• Title/Summary/Keyword: lipoxygenase activity

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Isolation of Lipoxygenase Inhibitor from Indonesian Herb

  • Alfi Khatib;Kim, Young-Chan;Chung, Shin-Kyo
    • Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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    • 2003.04a
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    • pp.111.2-112
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    • 2003
  • A total of 20 extracts derived from different plant family commonly used in Indonesian traditional inflammation medicine were screened for their inhibitory effect on soybean lipoxygenase (SBL) and hyaluronidase (HAse) activity. Three methanol extracts, the bark of Cinnamomum burmanni (CB), the leaves of Piper betel (PB), and fruit of Barringtonia acutangula (BA) were found to have high inhibitory effects, whereas the methanol extract of the leaves of Mimusops elengi (ME) have medium inhibitory effect. The IC50 of CB, PB, BA and ME were found to be 21.7, 16.9, 39.1 and 62.8 g/$m\ell$, respectively. Among the tested extracts, only CB inhibited HAse (IC50 = 27g/$m\ell$). CB was successively fractionated with n-hexane, ethyl acetate, butanol and water. The EtOAc fraction having the strongest activity was fractionated and some compounds were isolated and purified by a preparative HPLC(Develosil ODS-HG-5 column). Coumarin and 2-hydroxy cinnamaldehyde. were identified through the analyses of UV-Vis absorption 1H-NMR, 13C-NMR and FAB+-MS spectra.

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Effects of Sophoraflavanone G, a Prenylated Flavonoid from Sophora Flavescens, on Cyclooxygenase-2 and In Vivo Inflammatory Response

  • Kim, Dong-Wook;Chi, Yeon-Sook;Son, Kun-Ho;Chang, Hyeun-Wook;Kim, Ju-Sun;Kang, Sam-Sik;Kim, Hyun-Pyo
    • Archives of Pharmacal Research
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    • v.25 no.3
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    • pp.329-335
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    • 2002
  • Previously, several prenylated flavonoids having a C-8 lavandulyl moiety were found to inhibit cyclooxygenase-1 (COX-1) as well as 5-lipoxygenase (5-LOX), and sophoraflavanone G was the most potent inhibitor against these eicosanoid generating enzymes among 19 prenylated flavonoids tested. In this investigation, effects of sophoraflavanone G on COX-2 induction from RAW 264.7 cells and in vivo inflammatory response were studied. Sophoraflavanone G inhibited prostaglandin $E_2{\;}(PGE_2)$ production from lipopolysaccharide (LPS)-treated RAW cells by COX-2 down-regulation at 1-50 uM. Other prenylated flavonoids including kuraridin and sanggenon D also down-regulated COX-2 induction at 10-25 uM, while kurarinone and echinoisoflavanone did not. In addition, sophoraflavanone G showed in vivo anti-inflammatory activity against mouse croton oil-induced ear edema and rat carrageenan paw edema via oral (2-250 mg/kg) or topical administration (10-250 ug/ear). Although the potencies of inhibition were far less than that of a reference drug, prednisolone, this compound showed higher anti-inflammatory activity when applied topically, suggesting a potential use for several eicosanoidrelated skin inflammation such as atopic dermatitis.

Changes in Enzymatic Activities during Eoyukjang Fermentation (어육장 발효 시 생성되는 효소의 활성 변화)

  • Ham, Soo-Nam;Kim, Sang-Woo;Lee, Jae Hwan;Chang, Pahn-Shick
    • Korean Journal of Food Science and Technology
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    • v.40 no.3
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    • pp.251-256
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    • 2008
  • Eoyukjang is a traditional sauce-type of Korean food that is similar to a soybean sauce made from fermented soybeans, and it is produced from a fermented mixture of sea foods, meats, and meju (soybean paste). This study examined periodical changes in the enzymatic activities of ${\alpha}$-amylase, esterase, ${\beta}$-glucosidase, protease, lipase, and lipoxygenase within the culture broth and solids of eoyukjang during 1 year of fermentation. The eoyukjang solids had 234-532% higher protein content than the culture broth. The specific activities of ${\alpha}$-amylase, esterase, ${\beta}$-glucosidase, and protease increased in both the culture broth and solids. Particularly, in the culture broth, ${\alpha}$-amylase, esterase, ${\beta}$-glucosidase, and protease activities rapidly increased (3- to 8-fold) until 10 months of fermentation, and then drastically decreased. However, the activities of lipase and lipoxygenase in both the culture broth and solids were less than 0.05 unit/mg of protein, respectively, throughout fermentation; thus, their activity levels were low and changed little over the 12 months. Overall, while the solids had higher protein content than the culture broth, the broth had greater enzyme activity levels during eoyukjang preparation.

