• Journal of Applied Biological Chemistry
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• 제65권1호
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• pp.33-41
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• 2022

Esculetin (also known as 6, 7-dihydroxycoumarin) a type of coumarin, has been exhibited anti-inflammatory and anti-aging effects. Biorenovation is the microbe-mediated enhancement of biological efficacies and structurally diversified compounds relative to their substrate compounds. The production of different kinds of esculetin derivatives using Bacillus sp. JD3-7 and their effects on lipopolysaccharide (LPS)-triggered inflammatory response in RAW 26.7 cells were assessed. One of the biorenovation products, identified as esculetin 6-O-phosphate (ESP), at concentrations of 1.25, 2.5, and 5 μM inhibited the LPS-stimulated production of inflammation markers of nitric oxide synthase 2 and cyclooxygenase 2 as well as their respective enzymatic reaction products of nitric oxide and prostaglandin E2 in the order of increasing concentrations (1.25, 2.5, and 5 μM). Additionally, ESP treatment suppressed the LPS-stimulated secretion of pro-inflammatory cytokines of interleukin (IL)-1β, IL-6, and tumor necrosis factor- α. Furthermore, these anti-inflammatory effect of ESP was associated with the downregulation of mitogen-activated protein kinase signaling, that is, extracellular signal-regulated kinase, c-Jun NH2-terminal kinase, and p38 mitogen-activated protein kinase signaling pathways. This study would therefore provide interesting insights into the biorenovation-assisted generation of a novel anti-inflammatory compound. ESP may be used to develop treatments for inflammatory disorders.

• Journal of Applied Biological Chemistry
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• 제64권2호
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• pp.113-119
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• 2021

올리브 (Olea europaea L.) 잎은 풍부한 폴리 페놀을 함유하고 있으며, 이는 항산화, 항균, 항 바이러스, 항암 활성을 연관시키고 혈소판 활성화를 억제하는 것으로 보고 되어있다. 올리브 잎은 건강기능성식품 및 기능성 화장품의 원료로 상업적으로 사용되고 있으며, 이러한 건강상의 이점은 올리브 잎의 주요 생리활성 물질인 oleuropein와 관련이 있다. Oleuropein은 항산화, 항노화, 항염증, 항암, 항미생물, 항바이러스, 항죽상동맥혈증, 지질 저하 효과, 혈당 저하 효과, 피부 보호 효과가 보고되어 있다. oleuropein이 올리브 잎의 중요한 화합물 임에도 불구하고 상업용 제품에서 oleuropein 함량을 밝히는 정량적 접근 방법은 아직 없다. 본 연구에서는 UPLC-QTof/MS, PDA, CAD로 개발된 방법을 이용하여 10개의 상업용 올리브 잎 추출물의 성분 및 oleuropein 함량을 분석 하였다. Iridoids (1, 3, 4, 14, and 16-18), coumarin (2), phenylethanoids (5, 9, and 11), flavonoids (6-8, 10, 12, and 13), lignan (15)을 포함한 총 18종의 화합물이 올리브 잎에서 예상되었다. 총 10종의 올리브 잎 추출물 분석은 3종의 제품(A, G, H)에서 제품에서 제시한 oleuropein 함량보다 높게, 5종의 제품(B, E, H, I, J)에서 5-10%의 오차범위로 나타났고 C, D에서 함량 미달로 검출되었다. 본 연구에서 oleuropein의 함량분석이 올리브 잎의 품질 관리에 도움이 될 수 있음을 판단하였다.

• 생명과학회지
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• 제30권4호
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• pp.343-351
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• 2020

마가목은 한방에서 기침, 천식, 기관지 질환 등의 치료에 이용되고 있다. 본 연구에서는 마가목(sorbus commixta) 수피의 chloroform 분획물로부터 3종의 triterpenoid 화합물과 1종의 coumarin 화합물을 분리하였다. 분리된 화합물의 구조는 MS와 1D-, 2D-NMR분석에 의해 확인하였으며, 이들은 lupeol, β-sitosterol, ursolic acid와 scopoletin으로 구조동정되었다. 분리된 화합물 중 scopoletin은 마가목에서 처음으로 분리된 화합물이다. Scopoletin은 식물에 널리 분포하고 있는 물질로써 항염증 활성을 가진 기능성 화합물이다. 분리된 화합물들의 혈관 염증 억제에 대한 효과를 평가하기 위해 in vitro에서 LDL 산화 억제능을 평가한 결과, 분리된 화합물 중 scopoletin (IC₅₀=10.2 μM)이 강한 억제 활성을 나타내었다. TNF-α로 활성화된 인체혈관내피세포(EA.hy926)를 이용한 실험에서 scopoletin은 세포부착인자인 ICAM-1, VCAM-1, E-selectin의 발현을 저해하였고, THP-1 단핵구와 EA.hy926 혈관내피세포 간의 부착력도 약화시켰다. 뿐만 아니라, scopoletin은 TNF-α로 유도된 NF-κB 전사인자의 핵내 이동 및 IκBα의 인산화도 저해하였다. 따라서 마가목 추출물로부터 분리된 scopoletin은 NF-κB 신호전달의 억제를 통해 세포부착인자의 발현을 감소시키고, 단핵구의 혈관내피세포로의 부착을 억제시켜 혈관내 항염증 활성을 나타내었다. 이러한 실험결과, scopoletin은 혈관염증 반응으로부터 유도되는 죽상동맥경화증 치료를 위한 후보소재로서 이용될 가능성이 있다고 사료된다.

