Cordycepin production by Cordyceps militaris cultivation on spent brewery grains


  • Andrej Gregori



Cordyceps militaris, spent brewery grains, cordycepin, cultivation, medicinal mushrooms


This is a first report on C. militaris mycelia and fruiting bodies cultivation on solid-state containing spent brewery grains (SBG). Five different strains of C. militaris were cultivated on substrates containing rye grains and 0 to 60% SBG. Stromata formation on SBG-containing substrates was noticed with two C. militaris strains. All strains failed to grow on substrates containing SBG amounts higher than 50%. Highest (10.42 mg/g) cordycepin concentration in cultivating substrate was determined with strain CM2 on 50% SBG. One gram of CM11 strain fungal biomass was able to produce 787.11 mg/g of cordycepin. SBG as a byproduct represent a readily available, low price substrate for cordycepin solid-state production. Obtained concentrations of cordycepin are so far the highest reported concentrations obtained on solid-state substrates therefore we can talk about cordycepin hyperproduction.


Barbosa-Pereira, L., Bilbao, A., Vilches, P., Angulo, I., LLuis, J., Fité, B., Paseiro-Losada, P., Cruz, J.M., 2014. Brewery waste as a potential source of phenolic compounds: Optimisation of the extraction process and evaluation of antioxidant and antimicrobial activities. Food Chemistry, 145, 191-197. DOI:

Bhandari, A.K., Negi, J.S., Bisht, V.K., Rana, C.S., Bharti, M.K., Singh, N., 2010. Chemical Consti- tuent, Inorganic Elements and Properties of Cordyceps sinensis - a Review (Cordyceps sinensis - a Review). Nature and Science, 8 (9), 253-256.

Buenz, E.J., Bauer, B.A., Osmundson, T.W., Motley, T.J., 2005. The traditional Chinese medicine Cordyceps sinensis and its effects on apoptotic homeostasis. Journal of Ethnopharmacology, 96, 19–29. DOI:

Carvalheiro, F., Duarte, L.C., Lopes, S., Parajo, J.C., Pereira, H., Gı́rio, F.M., 2005. Evaluation of the detoxification of brewery’s spent grain hydrolysate for xylitol production by Debaryomyces hansenii CCMI 941. Process Biochemistry, 40 (3-4), 1215–1223. DOI:

Chen, S.Z., Wu, P.J., 1990. A brief introduction to bottle culture technique of Cordyceps militaris. Edible fungi, 4, 31-37.

Chen, Y.S., Liu, B.L., Chang, Y.N., 2011. Effects of light and heavy metals on Cordyceps militaris fruit body growth in rice grain-based cultivation. Korean Journal of Chemical Engineering, 28 (3), 875-879. DOI:

Choi, S.B., Park, C.H., Choi, M.K., Jun, D.W., Park, S., 2004. Improvement of insulin resistance and insulin secretion by water extracts of Cordyceps militaris, Phellinus linteus, and Paecilomyces tenuipes in 90% pancreatectomized rats. Bioscience, Biotechnology and Biochemistry, 68 (11), 2257–2264. DOI:

Das, S.K., Masuda, M., Sakurai, A., Sakakibara, M., 2010. Medicinal uses of the mushroom Cordyceps militaris: Current state and prospects. Fitoterapia 81 (8), 961–968. DOI:

Gao, S.Y., Wang, F.Z., 2008. Research of commercialized cultivation technology on Cordyceps mili- taris. Northern Horticulture, 9, 212-215.

Gao, X.H., Wu, W., Qian, G.C., 2000. Study on influence of abiotic factors on fruitbody differentiation of Cordyceps militaris. Acta Agriculture Shanghai, 16, 93-98.

