동충하초 균사체 및 동충하초 균사체로부터 분리된 코디세핀의 항암 활성 평가

하시영 ( Si Young Ha ),정지영 ( Ji Young Jung ),이동환 ( Dong Hwan Lee ),최민서 ( Min Seo Choe ),양재경 ( Jae-kyung Yang ) 한국목재공학회 2021 한국목재공학회 학술발표논문집 Vol.2021 No.1

Various medicinal fungi have been traditionally used in many countries for health maintenance, as well as for the prevention and treatment of various diseases. Dong-Chong-Xia-Cha in Chinese, which translates as “winter worm and summer grass”, is an entomogenous fungus that colonized the larvae or pupae of insect. Although various biological activities of crude extracts have been reported, the precise active compounds responsible for the anticancer activities of cultured Cordyceps are not well defined. Therefore, as part of our study program to identify potential anticancer agents from traditional medicines, we used cordycepin isolated from Cordyceps militaris obtained by submerged cultivation. The aim was to isolate the pure cordycepin from the extracts of Cordyceps militaris and evaluate its anticancer properties. Silica gel column chromatographic purification of Cordyceps militaris extracts resulted in the isolation of cordycepin. The isolated cordycepin and crude cordyceps were examined for their anti-proliferative effects on human cancer cell lines, HCT, MC5-7, U-87MG, AGS, A549, and MCF-7 cells were analyzed. The isolated cordycepin displayed potent growth inhibition on HCT, MC5-7, U-87MG, AGS, A549, and MCF-7 cells with IC50 values of 86.16, 28.13, 282.40, 78.41, 137.40, and 216.40 μg/ml, respectively. A similar inhibitory trend in these cancer cells was observed for crude cordyceps, with IC50 values ranging from 60.8 317.2. The present study provides scientific supporting information for the ethnopharmacological use of Cordyceps militaris as an anticancer agent.

열처리 우드 칩을 이용한 인공상토의 제조 및 상추의 생장 평가

이동환 ( Dong Hwan Lee ),정지영 ( Ji Young Jung ),하시영 ( Si Young Ha ),박재현 ( Jai Hyun Park ),최지원 ( Ji Won Choi ),양재경 ( Jae-kyung Yang ) 한국목재공학회 2019 한국목재공학회 학술발표논문집 Vol.2019 No.2

상토는 식물체를 지지하고 작물의 생육에 필요한 각종 양분과 수분을 공급해주는 물질로서 육묘에 있어 가장 기본이 되는 물질이다. 일반적인 상토로 가장 많이 이용되는 피트모스는 주로 캐나다와 북유럽 지역에서 채취되기 때문에 전량 수입에 의존하는 실정이다. 이에 따라 가격이 높아질 뿐만 아니라 피트모스 제조에 장기간이 소요된다는 단점이 있다. 따라서 최근 피트모스를 대체할 수 있는 원료를 개발하기 위한 연구가 주목받고 있다. 우드 칩은 사업장 또는 간벌 후 다량 발생되고 있으며 처리를 위해 대부분이 소각되는데, 이는 공기중에 유해가스를 유발시킨다. 따라서 본 연구에서는 피트모스를 대체하는 원료로서 우드 칩의 가능성을 평가하였다. 이를 위해 열처리 우드 칩을 이용해 인공상토를 제조하여 상추의 생장에 대한 평가를 하였다. 피트모스와 열처리 우드 칩을 10:0, 7:3, 5:5, 3:7, 0:10 (w/w) 비율로 혼합하였으며, 그 결과 피트모스와 열처리 우드 칩의 발아율은 유의성이 없었으나, 근장은 피트모스와 열처리 우드 칩의 혼합 비 3:7(w/w)과 0:10(w/w)에서 피트모스 보다 약 1.6배, 1.7배 높은 것으로 확인 되었다. 뿐만 아니라, 초장에서는 열처리 우드 칩에서 피트모스 보다 약 1.2배 높은 성장률을 보였다. 따라서 열처리 우드 칩이 피트모스의 대체 원료로서 이용 가능할 것으로 판단된다.

