP-1 with a MW of 210 kDa is
found to have strong antioxidative activity in rat pheochromocytoma PC12 cells, protecting against H2O2-induced cell
damage, lipid peroxidation, and activation of major antioxidant enzymes, including glutathione peroxidase and superoxide dismutase in cells. Very recently, an APS has been
isolated from Tolypocladium sinensis, another anamorph strain
of C. sinensis. This APS is found to markedly protect PC12 cells
from H2O2-induced cell injury by increasing glutathione peroxidase, catalase, and superoxide dismutase (SOD) activities,
as well as by reducing lactate dehydrogenase and malondialdehyde (MDA) levels (Shen, Song, Wu, & Zhang, 2011). In addition, IPSs also affect the antioxidative activity of H22-bearing
mice by enhancing the SOD activity of the liver, brain, and
serum, as well as the GSH-Px activity of the liver and brain

Hypoglycemic effect

Many research groups have evaluated the hypoglycaemic effects of natural products, including fungal polysaccharides
using the Streptozotocin (STZ)-induced and alloxan-induced
animal models (Hwang, & Yun, 2010; Hwang et al., 2005;
Yamac et al., 2008). IPSs extracted with hot water and alkalis
have significant hypoglycemic effects on normal and alloxandiabetic mice and STZ-diabetic rats in vivo by reducing the
plasma glucose level in both STZ-induced diabetic rats and alloxan-induced diabetic mice, thereby increasing the serum
insulin levels in diabetic animals (Li et al., 2006; Wang, &
Shiao, 2000; Zhang et al., 2006). IPSs also significantly lowered
the levels of plasma triglyceride and cholesterol in mice and
increase the activities of hepatic glucokinase, hexokinase,
and glucose-6-phosphate dehydrogenase (Kiho et al., 1993;
Kiho et al., 1996; Kiho et al., 1999).

5.5.

Other bioactivities

As aforementioned, IPSs and EPSs obtained from wild or cultured C. sinensis demonstrate immunoregulation, antitumour,
antioxidation, and hypoglycemic effects, as well as other
important bioactivities, including anti-fibrosis, anti-fatigue,
kidney protection, increasing plasma testosterone levels,
and radiation protection (Nie et al., 2013; Wang, Yin et al.,
2009, 2010; Wong, Wu, & Benzie, 2011; Yan, Zhang, & Wang,
2012; Yao et al., 2014; Zhang, Cheung, Al-Assaf, Phillips, &
Phillips, 2012; Zhong et al., 2009).

6.

Conclusions and future perspectives

C. sinensis is a well-known and precious medicinal fungus in

China for its ability to treat a broad spectrum of human diseases, especially those related to the functions of the lung
and kidney, the immune system, and for its ability to enhance
the quality of life and physical performance. Polysaccharides
have been identified as the major active components of C. sinensis with a wide range of bioactivities including immunomodulation, antitumour, antioxidation, and hypoglycemic
effects. Fermentation production, isolation, structural characterization, and the bioactivities of polysaccharides from

JOURNAL OF FUNCTIONAL FOODS

different wild or natural C. sinensis have been extensively

investigated in recent years, mainly in China, Japan, and Korea. However, the relationship between structural features,
solution behavior, space conformation, and their bioactivity
is unclear due to the structural diversity and complexity of
polysaccharide molecules. The fruiting bodies or mycelial biomass as the sources of IPSs, especially EPSs isolated from the
culture broth, have attained from different C. sinensis species
and in various conditions. Because of the variable properties
of raw material and the composition of polysaccharides, it
is difficult to maintain consistency, reproducibility and reliability of the results. There is a need to establish standard
protocols for collection and preparation of the source material and for the extraction, isolation and purification of polysaccharides. These will be useful for determination of the
chemical structures and chain conformations and the biological activities and for applications in food, medicine and
cosmetic products.

Acknowledgements
This work was supported by Grants from the Chinas Postdoctoral Science Fund Projects (2013M531292) and the Guangdong Education University-Industry Cooperation Project
(2008B090500002).

