석이로부터 분리한 페놀성 화합물의 phospholipase A_2 저해활성

김진우,송경식,유익동,장현욱,유승현,배강규,민태진 충남대학교 생물공학연구소 1997 생물공학연구지 Vol.5 No.-

호알카리성 진균 Cephalosporium sp. RYM-202가 생산하는 alkaline xylanase (CX-III)의 작용에 의해 xylan 기질로부터 생성되는 주요 가수분해 산물은 xylobiose와 중합도가 4 이상인 xylooligosaccharides이었다. 이 효소는 xylobiose에 대한 분해능을 가지고 있지 않지만 xylotriose로부터는 xylobiose를, xylotetraose로부터는 xylobiose와 xylotriose를 주산물로 형성하였다. 이러한 결과들은 CX-III가 transglycosidase 활성을 소유하는 전형적인 endo-type xylanase임을 보여준다. N-bromosuccinimide에 의한 CX-III의 화학적 변화 실험 결과 효소 1분자 당 2개의 tryptophan 잔기가 활성에 관여하는 것으로 나타났다. 그러나 iodoacetamide 및 diethylpyrocarbonate에 의한 효소활성의 저해효과는 나타나지 않음으로써 이 효소의 활성부위에 cysteine과 histidine 잔기가 필수적이지 않음이 확인되었다. The hydrolysis products formed from birchwood xylan by the action of an alkaline xylanase (CX-III) from alkalophilic Cephalosporium sp. RYM-202 were xylobiose and xylooligosaccharides polymerized with more than 4 sugar molecules. This enzyme was not active on xylobiose but readily attacked xylotriose accumulating xylobiose as a major product. The predominant end-products from xylotetraose by CX-III were xylobiose and xylotriose. These results indicate that the enzyme is typically endo-type xylanase possessing transglycosidase activity. Chemaical modification of CX-III with N-bromosuccinimide revealed that two tryptophan residues per molecule of CX-III were essential for its catalytic activity on xylan. On the other hand, iodoacetamide and diethylpyrocarbonate did not influence the activity of the enzyme, suggesting that cysteine and histidine residues are not involved in the active site of this alkaline xylanase.

Streptomyces tubercidicus ME-9189 균주가 생산하는 nucleoside계 제초 활성 물질

김원곤,김종평,김창진,유익동 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.1

방선균으로부터 제초활성 물질을 탐색하던 중 쇠비름과 바랭이에 대하여 강한 제초활성을 갖는 균주를 선발하여, 그 균주가 생산하는 활성물질을 분리 정제한 후 구조를 결정하고 정제된 물질에 대한 활성을 조사하였다. 선발된 ME-9189 균주의 미생물학적 특성을 조사한 결과 본 균주는 Streptomyces tubercidicus로 동정되어 본 균주를 Streptomyces tubercidicus ME-9189로 명명하였다. 또한 균 배양 상등액으로부터 활성탄, silica gel, sephadex LH-20 column chromatography 및 재결정화를 통하여 제초활성 물질 ME-9189 화합물을 정제한 후 UV, ^1H 및 ^13C-NMR, EIMS 등의 기기분석을 한 결과 본 물질을 tubercidin(4-amino-7β-D-ribofuranosyl-7H-pyrrolo[2,3-d]pyrimidine)으로 동정하였다. Tubercidin은 10ppm의 농도에서 무우의 발아후 생육을 50% 저해하였으며, herbicidin A보다 약 2배 강한 활성을, toyocamycin과는 비슷한 활성을 나타내었다. Three thousand microbial strains collected from different sources were screened for herbicidal activity. A strain of ME-9189 showed herbicidal activity against Digitaria sanguinalis and Portulaca oleracea was isolated from a mountainy soil. Based on taxonomic studies, the strain was identified as Streptomyces tubercidicus. The active compound of ME-9189 was purified from the culture broth by charcoal. silica gel, sephadex LH-20 column chromatography and crystalization, consecutively. The ME-9189 compound was identified as tubercidin by spectroscopic methods of UV, ^1H and ^13C-NMR, and EIMS. In the bioassay, growth of radish shoot and root was inhibited by 50% with tubercidin treatment of 10 ppm, showing 2 times higher activity than that of herbicidin A and similar to that of toyocamycin.