액체배양에서 배지 조성이 붉은사슴뿔버섯의 roridin E 함량에 미치는 영향

이동환 ( Dong Hwan Lee ),하시영 ( Si Young Ha ),정지영 ( Ji Young Jung ),양재경 ( Jae-kyung Yang ) 한국목재공학회 2020 한국목재공학회 학술발표논문집 Vol.2020 No.1

암은 우리나라 사망자들의 가장 큰 사망 원인 중 하나이며, 그 중 하나인 유방암은 5대 암 중의 하나이며, 여성암 중 가장 많이 발생하는 암이다. 이런 유방암을 치료하기 위한 기존의 치료 물질로 doxorubicin이 주로 이용되고 있으나, 작게는 구토, 모발 손실부터 크게 심장기능 약화 같은 심각한 부작용까지 유발하여 이를 대체하기 위한 새로운 치료 물질의 탐색이 필요하다. 붉은사슴뿔버섯에 함유되어 있는 roridin E는 doxorubicin 보다 강력한 항암활성을 가진 것으로 확인이 되었다. 붉은사슴뿔버섯 균사체는 PDA 배지에서 25℃, 30일간 배양 하여 사용하였으며, 액체배양을 위해 시판하는 PDB, MCM 2가지와 YMK(Glucose 20, Yeast extract 5, MgSO₄·7H₂O 1, KH₂PO₄ 2g/distilled water 1L), BSM(Glucose 30, Yeast extract 2.8, KH₂PO₄ 1, MgSO₄·7H₂O 0.5, KCl 0.5, FeSO₄·7H₂O 0.01, ZnSO₄·7H₂O 0.01, CuSO₄·5H₂O 0.005g/distilled water 1L)을 제조하여 사용하였다. 배양은 24℃, 100rpm, 암배양, 15일간 진행하였으며 3반복으로 배양하였다. 그 결과 균사체 생장량에선 유의미한 차이가 없었으나, HPLC분석을 한 결과, PDB배지를 사용하여 배양 했을 때 roridin E 함량이 0.41mg/g으로 0.15, 0.14, 0.32mg/g으로 나타나, 다른 배지에 비해 높은 것으로 확인 되었다.

붉은사슴뿔버섯으로부터 유방암세포에 항암활성을 가지는 roridin E의 함량 증대를 위한 조건 배양

이동환 ( Dong Hwan Lee ),하시영 ( Si Young Ha ),정지영 ( Ji Young Jung ),최민서 ( Min Seo Choe ),양재경 ( Jae-kyung Yang ) 한국목재공학회 2021 한국목재공학회 학술발표논문집 Vol.2021 No.1

Roridin E is a trichothecene that can be obtained from a poisonous mushroom known as Podostroma cornu-damae. Roridin E has anticancer activity against breast cancer cell lines; thus, it can be used to treat breast cancer. P. cornu-damae mycelia were cultured in submerged culture to obtain roridin E. By comparing the inside and outside of the mycelium, it was confirmed that more roridin E was present in the culture broth outside the mycelium. Variables were determined using the one-factor-at-a-time method. Response surface methodology (RSM), based on a three-level, three-variable Box-Behnken design (BBD), was employed to obtain an ideal combination of initial pH, incubation time, and rotational speed to roridin E production. The optimum culture conditions were as follows : initial pH as 4,; incubation time, 12.9 d; and rotational speed, 63 rpm. Under these conditions, the predicted production was 40.26 mg/L. The separated roridin E was found to have anti-cancer activity in five breast cancer cell lines: MCF-7, MDA-MB-231, HCC-70, T47D, and BT-474. The IC50 values were found to be 0.0004 - 0.002 mg/L.