R E F E R E N C E S

Akaki, J. J., Matsui, Y., Kojima, H., Nakajima, S., Kamei, K., &
Tamesada, M. (2009). Structural analysis of monocyte
activation constituents in cultured mycelia of Cordyceps
sinensis. Fitoterapia, 80, 182187.
Au, D., Wang, L., Yang, D., Mok, D. K., Chan, A. S., & Xu, H. (2012).
Application of microscopy in authentication of valuable
Chinese medicine I-Cordyceps sinensis, its counterfeits, and
related products. Micro Research Technology, 75, 5464.
Boukari, I., Putaux, J. L., Cathala, B., Barakat, A., Saake, B.,
Remond, C., ODonohue, M., & Chabbert, B. (2009). In vitro
model assemblies to study the impact of lignin-carbohydrate
interactions on the enzymatic conversion of xylan.
Biomacromolecules, 10, 24892498.
Brant, D. A. (1981). Solution properties of polysaccharides: Based on a
symposium. Division of Carbohydrate Chemistry: American
Chemical Society.
Buenz, E. J., Bauer, B. A., Osmundson, T. W., & Motley, T. J. (2005).
The traditional Chinese medicine Cordyceps sinensis and its
effects on apoptosis homeostasis. Journal of Ethnopharmacology,
96, 1929.
Cai, L. T., Li, P., & Lu, Z. H. (1999). Observation of the structure
morphology of Cordyceps polysaccharide by atomic force
microscope. Journal of Chinese Electron Microscopy Society, 18,
103105 (in Chinese).
Cha, S. H., Lim, J. S., Yoon, C. S., Koh, J. H., Chang, H. I., & Kim, S.
W. (2007). Production of mycelia and exo-biopolymer from
molasses by Cordyceps sinensis 16 in submerged culture.
Bioresource Technology, 98, 165168.
Chen, Y. J., Shiao, M. S., Lee, S. S., & Wang, S. Y. (1997). Effect of
Cordyceps sinensis on the proliferation and differentiation of
human leukemic U937 cells. Life Sciences, 60, 23492359.
Chen, S., Siu, K. C., Wang, W. Q., Liu, X. X., & Wu, J. Y. (2013).
Structure and antioxidant activity of a novel poly-N-

6 (2 0 14 ) 3 347

43

acetylhexosamine produced by a medicinal fungus.

Carbohydrate Polymers, 94, 332338.
Chen, P. X., Wang, S., Nie, S., & Marcone, M. (2013). Properties of
Cordyceps sinensis: A review. Journal of Functional Foods, 5,
550569.
Chen, W., Yuan, F., Wang, K., Song, D., & Zhang, W. (2012).
Modulatory effects of the acid polysaccharide fraction from
one of anamorph of Cordyceps sinensis on Ana-1 cells. Journal of
Ethnopharmacology, 142, 739745.
Chen, J. P., Zhang, W. Y., Lu, T. T., Li, J., Zheng, Y., & Kong, L. D.
(2006). Morphological and genetic characterization of a
cultivated Cordyceps sinensis fungus and its polysaccharide
component possessing antioxidant property in H22 tumourbearing mice. Life Sciences, 78, 27422748.
Chen, W., Zhang, W., Shen, W., & Wang, K. (2010). Effects of the
acid polysaccharide fraction isolated from a cultivated
Cordyceps sinensis on macrophages in vitro. Cellular Immunology,
262, 6974.
Cheung, J. K. H., Li, J., Cheung, A. W. H., Zhu, Y., Zheng, K. Y. Z., Bi,
C. W. C., Duan, R., Choi, R. C. Y., Lau, D. T. W., Dong, T. T. X.,
Lau, B. W. C., & Tsim, K. W. K. (2009). Cordysinocan, a
polysaccharide isolated from cultured Cordyceps, activates
immune responses in cultured T-lymphocytes and
macrophages: Signaling cascade and induction of cytokines.
Journal of Ethnopharmacology, 124, 6168.
Chihara, G. (1969). The antitumour polysaccharide lentinan: An
overview 222. In T. Aoki et al. (Eds.), Manipulation of host defense
mechanism (pp. 687694).
Choi, J. W., Ra, K. S., Kim, S. Y., Yoon, T. J., Yu, K. W., Shin, K. S., Lee,
S. P., & Suh, H. J. (2010). Ehancement of anti-complementary
and radical scavenging activities in the submerged culture of
Cordyceps sinensis by addition of citrus peel. Bioresource
Technology, 101, 60286034.
Cui, S. W. (2005). Structural analysis of polysaccharides. In Steve
W. Cui (Ed.), Food carbohydrates: Chemistry, physical properties and
applications (1 edition). Boca Raton, FL: CRC Press.
Ding, Q., Jiang, S., Zhang, L., & Wu, C. (1998). Laser light-scattering
studies of pachyman. Carbohydrate Research, 308, 339343.
Ding, C., Tian, P. X., Xue, W., Ding, X., Yan, H., Pan, X., Feng, X.,
Xiang, H., Hou, J., & Tian, X. (2011). Efficacy of Cordyceps sinensis
in long term treatment of renal transplant patients. Frontiers in
Bioscience (Elite Edition), 3, 301307.
Dong, C. H., & Yao, Y. J. (2005). Nutritional requirements of
mycelial growth of Cordyceps sinensis in submerged culture.
Journal of Applied Microbiology, 99, 483492.
Dong, C. H., & Yao, Y. J. (2008). In vitro evaluation of antioxidant
activities of aqueous extracts from natural and cultured
mycelia of Cordyceps sinensis. LWT-Food Science and Technology,
41, 669677.
Gong, M., Zhu, Q., Wang, T., Wang, X. L., Ma, J. X., & Zhang, W. J.
(1990). Molecular structure and immunoactivity of the
polysaccharide from Cordyceps sinensis (Berk) Sacc. Chinese
Biochemical Journal, 6, 486492 (in Chinese).
Guan, J., Zhao, J., Feng, K., Hu, D. J., & Li, S. P. (2011). Comparison
and characterization of polysaccharides from natural and
cultured Cordyceps using saccharide mapping. Analytical and
Bioanalytical Chemistry, 399, 34653474.
Gutteridge, J. M. C., & Halliwell, B. (1994). Antioxidants in nutrition,
health, and disease. Oxford, New York, NY: Oxford University
Press.
Hilliou, L., Freitas, F., Oliveira, R., Reis, M. A. M., Lespineux, D.,
Grandfils, C., & Alves, V. D. (2009). Solution properties of an
exopolysaccharide from a Pseudomonas strain obtained using
glycerol as sole carbon source. Carbohydrate Polymers, 78,
526532.
Holliday, J., & Cleaver, M. (2008). Medicinal value of the caterpillar
fungi species of the genus Cordyceps (Fr.) Link (Ascomycetes). A
review. International Journal of Medicinal Mushrooms, 10, 219234.

