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Purification and Characterization of Lipoxygenase Inhibitor Produced by Penicillium sp.
황지숙(Ji-Sook Hwang),정영기(Yong-Kee Jeong),이태호(Tae-Ho Lee) 한국식품영양과학회 1994 한국식품영양과학회지 Vol.22 No.6
토양으로 부터 분리한 lipoxygenase inhibitor 생산균주 Penicillium sp.를 배양하여, 배양액으로 부터 저해물질을 분리, 정제하였다. 배양액을 여과한 후 원심상등액을 농축하고, ethanol 침전에 의해 불순물을 제거하였다. Ethanol은 감압농축기로 제거한 후 Dowex 50W, Sephadex G-25, silica gel column 및 HPLC 등으로 single peak를 나타낼때까지 정제하였다. 정제된 분 저해제는 산성 영역에서 비교적 안정하였으며, 열 안정성이 특히 우수하여 100℃에서 2시간의 가열에 활성이 그대로 유지되었다. 분자량은 약 270으로 측정되었으며, 220℃~230℃ 부근에서 용융되지 않고 decomposition 되는 성질을 나타내었다. 저해제는 lipoxygenase와 complex를 형성하여 효소를 효율적으로 저해하였다. A strain of Penicillium sp. extracellularly produced an inhibitory substance for lipoxygenase. These purification procedures were followed: ethanol treatment, chromatographies on Dowex 50W, Sephadex G-25, silica gel column and HPLC. The inhibitor was stable in pH range from 3.0 to 5.0 at 25℃, and a treatment at 100°C for 2 hours didn‘t diminish its original activity. The purified inhibitor was charred at a temperature near 220℃~230℃ and decomposed. Molecular weight of the inhibitor was estimated to be approximately 270 by Sephadex G-25 column chromatography. The inhibitor rapidly formed EI complex with lipoxygenase and inhibited enzyme activity.
Polyozellus multiplex가 생산하는 지질괴산화 저해물질
황지숙,송경식,김양섭,석순자,이태호,유익동,Hwang, Ji-Sook,Song, Kyung-Sik,Kim, Yang-Sup,Seok, Soon-Ja,Lee, Tae-Ho,Yoo, Ick-Dong 한국미생물 · 생명공학회 1996 한국미생물·생명공학회지 Vol.24 No.5
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.
미세먼지 유도 기도염증에 대한 배암차즈기 추출물의 호흡기 보호 효과
송형우,지건영,김복규,양원경,한창균,신한재,박양춘,황지숙,강형식,김승형,Song, Hyeongwoo,Ji, Kon Young,Kim, Bok Kyu,Yang, Won Kyung,Han, Chang Kyun,Shin, Han Jae,Park, Yang Chun,Hwang, Ji Sook,Kang, Hyung Sik,Kim, Seung Hyung 한국약용작물학회 2017 한국약용작물학회지 Vol.25 No.5
Background: Small particles increase airway inflammation upon reaching the alveoli. Here, we investigated the protective or therapeutic effects of Salvia plebeia R. Br. (SP_R) extracts on airway inflammation. Methods and Results: To investigate the anti-inflammatory activity of SP_R extracts, we measured their inhibitory effect on the production of reactive oxygen species (ROS) expression of inflammatory mediators, and immune cell infiltration in MH-S alveolar macrophage cells and in the ambient particulate matter (APM)-exposed airway inflammation mice model. The SP_R extracts inhibited the production of ROS and expression of IL-4, IL-10, IL-15, and IL-17A mRNA in APM-stimulated MH-S cells. Oral administration of SP_R extracts suppressed APM-induced inflammatory symptoms, such as high alveolar wall thickness, excess collagen fibers, decreased mRNA expression of chemokines (Ccr9, Ccl5, Ccr3), inflammatory cytokines (IL-15, TNF-${\alpha}$), and IL-4 Th2 cytokine in the lung. The SP_R extracts also inhibited ROS production, granulocyte ($CD11b^+Gr-1^+$) infiltration, IL-17A, TNF-${\alpha}$, macrophage inflammatory protein (Mip-2), and chemokine (C-X-C motif) ligand 1 (Cxcl-1) production in the airway. The specific compounds in the SR-R extracts that mediate the anti-inflammatory effects were identified. Conclusions: In this study, SP_R extracts effectively inhibited airway inflammatory responses, such as ROS production and granulocyte infiltration into the airway, by regulating the expression of chemokines and inflammatory cytokines.
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.