< 구두-D-03 > LED (Light Emitting Diode) light가 Cordyceps militaris 균사체의 생장 및 cordycepin 함량 증대에 미치는 영향

하시영 ( Si Young Ha ),정지영 ( Ji Young Jung ),박재현 ( Jai Hyun Park ),이동환 ( Dong Hwan Lee ),최지원 ( Ji Won Choi ),양재경 ( Jae-kyung Yang ) 한국목재공학회 2019 한국목재공학회 학술발표논문집 Vol.2019 No.1

Mushroom is an abundant source of a wide range of useful natural products with biological activities. Because field cultivation of ushrooms takes a long time to grow and low productivity of bioactive compounds, submerged cultivation of mushrooms is viewed as a promising alterative for producing valuable substances. Cordyceps militaris (C. militaris), a caterpillar-shaped Chinese traditional medicinal mushroom, is usage as a crude drug, it has been extensively used as folk tonic food or invigorant since ancient times. This mushroom produces an important bioactive compound, cordycepin (3′-deoxyadenosine), which is a nucleoside analogue. Cordycepin is reported to possess many interesting biological and pharmacological activities, including immunological stimulating, anti-cancer, anti-virus and anti-infection activities. Previous work reported the isolation of cordycepin from liquid culture medium of C. militaris and its pharmacological functions. But, as far as we know, there is limited knowledge about the LED (Light Emitting Diode) condition for cordycepin production by C. militaris. In this paper, the effects of LED were focused in order to improve the cordycepin production by submerged cultivation of C. militaris. For this experiment, mycelial cultivation was performed in a shaking incubator at 24 ℃, 100 rpm for 5 days, and the medium was sabouraud dextrose broth (pH 5.6). The LED light mixed red and green, LED light mixed red and blue and LED light mixed green and blue were used for the LED, and dark culture and fluorescent lamp were used as the control. This results, C. militaris showed the highest mycelial weight when green light was irradiated, on the other hand, when irradiated with blue light, the content of cordycepin is about 7 times higher than that of fluorescent lamp in the cultures of C. militaris. In addition, the highest content of cordycepin was observed when irradiated for 6 h/day for 3 days. Interestingly, mycelial weight and cordycepin content were inversely related. The information obtained is considered fundamental and useful to the development of C. militaris cultivation process for efficient production of cordycepin on a large scale.

UVB 손상 세포에 대한 열처리 소나무 유래 추출물의 항산화 및 항염증 효과

정지영 ( Ji Young Jung ),하시영 ( Si Young Ha ),이동환 ( Dong Hwan Lee ),최민서 ( Min Seo Choe ),양재경 ( Jae-Kyung Yang ) 한국목재공학회 2021 한국목재공학회 학술발표논문집 Vol.2021 No.1

Oxidative stress and chronic inflammatory processes are major risk factors for aging and age-related diseases. The present study investigated the antioxidant and anti-inflammatory effects of extracts derived from steam-exploded pine in UVB-exposed human keratinocytes, HaCaT cells. The effects of extracts derived from steam-exploded pine on oxidative stress were examined in a UVB-induced aging model of human keratinocyte HaCaT cells. Extracts derived from steam-exploded pine protected cells from UVB-induced cytotoxicity (80 μg/mL) and reduced reactive oxygen species (ROS) production by UVB irradiation. Expression of genes associated with antioxidant effects, including superoxide dismutase 1 (SOD1) and catalase(CAT), increased in a dose-dependent manner. The expression of interleukin 6 (IL6), interleukin 6 (IL7), interleukin 8 (IL8), macrophage inflammatory protein-1β (MIP-1β), and monocyte chemoattractant Protein-1 (MCP-1) genes, which are associated with inflammation, also decreased in a dose-dependent manner. These results suggest that the extracts derived from steam-exploded pine may have an inhibitory effect on oxidative stress and inflammation.