Study on the Antioxidative Effects and Amino Acid Contents of the Roots of Cynanchum Wilfori (백하수오(白何首烏)의 항산화활성(抗酸化活性)과 amino acid의 분포(分布)에 관한 실험적(實驗的) 연구(硏究))

  • Han Ki-Sun;Shin Gil-Jo;Lee Won-Chul;Lee Jong-Hyung
    • The Journal of Internal Korean Medicine
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    • v.19 no.2
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    • pp.411-430
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    • 1998
  • The aim of the presents study is to investigate and compare the antioxidative effects and qualities of the cultivating root of Cynanchum Wilfori, which is increasingly used in recent days, with those of the wilding root, which has mainly been used in the past in oriental medicine for a tonic and also for prevention and treatment of various geriatric diseases including aging progress. For comparison of their antioxidative effects, the activities of the total extracts on lipid peroxidation and the activity of aldehyde oxidase(EC 1.2.3.1) as well as xanthine oxidase(EC 1.2.3.2) were investigated in vitro. In addition, their inhibitory effects on the activity of 5-lipoxygenase, which is known to induce inflammation and concerned with free radicals, were also determined in vitro. Furthermore, the amino acid contents of both roots were analyzed in order to compare their qualities. The results are as follows: 1. The wilding root inhibited significantly the activity of 5-lipoxygenase, showing five times more portent than the cultivating root. 2. Both of the wilding root and the cultivating root inhibited aldehyde oxidase activity in a dose-dependant manner. The wilding root was more effective than the other. 3. Both of the wilding root and the cultivating root dose-dependently suppressed lipid peroxidation in rat brain, kidney, and liver. 4. The anti-peroxidative effects of both roots appeared to be most strong in brain and least in liver. In particular, the cultivating root exhibited a significant inhibition on brain lipid peroxidation. 5. The cultivating root contained 15 amino acids including five essential amino acids in contrast with the less contents in the wilding root.

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Anti-Inflammatory Activity of Compounds from the Whole Plant of Patrinia saniculaefolia

  • An, Ren-Bo;Na, Min-Kyun;Min, Byung-Sun;Chang, Hyeun-Wook;Bae, Ki-Hwan
    • Natural Product Sciences
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    • v.17 no.2
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    • pp.90-94
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    • 2011
  • An in vitro bioassay-guide revealed that the methanol (MeOH) extract of the whole plant of Patrinia saniculaefolia (Valerianaceae) showed cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) dual inhibitory activity by assessing their effects on the production of prostaglandin $D_2$ ($PGD_2$) and leukotriene $C_4$ ($LTC_4$) in mouse bone marrow-derived mast cells (BMMCs). Phytochemical study of the MeOH extract of this plant led to the isolation of twelve compounds; ${\beta}$-farnesene (1), squalene (2), nardostachin (3), patridoid I (4), patridoid II (5), patridoid II-A (6), oleanolic acid (7), oleanonic acid (8), 23-hydroxyursolic acid (9), oleanolic acid 3-O-${\alpha}$-L-arabinopyranoside (10), oleanolic acid 3-O-${\beta}$-D-glucopyranoside (11), oleanolic acid 3-O-[${\beta}$-D-xylopyranosyl-(1${\rightarrow}$3)-${\beta}$-D-(6-O-butyl)glucuronopyranoside] (12). Among the compounds, 4 and 5 strongly inhibited both the COX-2-dependent $PGD_2$ generation with $IC_{50}$ values of 8.7 and 13.6 ${\mu}M$, respectively, and the generation of $LTC_4$ in the 5-LOX dependent phase with $IC_{50}$ values of 41.7 and 46.9 ${\mu}M$, respectively, which suggest that the anti-inflammatory activity of P. saniculaefolia might occur in part via the inhibition of both $PGD_2$ and $LTC_4$ generation by 4 and 5.