다양한 잡초로부터 생리활성물질의 탐색

김창진,강병화,유인자,박동진,이현선,김영호,유익동 충남대학교 약학대학 의약품개발연구소 1996 藥學論文集 Vol.12 No.-

다양한 잡초로부터 유용 생리활성물질을 탐색하고자 46종 잡초의 methanol 추출액을 대상으로 50~100 ㎍/㎖ 농도로 실험한 결과, 항균 활성에 있어서는 파리풀, 까실쑥부쟁이, 중머리대가리, 큰엉겅퀴, 부처꽃 등이 antobleb 활성에 있어서는 낙지다리, 밭뚝외풀, 까실쑥부쟁이, 술패랭이꽃 등이 항암활성에 있어서는 파리풀, 골풀, 밭뚝외풀, 까실쑥부쟁이, 술패랭이꽃, 겨우살이 등이 항산화활성에 있어서는 골풀, 물레나물, 청비녀골풀, 금불초, 방울고랭이, 좀고추나물 등이 비교적 강한 활성을 나타내었다. (1996년 4월 12일 접수, 1996년 8월 8일 수리) To search for bioactive compounds from plant resources, 80% methanol extracts of 46 species of weeds were screened for their activities of antimicrobial, antioxidative, antiblebing, antitumor and herbicidal. Among extracts tested, some showed activities at the concentration of 50 to 100 ㎍/㎖. Phryma leptostachya var. asiatica, Aster ageratoides, Centipeda minima, Cirsium pendulum. Lythrum anceps showed antibacterial activity. Penthorum chinense, Lindernia procumbens, Aster ageratoides, Dianthus superbus var. longicalycinus showed antiblebing activity. Phyma leptostachya var. asiatica, Juncus effusus var. decipiens, Lindernia procumbens, Aster ageratoides, Dianthus superbus var. longicalycinus, Viscum album var. coloratum showed antitumor activity. Juncus effusus var. decipiens, Hypericum ascyron, Juncus papillosus, Inula britannicar var. chinensis, Scirpus wichurae, Hypericum laxum showed antioxidant activity.

Umbilicaria esculenta가 생산하는 Depside계 화합물의 구조 및 Phospholipase A₂ 저해활성

김진우,송경식,장현욱,유승헌,유익동 영남대학교 약품개발연구소 1996 영남대학교 약품개발연구소 연구업적집 Vol.6 No.-

Phospholipase A₂(PLA₂) is lipolytic enzyme that has known to be involved in inflammation. In the course of our screening for antiinflammatory compounds from natural products, a compound having PLA₂ inhibitory activities was isolated from the methanol extract of Umbilicaria esculenta. The compound was identified as lecanoric acid based on various NMR studies including DEPT, HETERO-COSY and HMBC experiments. Lecanoric acid inhibited human rheumatoid synovial PLA₂ activity with IC_(50) of 0.17 mM and also exhibited antitumor activity (ED_(50)=2.7μg/ml) against skin tumor cell line (LOX-IMVI).

