• Journal of Microbiology and Biotechnology
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• 제27권7호
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• pp.1223-1232
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• 2017

Cordyceps militaris, a member of Ascomycota, a mushroom referred to as caterpillar Dong-chung-ha-cho, is commercially valuable because of its high content of bioactive substances, including cordycepin, and its potential for artificial cultivation. Cordycepin (3'-deoxyadenosine) is highly associated with the pharmacological effects of C. militaris. C. militaris is heterothallic in that two mating-type loci, idiomorph MAT1-1 and MAT1-2, exist discretely in two different spores. In this study, nine C. militaris strains were mated with each other to prepare newly bred strains that produced a larger amount of cordycepin than the parent strains. Nine strains of C. militaris were identified by comparing the internal transcribed spacer sequence, and a total of 12 single spores were isolated from the nine strains of C. militaris. After the MAT idiomorph was confirmed by PCR, 36 mating combinations were performed with six single spores with MAT1-1 and the others with MAT1-2. Eight mating combinations were successfully mated, producing stroma with perithecia. Cordycepin content analysis of all strains by high-performance liquid chromatography revealed that the KASP4-bred strain produced the maximum cordycepin among all strains, regardless of the medium and stroma parts. Finally, universal rice primer-PCR was performed to demonstrate that the bred strains were genetically different from the parental strains and new C. militaris strains. These results may be related to the recombination of genes during mating. The newly produced strains can be used to meet the industrial demand for cordycepin. In addition, breeding through mating suggests the possibility of producing numerous cordycepin-producing C. militaris strains.

• 한국버섯학회지
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• 제20권4호
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• pp.254-257
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• 2022

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.

• Mycobiology
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• 제40권2호
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• pp.100-106
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• 2012

Interest in commercial cultivation and product development of Cordyceps species has shown a recent increase. Due to its biochemical and pharmacological effects, Cordyceps militaris, commonly known as orange caterpillar fungus, is being investigated with great interest. Cultivation of C. militaris has been practiced on a large scale in order to fulfill a demand for scientific investigation and product development. Isolates of C. militaris can be easily established from both spores and tissue. For isolation of spores, ascospores released from mature stromata are trapped in sterile medium. Multi-ascospore isolates, as well as combinations of single ascospore strains, are used for production of fruiting bodies. Progeny ascospore strains can be isolated from artificial fruiting bodies, thus, the cycle of fruiting body production can be continued for a long period of time. In this study, we examined fruiting body production from multi-ascospore isolates and their progeny strains for three generations. $F_1$ progeny strains generally produced a larger number of fruiting bodies, compared with their mother multi-ascospore isolates; however, $F_2$ and $F_3$ progeny strains produced fewer fruiting bodies. Optimum preservation conditions could help to increase the vitality of the progeny strains. In order to retain the fruiting ability of the strains, further testing of various methods of preservation and different methods for isolation should be performed.

• 대한한방부인과학회지
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• 제25권3호
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• pp.16-26
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• 2012

Objectives: This study was performed to evaluate effects of Cordyceps militaris(CM) on osteoclast differentiation and its related gene expression. Methods: We used mouse myeloid cells RAW 264.7 stimulated by receptor activator of nuclear factor kappa-B ligand(RANKL) to induce osteoclast differentiation. There are four groups of which RAW 264.7 cells are not stimulated by RANKL (Normal), stimulated by RANKL without CM(Control), stimulated by RANKL with 0.1 ${\mu}g/ml$ of CM(CM 0.1), stimulated by RANKL with 1 ${\mu}g/ml$ of CM(CM 1). Osteoclastogenesis was measured by counting Tartrate-resistant acid phosphatase-positive multinucleated cells [TRAP(+) MNC]. RT-PCR was performed to evaluate the inhibitory effect of CM on gene expression(TRAP, AKT1, JNK1, NFATc1, c-Fos, MITF). Results: 1. CM decreased the number of TRAP(+) osteoclast in RANKL-stimulated RAW 264.7 cell at the concentration of 0.1 ${\mu}g/ml$ and 1 ${\mu}g/ml$. 2. CM decreased the expression of TRAP in osteoclast at the concentration of 1 ${\mu}g/ml$. 3. CM decreased the expression of AKT1, JNK1 in osteoclast at the concentration of 1 ${\mu}g/ml$. 4. CM didn't affect the expression of NFATc1, c-Fos, MITF in osteoclast. Conclusions: Cordyceps militaris has inhibitory effects on osteoclast differentiation and its related gene expression. These results suggest that Cordyceps militaris has a potential as a treatment of osteoporosis.

