Cordyceps militaris is widely used in China, Korea, and other Asian countries as both a traditional medicinal ingredient and an edible fungus. This study aimed to optimize the growth conditions and fruiting body production of C. militaris by investigating various culture media and physical parameters such as pH, aeration, illumination, temperature, spawn materials, and oat–sawdust-based substrate formulations. After a 7-day incubation period, oats with a pH of 6.0, under sealed and illuminated conditions at 32°C, demonstrated the most effective mycelial growth. Substrates consisting of 70% oat and 30% sawdust had the shortest incubation time of 30.5 days for fruiting body formation. The basidiospores showed a typical germination pattern where the sporidium produced a single germ tube that elongated, and branched to form monokaryotic primary mycelia. In conclusion, using oats as a substrate in the cultivation of C. militaris could reduce production costs and help protect the environment.

Cordyceps militaris (C. militaris) is a unique and valuable medicinal fungus belonging to the Cordyceps species. C. militaris is the only fungus that contains cordycepin which is a biologically active compound. In previous studies, light-emitting diodes (LEDs) are known to be effective in increasing cordycepin content, but metabolic profiling of LED-stimulated C. militaris has not been confirmed. Metabolic profiling is essential to understanding the metabolic regulation of cordycepin. This study studied the physiologically active secondary metabolites of C. militaris according to the presence or absence of stimulation of LEDs through GC-MS analysis. Most of the metabolites were detected in both samples, but there was a clear difference in the detected concentration. In particular, C. militaris had a significant difference in amino acid levels when stimulated with LEDs. Our results suggested that LEDs could stimulate amino acid synthesis in C. militaris mycelium to increase the cordycepin content.

This study aimed to verify the whitening effect of Cordyceps militaris, which is distributed in several countries worldwide, including Korea, Japan, and China, and has various medical effects. To screen the efficacy of C. militaris, the inhibitory activity of tyrosinase, which was 66% at a concentration of 1 mg/mL, was measured. Thereafter, the survival rate of melanoma cells was measured, and cell experiments were conducted at a concentration of 90% or more in which C. militaris was not toxic to cells. After measuring the inhibitory effect of TRP-1, TRP-2, tyrosinase protein, and mRNA expression, which are factors influencing melanin synthesis, C. militaris was found to decrease in all factors, with an expression level that was significantly lower compared to quercetin. This confirmed that C. militaris stimulated with LED has excellent whitening activity and can be used as a functional whitening cosmetics material.

To investigate the anti-inflammatory activity of submerged culture using Cordyceps militaris mycelium, culture-including mycelia was extracted and lyophilized into postbiotics (hot-water extract; CM-HW). HW was fractionated into crude polysaccharide (CM-CP) by ethanol precipitation, and CM-CP was further dialyzed into CM-DCP by dialysis with running water using 12~14 kDa dialysis tube. When the cytotoxicity of subfractions against cells was assessed, no subfraction had a cytotoxic impact that was substantially different from the control groups. In an inflammatory model using LPS-stimulated RAW 264.7 cells, CM-DCP significantly decreased IL-6 and MCP-1 production levels compared to the LPS-control group. CM-DCP also inhibited IL-6 and IL-8 secretion in HaCaT keratinocytes stimulated with TNF-α and IFN-γ. In the meanwhile, the neutral sugar content and mannose ratio of anti-inflammatory CM-DCP were higher than the other fractions, and CM-DCP contained β-1,3/1,6-glucan of 216.1 mg/g. High pressure size exclusion chromatography revealed that CM-DCP contained molecules with a molecular weight range of 5.6 to 144.0 kDa. In conclusion, postbiotics of C. militaris mycelium significantly promoted anti-inflammatory activity, suggesting that neutral polysaccharides including Glc and Man contribute to the anti-inflammation in RAW 264.7 or HaCaT cells.

