약용 및 야생식물로부터 트롬빈 저해물질의 탐색

손호용,권윤숙,권하영,권정숙,서영배,권기석,손건호 한국생명과학회 2003 한국생명과학회 심포지움 Vol.39 No.-

Regulation of Branched-Chain, and Sulfur-Containing Amino Acid Metabolism by Glutathione during Ultradian Metabolic Oscillation of Saccharomyces cerevisiae

손호용,Eun-Joo Kum,권기석,진익렬,Hiroshi Kuriyama 한국미생물학회 2005 The journal of microbiology Vol.43 No.4

Autonomous ultradian metabolic oscillation (T≅50 min) was detected in an aerobic chemostat culture of Saccharomyces cerevisiae. A pulse injection of GSH (a reduced form of glutathione) into the culture induced a perturbation in metabolic oscillation, with respiratory inhibition caused by H2S burst production. As the production of H2S in the culture was controlled by different amino acids, we attempted to characterize the effects of GSH on amino acid metabolism, particularly with regard to branched chain and sulfur-containing amino acids. During stable metabolic oscillation, concentrations of intracellular glutamate, aspartate, threonine, valine, leucine, isoleucine, and cysteine were observed to oscillate with the same periods of dissolved O2 oscillation, although the oscillation amplitudes and maximal phases were shown to differ. The methionine concentration was stably maintained at 0.05 mM. When GSH (100 M) was injected into the culture, cellular levels of branched chain amino acids increased ramatically with continuous H2S production, whereas the cysteine and methionine concentrations were noticeably reduced. These results indicate that GSH-dependent perturbation occurs as the result of the promotion of branched chain amino acid synthesis and an attenuation of cysteine and methionine synthesis, both of which activate the generation of H2S. In a low sulfate medium containing 2.5 mM sulfate, the GSH injections did not result in perturbations of dissolved O2, NAD(P)H redox oscillations without burst H2S production. This suggests that GSH-dependent perturbation is intimately linked with the metabolism of branched-chain amino acids and H2S generation, rather than with direct GSHGSSG redox control.

약용 및 야생식물로부터 트롬빈 저해물질의 탐색

손호용,권윤숙,김영숙,권하영,권기석,김경재,권정숙,손건호 한국생약학회 2004 생약학회지 Vol.35 No.1

Inhibitory activities of 264 methanol extracts, which were prepared from different parts of 210 kinds of wild and medicinal plants, against human thrombin were evaluated. Based on the anti-coagulation activity determined by thrombin time and activated partial thromboplastin time, the 14 extracts were screened. The fibrinolytic activity, heat stability and inhibition of other proteolytic digestive enzymes, such as pepsin, papain, trypsin and hymotrypsin, of the 14 extracts were further determined, and Ginko biloba (herba), Ephedra sinica (radix), Reynoutria elliptica (herba), Amomum tsao-ko Crevost (fructus), and Magnolia officinalis Rehd. et Wils (bark) were finally selected as possible plant sources for anti-thrombosis agent. These results suggested that medicinal and wild plants could be the potential source of thrombin inhibitor.

원유 누출 오염토양으로부터의 석유 탈황용 미생물 분리 ( Isolation of Microorganisms for Petroleum Desulfurization from Oil Contaminated Soil )

손호용,장제환,장용근,장호남,류희욱,조경숙 한국에너지학회 1996 한국에너지공학회 학술발표회 Vol.1996 No.-

약용 및 야생식물로부터 트롬빈 저해물질의 탐색 (II)

손호용,권정숙,권기석,김건우,손건호,류희영,권윤숙,금은주 한국생약학회 2005 생약학회지 Vol.36 No.4

− To develop safe and new anticoagulation agents from natural resources, the inhibitory activities of 291 methanol extracts, which were prepared from different parts of 197 kinds of medicinal and wild plants, against human thrombin were evaluated. Based on anti-coagulation activity determined by thrombin time and activated partial thromboplastin time, the extract of leaf of Myrica rubra was finally selected. The extract of M. rubra showed a strong thrombin inhibitory activity (above 1,819%) at 0.5 mg/ml as a final concentration, whereas aspirin showed 337% inhibition at concentration of 1.5 mg/ml. The activity of the extract remained more than 85% and 60% by heat treatment at 100 oC for 30 min, and acid treatment at pH 2 for 60 min, respectively. Our results suggested that the extract of Myrica rubra could be the potential source as thrombin inhibitor.

