당느릅나무로부터 Sesquiterpene o - Naphthoquinone 류 화합물 , Mansonone E , F 및 H 의 분리와 구조 결정

김종평(Jong Pyung Kim),김원곤(Won Gon Kim),박종희(Jong Hee Park),정진(Jin Jung),유익동(Ick Dong Yoo),(Hiroyuki Koshino) 한국응용생명화학회 1996 Applied Biological Chemistry (Appl Biol Chem) Vol.39 No.1

Three sesquiterpene ortho-naphthoquinones were isolated from the methanolic extract of root bark of Ulmus davidiana Planch whose stem and root bark have been used as an oriental medicine for the treatment of edema, mastitis, gastric cancer and inflammation. The structures of these compounds were established on the basis of spectral data obtained from UV-vis, IR, HR-EIMS and NMR spectrometry, including the pulse field gradient (PFG)-HMQC and HMBC techniques. Their structures were determined as 2,3-dihydro-3,6,9-trimethylnaphtho(1,8-b,c)pyran-7,8-dione, 3,6,9-trimethylnaphtho(1,8-b,c)pyran-7,8-dione and 2,3-dihydro-4-hedroxy-3,6, 9-trimethylnaphtho(1,8-b,c)pyran-7,8-dione, which were identified as mansonones E. F and H, respectively. These compounds have originally been isolated from Mansonia altissima Chev, but have never been isolated from Ulmus davidiana Planch. Especially, mansonone H was isolated for the first time from Ulmaceae. The mismatched carbon chemical shifts of mansonones E and F in the reported literature were corrected by the aid of the PFG-HMBC spectral data.

식물의 냉해에 대한 생체방어기구로서 항산소성 효소의 유도 : (II) $Mn^{+2}$이온에 의한 세포내 SOD의 활성화와 벼 유묘의 내냉성 향상

한창균,김종평,정진,Hahn, Chang-Kyun,Kim, Jong-Pyung,Jung, Jin 한국응용생명화학회 1991 Applied Biological Chemistry (Appl Biol Chem) Vol.34 No.2

벼 유묘에 의 한 $Mn^{+2}$(Mn-SOD의 cofactor)의 흡수는 유묘조직중 SOD 활성을 증가시킴과 아울러 유묘의 냉해저항성을 현저히 향상시키는 결과를 보였으며, SOD 활성 증가정도와 냉해 저 항성 향상정도간에는 정의 상관관계가 있었다. 이에 반하여, Fe-SOD와 Cu/Zn-SOD의 cofactor들인 $Fe^{+3},\;Cu^{+2},$ 및 $Zn^{+2}$의 흡수는 조직내 SOD활성이나 식물의 냉해저항성에 어떤 유의성 있는 영향도 미치지 않았다. 이러한 결과들이 시사하는 바는 아마도 superoxide에 의해 유도되고 $Mn^{+2}$의 존재에 의해 활성화된 Mn-SOD가 (최소한 벼의 경우에는) 저온 스트레스에 대항하는 생체방어 시스템의 중요한 子성분일 것이라는 점이다. 어느정도의 냉해억제효과가 있다고 인정된 Abscisic acid의 처리도 벼 유묘조직의 SOD 활성을 증가시켰다. 이 관찰결과도 식물의 냉해 유발상황 하에서 세포내 SOD가 담당하는 중요한 생체방어 역할을 부각시키는 또 하나의 정보를 제공한 것이다. The uptake of $Mn^{+2}$, a metal cofactor Mn-SOD, by rice seedings resulted in not only a substantial increase in SOD activity in leaf tissues of the plants, but also a significant enhancement of their cold tolerance : the relative extent of the cold tolerance appeared to accord with relative level of the SOD activity. In contrast, $Fe^{+3},\;Cu^{+2}$ and $Zn^{+2}$, which are the cofactors of Fe-SOD and Cu/Zn-SOD, were found to be ineffective for increasing the SOD activity as well as for improving the chilling-resistant capacity of the plants. The results suggest that Mn-SOD, which is most likely induced by its substrate(superoxide) and activated by the presence of $Mn^{+2}$a at high level, is the enzyme acting as an active component of the defense system against low temperature stress in rice plants. In addition, the application of abscisic acid which has been know to protect to some extent certain plants from chilling injury brought about an increase in SOD activity in rice tissues, providing another affirmative information for the crucial role of SOD under the circumstance of cold stress in plants.