44

JOURNAL OF FUNCTIONAL FOODS

Hsieh, C. Y., Tsai, M. J., Hsu, T. H., Chang, D. M., & Lo, C. T. (2005).
Medium optimization for polysaccharide production of
Cordyceps sinensis. Applied Biochemistry and Biotechnology, 120,
145157.
Hsu, T. H., Shiao, L. H., Hsieh, C., & Chang, D. M. (2002). A
comparison of the chemical composition and bioactive
ingredients of the Chinese medicinal mushroom
DongChongXiaCao, its counterfeit and mimic, and fermented
mycelium of Cordyceps sinensis. Food Chemistry, 78, 463469.
Huang, Q. L., Siu, K. C., Wang, W. Q., Cheung, Y. C., & Wu, J. Y.
(2013). Fractionation, characterization and antioxidant activity
of exopolysaccharides from fermentation broth of a Cordyceps
sinensis fungus. Process Biochemistry, 48, 380386.
Huang, J., Song, D., Yang, A., Yin, H., & Zhang, W. (2011).
Differentiation and maturation of human dendritic cells
modulated by an exopolysaccharide from an anamorph of
Cordyceps sinensis. Biomedicine & Preventive Nutrition, 1, 126131.
Hwang, H. J., Kim, S. W., Lim, J. M., Joo, J. H., Kim, H. O., Kim, H. M.,
& Yun, J. W. (2005). Hypoglycemic effect of crude
exopolysaccharides produced by a medicinal mushroom
Phellinus baumii in streptozotocin-induced diabetic rats. Life
Sciences, 76, 30693080.
Hwang, H. S., & Yun, J. W. (2010). Hypoglycemic effect of
polysaccharides produced by submerged mycelial culture of
Laetiporus sulphureus on streptozotocin-induced diabetic rats.
Biotechnology and Bioprocess Engineering, 15, 173181.
Jeffrey, G. (2012). The Viagra transforming local economies in
India. BBC News. Retrieved, 09(07), 2012.
Ji, D. B., Ye, J., Li, C. L., Wang, Y. H., Zhao, J., & Cai, S. Q. (2009).
Antiaging effect of Cordyceps sinensis extract. Phytotherapy
Research, 23, 116122.
Kashiwagi, Y., Norisuye, T., & Fujita, H. (1981). Triple helix of
Schizophyllum commune polysaccharide in dilute solution, 4.
Light scattering and viscosity in dilute aqueous sodium
hydroxide. Macromolecules, 14, 12201225.
Kiho, T., Hui, J., Yamane, A., & Ukai, S. (1993). Polysaccharide in
fungi. XXXII. Hypoglycemic activity and chemical properties of
a polysaccharide from the cultural mycelium of Cordyceps
sinensis. Biological and Pharmaceutical Bulletin, 16, 12911293.
Kiho, T., Ookubo, K., Usui, S., Ukai, S., & Hirano, K. (1999).
Structural features and hypoglycemic activity of a
polysaccharide (CS-F10) from the cultured mycelium of
Cordyceps sinensis. Biological and Pharmaceutical Bulletin, 22,
966970.
Kiho, T., Tabata, H., Ukai, S., & Hara, C. (1986). Polysaccharide in
fungi. 18. A minor, protein-containing galactomannan from a
sodium-carbonate extract of Cordyceps sinensis. Carbohydrate
Research, 156, 189197.
Kiho, T., Yamane, A., Hui, J., Usui, S., & Ukai, S. (1996).
Polysaccharides in fungi, XXXVI. Hypoglycemic activity of a
polysaccharide (CS-F30) from the cultural mycelium of
Cordyceps sinensis and its effect on glucose metabolism in
mouse liver. Biological and Pharmaceutical Bulletin, 19, 294296.
Kim, S. D. (2010). Isolation, structure and cholesterol esterase
inhibitory activity of a polysaccharide, PS-A, from C. sinensis.
Journal of the Korean Society from Applied Biological Chemistry, 53,
784789.
Kim, H. O., & Yun, J. W. (2005). A comparative study on the
production of exopolysaccharides between two
entomopathogenic fungi Cordyceps militaris and Cordyceps
sinensis in submerged mycelial cultures. Journal of Applied
Microbiology, 99, 728738.
Koh, J. H., Yu, K. W., Suh, H. J., Choi, Y. M., & Ahn, T. S. (2002).
Activation of macrophages and the intestinal immune system
by an orally administered decoction from cultured mycelia of
Cordyceps sinensis. Bioscience, Biotechnology, and Biochemistry, 66,
407411.