Streptomyces sp. 3D3 균주가 생산하는 항고추역병성 항생물질

윤봉식,김창진,이인경,히로유끼 고시노,유익동 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.1

미생물 배양액을 이용하여 고추역병방제를 위한 농업용 항생물질을 탐색하던 중 고추역병균인 Phytophthora capsici에 강한 항균활성을 나타내는 3D3 균주를 선발 하였다. 선발된 균주의 배양적, 형태적, 생리적 특성을 조사한 결과 Streptomyces속에 속하는 것으로 동정하였고 따라서 Streptomyces sp. 3D3으로 명명하였다. 배양액으로 부터 HP-20 column chromatography, EtOAc extraction, silica gel column chromatography 및 HPLC에 의하여 활성물질을 분리 정제하였으며, 발색반응, UV 흡수, ^1H-NMR등 각종 spectrum 분석을 통하여 본 활성물질을 bafilomycin C_1으로 동정하였다. Streptomyces sp. 3D3이 생산한 compound 3D3은 다양한 식물 병원성 곰팡이에 대하여 강한 항균활성을 나타내었다. During the screening for the antifungal compounds against Phytophthora capsici causing phytophthora blight of red pepper, we isolated a strong active compound, bafilomycin C_1, produced by strain 3D3. The producing organism was identified as Streptomyces sp. based on taxonomic studies. The antifungal compound was purified from culture broth by HP-20 column chromatography, ethylacetate extraction, silica gel column chromatography and HPLC, and was identified as bafilomycin C_1 by color reaction, UV and ^1H-NMR spectral data analysis. Bafilomycin C_1 showed strong antifungal activity against various phytopathogenic fungi.

Polyozellus multiplex가 생산하는 지질과산화 저해물질

황지숙,송경식,김양섭,석순자,이태호,유익동 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.5

담자균류로부터 지질과산화 저해물질을 탐색하던 중 강원도 오대산 지역에 자생하는 버섯의 한 종류로 부터, Sparague-Dawley계 rat의 간 mirosome 분획의 지질과산화를 강하게 억제하는 물질이 있음을 발견하였다. 이 버섯의 미생물학적 특성을 관찰한 결과 민주름버섯목, 굴뚝버섯과에 속하는 까치버섯(Polyzellus multiplex)으로 동정하였으며 본 버섯의 자실체 methanol 추출물을 benzene, ethylacetate로 순차적으로 추출하고 각종 chromatography를 이용하여 활성물질을 정제한 결과, ethylacetate층으로 부터 황산화활성을 나타내는 PM1을 benzene층으로 부터 지방산 계열의 PM2와 PM3 화합물을 순수하게 분리 정제하였다. PM1은 정색반응 및 각종 기기분석 결과 thelephoric acid계의 화합물로 동정되었으며 PM2 및 PM3는 각각 linoleic acid와 oleic acid로 동정되었다. 지질과산화 활성물질로 분리정제한 PM1 화합물의 IC_50 값은 1.96 ppm이었으며 LD_50 값은 500 mg/Kg 이상이었다. In the course of screening lipid peroxidation inhibitor from basidiomycetes, a mushroom, which was collected at O-Dae mountain in Kangweon-Do, was found to have active compound. The mushroom was identified as Polyzellus multiplex, which belongs to Aphylloporalles Thelephoraceae, on the basis of macroscopic and microscopic characteristics of the fruiting body. The methanol extract of fruiting body was extracted with benzene and ethylacetate, sequentially. By using various kinds of chromatographies, PM1, and PM2 and PM3, were purified from the ethylacetate extract and the benzene extract, respectively. Color reaction and analyses of IR, UV, and NMR spectra indicated that PM1 was a derivative of thelephoric acid, and PM2 and PM3 were linoleic acid and oleic acid, respectively. IC_50 of PM1 for inhibition of lipid peroxidation was 1.96 ppm and LD_50 was 500 mg/kg.

대두품종(大豆品種)과 선발(選拔) Rhizobium japonicum간의 숙주(宿主) 친화성(親和性)

김창진,이윤,김성훈,유익동,민태익,Kim, Chang-Jin,Rhee, Yoon,Kim, Sung-Hoon,Yoo, Ick-Dong,Mheen, Tae-Ick 한국토양비료학회 1986 한국토양비료학회지 Vol.19 No.4

