기존의 번데기동충하초의 자실체 생산은 자연계에서 채집된 버섯의 조직을 분리하거나 다중의 포자를 이용하는 방법이 대표적이었으나 본 연구를 수행한 결과, 단포자를 선발한 후 교배조합에 의한 F1균주를 이용한 배양이 안정적임을 확인 할 수 있었다. 단포자의 특성으로는 자실체의 형성이 양호하며 균주의 안정성이 우수한 균주를 선발하고 조합형성능이 우수한 균주를 선발하여야 한다. 본 연구를 통하여 06S05의 단포자를 획득하였으며 06S05x06S01, 06S05x06S33균주를 선발하였으며 06S05x06S33균주를 예당 3호로 명명하여 국립종자원에 품종보호 등록되었다.

Cordycepin (3’-deoxyadenosine) is a polyadenylation specific inhibitor, one of the components of Cordyceps militaris. It has been shown to possess many pharmacological activities including immunologically stimulating, anti-tumor, anti-virus, and anti-infection effects. However, its molecular mechanisms are poorly understood. In this study, the apoptotic effects by cordycepin were investigated in human leukemia cells. Cordycepin treatment inhibited leukemia cells growth a concentration-dependent manner by inducing apoptosis,as evidenced by morphological change and apoptotic cell death such as formation of apoptotic bodies, DNA fragmentation and increased populations of apoptoticsub G1 phase. Induction of apoptosis by cordycepin in leukemia cells were associated with modulation of Bcl-2 member and inhibitor of apoptosis (IAP) proteins expression. Cordycepin also increased ROS generation, activation of caspase-3, caspase-8, caspase-9, cleavage of poly(ADP-ribose) polymerase (PARP), -catenin and phospholipase (PLC)-1 protein. Both the this effect by cordycepin treatment were significantly inhibited by NAC, a ROS scavenger, demonstrating the important role of ROS in the observed cytotoxic effect. This results suggested that cordycepin may be a potential chemotherapeutic agent for the treatment of leukemia patients.

Cordyceps has been used in traditional Chinese medicine more than 2000 year ago. In this study, the new Cordyceps militaris was founded and isolated from O-dae mountain in Korea, and was identified its genetic characteristics. The newly isolation strain HB8 was most closet to Cordyceps militaris W141449 (99.82%), Cordyceps militaris JLCY-LI819 (99.82%) and Cordyceps militaris 4642 (99.81%), respectively. the genotypic result was show that train HB8 was belonging to the Cordyceps militaris genus, therefore, Cordycep militaris HB8 proposed with accession number MT835161. This study we find the optimal condition for production of cordycepin from Cordyceps militaris HB8 was 8 ㎎ /g (200 g of pupa, 1 g of KH2PO4, 0.5 g of K2HPO4, 20 g of glucose, 1 g of MgSO4, 0.05 g of vitamin B1, and 1 ㎎ of NAA per liter; light condition 300-700 Lux and day/night was 14 h/10 h) and the optimum condition for the production of adenosine was 2.6 ㎎/g (15 g of skim milk powder, 1 g of KH2PO4, 0.5 g of K2HPO4, 20 g of glucose, 1 g of MgSO4, 0.05 g of vitamin B1, and 1 ㎎ of NAA per liter; light condition 300-700 Lux and day/night was 14 h/10 h).

Background : Cordyceps militaris has been an wonder drug to anti-aging efficacy and called the three main drugs with ginseng and deer antler from the past. Cordycepin, cordycepic acid (d-mannitol) and adenosine are known as functional ingredients in Cordyceps militaris. Among them, cordycepin, the representative component, has been reported as antimicrobial substance containing immune enhancement, anti-cancer and anti-inflammatory effects. Methods and Results : After Cordyceps militaris produced from different types of medium mixed with 10-fold volume of purified water, the mixture were extracted at 70±5℃ for a hour and that extracts re-extracted using ultrasonics wave for 30 minutes. Qualitative analysis of the index component was determined by using the Q-TOF (A quadrupole time-of-flight mass spectrometer), and quantitative analysis was performed by using HPLC (high-performance liquid chromatography) with Xselect HSS T3 column (2.1 X 100 mm, 2.5㎛, Waters, USA) and ultrapure water and acetonitrile as mobile phase A and B. Detection column temperature, injection volume and the flow rate were 35℃, 2 μL and 0.3 mL / minute respectively. The cordycepin content of Cordyceps militaris produced from medium mixed with vegetable and animal ingredients higher than single ingredient. Moreover, through a variety of analyzes by varying the type and content of the medium additives, the cordycepin in Cordyceps militaris produced from medium mixed with animal ingredients highest. Furthermore, the cordycepin content of a fruit body was higher than those of the a mycelium. Conclusion : These results provide a method for producing an high cordycepin content of Cordyceps militaris as functional food ingredient.

세포 노쇠화(cell senescence)는 나이 듦에 따른 내인성 노화 및 질병들에서 나타날 수 있는 세포의 노화인자 발현, 세포분열 정지 등의 현상으로 일컬어진다. 피부세포의 경우, 노화 및 외부요인으로 인한 세포 노쇠화가 일어나 세포분열의 정지 및 기능 이상이 관찰되며 이는 피부노화를 가속화시키는 요인이 된다. 본 연구에서는, cordycepin을 이용하여 노화된 피부세포의 세포 노쇠화 억제 및 기능 향상을 유도하여 피부노화 개선의 가능성을 제시하였다. 사람에서 유래한 섬유아세포를 이용하여 세포의 β-galactosidase 활성 세포염색 결과, 많은 계대의 세포에서 발현이 높게 나타남을 알 수 있었다. 항산화 및 항염 효과가 알려진 cordycepin을 많은 계대의 세포에 처리하였을 때 β-galactosidase 활성이 확연히 떨어짐을 확인하였고 무혈청 배지 조건에서 많은 계대 세포의 증식 및 생존율을 높이는 결과를 보였으며 세포 노쇠화와 많은 연관성이 대두되고 있는 미토콘드리아의 기능관련 실험을 진행한 결과, 높은 ROS억제능이 나타났다. 본 연구를 통하여 노화된 사람 피부 섬유아세포에서의 cordycepin의 세포 노화 개선능을 알 수 있었으며, 피부 항노화소재로서의 가능성을 확인하였다.