6 ( 2 0 1 4 ) 3 3 4 7

Kuo, M. C., Chang, C. Y., Cheng, T. L., & Wu, M. J. (2007).

Immunomodulatory effect of exo-polysacchrides from
submerged cultured Cordyceps sinensis: Enhancement of
cytokine synthesis, CD11b expression, and phagocytosis.
Applied Microbiology and Biotechnology, 75, 769775.
Lang, J., Qi, X., Hou, Y., Zhao, S., & Jiang, G. (2009). Production of
exopolysaccharides by Cordyceps sinensis in liquid culture.
Journal of Dalian Polytechnic University, 28, 107110 (in Chinese).
Leung, P. H., & Wu, J. Y. (2007). Effects of ammonium feeding on
production of bioactive metabolites (cordycepin and
exopolysaccharides) in mycelial culture of a Cordyceps sinensis
fungus. Journal of Applied Microbiology, 103, 19421949.
Leung, P. H., Zhang, Q. X., & Wu, J. Y. (2006). Mycelium cultivation,
chemical composition and antitumour activity of a
Tolypocladium sp. fungus isolated from wild Cordyceps sinensis.
Journal of Applied Microbiology, 101, 275283.
Leung, P. H., Zhao, S. N., Ho, K. P., & Wu, J. Y. (2009). Chemical
properties and antioxidant activity of exopolysaccharides
from mycelial culture of Cordyceps sinensis fungus Cs-HK1. Food
Chemistry, 114, 12511256.
Li, S. P., & Tsim, K. W. K. (2004). The biological and
pharmacological properties of Cordyceps sinensis, a
traditional Chinese medicine that has broad clinical
applications. In L. Packer, C. N. Ong, & B. Halliwell (Eds.), Herbal
and traditional medicine: Molecular aspects of health (pp. 657683).
New York: Marcel Dekker.
Li, R., Jiang, X. L., & Guan, H. S. (2010). Optimization of mycelium
biomass and exopolysaccharides production by Hirsutella sp in
submerged fermentation and evaluation of
exopolysaccharides antibacterial activity. African Journal of
Biotechnology, 9, 196203.
Li, S. P., Li, P., Dong, T. T. X., & Tsim, K. W. K. (2001). Anti-oxidation
activity of different types of natural Cordyceps sinensis and
cultured Cordyceps mycelia. Phytomedicine, 8, 207212.
Li, S. P., Su, Z. R., Dong, T. T. X., & Tsim, K. W. K. (2002). The fruiting
body and its caterpillar host of Cordyceps sinensis show close
resemblance in main constituents and anti-oxidation activity.
Phytomedicine, 9, 319324.
Li, Y., Wang, X. L., Jiao, L., Jiang, Y., Li, H., Jiang, S. P.,
Lhosumtseiring, N., Fu, S. Z., Dong, C. H., Zhan, Y., & Yao, Y. J.
(2011). A survey of the geographic distribution of
Ophiocordyceps sinensis. The Journal of Microbiology, 49, 913919.
Li, S. P., Zhang, G. H., Zeng, Q., Huang, Z. G., Wang, Y. T., Dong, T. T.
X., & Tsim, K. W. K. (2006). Hypoglycemic activity of
polysaccharide, with antioxidation, isolated from cultured
Cordyceps mycelia. Phytomedicine, 13, 428433.
Li, S. P., Zhao, K. J., Ji, Z. N., Song, Z. H., Dong, T. T. X., Lo, C. K.,
Cheung, J. K. H., Zhu, S. Q., & Tsim, K. W. K. (2003). A
polysaccharide isolated from Cordyceps sinensis, a traditional
Chinese medicine, protects PC12 cells against hydrogen
peroxide-induced injury. Life Sciences, 73, 25032513.
Liu, Z., Li, P., Zhao, D., Tang, H., & Guo, J. (2010). Protective effect of
extract of Cordyceps sinensis in middle cerebral artery
occlusion-induced focal cerebral ischemia in rats. Behavioral
and Brain Functions, 6, 6168.
Liu, Y., Liu, N., Han, B., Tian, X., Su, J., & Zhao, D. (2009). Effect of
crude polysaccharides from mycelium of Cordyceps sinensis on
expression of p53 gene and induction of apoptosis in SP2/0
cells. Acta Veterinaria et Zootechnica Sinica, 40, 117121 (in
Chinese).
Liu, Y. S., & Wu, J. Y. (2012). Effects of Tween 80 and pH on mycelial
pellets and exopolysaccharide production in liquid culture of a
medicinal fungus. Journal of Industrial Microbiology and
Biotechnology, 39, 623628.
Lo, H. C., Hsieh, C., Lin, F. Y., & Hsu, T. H. (2013). A Systematic
Review of the Mysterious Caterpillar Fungus Ophiocordyceps
sinensis in DongChongXiaCao and Related Bioactive

