• Mycobiology
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• v.38 no.2
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• pp.128-132
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• 2010

Injection inoculation protocols for fruit body formation of Cordyceps militaris (C. militaris) were investigated to improve the incidence of infection in the silkworm species Bombyx mori (B. mori). Injection, with suspensions of C. militaris hyphal bodies into living silkworm pupae, was used to test for fruit body production. Use of Daeseungjam rather than Baegokjam or Keumokjam varieties of B. mori is thought to be suitable for infection by C. militaris. From mounting, nine-day-old to 11-day-old pupae showed the best incidence of infection with a $100\;{\mu}L$ injection volume. Silkworm pupae injected with a hyphal suspension concentration of more than $2\;{\times}\;10^5$ colony-forming unit (cfu) recorded a greater than 96% incidence of infection. Also, fruit bodies of C. militaris were induced and produced at a light intensity between 500 and 1,000 lx.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.4
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• pp.1102-1106
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• 2004

To investigate the anti-proliferative effects of an aqueous extract of Cordyceps militaris (AECM) on the growth of human lung carcinoma cell line A549, we performed various biochemical experiments such as the effects of AECM on the cell proliferation and viability, the morphological changes, the effects on expression of apoptosis and cell growth-regulatory gene products. Results obtained are as follow; AECM treatment declined the cell viability and proliferation of A549 cells in a concentration-dependent manner. The anti-proliferative effect by AECM treatment in A549 cells was associated with morphological changes such as membrane shrinking and cell rounding up. Taken together, these findings suggest that AECM-induced inhibition of human lung cancer cell proliferation is associated with the induction of apoptotic cell death via regulation of several major growth regulatory gene products, and C. militaris may have therapeutic potential in human lung cancer.

• Mycobiology
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• v.43 no.1
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• pp.37-42
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• 2015

A mutant library of Cordyceps militaris was constructed by improved Agrobacterium tumefaciens-mediated transformation and screened for degradation features. Six mutants with altered characters in in vitro and in vivo fruiting body production, and cordycepin formation were found to contain a single copy T-DNA. T-DNA flanking sequences of these mutants were identified by thermal asymmetric interlaced-PCR approach. ATP-dependent helicase, cytochrome oxidase subunit I and ubiquitin-like activating enzyme were involved in in vitro fruiting body production, serine/threonine phosphatase involved in in vivo fruiting body production, while glucose-methanol-choline oxidoreductase and telomerase reverse transcriptase involved in cordycepin formation. These genes were analyzed by bioinformatics methods, and their molecular function and biology process were speculated by Gene Ontology (GO) analysis. The results provided useful information for the control of culture degeneration in commercial production of C. militaris.

• Journal of Microbiology and Biotechnology
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• v.18 no.12
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• pp.1997-2003
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• 2008

Cordyceps militaris is well known as a traditional medicinal mushroom and has been shown to exhibit immunostimulatory and anticancer activities. In this study, we investigated the apoptosis induced by an aqueous extract of C. militaris (AECM) via the activation of caspases and altered mitochondrial membrane permeability in human breast cancer MDA-MB-231 cells. Exposure to AECM induced apoptosis, as demonstrated by a quantitative analysis of nuclear morphological change and a flow cytometric analysis. AECM increased hyperpolarization of mitochondrial membrane potential and promoted the activation of caspases. Both the cytotoxic effect and apoptotic characteristics induced by AECM treatment were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor, which demonstrates the important role of caspase-3 in the observed cytotoxic effect. AECM-induced apoptosis was associated with the inhibition of Akt activation in a time-dependent manner, and pretreatment with LY294002, a PI3K/Akt inhibitor, significantly increased AECM-induced apoptosis. The results indicated that AECM-induced apoptosis may relate to the activation of caspase-3 and mitochondria dysfunctions that correlate with the inactivation of Akt.

• Journal of Life Science
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• v.23 no.8
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• pp.1032-1040
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• 2013

The comparative effects of fibrinolytic, antioxidative activity and electrophoretical protein patterns with Cordyceps militaris powder (CM) and cordycepin-enriched Cordyceps militaris JLM0636 powder (CCM) and fermented cordycepin-enriched Cordyceps militaris JLM0636 powder by several microscopic organisms were investigated. In addition, nutritional materials such as mineral, protein, and fatty acids were also measured. The protein concentration was higher in CCM than that in CM. The protein concentration in fermented CCM was the highest in CCM treated by Aspergillus kawachii among the various samples. When total protein patterns of CM, CCM, and both fermented CCMs were analyzed by native- and SDS-PAGE, there were slightly varietal differences in electrophoretical protein patterns. Major minerals were K, Ca, Mg, and Zn. Major fatty acids were palmitic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid. Fibrinolytic activity was the highest in the fermented CCM by Bs treatment among the various samples. The ${\alpha},{\alpha}^{\prime}$-diphenyl-${\beta}$-picrylhydrazyl (DPPH) radical scavenging activity was slightly stronger in the CCM treated with Aspergillus kawachii among the various samples; however, these samples all exhibited relatively low levels of activity compared with the butylated hydroxytoluene (BHT). These results may provide the basic data necessary to understand the biological activities and chemical characteristics of Cordyceps militaris JLM0636 powder fermented by several microscopic organisms to develop functional foods.