The Rhizomes of Acorus gramineus and the Constituents Inhibit Allergic Response In vitro and In vivo

  • Lim, Hyun;Lee, Seung-Young;Lee, Kang-Ro;Kim, Yeong-Shik;Kim, Hyun-Pyo
    • Biomolecules & Therapeutics
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    • v.20 no.5
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    • pp.477-481
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    • 2012
  • The rhizomes of Acorus gramineus have frequently been used in traditional medicine mainly for sedation as well as enhancing brain function. In this study, the anti-allergic activity of A. gramineus was investigated. The 70% ethanol extract of the rhizomes of A. gramineus was found to inhibit the allergic response against 5-lipoxygenase (5-LOX)-catalyzed leukotriene (LT) production from rat basophilic leukemia (RBL)-1 cells and ${\beta}$-hexosaminidase release from RBL-2H3 cells with $IC_{50}$'s of 48.9 and > $200{\mu}g/ml$, respectively. Among the 9 major constituents isolated, ${\beta}$-asarone, (2R,3R,4S,5S)-2,4-dimethyl-1,3-bis (2',4',5'-trimethoxyphenyl)tetrahydrofuran (AF) and 2,3-dihydro-4,5,7-trimethoxy-1-ethyl-2-methyl-3-(2,4,5-trimethoxyphenyl)indene (AI) strongly inhibited 5-LOX-catalyzed LT production in A23187-treated RBL-1 cells, AI being the most potent ($IC_{50}=6.7{\mu}M$). Against ${\beta}$-hexosaminidase release by antigen-stimulated RBL-2H3 cells, only AI exhibited strong inhibition ($IC_{50}=7.3{\mu}M$) while ${\beta}$-asarone and AF showed 26.0% and 39.9% inhibition at $50{\mu}M$, respectively. In addition, the ethanol extract of A. gramineus showed significant inhibitory action against the hapten-induced delayed hypersensitivity reaction in mice by oral administration at 200 mg/kg. Therefore, it is suggested that A. gramineus possesses anti-allergic activity and the constituents including ${\beta}$-asarone and AI certainly contribute to the anti-allergic activity of the rhizomes of A. gramineus.

The N-Terminal α-Helix Domain of Pseudomonas aeruginosa Lipoxygenase Is Required for Its Soluble Expression in Escherichia coli but Not for Catalysis

  • Lu, Xinyao;Wang, Guangsheng;Feng, Yue;Liu, Song;Zhou, Xiaoman;Du, Guocheng;Chen, Jian
    • Journal of Microbiology and Biotechnology
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    • v.26 no.10
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    • pp.1701-1707
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    • 2016
  • Lipoxygenase (LOX) is an industrial enzyme with wide applications in food and pharmaceutical industries. The available structure information indicates that eukaryotic LOXs consist of N terminus β-barrel and C terminus catalytic domains. However, the latest crystal structure of Pseudomonas aeruginosa LOX shows it is significantly different from those of eukaryotic LOXs, including the N-terminal helix domain. In this paper, the functions of this N-terminal helix domain in the soluble expression and catalysis of P. aeruginosa LOX were analyzed. Genetic truncation of this helix domain resulted in an insoluble P. aeruginosa LOX mutant. The active C-terminal domain was obtained by dispase digestion of the P. aeruginosa LOX derivative containing the genetically introduced dispase recognition sites. This functional C-terminal domain showed raised substrate affinity but reduced catalytic activity and thermostability. Crystal structure analyses demonstrate that the broken polar contacts connecting the two domains and the exposed hydrophobic substrate binding pocket may contribute to the insoluble expression of the C terminus domain and the changes in the enzyme properties. Our data suggest that the N terminus domain of P. aeruginosa LOX is required for its soluble expression in E. coli, which is different from that of the eukaryotic LOXs. Besides this, this N-terminal domain is not necessary for catalysis but shows positive effects on the enzyme properties. The results presented here provide new and valuable information on the functions of the N terminus helix domain of P. aeruginosa LOX and further improvement of its enzyme properties by molecular modification.

Biosynthesis of (R)-(-)-1-Octen-3-ol in Recombinant Saccharomyces cerevisiae with Lipoxygenase-1 and Hydroperoxide Lyase Genes from Tricholoma matsutake