JOURNAL OF FUNCTIONAL FOODS

Ingredients. Journal of Traditional and Complementary Medicine,

3, 1632.
Marchbank, T., Ojobo, E., Playford, G., & Playford, R. (2011).
Reparative properties of the traditional Chinese medicine
Cordyceps sinensis (Chinese caterpillar mushroom) using HT29
cell culture and rat gastric damage models of injury. British
Journal of Nutrition, 105, 13031310.
Medzhitov, R., & Janeway, C. (2000). Innate immune recognition:
mechanisms and pathways. Immunological Reviews, 173, 8997.
Miyazaki, T., Oikawa, N., & Yamada, H. (1977). Studies on fungal
polysaccharides. 20. Galactomannan of Cordyceps sinensis.
Chemical & Pharmaceutical Bulletin, 25, 33243328.
Moradali, M., Mostafavi, H., Ghods, S., & Hedjaroude, G. (2007).
Immunomodulating and anticancer agents in the realm of
macromycetes fungi (macrofungi). International
Immunopharmacology, 7, 701724.
Negre-Salvayre, A., Coatrieux, C., Ingueneau, C., & Salvayre, R.
(2008). Advanced lipid peroxidation end products in oxidative
damage to proteins. Potential role in diseases and therapeutic
prospects for the inhibitors. British Journal of Pharmacology, 153,
620.
Nie, S. P., Cui, S. W., Phillips, A. O., Xie, M. Y., Phillips, G. O., AlAssaf, S., & Zhang, X. L. (2011). Elucidation of the structure of a
bioactive hydrophobic polysaccharide from Cordyceps sinensis
by methylation analysis and NMR spectroscopy. Carbohydrate
Polymers, 84, 894899.
Nie, S. P., Cui, S. W., Xie, M., Phillips, A. O., & Phillips, G. O. (2013).
Bioactive polysaccharides from Cordyceps sinensis: isolation,
structure features and bioactivities. Bioactive Carbohydrates and
Dietary Fiber, 1, 3852.
Nie, S. P., & Xie, M. Y. (2011). A review on the isolation and
structure of tea polysaccharides and their bioactivities. Food
Hydrocolloids, 25, 144149.
Panagiotopoulos, C., Sempere, R., Lafont, R., & Kerherve, P. (2001).
Sub-ambient temperature effects on the separation of
monosaccharide by high-performance anion-exchange
chromatography with pulse amperometric detection:
application to marine chemistry. Journal of Chromatography A,
920, 1322.
Paterson, R. R. M. (2008). Cordyceps a traditional Chinese
medicine and another fungal therapeutic biofactory?
Phytochemistry, 69, 14691495.
Perez, S., Kouwijzer, M., Mazeau, K., & Engelsen, S. B. (1996).
Modeling polysaccharides: present status and challenges.
Journal of Molecular Graphics, 14, 307321.
Pol-Fachin, L., Fernandes, C. L., & Verli, H. (2009). GROMOS96 43a1
performance on the characterization of glycoprotein
conformational ensembles through molecular dynamics
simulations. Carbohydrate Research, 344, 491500.
Quan, W., Wang, J., Du, S., & Liu, G. (2007). Studies on the
production of exopolysaccharides by liquid culture of
Cordyceps sinensis. Fajiao Keji Tongxun, 36, 24 (in Chinese).
Sato, T., Norisuye, T., & Fujita, H. (1984). Double-stranded helix of
xanthan: dimensional and hydrodynamic properties in 0.1 M
aqueous sodium chloride. Macromolecules, 17, 26962700.
Schlesier, K., Harwat, M., Bohm, V., & Bitsch, R. (2002). Assessment
of antioxidant activity by using different in vitro methods. Free
Radical Research, 36, 177187.
Senti, F. r., Hellman, N. N., Ludwig, N. H., Babcock, G. E., Tobin, R.,
Glass, C. A., & Lamberts, B. L. (1955). Viscosity, sedimentation,
and light-scattering properties of fraction of an aciDhydrolyzed dextran. Journal of Polymer Science, 17, 527546.
Shashidhar, M. G., Giridhar, P., Sankar, K. U., & Manohar, B. (2013).
Bioactive principles from Cordyceps sinensis: A potent food
supplement A review. Journal of Functional Foods, 5, 1013
1030.
Shen, Y., Shao, X., Ni, Y., Xu, H., & Tong, X. (2009). Cordyceps
sinensis polysaccharide enhances apoptosis of HL-60 cells