우리나라 벼 근권으로부터 분리한 Azospirillum 균주의 미생물학적 특성

김원곤,서현창,김종평,김창진,이계호,유익동,Kim, Won-Gon,Seo, Hyun-Chang,Kim, Jong-Pyung,Kim, Chang-Jin,Lee, Ke-Ho,Yoo, Ick-Dong 한국미생물학회 1994 미생물학회지 Vol.32 No.2

경기도, 충청남도 일원의 벼 근권에서 400~900nmol $C_2H_4/hr/vial$의 아세틸린 환원 활성을 갖는 질소고정균인 Azospirillum 15균주를 분리하였다. 이 분리균주들은 $1.0{\times}3.0{\mu}m$크기를 갖는 vibrioid 형태이었으며 액체배양시 monopolar single 편모를 가졌다. 이 분리균주들은 생리적, 형태적 특성에 따라 두 그룹으로 분류되었는데, 그룹 I 균주들은 탄소원으로 glucose를 잘 이용하였고 biotin 요구성인 반면, 그룹 II 균주들은 glucose를 전혀 이용하지 못하였고 biotin 비요구성이었다. 또한 semisolid 무질소 배지에서 48시간 배양할 때 그룹 I 균주들은 원래의 vibrioid 형태에서 좀 더 길어지면서 S자 모양으로 변하는 pleomorphic 특성을 보인 반면, 그룹 II 균주들은 원래의 모양과 운동성을 유지하였다. 이상의 결과로부터 그룹 I 균주들은 Azospirillum lipoferum으로, 그룹 II 균주들은 Azospirillum brasilense으로 동정하였다. Fifteen strains of the nitrogen fixer Axospirillum were isolated from the rhizosphere of rice collected from Kyonggi-do and Chungcheongnam in Korea. They had strong acetylene-reducing activity of 400 of 900 nmol $C_2H_4$ per hour vial had a similar morphology in succinate-malate medium: vibrioid cells having a diameter of 1.0 ${\mu}m$ and a monopolar single flagellum in liquid media. According to their physiological and morphological characteristics, they were divided into two distinct groups, group I and group II. Group I strain were, unlike group II, distinguished by their ability to use glucose as a sole carbon source in nitrogen-free medium, requirement for biotin, and formation of wider, longer, and S-shaped cells in semisolid nitrogen-free malate medium. On the basis of their characteristics, strains belonging to group I were identified as Azospirillum lipoferum, while those belonging to group II were identified as Azospirillum brasilense.

유칼리나무의 수피로부터 분리한 항산화활성 물질

이인경,윤봉식,김종평,정성현,심규섭,유익동,Lee, In-Kyoung,Yun, Bong-Sik,Kim, Jong-Pyung,Chung, Sung-Hyun,Shim, Gyu-Seop,Yoo, Ick-Dong 한국생약학회 1998 생약학회지 Vol.29 No.3

Seven antioxidative compounds were isolated from chloroform and ethyl acetate extracts of the stem bark of Eucalyptus globulus (Myrtaceae). They were identified as rhamnazin (1), rhamnetin (2), naringenin (3), eriodictyol (4), quercetin (5), taxifolin (6) and dihydrokaempferol-3-rhamnoside (7) on the basis of various spectroscopic analyses. These compounds inhibited lipid peroxidation with $IC_{50}$ values of 0.08-30 ${\mu}g/ml$.

Streptomyces sp . ME - 13 균주가 생산하는 anisomycin 계 AB3217 화합물의 제초활성

김원곤(Won Gon Kim),김종평(Jong Pyung Kim),박동진(Dong Jin Park),김창진(Chang Jin Kim),곽상수(Sang Soo Kwak),유익동(Ick Dong Yoo) 한국응용생명화학회 1996 Applied Biological Chemistry (Appl Biol Chem) Vol.39 No.2

During the screening of herbicidal substances from microbial secondary metabolites using photoautotrophic cells, a strain of ME-13 with strong herbicidal activity was isolated from soil. Based on the taxonomic studies, the strain was identified as Streptomyces. Two active compounds were purified from the culture broth through the column chromatographies using active charcoal, silica gel, MCI gel, and ODS HPLC. The compounds were identified as AB3217-A and B, respectively, related to anisomycin by spectroscopic method. AB3217-A and B completely suppressed the germination of radish and barnyard grass at 25 ppm. In comparison to anisomycin, they showed the 6 times higher inhibitory activities against the growth of shoot and root of radish and barnyard grass with EC50 of around 6 ppm.

