• Title/Summary/Keyword: 5-lipoxygenase inhibitor

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Purification and Characterization of Lipoxygenase Inhibitor Produced by Penicillium sp. (Penicillum sp. 에 의해 생산되는 Lipoxygenase Inhibitor의 정제 및 성질)

  • 황지숙;이태호;정영기
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.6
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    • pp.833-838
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    • 1993
  • 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 $25^{\circ}C$, and a treatment at 10$0^{\circ}C$ for 2 hours didn't diminish its original activity. The purified inhibitor was charred at temperature near 22$0^{\circ}C$~23$0^{\circ}C$ 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.

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A New Soybean Cultivar "Gaechuck#1" : Black Soybean Cultivar with Lipoxygenase2,3-free, Kunitz Trypsin Inhibitor-free and Green Cotyledon

  • Chung, Jong Il
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.603-606
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    • 2009
  • Lipoxygenase and Kunitz trypsin inhibitor protein of mature soybean [Glycine max (L.) Merr.] seed are main anti-nutritional factors in soybean seed. A new soybean cultivar, "Gaechuck#1" with the traits of black seed coat, green cotyledon, lipoxygenase2,3 and Kunitz trypsin inhibitor protein free was developed. It was selected from the population derived the cross of "Gyeongsang#1" and C242. Plants of "Gaechuck#1" have a determinate growth habit with purple flowers, brown pubescence, black seed coat, black hilum, oval leaflet shape and brown pods at maturity. Seed protein and oil content on dry weight basis have averaged 39.1% and 16.2%, respectively. It has shown resistant reaction to soybean necrosis, soybean mosaic virus, Cercospora leaf spot and blight, black root rot, pod and stem blight, and soybean pod borer. "Gaechuck#1" matured on 5-10 October with a plant height of 50 cm. The 100-seed weight of "Gaechuck#1" was 23.2g. Yield of "Gaechuck#1" was averaged 2.2 ton/ha from 2005 to 2007.

An 5-Lipoxygenase Inhibitor Isolated from the Roots of Cynanchum wilfordi Hemsley (은조롱뿌리의 5-Lipoxygenase활성 억제성분)

  • Lee, Won-Churl;Lee, Dong-Ung
    • Korean Journal of Pharmacognosy
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    • v.28 no.4
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    • pp.247-251
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    • 1997
  • The effects of the extract of the root of Cynanchum wilfordi (Asclepiadaceae), the alkaloid fraction, and the isolated main constituent (gagaminine) on 5-lipoxygenase(5-LOEC 1.13.11.34) in bovine PMNL have been studied. The effect of the crude extracts of wildand cultivated plant was also compared each other. Furthermore, the inhibitory effect of gagaminine on 5-Lo was compared with those of the standard drugs. Gagaminine inhibited 5-Lo activity with an $IC_{50}$ value of $26\;{\mu}M$, this result indicates that gagaminine may be useful for in vivo experiments as 5-LO inhibitor.

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Selection of Lipoxygenase, Kunitz Trypsin inhibitor and 7Sα′-subunit Protein Free Soybean Strain (Lipoxygenase, Kunitz Trypsin inhibitor, 7Sα′-subunit 단백질이 결핍된 콩 계통의 선발)

  • Sung, Mi-Kyung;Kim, Kyung-Roc;Park, Jung-Soo;Han, Eun-Hui;Nam, Jin-Woo;Chung, Jong-Il
    • Journal of agriculture & life science
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    • v.44 no.5
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    • pp.29-33
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    • 2010
  • Soybean (Glycine max (L.) Merr.) seed is the main source of protein and oil for human and animal. The use of soybean protein has been expanded in the food industry due to their excellent nutritional benefits. But, antinutritional and allergenic factors are exist in the raw mature soybean. Lipoxygenase, Kunitz trypsin inhibitor (KTI) protein, and ${\alpha}^{\prime}-subunit$ of 7S globulin are main antinutritional factors in soybean seed. Breeding of a new soybean strain with lacking these components is needed. The objective of this research was to select new soybean line with lipoxygenase-free, KTI-free, and ${\alpha}^{\prime}-subunit$ free (lx1lx1lx2lx2lx3lx3titicgy1cgy1 genotype). Total 434 $F_2$seeds were obtained from the cross of cultivar, "Gaechuck#2" and PI506876. Presence and absence of lipoxygenase, KTI protein, and ${\alpha}^{\prime}-subunit$ of 7S globulin was tested by SDS electrophoresis using a partial seed of each $F_2$seed. Only one $F_2$seed with lacking these three components was selected and was planted to $F_2$plant. Absence of lipoxygenase, KTI, and ${\alpha}^{\prime}-subunit$ protein was confirmed on the $F_3$seeds harvested. Selected line with lx1lx1lx2lx2lx3lx3titicgy1cgy1 genotype might be useful for soybean breeding.