HepG2 인간 세포주, Lumbricus rubellus 및 Saccharomyces cerevisiae를 이용한 내분비교란물질의 독성평가

손호용,김홍주,금은주,조민섭,이중복,김종식,권기석,Sohn, Ho-Yong,Kim, Hong-Ju,Kum, Eun-Joo,Cho, Min-Seop,Lee, Jung-Bok,Kim, Jong-Sik,Kwon, Gi-Seok 한국생명과학회 2006 생명과학회지 Vol.16 No.6

다양한 독성물질 및 이들의 대사산물의 효율적인 생물독성 평가시스템이 지속적으로 개발되고 있다. 본 연구에서는 대분비계 교란물질인 endosulfan, bisphenol A, vinclozolin 및 3,5-dichloroaniline을 대상으로 인간 간암세포주, 지렁이, 효모를 이용한 세포독성 및 성장억제 효과를 평가하였다. 인간 간암세포주 독성평가에서는 endosulfan, 3,5-dichloroaniline, bisphenol A의 순으로 독성이 나타났으며, 지렁이 독성평가에서는 endosulfan, bisphenol A, 3,5-dichloroaniline의 순으로 독성이 나타났다. 효모를 이용한 독성평가에서는 3,5-dichloroaniline, endosulfan, bisphenol A의 순으로 독성이 나타나 다른 시스템과는 부분적인 차이가 나타났으며, vinclozolin의 경우 3가지 독성평가 시스템에서 모두 독성이 나타나지 않았다. 이러한 결과는, 동일한 물질을 서로 다른 생물 독성평가 시스템을 사용하여 평가하는 경우, 부분적인 오류가 나타날 수 있음을 암시하고 있으며, 독성 유무 판단은 가능하더라도, 독성 정량평가 및 독성 정도를 비교하는 것은 어렵다는 것을 제시하고 있다. 또한 본 결과는, 다양한 물질 및 이들의 대사산물의 일차적 독성평가에는 지렁이 및 효모시스템이 빠르고 경제적임을 암시하고 있으며, 독성이 인정될 경우 인간세포주 및 동물시험에 의한 검증이 효율적이라고 판단된다. Toxicity evaluation systems for various chemicals and their metabolites have been developed during last decades. In this study, the acute toxicity of endocrine disrupting chemicals, such as endosulfan, bisphenol A, vinclozolin, and 3,5-dichloroaniline, was evaluated using HepG2 cell line, Lumbricus rubellus and Saccharomyces cerevisiae, respectively. The extents of toxicity of the chemicals in different bioassay systems varied substantially, such as endosulfan>3,5-dichloroaniline> bisphenol A in HepG2 cell line system, endosulfan>bisphenol A>3,5-dichloro aniline in L. rubellus system, and 3,5-dichloroaniline>endosulfan>bisphenol A in S. cerevisiae system. Meanwhile, no cytotoxicity was observed by treatment of vinclozolin in the evaluation systems. Our results suggest that earthworm and yeast are useful to evaluate acute toxicity of endocrine disrupting chemicals, and direct comparison of toxicity data from different bioassay systems is unattainable. Based on our results, we propose that the bioassay system with earthworm or yeast, a rapid, simple and economic system, could be applied as pre-test for the toxicity evaluation using human cell line or animals.