6 (2 0 14 ) 3 347

45

induced by triptolide. Journal of Zhejiang University (Medical

Sciences), 38, 158162 (in Chinese).
Shen, W., Song, D., Wu, J., & Zhang, W. (2011). Protective effects of
a polysaccharide isolated from a cultured Cordyceps mycelia on
hydrogen peroxide-induced oxidative damage in PC12 cells.
Phytotherapy Research, 25, 675680.
Sheng, L., Chen, J., Li, J., & Zhang, W. (2011). An exopolysaccharide
from cultivated Cordyceps sinensis and its effects on cytokine
expression of immunocytes. Applied Biochemistry and
Biotechnology, 163, 669678.
Song, D., He, Z., Wang, C., Yuan, F., Dong, P., & Zhang, W. (2013).
Regulation of the exopolysaccharide from an anamorph of
Cordyceps sinensis on dendritic cell sarcoma (DCS) cell line.
European Journal of Nutrition, 52, 687694.
Song, D., Lin, J., Yuan, F., & Zhang, W. (2011). Ex vivo stimulation of
murine dendritic cells by an exopolysaccharide from one of
the anamorph of Cordyceps sinensis. Cell Biochemistry and
Function, 29, 555561.
Song, L. Q., Ming, Y. S., Peng, M. X., & Xia, J. L. (2010). The
protective effects of Cordyceps sinensis extract on extracellular
matrix accumulation of glomerular sclerosis in rats. African
Journal of Pharmacy and Pharmacology, 4, 471478.
Strlegel, A. M., Plattner, R. D., & Willett, J. L. (1999). Dilute solution
behavior of dendrimers and polysaccharides: SEC, ESI-MS, and
computer modeling. Analytical Chemistry, 71, 978986.
Velickovic, D. T., Milenovic, D. M., Ristic, M. S., & Veljkovic, V. B.
(2006). Kinetics of ultrasonic extraction of extractive
substances from garden (Salvia officinalis L.) and glutinous
(Salvia glutinosa L.) sage. Ultrasonics Sonochemistry, 13, 150156.
Wang, Z. M., Cheung, Y. C., Leung, P. H., & Wu, J. Y. (2010).
Ultrasonic treatment for improved solution properties of a
high-molecular weight exopolysaccharide produced by a
medicinal fungus. Bioresource Technology, 101, 55175522.
Wang, X. L., Liu, G. Q., Zhu, C. Y., & Kuang, S. M. (2011). Enhanced
production of mycelial biomass and extracellular
polysaccharides in caterpillar-shaped medicinal mushroom
Cordyceps sinensis CS001 by the addition of palmitic acid.
Journal of Medicinal Plants Research, 5, 28732878.
Wang, Z. M., Peng, X., Lee, K. L. D., Tang, J. C., Cheung, P. C. K., &
Wu, J. Y. (2011). Structural characterisation and
immunomodulatory property of an acidic polysaccharide from
mycelial culture of Cordyceps sinensis fungus Cs-HK1. Food
Chemistry, 125, 637643.
Wang, S. Y., & Shiao, M. S. (2000). Pharmacological functions of
Chinese medicinal fungus Cordyceps sinensis and related
species. Journal of Food and Drug Analysis, 8, 248257.
Wang, Y., Wang, M., Ling, Y., Fan, W. Q., Wang, Y. F., & Yin, H. P.
(2009). Structural determination and antioxidant activity of a
polysaccharide from the fruiting bodies of cultured Cordyceps
sinensis. The American Journal of Chinese Medicine, 37, 977989.
Wang, L., Wang, G., Zhang, J., Zhang, G., Jia, L., Liu, X., Deng, P., &
Fan, K. (2011). Extraction optimization and antioxidant activity
of intracellular selenium polysaccharide by Cordyceps sinensis
SU-02. Carbohydrate Polymers, 86, 17451750.
Wang, Y., Yin, H., Chen, T., & Wang, M. (2009). Preliminary
structural identification and protection on renal cell injury of
acidic polysaccharide from Cordyceps sinensis. Journal of China
Pharmaceutical University, 40, 559564 (in Chinese).
Wang, Y., Yin, H., Lv, X., Wang, Y., Gao, H., & Wang, M. (2010).
Protection of chronic renal failure by a polysaccharide from
Cordyceps sinensis. Fitoterapia, 81, 397402.
Wasser, S. P. (2002). Medicinal mushrooms as a source of
antitumour and immunomodulating polysaccharides. Applied
Microbiology and Biotechnology, 10, 1332.
Wasser, S. P., & Weis, A. L. (1999). Medicinal properties of
substances occurring in higher basidiomycetes mushroom:
current perspective. International Journal of Medicinal
Mushrooms, 1, 3151.

