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Nutritional requirements of mycelial growth of Cordyceps
sinensis in submerged culture
C.-H. Dong
and Y.-J. Yao
Systematic Mycology and Lichenology Laboratory, Institute of Microbiology, Chinese Academy of Sciences, Beijing, and
School of the Chinese Academy of Sciences, Beijing, China
2004/1207: received 19 October 2004, revised 2 March 2005 and accepted 7 March 2005
C . - H . D O N G A N D Y . - J . Y A O . 2005.
Aims: The nutritional requirements for mycelial growth of Cordyceps sinensis in semi-synthetic liquid media were
investigated. The results provide a basis for further physiological study and industrial fermentation of the fungus.
Methods and Results: Nutritional requirements, including 17 carbohydrates, 16 nitrogen compounds, nine
vitamins, four macro-elements, four trace-elements and eight ratios of carbon to nitrogen, were studied for their
effects on the mycelial growth in submerged cultures of C. sinensis by using one-factor-at-a-time and orthogonal
matrix methods. Among these variables, sucrose, peptone, folic acid, calcium, zinc and a carbon to nitrogen
ratio 12 : 1 were identified as the requirements for the optimum mycelial growth. The concentrations of
sucrose, peptone and yeast extract were optimized and the effects of medium composition on mycelial growth
were found to be in the order sucrose > yeast extract > peptone. The optimal concentration for mycelial
growth was determined as 50 g l
sucrose, 10 g l
peptone and 3 g l
yeast extract.
Conclusions: Under optimal culture conditions, over 22 g l
of mycelial biomass could be obtained after
40 days in submerged cultures.
Significance and Impact of the Study: Cordyceps sinensis, one of the most valued medicinal fungi, is shown
to grow in axenic culture. This is the first report on nutritional requirements and design of a simplified semi-
synthetic medium for mycelial growth of this psychrophilic species, which grows slowly below 20°C. The
results of this study will facilitate research on mass production of the fungus under defined culture conditions.
Keywords: Cordyceps sinensis, mycelial growth, optimization, orthogonal design, semi-synthetic medium,
submerged culture.
Cordyceps sinensis (Berk.) Sacc., an ascomycete, has been
used as one of the most valued traditional Chinese
medicines. It is popularly referred to as the Chinese
Caterpillar Fungus or ÔDong Chong Xia CaoÕ (summer-
plant, winter-worm) in Chinese, or ÔHia Tsao Tong TchongÕ
and ÔHea Tsaon Tsong ChungÕ in early English translations
(Pegler et al. 1994). The fungus parasitizes larvae of moths
(Lepidoptera), especially Hepialus armoricanus Oberthur, and
converts each larva into a sclerotium, from which the fruit-
body grows.
The use of C. sinensis as a medicine and tonic has been
appreciated for hundreds of years in China. It is used
commonly in the treatment of kidney and lung problems in
the traditional Chinese medicines. Recent studies have
shown its multiple pharmacological effects, including
immunomodulating activity (Chiu et al. 1998; Gao et al.
2000; Koh et al. 2002), hypocholesterolaemic (Koh et al.
2003), antitumour (Chen et al. 1997; Bok et al. 1999) and
Correspondence to: Y. -J. Yao, Systematic Mycology and Lichenology Laboratory,
Institute of Microbiology, Chinese Academy of Sciences, PO Box 2714, Beijing
100080, China (e-mail: yaoyj@sun.im.ac.cn).
ª 2005 The Society for Applied Microbiology
Journal of Applied Microbiology 2005, 99, 483–492 doi:10.1111/j.1365-2672.2005.02640.x
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