• 한국식품영양과학회지
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• 제31권5호
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• pp.743-748
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• 2002

식빵에 단백질을 보강할 목적으로 눈꽃 동충하초와 번데기 동충하초의 분말을 첨가하여 제조한 식빵의 관능적 특성을 측정한 결과는 대조군과 눈꽃 동충하초 첨가군에서 구수한 맛, 입안에서의 감촉, 향의 기호도가 높게 평가되었다 수분함량은 대조군에 비해 J2군이 가장 높았으며 M4군이 가장 낮았다. 부피는 대조군에 비해 눈꽃 동충하초 첨가군은 커졌고 번데기 동충하초 첨가군은 감소하였으며, 동충하초 첨가량이 증가될수록 감소하였다. 색도에서 명도 값(L)은 대조군을 기준으로 동충하초 첨가군이 모두 낮았으며 특히 M4군이 유의적으로 낮았다. 적색도 값(a)은 대조군을 기준으로 번데기 동충하초의 첨가량이 많을수록 양의 값을 가졌으며 눈꽃 동충하초의 첨가량이 많을수록 음의 값을 나타내 상반된 결과를 가져왔다. 황색도 값(b)은 대조군과 눈꽃 동충하초 천가군에 비해 M4군이 유의적으로 높았다. 경도(Hardness)는 M4군이 가장 높았으며(p<0.001). 탄력성은 대조군에 비해 눈꽃과 번데기 동충하초 첨가 식빵의 유의적인 차이가 없었다. 관능과 기계적 검사의 상관관계에서, 명도는 관능항목의 색, 경도와 부의 상관관계(p<0.001), 적색도와 황색도, 경도는 관능항목의 색, 묵은 곡식가루 냄새, 경도를 제외한 모든 항목에서 부의 상관관계(p<0.001)를 보였다. 또한 기계적 검사의 응집성과 탄력성은 관능 항목의 색과 묵은 곡식가루 냄새, 경도와 부의 상관관계를 나타냈다. 이상의 결과 눈꽃 동충하초의 경우 2% 첨가군이 대조군과 유사한 경향을 나타냈으며 특히 기호도, 수분 함량과 부피에서 좋은 특성을 가졌다. 동충하초의 첨가량은 눈꽃 동충하초의 경우 4%까지도 무방하며, 번데기 동충하초의 경우 2%가 적절하였다.

• 한국균학회지
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• 제37권2호
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• pp.161-166
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• 2009

The purpose of the present study is to develop a simple, fast and sensitive LC/MS method for simultaneous separation and the determination of an active component in the oriental medicinal mushroom Cordyceps militaris. Based on this work, the contents of cordycepin in Cordyceps militaris fruiting cultivated on various media were determined and compared. And also, the nutritional components such as minerals and vitamins were determined in order to provide useful information to consumer as a food material. The analysis methods of nutritional components were chosen on the basis of AOAC. The optimum separation for cordycepin was achieved using a solvent gradient consisting of the mixture of 0.1% formic acid in methanol (solvent B) in a background of 0.1% formic acid in water (solvent A) as a mobile phase and a 3.0${\times}$150 Waters XTera column. Selective ion monitoring (SIR) mode ([M+H]+ at m/z 252) was used for quantitative analysis of cordycepin. The cultivated Cordyceps militaris on various media contained 1~14 /g of cordycepin, 0.65~1.08% of thiamine, 0.86~7.17% of riboflavin, and 3.01~5.26% of niacin. The content of mineral components varied on categories, especially contained 500~3500% of potassium as a major mineral. Cordycepin, niacin and potassium were found much higher in the fruiting cultivated with soy power media (gold 10) than other media.