능이버섯(Sarcodon aspratus)으로부터 분리한 diketopiperazine계 화합물의 화학구조 및 항산화활성

김진우 ( Jin Woo Kim ),문병식 ( Byung Sik Moon ),박영민 ( Young Min Park ),유남희 ( Nam Hee Yoo ),유인자 ( In Ja Ryoo ),유익동 ( Ick Dong Yoo ),김종평 ( Jong Pyung Kim ),( Nguyen Thi Chinh ) 한국응용생명화학회 2005 Applied Biological Chemistry (Appl Biol Chem) Vol.48 No.1

천연물 유래 항산화 기능성 화장품 신소재 개발

유익동 ( Ick Dong Yoo ),김종평 ( Jong Pyung Kim ),김원곤 ( Won Gon Kim ),윤봉식 ( Bong Sik Yun ),유인자 ( In Ja Ryoo ) 대한화장품학회 2005 대한화장품학회지 Vol.31 No.4

미생물, 버섯류, 약용식물 등 천연물 대사산물로부터 미백, 주름개선용 화장품 신소재로 활용 가능한 항산화 신소재를 탐색하였다. 그 결과 Eupenicillium shearii 균주 배양액으로부터 4개의 phenol계 항산화 활성물질을 순수하게 분리하고 화학구조를 결정하여 melanocin A-D로 명명하였다. Melanocin A~D 화합물의 DPPH 라디칼 소거활성은 EC₅₀ 21~94 μM, superoxide 소거활성은 EC₅₀ 7~84 μM로 arbutin 및 BHA보다 우수한 활성을 나타냈으며, 주름개선 효과도 매우 우수하였다. 기와층버섯(Inonotus xeranticus) 자실체 추출물로부터 강력한 항산화 활성물질인 히스피딘계 신물질을 얻어 inoscavin A로 명명하였다. Inoscavin A 화합물의 항산화 활성을 측정한 결과, superoxide radical 소거활성은 EC₅₀ 0.03 μg/mL, rat의 간 microsome의 지질과산화 억제활성 EC₅₀은 0.3 μg/mL로서 α-tocopherol의 1.5 μg/mL, BHA의 4.9 μg/mL 보다 우수하였다. Streptomyces nitrosporeus 균주 배양액으로부터 신규 지질과산화 억제 활성물질을 benzastatin A~G를 분리하였다. Benzastatin A~G의 지질과산화 억제활성 EC₅₀는 3~30 μM로 매우 우수하였다. Penicillium sp. 균주로부터 미백활성물질을 탐색하여 cyclopentene계 화합물인 terrein을 얻었다. Terrein은 kojic acid 보다 약 10배 이상 강력한 미백활성을 나타낸 반면, 100 μM 농도에서도 비교적 안전한 화합물로 확인되었으며, 작용기전은 MITF 단백질을 조절함으로 활성을 나타내는 것으로 확인되었다. New antioxidative substances for cosmeceuticals were screened from natural resources such as microbial metabolites, mushrooms, and medicinal plants. Four antioxidants were isolated from the fungal metabolite of Eupenicillium shearii and their structures were determined to be new phenolic compounds. The compounds were designated as melanocins A, B, C, and D. Melanocins A~D exhibited free radical scavenging activity on DPPH and superoxide with EC₅₀ values of 21~94 and 7~84 μM, respectively, which were stronger activity than those of α-tocopherol and BHA. Melanocin A showed anti-wrinkle effects on the UV-irrated hairless mouse skin. A novel hispidin antioxidative compound designated as inoscavin A was isolated from the fruiting body of the mushroom, Inonotus xeranticus. Inoscavin A scavenged superoxide radical with EC₅₀ values of 0.03 μg/mL, and inhibited rat liver microsomal lipid peroxidation with EC₅₀ values of 0.3 μg/mL. Benzastatins A~G, the novel antioxidants isolated from the culture of Streptomyces nitrosporeus showed potent lipid peroxidation inhibitory activity with EC₅₀ values of 3~30 μM. A cyclopentene compound with strong hypopigmentary effect was isolated from the fungal metabolite of Penicillium sp. and identifed as terrein. Terrein significantly reduced melanin levels in a melanomacyte cell line, Mel-Ab. It showed 10 times stronger activity than kojic acid, but exhibited no cytotoxic effect even in 100 μM. It was suggested that terrein reduced melanin synthesis by reducing tyrosinase production by MITF down-regulation.

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.