ICT 기술을 이용한 토종다래 재배지의 토양환경인자 분석 및 결정트리 적용

하시영 ( Si Young Ha ),정지영 ( Ji Young Jung ),박재현 ( Jai Hyun Park ),이동환 ( Dong Hwan Lee ),최지원 ( Ji Won Choi ),양재경 ( Jae-kyung Yang ) 한국목재공학회 2019 한국목재공학회 학술발표논문집 Vol.2019 No.2

The other edible and very promising Actinidia species is Actinidia arguta (Siebold & Zucc.) Planch. ex Miq., also known as kiwiberry, hardy kiwi, baby kiwi, or mini kiwi. This exotic species is very interesting and promising given the horticultural advantages it has over kiwifruit, especially its high frost hardiness (down to -30 ℃ in midwinter) and relatively short vegetation period. To the best of our knowledge, there have been few experiments studying the effect of soil properties on mini kiwifruit productivity as well as the sugar content of A. arguta fruit. The variety of cultivar and expansion in arable farming has emphasized a need for more specific information related to factors that affect A. arguta productivity and sugar content of the fruit in the region. We analyzed various soil properties at experimental sites in South Korea. A Pearson’s correlation analysis was performed between the soil properties and sugar content or productivity of A. arguta. Further, a decision tree was used to determine the optimal soil conditions. The difference in the fruit size, sugar content, and productivity of A. arguta across sites was significant, confirming the effects of soil properties. The decision tree analysis showed that a soil C/N ratio of over 11.49 predicted a sugar content of more than 7 °Bx at harvest time, and soil electrical capacity below 131.83 μS/cm predicted productivity more than 50 kg/vine at harvest time. Our results present the soil conditions required to increase the sugar content or productivity of A. arguta, a new A. arguta cultivar in South Korea.

< 전시-P-103 > 목질 바이오매스를 이용한 옥상 녹화용 식생기반재의 물리·화학적 특성 및 식물 독성 평가

박재현 ( Jai Hyun Park ),하시영 ( Si Young Ha ),정지영 ( Ji Young Jung ),이동환 ( Dong Hwan Lee Jae-kyung Yang ),양재경 한국목재공학회 2019 한국목재공학회 학술발표논문집 Vol.2019 No.1

1990년대 후반부터 옥상녹화에 대한 연구가 시작되었으며, 현재에 이르러서는 경량의 식생기반재를 사용하여 건물 옥상에 실제 숲과 같은 ‘Eco-region’을 만드는 것에 대한 관심이 모아지고 있다. 이와 함께, 하수 슬러지, 농업 폐기물 등을 이용한 옥상 녹화용 식생기반재 개발에 대한 연구들이 많이 이루어지고 있다. 본 연구에서는 목질 자원을 이용하여 식생기반재를 제조하였고, 제조된 식생기반재의 옥상 녹화 이용가능성을 알아보기 위해 물리·화학적 특성 및 식물 독성을 평가하였다. 식생기반재는 목질원료, 피트모스, 펄라이트, 녹화용 비료, 녹화용 접착제, 액상 첨가제를 혼합하여 제조하였다. 토양을 대조구로 하여 시판 제품과 제조된 식생기반재의 물리·화학적 특성을 비교하고, 2종의 식물을 (벌노랑이, 월동춘채) 사용하여 제조된 식생기반재의 식물 독성 평가 (Germination index) 를 확인하였다. 실험 결과, 제조된 식생기반재의 물리적 특성 중 수분함량, 가비중, 공극률에서 (각각 26.5%, 0.1 g/cm3, 78.5%) 토양과 시판제품보다 식물 생장에 적합하였고, 화학적 특성에서는 유기물 함량, CEC, K 의 함량이 (각각 82.5%, 16.9 cmolc/kg, 88.9 cmolc/kg) 토양과 시판제품보다 식물 생장에 적합한 것을 확인하였다. 그리고 식물 독성 평가 결과, 벌노랑이, 월동춘채의 germination index는 각각 84.0%, 76.7%, 65.7% 인 것을 확인하여 식물에 독성을 미치지 않는 것으로 확인되었다. 결론적으로, 목질 자원을 이용하여 제조된 식생기반재는 식물 생장에 적합한 물리·화학적 특성을 가지고 있으며, 식물에 독성을 미치지 않으므로 옥상 녹화를 위해 사용할 수 있을 것으로 확인되었다.

