Isolation and Identification of Secondary Metabolites from Cordyceps cardinalis / 中国药房

OBJECTIVE: To isolate and identify the secondary metabolites of Cordyceps cardinalis for providing reference for the further exploration of the active substance of C. cardinalis and the natural product library of Cordyceps. METHODS: The compounds from C. cardinalis fermentation broth were isolated by silica gel column chromatography, C18 reverse phase silica gel chromatography, Sephadex LH-20 gel column chromatography, TLC and so on. The structures of isolated compound were identified by 1H-NMR, 13C-NMR and MS spectra. RESULTS: Six compounds were isolated from the ethyl acetate extract of C. cardinalis fermentation broth as 5-methyl-1, 3-benzenediol (compound 1), 4-hydroxy-17R-methylincisterol (compound 2), 5α, 8 α-epidioxy-(22E,24R)-ergosta-6,22-dien-3β-ol (compound 3),3β,5α,6β-trihydroxy-(22E,24R)-ergosta-7,22-dien (compound 4), uridine (compound 5), oosporein (compound 6). CONCLUSIONS: Six compounds are isolated from C. cardinalis strain, among which compound 1 and compound 2 are isolated from the genus Cordyceps for the first time.

Optimum Conditions for Artificial Fruiting Body Formation of Cordyceps cardinalis

Stromatal fruiting bodies of Cordyceps cardinalis were successfully produced in cereals. Brown rice, German millet and standard millet produced the longest-length of stromata, followed by Chinese pearl barley, Indian millet, black rice and standard barley. Oatmeal produced the shortest-length of fruiting bodies. Supplementation of pupa and larva to the grains resulted in a slightly enhanced production of fruiting bodies; pupa showing better production than larva. 50~60 g of brown rice and 10~20 g of pupa mixed with 50~60 mL of water in 1,000 mL polypropylene (PP) bottle was found to be optimum for fruiting body production. Liquid inoculation of 15~20 mL per PP bottle produced best fruiting bodies. The optimal temperature for the formation of fruiting bodies was 25degrees C, under conditions of continuous light. Few fruiting bodies were produced under the condition of complete darkness, and the fresh weight was considerable low, compared to that of light condition.

Growth and Cultural Characteristics of Cordyceps cardinalis Collected from Korea

Cordyceps cardinalis was reported in Japan and the USA in 2004, and its fruiting bodies have recently been cultured in Korea. Herbarium specimens preserved at the Cordyceps Research Institute, Mushtech, Korea were revised and identified as C. cardinalis, based on morphological characters and conidial structures. Most of the C. cardinalis specimens were collected from Mt. Halla in Jeju-do. The effects of various nutritional sources and environmental conditions such as temperature and pH on mycelial growth of C. cardinalis were studied. Oatmeal agar, Martin's peptone dextrose agar, and Schizophyllum (mushroom) genetics complete medium plus yeast extract resulted in the best mycelial growth. Among carbon sources, cereals, and nitrogen sources, maltose, oatmeal, and peptone resulted in the best mycelial growth respectively. Mineral salts helped to increase growth rate but only resulted in thin mycelial density, similar to water agar. A temperature of 25degrees C and a pH of 7 resulted in the highest mycelial growth. Based on these results, a Cordyceps cardinalis composite medium (CCM) was formulated with 1% maltose, 2% oatmeal, 1% peptone, and 2% agar. Use of the CCM resulted in slightly better mycelial growth than that of other commonly used agar media. Only organic nitrogen sources imparted a reddish pigmentation to the agar media, but this character diminished after several subcultures. A 7 day culture duration resulted in the best mycelial growth.