식물세포 미토콘드리아막에서 일어나는 청색광 Photosensitization

김경현,정진,김종평 한국환경농학회 1987 한국환경농학회지 Vol.6 No.2

미토콘드리아는 가시광선의 조사에 의해 그 고유한 생화학적 기능에 저해를 받게 되며 그것은 주로 파장 영역 350∼500㎚의 청색광이 유발하는 광역학적 작용(photodynamic action)의 결과라는 가정을 입증하는 자료를 수집하였다. 미토콘드리아막에 결합되어 있는 전자전달계효소들 중에서 NADH dehydrogenase, succinate dehydrogenase, 및 cytochrome c oxidase의 광저해(photoinhibition)를 조사하였던 바, 모든 효소들이 청색광에 의해 상대적으로 심한 활성상실을 보였다. NADH dehydrogenase의 FMN과 cytochrome c oxidase의 heme group은 산소가 관여하는 photosensitizer(photodynamic sensitizer)임에 반해, succinate dehydrogenase의 FAD는 sensitizer로서의 기능을 보이지 않는 대신 Fe-S center가 산소와 무관한 photosensitizer일 것이라고 해석되었다. heme group에 들어 있는 Fe도 역시 산소와 무관한 광화학반응에 어느 정도 기여하리라고 추정되는 결과도 얻었다. 미토콘드리아 전체로 볼 때 생리적 활성저해에 가장 크게 기여하는 가시광은 산소존재 조건하의 청색광이였고, 그 저해기작에는 active oxygens가 관여되어 있다는 것을 O₂의 분석을 통해 확인하였다. 한편 active oxygens의 생성은 미토콘드리아막의 과산화를 초래하였으며, 역시 청색광/O₂조건에서 그 정도가 가장 심하였다. Plant mitochondria, irradiated with blue-colored sunlight(350∼500㎚) under aerobic and anaerobic conditions, were assayed as to the electron transfer activity of respiratory enzyme system, and compared with those irradiated with orange-colored light(white sunlight minus blue-colored light). The respiratory activity of mitochondria was most seriousely inhibited by illumination with blue-colored light under aerobic condition. Deaeration of mitochondrial suspension resulted in substantial decrease of the photoinhibition by blue-colored light. Meanwhile, orange-colored light demonstrated much less effectiveness-almost ineffectiveness-in causing the inhibition of mitochondrial respiration system. The results of enzymatic assay revealed a strong possibility that FMN in NDH and heme group at least in cytochrome c oxidase, but not FAD in SDH, are the photodynamic sensitizers in mitochondrial inner membrane. Also worthwhile to note is the significant difference from the others of SDH in its photoinhibitory response to the light quality of visible light; that the inhibition of SDH by irradiation was not affected by atmospheric condition and that orangecolored light gave rise to considerable extents of inhibition to the enzyme. This observation was tentatively interpreted in terms of photosensitized reaction not involving molecular oxygen possibly catalyzed by Fe-S centers in the enzyme. The superoxide production and the membrane peroxidation of mitochondria under various treatments also indicated that there was blue-light photodynamic reaction in mitochondria involving active oxygens.

식물의 냉해에 대한 생체방어기구로서 항산소성 효소의 유도 :(Ⅱ) : Mn+2 이온에 의한 세포내 SOD 의 활성화와 벼 유모의 내냉성 향상 Mn+2 - induced SOD activation and enhancement of cold tolerance in rice seedlings

정진,김종평,한창균 한국농화학회 1991 Applied Biological Chemistry (Appl Biol Chem) Vol.34 No.2

The uptake of Mn^(+2), a metal cofactor Mn-SOD, by rice seedings resulted in not only a substantial increase in SOD activity in leaf tissues of the plants, but also a significant enhancement of their cold tolerance : the relative extent of the cold tolerance appeared to accord with relative level of the SOD activity. In contrast, Fe^(+3). Cu^(+2) and Zn^(+2). which are the cofactors of Fe-SOD and Cu/Zn-SOD, were found to be ineffective for increasing the SOD activity as well as for improving the chilling-resistant capacity of the plants. The results suggest that Mn-SOD, which is most likely induced by its substrate(superoxide) and activated by the presence of Mn^(+2) at high level, is the enzyme acting as an active component of the defense system against low temperature stress in rice plants. In addition, the application of abscisic acid which has been know to protect to some extent certain plants from chilling injury brought about an increase in SOD activity in rice tissues, providing another affirmative information for the crucial role of SOD under the circumstance of cold stress in plants.