액체배양된 Paecilomyces japonica의 코디세핀 함량에 대한 LED와 우드밀이 포함된 배지의 영향

하시영 ( Si Young Ha ),정지영 ( Ji Young Jung ),박재현 ( Jai Hyun Park ),이동환 ( Dong Hwan Lee ),최지원 ( Ji Won Choi ),양재경 ( Jae-kyung Yang ) 한국목재공학회 2020 한국목재공학회 학술발표논문집 Vol.2020 No.1

Paecilomyces japonica has been widely used based on folklore, not scientific studies. Cordycepin, a metabolite of P. japonica, has been showed to inhibit the growth of various tumor cells. Previous work reported the isolation of cordycepin from a liquid culture medium of P. japonica and its pharmacological functions. However, to the best of our knowledge, information on the light-emitting diode (LED) condition and wood meal-based media for cordycepin production by P. japonica is lacking. This study focused on the effects of a LED and wood meal based-media to improve cordycepin production by the submerged cultivation of P. japonica. For this experiment, mycelial cultivation was performed in a shaking incubator at 24 ℃ and 100 rpm for 5 days, with a medium of sabouraud dextrose broth (pH 5.6) with wood meal (P. densiflora). Red-green, red-blue, and green-blue LEDs were used for the experiments; a dark culture and fluorescent lamp were used as the control. P. japonica showed the highest cordycepin content when red-blue light was irradiated, the content of cordycepin was approximately 6 times higher than that with the fluorescent lamp in the cultures of P. japonica. In addition, the highest content of cordycepin was observed when cultivated in media with 20 g/L of 1% NaOH pretreated wood meal (100 mesh) under red-blue light.The combined application of LED and wood meal based-media has been shown to show synergistic effect for increasing cordycepin content from P. japonica. The information obtained is fundamental and useful to the development of P. japonica cultivation processes for the efficient production of cordycepin on a large scale.

열처리된 목질계바이오매스로부터 페놀성 화합물 분리를 위한 공융 용매 기반 추출 최적화에 있어서 Box-behnken과 Central composite design 설계의 비교

정지영 ( Ji Young Jung ),하시영 ( Si Young Ha ),박재현 ( Jai Hyun Park ),이동환 ( Dong Hwan Lee ),최지원 ( Ji Won Choi ),양재경 ( Jae-Kyung Yang ) 한국목재공학회 2020 한국목재공학회 학술발표논문집 Vol.2020 No.1

Deep eutectic solvents, environmentally friendly “green”solvents, have recently been applied extensively in several areas of chemistry, including the preparation of inorganic materials, organic synthesis, biochemistry, and analytical chemistry. There are an increasing number of studies in extraction and separation media for bioactive plant compound, including flavonoids, catechin, phenolic acid, terpenoids, and saponin. For the optimization of liquid-solid extraction processes, the response surface methodology is usually applied in combination with the full factorial design (FFD), central composite design (CCD) or Box-Behnken design (BBD) serving for the data collection. The present study deals with comparing the performances of CCD and BBD, which are used in combination with the response surface methodology (RSM) for the optimization of the phenolic compound extraction by deep eutectic solvent with respect to deep eutectic solvent concentration (1.6-3.2 g), time (120-240 min), solvent-to-sample ratio (20:1-60:1) and temperature (40-80℃). The optimum range condition was deep eutectic solvent concentration (2.1-2.2 g), time (182-200 min), solvent-to-sample ratio (36-47) and temperature (60-70℃). All two methods were efficient in the statistical modeling and optimization of the influential process variables and led to almost the same optimal process conditions and predicted phenolic compound content. Having better statistical performances and being economically advantageous over the CCD with repetition, the BBD combined with the RSM is recommended for the optimization of liquid-solid extraction processes.