46

JOURNAL OF FUNCTIONAL FOODS

Winkler, D. (2009). Caterpillar fungus (Ophiocordyceps sinensis)

production and sustainability on the Tibetan Plateau and in
the Himanayas. Asian Medicines, 5, 291316.
Winkler, D. (2010). Cordyceps sinensis a previous parasitic fungus
infecting Tibet. Field Mycology, 11, 6067.
Wong, W. C., Wu, J. Y., & Benzie, I. F. F. (2011). Photoprotective
potential of Cordyceps polysaccharides against ultraviolet B
radiation-induced DNA damage to human skin cells. British
Journal of Dermatology, 164, 980986.
Wu, C., Chen, Y., & Hao, Y. (2009). Production of mycelia and
polysaccharides by liquid fermentation of Cordyceps sinensis.
Food Science, 30, 171174 (in Chinese).
Wu, Y. L., Hu, N., Pan, Y. J., Zhou, L. J., & Zhou, X. X. (2007).
Isolation and characterization of a mannoglucan from edible
Cordyceps sinensis mycelium. Carbohydrate Research, 342,
870875.
Wu, Y. L., Ishurd, O., Sun, C. R., & Pan, Y. J. (2005). Structure
analysis and antitumour activity of (1 ! 3)-beta-D-glucans
(Cordyglucans) from the mycelia of Cordyceps sinensis. Planta
Medica, 71, 381384.
Wu, Y. L., Sun, C. R., & Pan, Y. J. (2005). Structural analysis of a
neutral (1 ! 3), (1 ! 4)-b-D-glucan from the mycelia of
Cordyceps sinensis. Journal of Natural Products, 68, 812814.
Wu, Y. L., Sun, C. R., & Pan, Y. J. (2006). Studies on isolation and
structural features of a polysaccharide from the mycelium of a
Chinese edible fungus (Cordyceps sinensis). Carbohydrate
Polymers, 63, 251256.
Wu, Y. L., Sun, H. X., Qin, F., Pan, Y. J., & Sun, C. R. (2006). Effect of
various extracts and a polysaccharide from the edible mycelia
of Cordyceps sinensis on cellular and humoral immune
response against ovalbumin in mice. Phytotherapy Research, 20,
646652.
Xiao, J. H. (2008). Current status and ponderation on preparations
and chemical structures of polysaccharide in fungi of
Cordyceps (Fr.) Link. Chinese Traditional and Herbal Drugs, 39,
454460.
Xie, J., Shan, B., & Zhang, W. (2009). Microwave-assisted extraction
of polysaccharides from fermented Cordyceps sinensis
mycelium optimized by response surface analysis. Chinese
Journal of Bioprocess Engineering, 7, 3438 (in Chinese).
Yamac, M., Kanbak, G., Zeytinoglu, M., Bayramoglu, G., Senturk,
H., & Uyanoglu, M. (2008). Hypoglycemic effect of Lentinus
strigosus (Schwein.) fr. crude exopolysaccharide in
streptozotocin-induced diabetic rats. Journal of Medicinal Food,
11, 513517.
Yamada, H., Kawaguchi, N., Ohmori, T., Takeshita, Y., Taneya, S., &
Miyazaki, T. (1984). Strucrual and antitumour activity of an
alkali-soluble polysaccharide from Cordyceps ophioglossoides.
Carbohydrate Research, 125, 107115.
Yan, J. K., Li, L., Wang, Z. M., Leung, P. H., Wang, W. Q., & Wu, J. Y.
(2009). Acidic degradation and enhanced antioxidant activities
of exopolysaccharides from Cordyceps sinensis mycelial culture.
Food Chemistry, 117, 641646.
Yan, J. K., Li, L., Wang, Z. M., & Wu, J. Y. (2010). Structural
elucidation of an exopolysaccharide form mycelial
fermentation of a Tolypocladium sp. fungus isolated from wild
Cordyceps sinensis. Carbohydrate Polymers, 79, 125130.
Yan, J. K., Wang, W. Q., Li, L., & Wu, J. Y. (2011). Physiochemical
properties and antitumour activities of two a-glucans isolated
from hot water and alkaline extracts of Cordyceps (Cs-HK1)
fungal mycelia. Carbohydrate Polymers, 85, 753758.
Yan, J. K., Wang, W. Q., Ma, H. L., & Wu, J. Y. (2013). Sulfation and
enhanced antioxidant capacity of an exopolysaccharide
produced by the medicinal fungus Cordyceps sinensis. Molecules,
18, 167177.
Yan, F., Zhang, Y., & Wang, B. (2012). Effects of polysaccharides
from Cordyceps sinensis mycelium on physical fatigue in mice.
Bangladesh Journal of Pharmacology, 7, 217221.