비스페놀 에이의 세포독성과 산화적 스트레스의 연관성

손호용,금은주,이중복,권기석 한국작물학회 2006 한국작물학회 학술발표대회 논문집 Vol.2006 No.-

Lumbricus rubellus를 이용한 endosulfan의 간편, 신속 독성 평가 및 endosulfan 분해 미생물의 선별

손호용,김홍주,금은주,이중복,권기석,Sohn Ho-Yong,Kim Hong-Ju,Kum Eun-Joo,Lee Jung-Bok,Kwon Gi-Seok 한국생명과학회 2006 생명과학회지 Vol.16 No.1

독성물질의 무독화 연구의 화학적 분석방법의 문제점을 보완하고, 난분해성 독성물질의 분해 미생물을 효율적으로 탐색하기 위해, 지렁이와 microplate를 이용한 빠르고 간편한 독성평가 시스템을 구축하였다. 본 방법은 지렁이의 사육이나 특수시설이 필요하지 않으며, 치사 및 무게감소가 현저하게 나타나지 않는 저농도, 단기간 처리시에도 신속하게 독성의 정량평가가 가능하다 실제 endosulfan 및 다양한 endosulfan 유도체, 구조적 유사체 등을 대상으로 독성 평가한 결과, endosulfan, endosulfan sulfate, aldrin 및 dieldrin 에서만 독성이 나타나 기존의 연구결과와 잘 일치하였다. 이를 이용하여 endosulfan분해 및 무독화 균주 선별을 시도한 결과, 기존 endosulfan분해 및 무독화 균주로 선별된 KE-1, KE-8, KS-2P균주 처리의 경우에는 지렁이의 독성이 거의 나타나지 않았으나, E. coli, B. subtilis, 및 YSU 균주들을 처리한 경우에는, 지렁이 사멸과 함께 심각한 독성이 나타났다. 따라서, 본 시스템은 독성물질의 독성평가뿐 아니라, 독성물질의 분해 및 무독화 미생물 선별에 매우 유용함을 확인하였다. To compensate the problems of chemical assay in detoxification of recalcitrant and a practical approach in selection of bioremediation bacteria, a simple and rapid toxicity evaluation system was constructed based on Lumbricus rubellus. Long term-culture and specific equipment are not necessary, and semi-quantitative analysis of toxicity at sub-lethal concentration is possible by measuring of dose-dependent increased yellowish secreted compounds. When the toxicity of endosulfan, its metabolites and structurally related chemicals were measured for 24 h, the results were coincided with previous reports. Toxicity was found in endosulfan, endosulfan sulfate, aldrin, and dieldrin, respectively. Rapid and economic selection of endosulfan-detoxifying bacteria was possible using our system. Klebsiella pneumoniae KE-1, K. oxytoca KE-8 and Pseudomonas sp. KS-2P, reported endosulfan degrading bacteria, ameliorated the endosulfan toxicity, whereas E. coli, B. subtilis and other bacteria failed to protect the toxicity of endosulfan in L. rubellus. Our results suggest that the constructed system is useful to selection of microorganism as well as toxicity evaluation against toxic recalcitrants.

