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낙우송(Metasequoia glyptostroboides)으로부터 분리한 flavonoid의 금속단백분해효소-9 발현 억제 활성
양재영,이호재,고영희,권병목,전효곤,Yang Jae-Young,Lee Ho-Jae,Kho Yung-Hee,Kwon Byoung-Mok,Chun Hyo Kon 한국생명과학회 2005 생명과학회지 Vol.15 No.2
기질 금속단백분해효소(MMP)는 기저막이나 간질성 조직 등에 있는 세포외기질 성분을 분해하여 상처 치유, 태아의 발생, 종양세포의 침윤과 전이 등을 포함하여 조직이 재구성되는 과정에서 생리학적 및 병리학적인 과정 양쪽 모두에 매우 중요한 역할을 한다. 본 연구에서는 MMP-9 유전자의 발현을 억제하는 물질을 천연물에서 탐색하였고, 탐색된 시료 중에서 높은 역가를 가진 낙우송과의 Metasequoia glyptostroboides가 선택되었다. 여러 가지 flavonoid 성분을 가지고 있는 Metasequoia glyptostroboides를 이용하여 4개의 biflavonoid와 2개의 monoflavonoid 구조의 물질을 분리하였고, 이 flavonoid들은 sciadopitysin, isoginkgetin, bilobetin, 2, 3-dihydrohino-kiflavone, luteolin, apigenin으로 정제하여 구조를 밝히고 zymography, WST-1을 이용한 세포독성시험, northern blot 등의 실험을 통하여 각 화합물의 구조적인 특성과 함께 MMP-9 유전자 발현을 억제하는 효과가 있는 것을 확인하였다. Matrix metalloproteinases (MMPs) are a family of structurally and functionally related zinc-dependent enzymes responsible for proteolytic degradation of extracellular matrix components such as base membrane or interstitial stroma. MMPs play an important role in a variety of physiological and pathological tissue remodeling processes, including wound healing, embryo implantation, tumor invasion and metastasis. Since MMP-9 (gelatinase B) has unique ability to cleave type IV collagen, gene expression of MMP-9 has been focused on as a pharmacological target. Flavonoids are a class of compounds that are widely spread in plants. In the coures of screening for the suppressors of MMP-9 gene expression from natural products, Metasequoia glyptostroboides was selected. Six flavonoids, sciadopitysin, isoginkgetin, bilobetin, 2,3-dihydrohinokiflavone, luteolin and apigenin were purified as suppressors of MMP-9 gene expression from M. glyptostroboides. The suppressing activity of the isolated flavinoids on the MMP-9 gene expression was measured by gelatin zymography and Nothern blot analysis.
무릇에서 분리한 nortriterpenoid glycoside의 암세포에 대한 세포독성 및 함량 분석
이상명,전효곤,이충환,이호재,강진정,맹학영,고영희,Lee, Sang-Myung,Chun, Hyo-Kon,Lee, Choong-Hwan,Lee, Ho-Jae,Kang, Shin-Jyung,Maeng, Hack-Young,Kho, Yung-Hee 한국생약학회 2001 생약학회지 Vol.32 No.3
Scillascilloside E-3 (1) and E-1 (2) were isolated from the bulbs of Scilla scilloides. The cytotoxic activities of these compounds were tested against murine (B16/F-10, 3LL) and human cancer cell lines (MCF7, PC-3, HT29, LOX-IMVI, A549 and HT1080). These compounds exhibited a significant cytotoxic activities against all tested cancer cells. Futhermore, the contents of 1 and 2 in S. scilloides are 43.2 and 27.9 mg/kg, respectively.