• 임산에너지
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• 제17권1호
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• pp.56-70
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• 1998

This study was carried out to investigate the structural characteristics of kraft lignin and the wood degrading characteristics, the productivity of ligninolytic enzymes and the enzymatic degradation of kraft lignin by white-rot fungi. To purify kraft lignin, precipitation of kraft pulping black liquors of pitch pine meal was done by titration with lN $H_{2}SO_{4}$ reaching to pH 2, and isolation of the precipitates done by centrifugation. The isolated precipitates from pitch pine were redissloved in lN NaOH, reprecipitated by titration with lN $H_{2}SO_{4}$, washed with deionized water, and kept ofr analysis after freeze drying. Fractionation of the precipitates in solution by successive extraction with $CH_{2}Cl_{2}$ and MeOH, and the fractionates were named SwKL, SwKL I, SwKL II, and SwKL III for pitch pine kraft lignin. The more molecular weights of kraft lignin increased, the less phenolic hydroxyl groups and the more aliphatic hydroxyl groups. Because as the molecular weights increased, the ratio of etherified guaiayl/syringyl(G/S ratio) and the percentage were increased. The spectra obtained by 13C NMR and FTIR assigned by comparing the chemical shifts of various signals with shifts of signals from autherized ones reported. The optimal growth temperature and pH of white-rot fungi in medium were $28^{\circ}C$ and 4.5-5.0, respectively. Especially, in temperature and pH range, and mycelial growth, the best white-rot fungus selected was Phanerochaete chrysosporium for biodegradation. For the degradation pathways, the ligninolytic fungus jcultivated with stationary culture using medium of 1% kraft lignin as a substrate for 3 weeks at $28^{\circ}C$. The weight loss of pitch pine kraft lignin was 15.8%. The degraded products extracted successively methoanol, 90% dioxane and diethyl ether. The ether solubles were analyzed by HPLC. Kraft lignin degradation was initiated in $\beta$-O-4 bonds of lignin by the laccase from Phanerochaete chrysosporium and the degraded compounds were produced from the cleavage of $C\alpha$-$C\beta$ linkages at the side chains by oxidation process. After $C\alpha$-$C\beta$ cleavage, $C\alpha$-Carbon was oxidized and changed into aldehyde and acidic compounds such as syringic acid, syringic aldehyde and vanilline. And the other compound as quinonemethide, coumarin, was analyzed. The structural characteristics of kraft lignin were composed of guaiacyl group substituted functional OHs, methoxyl, and carbonyl at C-3, -4, and -5 and these groups were combinated with $\alpha$ aryl ether, $\beta$ aryl ether and biphenyl. Kraft lignin degradation pathways by Phanerochaete chrysosporium were initially accomplished cleavage of $C\alpha$-$C\beta$ linkages and $C\alpha$ oxidation at the propyl side chains and finally cleavage of aromatic ring and oxidation of OHs.

• 생명과학회지
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• 제19권2호
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• pp.249-255
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• 2009

Esculetin, a coumarin compound, has been known to inhibit proliferation and induce apoptosis in several types of human cancer cells. However, the molecular mechanisms involved in esculetin-induced apoptosis are still uncharacterized in human leukemia cells. In this study, we have investigated whether esculetin exerts anti-proliferative and apoptotic effects on human leukemia U937 cells. It was found that esculetin could inhibit cell viability in a time-dependent manner, which was associated with the induction of apoptotic cell death such as increased populations of apoptotic- sub G1 phase. Apoptosis of U937 cells by esculetin was associated with an inhibition of Bcl-2/Bax binding activity, formation of tBid, down-regulation of X-linked inhibitor of apoptotic protein (XIAP) expression, and up-regulation of death receptor 4 (DR4) and FasL expression. Esculetin treatment also induced the degradation of ${\beta}$-catenin and DNA fragmentation factor 45/inhibitor of caspase-activated DNase (DFF45/ICAD). Furthermore, a caspase-3 specific inhibitor, z-DEVD-fmk, significantly inhibited sub-G1 phase DNA content, morphological changes and degradation of ${\beta}$-catenin and DEE45/ICAD. These results indicated that a key regulator in esculetin-induced apoptosis was caspase-3 in human leukemia U937 cells.