6 ( 2 0 1 4 ) 3 3 4 7

Yang, L. Y., Chen, A., Kuo, Y. C., & Lin, C. Y. (1999). Efficacy of a pure
compound H1-A extracted from Cordyceps sinensis on
autoimmune disease of MRL LPR/LPR mice. The Journal of
Laboratory and Clinical Medicine, 134, 492500.
Yang, L. Y., Huang, W. J., Hsieh, H. G., & Lin, C. Y. (2003). H1-A
extracted from Cordyceps sinensis suppresses the proliferation
of human mesangial cells and promotes apoptosis, probably
by inhibiting the tyrosine phosphorylation of Bcl-2 and
Bcl-XL. The Journal of Laboratory and Clinical Medicine, 141,
7483.
Yang, L., & Zhang, L. M. (2009). Chemical structural and chain
conformational characterization of some bioactive
polysaccharides isolated from natural sources. Carbohydrate
Polymers, 76, 349361.
Yang, J. Y., Zhang, W. Y., Shi, P. H., Chen, J. P., Han, X. D., & Wang, Y.
(2005). Effects of exopolysaccharide fraction (EPSF) from a
cultivated Cordyceps sinensis fungus on c-Myc, c-Fos, and VEGF
expression in B16 melanoma-bearing mice. Pathology, Research
and Practice, 201, 745750.
Yao, Y. J. (2004). Conservation and rational use of the natural
resources of Cordyceps sinensis. Science News, 15, 2829 (in
Chinese).
Yao, X., Meran, S., Fang, Y., Martin, J., Midgley, A., Pan, M. M., Liu,
B. C., Cui, S. W., Phillips, G. O., & Phllips, A. O. (2014). Cordyceps
sinensis: in vitro anti-fibrotic bioactivity of natural and cultured
preparations. Food Hydrocolloids, 35, 444452.
Yin, H., Qiao, C., Qin, S., Tang, W., & Jia, S. (2013). Optimization of
fermentation medium for Cordyceps sinensis. Food Research &
Development, 34, 58 (in Chinese).
Yin, D. H., & Tang, X. M. (1995). Progresses of cultivation research
of Cordyceps sinensis. China Journal of Chinese Materia Medica, 20,
707709.
Yoon, T. J., Yu, K. W., Shin, K. S., & Suh, H. J. (2008). Innate immune
stimulation of exo-polymers prepared from Cordyceps sinensis
by submerged culture. Applied Microbiology and Biotechnology,
80, 10871093.
Yue, K., Ye, M., Zhou, Z., Sun, W., & Lin, X. (2013). The genus
Cordyceps: a chemical and pharmacological review. Journal of
Pharmacy and Pharmacology, 65, 474493.
Zhang, X. L., Cheung, L. B., Al-Assaf, S., Phillips, G. O., & Phillips,
A. O. (2012b). Cordyceps sinensis decreases TGF-b1 dependent
epithelial to mesenchymal transdifferentiation and attenuates
renal fibrosis. Food Hydrocolloids, 28, 200212.
Zhang, M., Cui, S., Cheung, P., & Wang, Q. (2007). Antitumour
polysaccharides from mushrooms: a review on their isolation
process, structural characteristics and antitumour activity.
Trends in Food Science & Technology, 18, 419.
Zhang, G. Q., Huang, Y. D., Bian, Y., Wong, J. H., Ng, T. B., & Wang,
H. X. (2006). Hypoglycemic activity of the fungi Cordyceps
militaris, Cordyceps sinensis, Tricholoma mongolicum, and
Omphalia lapidescens in streptozotocin-induced diabetic rats.
Applied Microbiology and Biotechnology, 72, 11521156.
Zhang, W., Li, J., Qiu, S., Chen, J., & Zheng, Y. (2008). Effects of the
exopolysaccharide fraction (EPSF) from a cultivated C. sinensis
on immunocytes of H22 tumour bearing mice. Fitoterapia, 79,
168173.
Zhang, Y., Li, E., Wang, C., Li, Y., & Liu, X. (2012a). Ophiocordyceps
sinensis, the flagship fungus of China: terminology, life strategy
and ecology. Mycology, 3, 210.
Zhang, Z., Wang, X., Zhang, Y., & Ye, G. (2011). Effect of Cordyceps
sinensis on renal function of patients with chronic allograft
nephropathy. Urologia Internationalis, 86, 298301.
Zhang, L., & Yang, L. (1995). Solution properties of pachyman from
Poria cocos mycelia in dimenthyl sulfoxide. Biopolymers, 36,
695700.
Zhang, W. Y., Yang, J. Y., Chen, J. P., Hou, Y. Y., & Han, X. D. (2005).
Immunomodulatory and antitumour effects of an
exopolysaccharide fraction from cultivated Cordyceps sinensis

JOURNAL OF FUNCTIONAL FOODS

(Chinese caterpillar fungus) on tumour-bearing mice.

Biotechnology and Applied Biochemistry, 42, 915.
Zhao, J., Xie, J., Wang, L. Y., & Li, S. P. (2013). Advanced
development in chemical analysis of Cordyceps. Journal of
Pharmaceutical and Biomedical Analysis. http://dx.doi.org/
10.1016/j.jpba.2013.04.025.
Zhong, S., Pan, H. J., Fan, L. F., Lv, G., Wu, Y., Parmeswaran, B.,
Pandey, A., & Soccol, C. R. (2009). Advances in research of
polysaccharides in Cordyceps species. Food Technology and
Biotechnology, 47, 304312.

6 (2 0 14 ) 3 347

47

Zhou, X. W., Gong, Z. H., Su, Y., Lin, J., & Tang, K. (2009). Cordyceps
fungi: natural products, pharmacological functions and
developmental products. Journal of Pharmacy and Pharmacology,
61, 279291.
Zhu, J. S., Halpern, G. M., & Jones, K. (1998). The scientific
rediscovery of an ancient Chinese herbal medicine: Cordyceps
sinensis. Journal of Alternative and Complementary Medicine, 4,
289303 (Part I); 4 429457 (Part II).