• Journal of Life Science
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• v.23 no.12
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• pp.1516-1524
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• 2013

To find a novel skin whitening agent, the effect of cordycepin-enriched Cordyceps militaris (CM${\alpha}$) extract fermented by fungi on anti-melanogenesis in B16F0 mouse melanoma cells was investigated. Fermented CM${\alpha}$ was prepared with fungi, including Monascus purpureus (Mp), Aspergillus oryzae (Ao), Aspergillus kawachii (Ak), and Rhizopus oryzae (Ro), respectively. When the content of the phenolics and the flavonoids and the activities of the antioxidant and the mushroom tyrosinase inhibition were measured in the CM fermented by Ak (AkF-CM), the highest content of the phenolics was 46 mg/g dry weight and the highest content of the flavonoids was 0.93 mg/g; the highest activity of the DPPH radical scavenging was 62.74% and the highest activity of the mushroom tyrosinase inhibition was 79.97% CM${\alpha}$CM${\alpha}$. From this result, AkF-CM${\alpha}$ exhibited the highest mushroom tyrosinase inhibitory activity and so it was used in subsequent anti-melanogenesis. B16F0 melanoma cells were treated with 1-10 mg/ml concentrations of AkF-CM${\alpha}$ and 200 ${\mu}M$ arbutin as the positive control. The melanin content and cell viability of the melanoma cells by arbutin treatment decreased to 43% and 92% of the control, respectively. AkF-CM${\alpha}$ treatment at 1, 3, and 5 mg/ml concentrations decreased the extracellular melanin release induced by IBMX treatment by 35%, 45%, and 53%, respectively. AkF-CM${\alpha}$ showed inhibitory activity against both intracellular tyrosinase in melanoma cells and mushroom tyrosinase. AkF-CM${\alpha}$ reduced the protein level of tyrosinase in the IBMX-stimulated cells. These results indicate that AkF-CM${\alpha}$ suppressed the activity and protein content of cellular tyrosinase and decreased the total melanin content in cultured B16F0 melanoma cells.

• The Korean Journal of Mycology
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• v.28 no.2
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• pp.112-117
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• 2000

A similarity coefficient were analyzed by RFLP of fourteen species of entomopathogenic fungi, isolated from inhabiting pupa and adult insect at forest. Each rDNA ITS I and ITS II with primers of ITS 1 and ITS 4 was amplified by PCR. The amplified products were conserved to 500 bp were not demarcated between genus and species. Four Paeciliomyces tenuipes, two Beauveria bassiana and six Cordyceps militaris were treated by seven restriction enzymes and confirmed in species except JB3 by electrophoresis band. However, the band of C. scarabaeicola showed the identity with B. bassiana. The result of this experiment indicated that the teleomorph of C. scarabaeicola was the same as that of B. bassiana. CfoI and HpaII of seven restricted enzymes were easily discriminating in the genus between Paecilomyees and Cordyceps. Especially, CfoI was more effective to classify the genera of Paecilomyees, Cordyceps and Beauveria than other restriction enzymes. The band patterns of RFLP of P. tenuipes, C. militaris, C. scarabaeicola and B. bassiana were also analyzed by UPGMA program of NTSYS-pc and showed 100% significance. Thus, the similarity coefficient tended to be lower between genera by RFLP analysis, but was higher between species.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.243-245
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• 2003

The continuous fermentation process for exe-biopolymer production with Cordyceps militaris has been developed in this research. With the change of dilution rate, biopolymer production has been changed and the maximal biopolymer production has been achieved when the dilution rate was 0.0225/hr. The maximum mycelium concentration and biopolymer concentration were 8 g/L and 4.2 g/L, respectively.

• Plant Resources
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• v.3 no.2
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• pp.118-122
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• 2000

Many ascomycetes were collected at Mt.Unjang, Pyonsan penisula national park, Mt.Moak Chonbuk provincial park, Daea-ri hebarium and Mt.Whaam from August 1996 to October 1999. They were identified. According to the resulting, Lanzia echinophila, Discina parma, Hypomyces auranticus, Helotium versicolor, Cordyceps militaris f. albino, Isaria sinclairii are newly to Korea. They were designed Korean common names by author.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.2
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• pp.452-458
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• 2005

Cordyceps militaris is a medicinal fungus, which has been used for patient suffering from cancer in Oriental medicine. It was reported previously that C. militaris extracts are capable of inhibiting tumor growth, however, the anti-poliferative effects of human cancer cells have not been poorly understood. In this study, to elucidate the growth inhibitory mechanisms of human cancer cells by treatment of aqueous extract of C. militaris (AECM) we investigated the anti-proliferative effects of AECM in human leukemia U937 cell line. AECM treatment inhibited the growth of U937 cells and induced the apoptotic cell death in a concentration-dependent manner, which was associated with morphological changes. We observed the up-regulation of cyclin-dependent kinase (Cdk) inhibitor p21(WAF1/CIP1) by p53-independent manner and activation of caspase-3 in AECM-treated U937 cells, however, the activity of caspase-9 was remained unchanged. Additionally, AECM treatment caused a dose-dependent inhibition of the expression of telomere regulatory gene products such as human telomere reverse transcriptase (hTERT) and telomerase-associated protein-1 (TEP-1). Taken together, these findings suggest that AECM-induced inhibition of human leukemic cell proliferation is associated with the induction of apoptotic cell death via modulation of several major growth regulatory gene products, and C. militaris may have therapeutic potential in human lung cancer.