• Applied Biological Chemistry
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• 제51권2호
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• pp.136-141
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• 2008

곧은 나무이끼의 전초에서 물질을 분리 및 동정하기 위해 곧은 나무이끼의 MeOH 추출물을 $CHCl_3$, EtOAc 및 BuOH로 용매 분획하였다. $CHCl_3$ 분획으로부터 2-chromenone(coumarin)을 분리 하였고 EtOAc 분획에서 3,4-dihydroxy-cinnamic acid (caffeic acid), 3,3',4',5,7-pentahydroxy-2-phenyl-chromen-4-one (quercetin)과 kaemperol-3-O-${\beta}$-D-glucopyranoside(astragalin)을 분리하였다. BuOH 분획으로부터 3-[[3-(3,4-Dihydr oxyphenyi)-1-oxo-2-propenyl]oxy]-1,4,5-trihydroxycyclohexanecarboxylic acid(chlorogenic acid)와 quercetin-3-O-rutinoside(rutin)를 분리하여 IR, UV, Mass, NMR과 이화학적인 방법으로 그 구조를 규명하였다. 곧은 나무이끼에서 이 물질들이 분리된 것은 처음으로 보고되는 것이다.

• 한국잡초학회지
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• 제30권4호
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• pp.371-379
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• 2010

피복작물의 생육 단계에 따른 allelopathy 효과의 차이를 알아보기 위해, 각 피복작물의 영양생장기, 개화기, 결실기에 채취한 시료로 부터 추출한 추출물을 이용해 콩, 무, 유채, 상추의 발아에 미치는 영향을 확인하였다. 콩과 무에서는 각 피복작물 추출물의 처리구에서 발아율에 별다른 차이를 나타내지 않았으나, 유채에서는 크림손클로버의 개화기 20% 추출물, 헤어리베치의 영양생장기, 개화기, 결실기 20% 추출물, 호밀의 영양생장기 20% 추출물들에서 20% 이상의 발아 억제가 나타났으며, 상추는 영양생장기의 크림손클로버와 헤어리베치의 20% 추출물 처리에서 발아세가 낮게 나타났다. 피복작물의 영양생장기 추출물에서 유근 생장 억제율이 가장 높았으며 $GR_{50}$ 값도 낮게 나타났다. 피복작물 추출물의 총 페놀화합물 함량도 영양생장기 때 가장 높았고, 결실기로 생육이 진전되면서 감소되었다. HPLC 분석 결과 크림손클로버 추출물에서 검출 된 p-hydroxybenzoic acid를 제외한 나머지 phenolic acid 함량들은 총 페놀화합물의 함량과 다른 경향을 보였으며, 헤어리베치 추출물에서는 coumarin, 호밀 추출물에서는 BOA와 caffeic acid를 제외한 나머지 물질들이 개화기에서 가장 높은 함량을 나타냈다. 그러나 크림손클로버와 호밀 추출물에서 분석 물질들의 총 함량은 영양생장기 추출물에서 가장 높게 나타났다.

• Journal of Microbiology and Biotechnology
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• 제22권12호
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• pp.1659-1664
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• 2012

TSAHC [4'-(p-toluenesulfonylamido)-4-hydroxychalcone] is a promising antitumorigenic chalcone compound, especially against TM4SF5 (four-transmembrane L6 family member 5)-mediated hepatocarcinoma. We evaluated the potential of TSAHC to inhibit the catalytic activities of nine cytochrome P450 isoforms and of P-glycoprotein (P-gp). The abilities of TSAHC to inhibit phenacetin O-deethylation (CYP1A2), coumarin 6-hydroxylation (CYP2A6), bupropion hydroxylation (CYP2B6), amodiaquine N-deethylation (CYP2C8), diclofenac 4-hydroxylation (CYP2C9), omeprazole 5-hydroxylation (CYP2C19), dextromethorphan O-demethylation (CYP2D6), chlorzoxazone 6-hydroxylation (CYP2E1), and midazolam 1'-hydroxylation (CYP3A) were tested using human liver microsomes. The P-gp inhibitory effect of TSAHC was assessed by [$^3H$]digoxin accumulation in the LLCPK1-MDR1 cell system. TSAHC strongly inhibited CYP2C8, CYP2C9, and CYP2C19 isoform activities with $K_i$ values of 0.81, 0.076, and $3.45{\mu}M$, respectively. It also enhanced digoxin accumulation in a dose-dependent manner in the LLCPK1-MDR1 cells. These findings indicate that TSAHC has the potential to inhibit CYP2C isoforms and P-gp activities in vitro. TSAHC might be used as a nonspecific inhibitor of CYP2C isoforms based on its negligible inhibitory effect on other P450 isoforms such as CYP1A2, CYP2A6, CYP2B6, CYP2D6, CYP2E1, and CYP3A.