오희목,구영환,안극현,장감용,고영희,권기석,윤병대 한국산업미생물학회 1995 한국미생물·생명공학회지 Vol.23 No.6
고농도의 phenol과 formaldehyde를 주성분으로 하는 phenol계 산업폐수를 단계적으로 희석하면서, 혼합균주인 GE2와 세라믹담체를 적용하여 80일간에 걸쳐 생물학적으로 처리하였다. 실험구는 제지폐수 슬러지로 30일간 회분식 운전에 의해 적응시킨 대조구, 이것에 GE2를 첨가한 GE2 처리구 그리고 세라믹담체를 적용한 세라믹처리구로 구분하였다. 원폐수의 80배 희석으로 유입수의 COD_Mn이 1,140mg/l 인 경우 각 실험구의 유출수 COD_Mn은 22∼35mg/l로 큰 차이가 없었으나, 20배 희석의 경우 유입수의 COD_Mn이 4,800mg/l일 때, 대조구, GE2 처리구 그리고 세라믹처리구의 유출수 COD_Mn은 179, 128 그리고 94mg/l로서 세라믹처리구의 처리효율이 98.0%로 가장 높았다. 이때 유출수의 phenol 농도는 각기 10.71, 7.93 그리고 5.60mg/l으로 GE2 및 세라믹담체의 적용에 의하여 처리효율이 향상되었다. 한편, 희석배율이 낮아짐에 따라 COD_Mn과 phenol 처리효율에는 큰 변화가 없으나, 유출수의 COD_Mn과 phenol농도가 높아졌다. 따라서, 원폐수를 40배 희석하여 GE2와 세라믹담체를 첨가하여 처리할 경우 유출수의 COD_Mn과 phenol 농도가 32와 1.02mg/l로 수계의 방류수 수질기준에 충분히 적합한 생물학적 처리로 사료된다. A phenolic resin industrial wastewater containing about 41,000 mg/l of phenol and 2,800 mg/l of formaldehyde was biologically treated by a mixed culture GE2 immobilized on ceramic beads. This study was carried out with three experimental groups : Control-only added the sludge of papermill wastewater ; GE2 treatment-added GE2 to Control ; Ceramic treatment-applied ceramic carrier to GE2 treatment. When the original wastewater was diluted 80 times with aerated tap-water, influent COD_Mn was 1,140 mg/l and that of the effluent was in the range of 22∼35 mg/l, which was not much different among the experimental groups. However, at 20-times dilution, influent COD_Mn was 4,800 mg/l and the effluent COD_Mn of control, GE2 treatment and Ceramic treatment was 179, 128 and 94 mg/l, respectively. COD_Mn removal efficiency by Ceramic treatment was the highest, at 98.0%. At this time, the effluent phenol concentration of Control, GE2 treatment and Ceramic treatment was 10.71, 7.93 and 5.60, respectively. As the dilution times decreased, the removal efficiency of COD_Mn and phenol did not change much, but COD_Mn and phenol concentration of the effluent increased. Consequently, it is likely that the phenolic industrial wastewater containing phenol and formaldehyde can be biologically treated using a GE2 and ceramic carrier and that at 40-times dilution, the effluent completely meets the effluent standards for industrial wastewater treatment plant.
Rhizoctonia solani가 생산하는 Trehalase의 정제 및 특성
오태광,서영배,고영희 한국산업미생물학회 1992 한국미생물·생명공학회지 Vol.20 No.1
벼 문고병원균인 Rhizoctonia solani의 균사체에서 비조절 trehalase을 초음파처리, gel filtration 및 Fast protein liquid chromatography을 통해서 분리하여 단일 단백질임을 확인하였다. 분리된 trehalase는 분자량이 54000, pI가 5.1인 단백질로 최대 역가가 pH 5.4, 45℃에서 나타났다. Trehalase에 대한 Km치는 3.llmM, Vmax는 105.3μmolmin^1×㎎^1로 나타났고 벼 문고병 농용항생제인 validamycin은 trehalase에 대해서 non-competitive inhibition을 하였다. Nonregulatory trehalase has been purified from mycelia of Rhizoctonia solani, a pathogen of rice sheath blight. Purification procedures involved sonification, gel filtration and fast protein liquid chromatography. Purity was confirmed by isoelectric focusing with silver staining. The purified trehalase was estimated to have a molecular weight of 54,000 and pI point of 5.1. Maximal activity was observed at pH 5.4 and temperature 45℃. The purified trehalase exhibited on apparent Km for trehalose of 3.11mM and a Vmax of 105.3 μmol min^-1×㎎^-1. Validamycin, a commercial antibiotics of rice sheath blight, was a non-competitive inhibitor of Rhizoctonia solani trehalase.
Streptomyces sp. 2074이 생산하는 Tyrosinase-Inhibiting Isoflavonoids
이충환,전효곤,서영배,고영희 한국산업미생물학회 1993 한국미생물·생명공학회지 Vol.21 No.2
Streptomyces sp. 20747의 배양 상등액에서 tyrosinase를 저해하는 세개의 isoflavonoid 물질을 분리하였다. 이들의 구조는 UV, EI-MS, ^1H-NMR, ^(13)C-NMR 분석결과 daidzein, daidzein 7-rhamnoside 그리고 genistein 7-rhamnoside로 결정되었다. 이 물질들은 mushroom tyrosinase를 기질과 경쟁적으로 저해하였으며, 각각 14, 19, 16 ㎍/ ㎖의 IC_(50) 값을 나타내었고, Streptomyces bikiniensis의 melanin 생성은 억제하지 않았다. Soybean meal 뿐만 아니고 bacto-peptone도 tyrosinase를 저해하는 isoflavonoid 생산에 효과적이라는 사실은 soybean meal이 Streptomyces sp. 20747이 생산하는 isoflavonoids의 기원물질이 아니라는 것을 보여준다. Three isoflavonoids having tyrosinase-inhibiting activity were isolated from the culture filtrate of Streptomyces sp. 20747. Their structures were determined by UV, EI-MS, ^1H-NMR, ^(13)CNMR to be daidzein, daidzein 7-rhamnoside, and genistein 7-rhamnoside, which were competitive with substrate and had IC_50 value of 14,19, and 16 ㎍/㎖, respectively to mushroom tyrosinase and did not inhibit melanin production of Streptomyces bikiniensis. Soybean meal as well as peptone were found to be a good nitrogen source for tyrosinase-inhibiting isoflavonoids production, susgesting that soybean meal is not the origin of tyrosinase inhibiting isoflavonoids formation in Streptomyces sp. 20747 strain.