Gregori, A., Švagelj, M., Pahor, B., Berovič, M., Pohleven, F., 2008. The use of spent brewery grains for Pleurotus ostreatus cultivation and enzyme production. New Biotechnology, 25 (2-3), 157-161. DOI:

Gupta, S., Jaiswal, K. A., Abu-Ghannam, N., 2013. Optimization of fermentation conditions for the utilization of brewing waste to develop a nutraceutical rich liquid product. Industrial Crops and Products, 44, 272–282. DOI:

Holliday, J.C., 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 (3), 219–234. DOI:

Holliday, J.C., Cleaver. P., Loomis-Powers, M., Patel, D., 2004. Analysis of quality and techniques for hybridization of medicinal fungus Cordyceps sinensis. International Journal of Medicinal Mushrooms, 6, 151-164. DOI:

Kobayashi, Y., 1941. The genus Cordyceps and its allies. Science reports of the Tokyo Bunrika Dai- gaku, 84, 53-260.

Li, C.B., Tong, X.D., Bai, J., Fan, S.D., 2004. Artificial stromata production of Cordyceps militaris. Journal of Dalian National University, 6 (5), 29-31.

Li, X., 2002. Manmade cultivates of Cordyceps militaris (L) Link. Journal of microbiology. 22 (6), 56-57.

Mahmooda, A.S.N., Brammera, J.G., Hornunga, A., Steeleb, A., Poulstonb, S., 2013. The intermediate pyrolysis and catalytic steam reforming of Brewers spent grain. Journal of Analytical and Applied Pyrolysis, 103, 328–342. DOI:

Martin, F., Delaruelle C., Hilbert, J.L., 1990. An improved ergosterol assay to estimate fungal biomass in ectomycorrhizas. Mycological Research, 94, 1059-1064. DOI:

Mussatto, S.I., Dragone, G., Roberto, I.C., 2006. Brewers’ spent grain: generation, characteristics and potential applications. Journal of Cereal Science, 43, 1–14. DOI:

Nan, J.X., Park, E.J., Yang, B.K., Song, C.H., Ko, G., Sohn, D.H., 2001. Antibiotic effect of extracellular biopolymer from submerged mycelial cultures of Cordyceps militaris on liver fibrosis induced by bile duct ligation and scission in rats. Archives of Pharmacal Research, 24 (4), 327–332. DOI:

Ni, H., Zhou, X.H., Li, H.H., Huang, W.F., 2009. Column chromatographic extraction and prepara- tion of cordycepin from Cordyceps militaris waster medium. Journal of Chromatography B, 877, 2135-2141. DOI:

Nylund, J.E., Wallander, H., 1992. Ergosterol analysis as a means of quantifying mycorrhizal biomass. Methods in Microbiology, 24, 77-88. DOI:

Ogris, N., 2013. Podatkovna zbirka gliv Slovenije Boletus informaticus.

Plessas, S., Trantallidi, M., Bekatorou, A., Kanellaki, M., Nigam, P., Koutinas, A.A., 2007. Immobili- zation of kefir and Lactobacillus casei on brewery spent grains for use in sourdough wheat bread making. Food Chemistry, 105 (1), 187–194. DOI:

Poerschmanna, J., Weinera, B., Wedwitschkab, H., Baskyra, I., Koehlera, R., Kopinkea, F.D., 2014. Characterization of biocoals and dissolved organic matter phases obtained upon hydrothermal carbonization of brewer’s spent grain. Bioresource Technology, 164, 162–169. DOI:

Raoa, Y.K., Fangb, S.H., Wuc, W.S., Tzenga, Y.M., 2010. Constituents isolated from Cordyceps mili- taris suppress enhanced inflammatory mediator’s production and human cancer cell proliferation. Journal of Ethnopharmacology, 131, 363–367. DOI:

Reis, F.S., Barros, L., Calhelha, R.C., Ćirić, A., van Griensven, L., Soković, M., Ferreira, I., 2013. The methanolic extract of Cordyceps militaris (L.) Link fruiting body shows antioxidant, antibacterial, antifungal and antihuman tumor cell lines properties. Food and Chemical Toxicology, 62, 91–98. DOI:

Robertson, J.A., I’Anson, K.J.A., Treimo, J., Faulds, C.B., Brocklehurstb T.F., Eijsinkc, V.G.H., Waldrona, K.W., 2010. Profiling brewers’ spent grain for composition and microbial ecology at the site of production. LWT - Food Science and Technology, 43 (6), 890–896. DOI:

Shrestha, B., Han, S. K., Sung, J. M., Sung, G. H., 2012. Fruiting Body Formation of Cordyceps mi- litaris from Multi-Ascospore Isolates and Their Single Ascospore Progeny Strains. Mycobiology, 40 (2), 100-106. DOI:

Song, C.H., Jeon, Y.J., Yang, B.K., Ra, K.S., Sung, J.M., 1998. Anti-complementary activity of exo- polymers produced from submerged mycelial cultures of higher fungi with particular reference to Cordyceps militaris. Journal of Microbiology and Biotechnology, 8, 536–539.

Sung, J.M., 1996. The insect-borne fungus of Korea in color. Kyohak Publishing Co. Ltd., Seul. Wei, Q., Huang, M.Q., 2009. Effects of nutrient ingredient in culture medium on the growth of Cordyceps militaris. Beijing agriculture, 27, 36-38.

Wen, T.C., Li, G.R., Kang, J.C., Kang, C., Hyde, K.D., 2014. Optimization of Solid-state Fermentation for Fruiting Body Growth and Cordycepin Production by Cordyceps militaris. Chiang Mai Journal of Science, 41 (4), 858-872.

Won, S.Y., Park, E.H., 2005. Anti-inflammatory and related pharmacological activities of cultured mycelia and fruiting bodies of Cordyceps militaris. Journal of Ethnopharmacology, 96, 555–561. DOI:

Wu, F.C., Chen, Y.L., Chang, S.M., Shih, I.L., 2013. Cultivation of medicinal caterpillar fungus, Cordyceps militaris (Ascomycetes), and production of cordycepin using the spent medium from levan fermentation. International Journal Of Medicinal Mushrooms, 15 (4), 393-405. DOI:

Xie, C.Y., Gu, Z.X., Fan, G.J., 2009. Production of cordycepin and mycelia by submerged fermenta- tion of Cordyceps militaris in mixture natural culture. Applied Biochemistry and Biotechnology, 158, 483-492. DOI:

Yang, S., Jin, L., Ren, X., Lu, J., Meng, Q., 2014. Optimization of fermentation process of Cordyceps militaris and antitumor activities of polysaccharides in vitro. Journal of Food and Drug Analysis, In Press. DOI:

Yi., Z.L., Huang, W.F., Ren, Y., Onac, E., Zhou, G.F., Peng, S., Wang, X.J., Li, H.H., 2014. LED

lights increase bioactive substances at low energy costs in culturing fruiting bodies of Cordyceps militaris. Scientia Horticulturae, 175, 139-143. DOI:

Ying, J., Mao. X., Mao, Q., Zong, Y., Wen, H., 1987. Icons of Medicinal Mushroom from China, Science Press, Beijing, 151–155.

Yoo, H.S., Shin, J.W., Cho, J.H., Son, C.G., Lee, Y.W., Park, S.Y., Cho, C.K., 2004. Effects of Cordyceps militaris extract on angiogenesis and tumor growth. Acta Pharmacologica Sinica, 25 (5), 657-65.

Zhang, X.K., Liu, W.X., 1997. Experimental studies on planting Cordyceps militaris (L. ex Fr.) Link with different culture materials. Edible fungi of China, 16 (2), 21-22.

Zhao, C.Y., Li, H., Zhang, M., 2006. Optimization on conditions of artificial cultivation of Cordyceps militaris. Journal of Shenyang Agricultural University, 37, 209-212.






Original Research Paper

How to Cite

Gregori, A. (2014). Cordycepin production by Cordyceps militaris cultivation on spent brewery grains. Acta Biologica Slovenica, 57(2), 45-52.

Similar Articles

1-10 of 13

You may also start an advanced similarity search for this article.