Candidiosis 치료제 개발을 위한 약용 및 야생 식물의 항진균 활성의 검색

손호용,금은주,권윤숙,권기석,진익렬,권하영,권정숙,손건호 한국생명과학회 2003 생명과학회지 Vol.13 No.5

안전성이 확보된 식용, 또는 약용식물로부터 항캔디다 활성이 우수한 항진균 후보물질을 개발하기 위해, 298종의 천연물로부터 추출부위 및 추출 용매를 달리하여 384종의 추출물을 제조하였다. 제조된 추출물의 항캔디다 활성을 disc-paper를 이용한 생육 저지환의 유무 및 크기 판정과 microbroth dilution법을 이용하여 C. albicans KCTC 1940과 S. cerevisiae IFO 0233를 대상으로 평가하였다. 그 결과 20종 식물로부터 조제된 25종의 추출물이 우수한 항캔디다 활성을 나타냄을 확인하였고, 그 중에서도 멍석딸기, 정향, 천황련, 작약, 모란, 대극잎 추출물은 강력 한 항캔디다 활성$(MIC=50 \mug/m\ell)$을 보였다. 특히, 멍석딸기(에틸아세테이트 추출물), 정향(부탄올 추출물), 호장근(에틸아세테이트 추출물), 작약(부탄올 추출물), 대극잎(메탄올 추출물)의 경우에는 C. albicans에 높은 선택독성을 나타내어, 안정성이 확보된 식물 추출물로부터 새로운 진균 감염증 치료제 개발이 가능함을 제시하였다. Candida albicans is one of the most common etiological agents in fungi-associated skin infections. There is an increase of candidiosis especially in the patient of acquired or induced immunodeficiency syndromes or in the event of long-term antibiotics and immuno-suppressor or cytotoxic therapies. To screen out reliable and effective anti-candidiosis agent, in this study, we have evaluated antifungal activity of 298 plant extracts against C. albicans. Based on the results of disc-paper method and determination of minimal inhibitory concentration, fifteen extracts were finally selected as possible sources of anti-candidiosis agent. Especially, six different plant extracts, such as Rubus parvifolius, Euphorbia pekinensis, Coptis chinensis, Eugenia aromaticum, Paeonia lactiflora var. hortensis and Paeonia suffruticosa showed strong antifungal activity against C. albicans, not to S. cerevisiae. These results suggested that medicinal and wild plants could be the potential source of antifungal agent.

Antifungal Activity of Fistulosides, Steroidal Saponins, from Allium fistulosum L

손호용,금은주,류희영,전수진,김남순,손건호,Sohn Ho-Yong,Kum Eun-Joo,Ryu Hee-Young,Jeon Su-Jin,Kim Nam-Soon,Son Kun-Ho Korean Society of Life Science 2006 생명과학회지 Vol.16 No.2

대파(Allium fistulosum L)는 백합과 다년초로서 한국, 일본 중국 등 전 세계적으로 식용으로 재배되어 왔다. 현재까지 대파의 다양한 생리활성이 보고되어 왔으나, 항균 활성에 대해서는 거의 보고되어 있지 않다. 본 연구에서는 대파 식용부위로부터 강력한 항미생물성 steroidal saponins (fistuloside A, B, 및 C)을 분리하였고, 이들의 항균력을 병원성균 및 식품부패균을 대상으로 평가하였다. Fistuloside A 및 C는 강력한 항진균 활성과 항Proteus 세균활성을 나타낸 반면, fistuloside B는 항진균 활성만 인정되었다. 특히 fistuloside C는 $3.1{\sim}6.2{\mu}g/ml$의 MIC (minimal inhibitory concentration) 및 MFC (minimal fungicidal concentration)를 나타내어 강력한 항진균 활성이 진균사멸에 의한 항균효과임을 확인하였다. 본 연구결과는 대파가 미생물 감염증 제어용으로 사용되어 온 근거를 제시하며, 또한 fistuloside C의 항진균제로서의 이용 가능성을 제시하고 있다. Allium fistulosum L. (Welsh onion) is a perennial herb that is widely cultivated throughout the world, especially in China, Japan and Korea. Although, various activities were reported, little is known about antimicrobial activity of A. fistulosum L.. In this study, strong antimicrobial substances, fistuloside A, B, and C were isolated from the edible parts of A. fistulosum L. and their antimicrobial activity was evaluated with pathogenic- or food-spoilage microorganism based on disk-diffusion assay, minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) determination. Fistuloside A and fistuloside C showed strong antifungal activity and anti-Proteus activity, while fistuloside B is effective to only fungi. The fistuloside C showed a prominent antifungal activity with $3.1{\sim}6.2{\mu}g/ml$ of MIC and MFC. Our results indicated that fistuloside C has a prominent antifungal activity and support the use of A. fistulosum to treat microbial infection.