우수(優秀) 대두접종제(大豆接種劑)로 이용할 균주(菌株)의 선발을 목적으로 국내(國內)에서 선발한 R. japonicum 균주(菌株)들과 현재 대두장려품종으로 보급되고 있는 대두품종들과의 상호(相互) 접종실험(接種實驗)을 통하여 숙주친화성(宿主親和性) 정도를 조사하였다. 얻어진 결과는 다음과 같다. 1. 공시(供試) 근류균주별(根瘤菌株別) 대두품종(大豆品種)과의 친화성(親和性) 정도를 보면 R. japonicum R-214, R-138 균주(菌株)가 근류형성능(根瘤形成能), 질소고정활성(窒素固定活性) 및 총(總) 질소흡수량(窒素吸收量)도 높아 대두품종(大豆品種)과 친화성(親和性) 범위가 비교적 넓었으며 R. japoncum R-224 균주(菌株)는 아주 낮은 것으로 판단되었다. 2. 대두품종별(大豆品種別) 근류균(根瘤菌)과의 친화성(親和性)은 황금(黃金)콩, 장백(長白)콩 등이 높았으며 그 다음이 장엽(長葉)콩, 단엽(短葉)콩, 백운(白雲)콩, 동북태(東北太) 등의 순이었고 방사(放射)콩이 제일 낮았다. 3. 일반적으로 질소고정활성(窒素固定活性)이 우수(優秀)한 균주(菌株)가 열등한 균주(菌株)보다 숙주친화성(宿主親和性) 범위가 넓은 것으로 판단되었다. In order to select suitable strains for utilization as soybean inoculants, the affinity between Rhizobium japonicum strains isolated from the domestic field and the several soybean varieties being distributed as recommended species was investigated by cross-inoculation experiments. The results obtained are as follows: 1. Rhizobium japonicum R-214 and R-138 showed broad affinity range to varies soybean species in terms of the nodule forming ability, the nitrogen fixing activity and the amount of total nitrogen absorption. In contrast, Rhizobium japonicum R-224 showed very low affinity to the same soybean groups. 2. The affinity of Hwang-Keum Kong and Jang-Paeck Kong to the rhizobium strains was the most signigicant, and that of Jang-Yeop Kong, Dong-Buk Tae, Paeck-Woon Kong and Dan-Yeop Kong was also significant. On the other hand, Pang-Sa Kong showed low affinity to the strains. 3. In general, the strains having high nitrogen-fixing activity seemed to possess the broad affinity range to the soybean hosts.

Status and Perspectives of Microbial Herbicide Development

Ick-Dong Yoo 韓國雜草學會 1991 Weed&Turfgrass Science Vol.11 No.5

EFFECT OF DUAL INOCULATION WITH GLOMUS MOSSEAE AND BRADYRHIZOBIUM SP . R938 ON NITROGENASE ACTIVITY OF PEANUT IN POT EXPERIMENT

( Ick Dong Yoo,Hyun Chang Seo,Chang Jin Kim,Shin Duk Kim ) 한국토양비료학회 1990 한국토양비료학회 학술발표회 Vol.1990 No.45

Phosphorylation of the Nucleocapsid Protein of Bovine Coronavirus Expressed with a Recombinant Baculovirus Vector

YOO, DONG WAN,COX, GRAHAM J.,YOO, ICK DONG 한국미생물 · 생명공학회 1992 Journal of microbiology and biotechnology Vol.2 No.2

Post-translational modifications of the nucleocapsid protein of bovine coronavirus (Quebec strain) were investigated. Coronavirions were radiolabelled in vivo with inorganic [^32P]orthophosphate and analysed by SDS-PAGE, followed by autoradiography. A single polypeptide with a migration rate of 55 KDa was identified by metabolic phosphate labelling, demonstrating that the nucleocapsid protein of bovine coronavirus was a phosphoprotein. A gene encoding the nucleocapsid protein was inserted immediately downstream from the polyhedrin promoter of Autographa califomica nuclear polyhedrosis baculovirus. Spodoptera frugiperda cells infected with this recombinant baculovirus synthesized a 55 KDa polypeptide, as demonstrated by immunoprecipitation with anti-nucleocapsid monoclonal antibody. The recombinant nucleocapsid protein synthesized in Spodoptera cells could also be labelled by [^32P]orthophosphate. Phosphoamino acid analysis showed that both satin and threonine residues were phosphorylated in authentic, as well as in recombinant nucleocapsid proteins, with a relative phosphorylation ratio of 7:3. Our studies demonstrated that the nudeocapsid protein of bovine coronavirus was a serine and threoninephosphorylated protein and that Spodoptera insect cells were able to properly phosphorylate the relevant foreign proteins.