Owing to its excellent nutritional value, eggs are among the most important components of the human diet. Gender and environmental factors, such as feed composition, may alter the nutritional profile and quality of eggs. Feed additives have recently been used to enhance the health and productivity of hens, which has resulted in the production of higher-quality eggs. The fungus Cordyceps militaris, a well-established source of traditional medicines, contains potential bioactive metabolites, which prompted us to examine the effects of C. militaris-supplemented diets on the quality of hens’ eggs. The hens of two species (Gallus gallus domesticus and Araucana) were fed with one of three different diets: a control diet and diets supplemented with 2% or 5% of C. militaris. Egg quality was determined by measuring the Haugh Unit, yolk color, and shell thickness. In addition, egg and shell densities together with the ratio of yolk to albumen were calculated. Eggshell thickness and yolk color were both enhanced by the addition of C. militaris, whereas Haugh Unit values were somewhat reduced. Egg size, eggshell weight, and yolk and albumen production were all enhanced by C. militaris supplementation. Notably, in hens fed the 2% C. militaris-supplemented diet, enhancement was more evident in the yolk than in the albumen. The overall quality of the egg yolk was enhanced when 2% C. militaris was added to the hens' diet, which led to increases in both yolk color and quantity. Eggshell thickness and weight were also higher among eggs laid by hens fed the supplemented diets. Although these effects differed depending on the chicken species, we established that, in general, C. militaris contributes to improving egg quality.

Cordyceps militaris mycelium extracts containing high amounts of cordycepin were evaluated in vitro for their antiinflammatory and tumor cell growth-inhibitory activities. All extracts dose dependently inhibited the increased production of inflammatory mediators including reactive oxygen species (ROS), nitric oxide (NO), and β-hexosaminidase in lipopolysaccharide (LPS)-stimulated inflammatory cells. All extracts were evaluated for anti-proliferative activity against normal RBL-2H3 cells and diverse types of cancer cell lines, including HCT, MC5-7, U-87MG, AGS, and A549 cells. The extract showed the strongest growth inhibition (IC50 = 28.13 μg/mL) relative to vehicle-treated control cells against fibrosarcoma (MC5-7). We have demonstrated anti-inflammatory activity of C. militaris via inhibition of NO, ROS production, and β-hexosaminidase release in activated cells. C. militaris mycelium extract was also evaluated mechanistically and found to exert six types of anti-cancer activity, confirming its pharmacological potential. Our study suggests C. militaris use as a potential source of anti-inflammatory and anticancer agents. C. militaris may also be considered a functional food.

Cordyceps militaris, a well-known traditional Chinese medicine, has multiple health-promoting effects. It is used as a herbal remedy and health food in Asian countries. Cultured mycelia are often used as a substitute for natural C. militaris. In the present study, the mycelia and fruiting bodies of artificially cultured C. militaris were analyzed using a metabolomics approach. The protein and crude fat contents of the mycelia were substantially higher than those of the fruiting bodies. The top three abundant amino acids in the mycelia were proline (3.9 g/100 g), aspartic acid (2.9 g/100 g), and glutamic acid (2.7 g/100 g). The carbohydrate content was similar in the fruiting bodies and mycelia. Analysis revealed that both the fruiting bodies and mycelia are rich in phenolic compounds and exhibit antioxidant activity. Further, six metabolites were significantly different between the mycelia and fruiting bodies. The levels of Ca, glucose, Mg, and Se were higher in the mycelia than in the fruiting bodies. In contrast, mannitol and Zn were more abundant in the fruiting bodies. The current study provides a comprehensive metabolic profile of the mycelia and fruiting bodies of artificially cultured C. militaris. Such an exercise is potentially important for understanding the metabolism of C. militaris and facilitating the use of cultured mycelia as a supplement to C. militaris fruiting bodies in traditional Chinese medicine.

The ascomycete fungus Cordyceps militaris infects lepidopteran insect pupae, forming characteristic fruiting bodies called “Dong Chung Ha Cho” in Korean. They have been used as medicines owing to their anti-allergic, anti-inflammatory, and immune-enhancing effects. This fungus can be grown on the geminated soybeans Rhynchosia nulubilis, which also contains several novel isoflavones. We performed a comparative transcriptome analysis to determine core gene sets or pathways contributing to biologically active products such as isoflavone. Initially, we sequenced 2-week-old fungal cultures on different soybean agar media, where different amounts of water agar were implemented to show different surface topology. We selected 830 upregulated and 188 downregulated genes by comparing linear models of the samples (two-fold change threshold). Gene ontology analysis identified that the “IMP biosynthesis” term was significantly found in the upregulated gene sets. The pathway is involved in the synthesis of cordycepin, the reference chemical for C. militaris. This finding in the transcriptome data is consistent with the previous observation: increased cordycepin concentrations in the C. militaris cultured on germinated soybean.