The Increases of Proenkephalin A mRNA Levels and the Secretion of $[Met^{5}]-Enkephalin$ Induced by Long-term Stimulation with Nicotine are Mediated by a Lipoxygenase Pathway in Bovine Adrenal Medullary Chromaffin Cells (소 부신수실 크롬친화성 세포에서 Nicotine의 장기간 자극으로 유발된 Proenkephalin A mRNA의 증가 및 $[Met^{5}]-enkephalin$의 분비 증가가 Lipoxygenase 경로에 의해 매개됨)

  • Suh, Hong-Won;Kim, Yung-Hi
    • The Korean Journal of Pharmacology
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    • v.29 no.2
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    • pp.237-244
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    • 1993
  • The effect of nicotine on the secretion of $[Met^{5}]-enkephalin$ (ME) in addition to proenkephalin A (proENK) mRNA levels and effects of indomethacin, nordihydroguaiaretic acid (NDGA), and captopril on nicotine-induced responses were studied in bovine adrenal medullary chromaffrin (BAMC) cells. Long-term exposure of BAMC cells to nicotine at a concentration of $10{\mu}M$ significantly increased proENK mRNA level and the secretion of ME into the medium. Treatment of BAMC cells with NDGA (a lipoxygenase inhibitor, $10{\mu}M$), indomethacin (a cycloooxygenase inhibitor) or captopril (an angiotensin converting enzyme inhibitor) alone did not affect ME secretion and proENK mRNA levels. The pretreatment of BAMC cells with NDGA inhibited the increased ME secretion and proENK mRNA level induced by nicotine. However, indomethacin and captopril did not affect nicotine-induced responses. Our results indicate that neuronal regulations of ME secretion and proENK mRNA level induced by nicotine in BAMC cells are in part mediated by a lipoxygenase-but not cyclooxygenase-and endogenous renin-angiotensin pathway.

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Screening for Potato Lipoxygenase-1 Inhibitor in Unused MArine Resources by the Polarographic Method (수산 미이용자원 중에 존재하는 효소적 산화 억제제의 검색 1. 감자 Lipoxygenase-1에 의한 효소적 산화에 대한 억제)

  • 조순영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.6
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    • pp.959-963
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    • 1994
  • To detect bioactive compounds present in unused marine resources, the screening for the 5-lipoxygenase inhitors in Asterina pectinifera, Halocynthia roretzi skin, Nototodarus sloani ink, Anthocidaris crassispina skin, SArgassum horneri, Agarum cribrosum, Odonthalia corymbifera and Desmarestia ligulata was carried out. THe ether and acetone extracts of Sargassum horneri had the strongest antioxygnic activityon lipid oxidation by potato lipoxygenase-1 (one of 5-lipoygenase) among the tested marine samples and their $IC_{50}$ were 0.3 and 1.1g/ml, respectively. The ether and acetone extracts of Asterina pectinifera, the acetone extracts of Halocynthia roretizi, and the acetone extracts of Nototodarus sloani ink had strong inhibitory activity and their $IC_{50}$ were 72.5, 65, 13.3 and $13.3\mu\textrm{g}/ml$, respectively. In addition, the $IC_{50}$ of the acetone extracts of Agarum cribrosum and Desmarestia ligulata, and the ether extracts of Desmarestia ligulata were 15.5, 35 and $30.5\mu\textrm{g}/ml$, respectively. The nonpolar solvent (ether, acetone) extracts of tested marine organism had more antioxigenic effect against 5-lipoxygenase than the polar solvent(water) extracts.

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Chemical Modification of 5-Lipoxygenase from the Korean Red Potato

  • Kim, Kyoung-Ja
    • BMB Reports
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    • v.33 no.2
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    • pp.172-178
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    • 2000
  • The lipoxygenase was purified 35 fold to homogeneity from the Korean red potato by an ammonium sulfate precipitation and DEAE-cellulose column chromatography. The simple purification method is useful for the preparation of pure lipoxygenase. The molecular weight of the enzyme was estimated to be 38,000 by SDS-polyacrylamide gel electrophoreses and Sepharose 6B column chromatography. The purified enzyme with 2 M $(NH_4)_2SO_4$ in a potassium phosphate buffer, pH 7.0, was very stable for 5 months at $-20^{\circ}C$. Because the purified lipoxygenase is very stable, it could be useful for the screening of a lipoxygenase inhibitor. The optimal pH and temperature for lipoxygenase purified from the red potato were found to be pH 9.0. and $30^{\circ}C$, respectively. The Km and Vmax values for linoleic acid of the lipoxygenase purified from the red potato were $48\;{\mu}M$ and $0.03\;{\mu}M$ per minute per milligram of protein, respectively. The enzyme was insensitive to the metal chelating agents tested (2 mM KCN, 1 and 10mM EDTA, and 1 mM $NaN_3$), but was inhibited by several divalent cations, such as $Cu^{++}$, $Co^{++}$ and $Ni^{++}$. The essential amino acids that were involved in the catalytic mechanism of the 5-lipoxygenase from the Korean red potato were determined by chemical modification studies. The catalytic activity of lipoxygenase from the red potato was seriously reduced after treatment with a diethylpyrocarbonate (DEPC) modifying histidine residue and Woodward's reagent (WRK) modifying aspartic/glutamic acid. The inactivation reaction of DEPC (WRK) processed in the form of pseudo-first-order kinetics. The double-logarithmic plot of the observed pseudo-first-order rate constant against the modifier concentration yielded a reaction order 2, indicating that two histidine residues (carboxylic acids) were essential for the lipoxygenase activity from the red potato. The linoleic acid protected the enzyme against inactivation by DEPC(WRK), revealing that histidine and carboxylic amino acids residues were present at the substrate binding site of the enzyme molecules.