• 한국균학회지
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• 제1권1호
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• pp.13-16
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• 1973

맥각균과(麥角菌科)의 Cordyceps속(屬)은 오래전부터 약용(藥用)으로 쓰여졌으며 현재(現在)까지 한국(韓國)에서 Cordyceps sobolifera, C. militaris 및 C. nutans의 3종(種)만이 보고(報告)되어 있다. 저자(著者)들은 Cordyceps tricentri Yasuda 및 C. sphecocephala(Klotz) Mass의 미기록(未記錄) 2종(種)을 한국(韓國)에서 채집하여 보고(報告)한다.

• 생약학회지
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• 제44권4호
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• pp.336-343
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• 2013

In a continuation of investigation on Cordyceps militaris, thirteen compounds were isolated from the $CH_2Cl_2$ and n-BuOH-soluble fraction of C. militaris. They were identified as twelve diketopiperazines such as cyclo($\small{L}$-Gly-$\small{L}$-Pro) (1), cyclo($\small{L}$-Ala-$\small{L}$-Pro) (2), cyclo($\small{L}$-Ser-$\small{L}$-Pro) (3), cyclo($\small{L}$-Val-$\small{L}$-Pro) (4), cyclo($\small{L}$-Thr-$\small{L}$-Pro) (5), cyclo($\small{L}$-Pro-$\small{L}$-Pro) (6), cyclo($\small{L}$-Thr-$\small{L}$-Leu) (7), cyclo($\small{L}$-Tyr-$\small{L}$-Ala) (8), cyclo($\small{L}$-Phe-$\small{L}$-Ser) (9), cyclo($\small{L}$-Phe-$\small{L}$-Pro) (10), cyclo($\small{L}$-Tyr-$\small{L}$-Pro) (11) and brevianamide F (13), and an amino acid, tryptophan (12). Their structures were identified on the basis of chemical evidences and spectroscopic analysis including 1D-NMR ($^1H$, $^{13}C$), 2D-NMR (HSQC, HMBC) and MS spectral data. Among the isolated compounds, compounds 1, 2, 6-11 are first reported from C. militaris.

• Mycobiology
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• 제34권2호
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• pp.83-91
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• 2006

Characteristic growth patterns of Cordyceps militaris isolates on various media, under varying light conditions and at varying incubation periods were examined. Light was found to be the most critical single factor in determining the density, texture, and pigmentation of the mycelial culture of the fungus. However, under the light condition, the degree of pigmentation and mycelial density were found to be affected by the incubation period and type of medium. Irrespective of the variations in medium type or incubation period, there was no pigmentation of the mycelium under dark condition. Radial growth of the mycelium was faster under dark incubation rather than under light incubation. Abundant mycelial density and darkest pigmentation of C. militaris isolates were produced in nutritionally rich media like SDAY, SMAY and CZYA, suggesting that these media may fulfill all the requirements for vegetative growth of the fungus. Growth characteristics of C. militaris isolates could be easily observed by the simple agar culture method, which would be useful to characterize the phenotypic characteristics of large number of pure cultures of the fungus under given conditions of growth factors such as medium, light and temperature.

• Mycobiology
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• 제50권1호
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• pp.1-11
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• 2022

The ascomycete fungus Cordyceps militaris infects lepidopteran larvae and pupae and forms characteristic fruiting bodies. Owing to its immune-enhancing effects, the fungus has been used as a medicine. For industrial application, this fungus can be grown on geminated soybeans as an alternative protein source. In our study, we performed a comprehensive transcriptomic analysis to identify core gene sets during C. militaris cultivation on germinated soybeans. RNA-Seq technology was applied to the fungal cultures at seven-time points (2, 4, and 7-day and 2, 3, 5, 7-week old cultures) to investigate the global transcriptomic change. We conducted a time-series analysis using a two-step regression strategy and chose 1460 significant genes and assigned them into five clusters. Characterization of each cluster based on Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases revealed that transcription profiles changed after two weeks of incubation. Gene mapping of cordycepin biosynthesis and isoflavone modification pathways also confirmed that gene expression in the early stage of GSC cultivation is important for these metabolic pathways. Our transcriptomic analysis and selected genes provided a comprehensive molecular basis for the cultivation of C. militaris on germinated soybeans.