  • Lee, Nan-Yeong;Choi, Doo-Ho;Kim, Mi-Gyeong;Jeong, Min-Ji;Kwon, Hae-Jun;Kim, Dong-Hyun;Kim, Young-Guk;Luccio, Eric di;Arioka, Manabu;Yoon, Hyeok-Jun;Kim, Jong-Guk
    • Journal of Microbiology and Biotechnology
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    • v.30 no.2
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    • pp.296-305
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    • 2020
  • Tricholoma matsutake is an ectomycorrhizal fungus, related with the host of Pinus densiflora. Most of studies on T. matsutake have focused on mycelial growth, genes and genomics, phylogenetics, symbiosis, and immune activity of this strain. T. matsutake is known for its unique fragrance in Eastern Asia. The most major component of its scent is (R)-(-)-1-octen-3-ol and is biosynthesized from the substrate linoleic acid by the sequential reaction of lipoxygenase and peroxide lyase. Here, we report for the first time the biosynthesis of (R)-(-)-1-octen-3-ol of T. matsutake using the yeast Saccharomyces cerevisiae as a host. In this study, cDNA genes correlated with these reactions were cloned from T. matsutake, and expression studies of theses genes were carried out in the yeast Saccharomyces cerevisiae. The product of these genes expression study was carried out with Western blotting. The biosynthesis of (R)-(-)-1-octen-3-ol of T. matsutake in recombinant Saccharomyces cerevisiae was subsequently identified with GC-MS chromatography analysis. The biosynthesis of (R)-(-)-1-octen-3-ol with S. cerevisiae represents a significant step forward.

Anti-inflammatory Effects of Prescription Extracts Containing Forsythia viridissima L. (연교를 함유한 처방단 추출물들의 항염증 효과)

  • Kim, Mi-Jin;Im, Kyung-Ran;Yoon, Kyung-Sup
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.4
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    • pp.277-285
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    • 2009
  • Forsythia fructus has been shown to have antioxidative, anti-inflammatory, antiviral, antitumor, antibacterial, antipyretic and anti-aging activities. This work was carried out to investigate the anti-inflammatory effects of the Korean traditional medicinal prescriptions containing Forsythia viridissima extract. The prescriptions containing Forsythia fructus were evaluated for antioxidative effects, inhibitory effects on lipoxygenase activity and on LPS-induced NO and $PGE_2$ production. In human irritation test, they did not show any adverse effect. Based on these results, we suggest that the se prescriptions hold great promise for application as an anti-inflammatory agent for trouble skins such as atopic dermatitis and acne.

An Arachidonic Acid Metabolizing Enzyme, 8S-Lipoxygenase, in Mouse Skin Carcinogenesis

  • Kim Eun-Jung
    • Nutritional Sciences
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    • v.9 no.3
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    • pp.212-226
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    • 2006
  • The involvement of arachidonic acid (AA) metabolizing enzyme, lipoxygenase (LOX), in the development of particular tumors in humans has gradually been acknowledged and LOX has emerged as a novel target to prevent or treat human cancers. In the mouse skin carcinogenesis model, which provides an excellent model to study multistage nature of human cancer development, many studies have shown that some of the LOXs are constitutively upregulated in their expression. Moreover, application of LOX inhibitors effectively reduced tumor burdens, which implicates the involvement of LOX in mouse skin tumor development as well. 8S-LOX is a recently cloned LOX, which is specifically expressed in mouse skin after 12-O-tetradecanoyl-phorbol-13-acetate (TPA) treatment but not in normal skin. Unlike other members of the LOX 'family' expressed in mouse skin, this TPA-induced expression of 8S-LOX is prominent only in the skin of the TPA tumor promotion-sensitive strains of mice (SENCAR, CD-1, and NMRI) but not in the promotion-resistant C57BL/6J mice. This is a very unique phenomenon among strains of mice. Constitutive upregulation of 8S-LOX was also found in early stage papillomas and the expression was gradually reduced as the tumors became malignant. Based on these observations, it has been thought that 8S-LOX is involved in TPA-induced tumor promotion as well as in tumor conversion from papillomas to carcinomas. In accordance with this hypothesis, several studies have suggested possible roles of 8S-hydroxyeicosatetraenoic acid (HETE), an AA metabolite of 8S-LOX, in mouse skin tumor development. A clastogenic activity of 8S-HETE was demonstrated in primary keratinocytes and a close correlation between the levels of etheno-DNA adducts and 8S-HETE during skin carcinogenesis was also reported. On the other hand, it has been reported that 8S-LOX protein expression is restricted to a differentiated keratinocyte compartment Moreover, reported findings on the ability of 8S-HETE to cause keratinocyte differentiation appear to be contrary to the procarcinogenic features of the 8S-LOX expression, presenting a question as to the role of 8S-LOX during mouse skin carcinogenesis. In this review, molecular and biological features of 8S-LOX as well as current views on the functional role of 8S-LOX/8S-HETE during mouse skin carcinogenesis are presented.