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Screening for Potato Lipoxygenase-II Inhibitor in Unused Marine Resources by the Polarographic Method- (수산 미이용자원 중에 존재하는 효소적 산화 억제제의 검색 -2. 감자 Lipoxygenase-II에 의한 효소적 산화에 대한 억제-)

  • Cho, Soon-Yeong;You, Byeong-Jin;Chang, Mi-Hwa;Lee, Soo-Jung;Sung, Nak-Ju;Lee, Eung-Ho
    • Applied Biological Chemistry
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    • v.37 no.6
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    • pp.451-455
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    • 1994
  • To detect naturally occurring bioactive compounds from unused marine resources, the screening for the 5-lipoxygenase(potato lipoxygenase-II) inhibitors in Asterina pectinifera, Halocynthia roretzi skin, Nototodarus sloani ink, Anthocidaris crassispina skin, Sargassum horneri, Agarum cribrosum, Odonthalia corymbifera and Desmarestia ligulata was carried out. Water, ether, acetone and methanol fractions extracted from Sargassum horneri had strong inhibitory effect on enzymatic lipid oxidation by potato lipoxygenase-II, and their $IC_{50}$ were 320, 18, 9.5 and $100\;{\mu}g/mL$, respectively. The $IC_{50}$ of ether fraction extracted from Asterina pectinifera and acetone fraction extracted from Nototodarus sloani ink were 29.5 and $34.3\;{\mu}g/mL$, and these extracts showed relatively excellent inhibitory activity. Nonpolar solvent (ether, acetone) extracts of tested marine organisms had more inhibitory effect against 5-lipoxygenase than the polar solvent(water) extracts.

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The Protective Effect of Eupatilin against Hydrogen Peroxide-Induced Injury Involving 5-Lipoxygenase in Feline Esophageal Epithelial Cells

  • Lim, Jae-Chun;Park, Sun-Young;Nam, Yoon-Jin;Nguyen, Thanh Thao;Sohn, Uy-Dong
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.5
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    • pp.313-320
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    • 2012
  • In this study, we focused to identify whether eupatilin (5,7-dihydroxy-3',4',6-trimethoxyflavone), an extract from Artemisia argyi folium, prevents $H_2O_2$-induced injury of cultured feline esophageal epithelial cells. Cell viability was measured by the conventional MTT reduction assay. Western blot analysis was performed to investigate the expression of 5-lipoxygenase by $H_2O_2$ treatment in the absence and presence of inhibitors. When cells were exposed to 600 ${\mu}M$ $H_2O_2$ for 24 hours, cell viability was decreased to 40%. However, when cells were pretreated with 25~150 ${\mu}M$ eupatilin for 12 hours, viability was significantly restored in a concentration-dependent manner. $H_2O_2$-treated cells were shown to express 5-lipoxygenase, whereas the cells pretreated with eupatilin exhibited reduction in the expression of 5-lipoxygenase. The $H_2O_2$-induced increase of 5-lipoxygenase expression was prevented by SB202190, SP600125, or NAC. We further demonstrated that the level of leukotriene $B_4$ ($LTB_4$) was also reduced by eupatilin, SB202190, SP600125, NAC, or nordihydroguaiaretic acid (a lipoxygenase inhibitor) pretreatment. $H_2O_2$ induced the activation of p38MAPK and JNK, this activation was inhibited by eupatilin. These results indicate that eupatilin may reduce $H_2O_2$-induced cytotoxicity, and 5-lipoxygenase expression and $LTB_4$ production by controlling the p38 MAPK and JNK signaling pathways through antioxidative action in feline esophageal epithelial cells.

Changes of Enzyme Activity, Trypsin Inhibitor, Tannin and Phytic Acid during Heat Treatment of Soybean (콩의 열처리 중 효소, 트립신 저해제, 탄닌, 피트산의 함량 변화)

  • Kim, Young-Mi;Kim, Yong-Wook
    • Korean Journal of Food Science and Technology
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    • v.30 no.5
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    • pp.1012-1017
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    • 1998
  • This study was performed to investigate the change of lipoxygenases and urease activities, trypsin inhibitor, tannin and phytic acid contents during heat treatment of Jinpum soybean. The lipoxygenase-1 and urease possessed their activities after heating at $60^{\circ}C$ for 100 min, but their activities disappeared rapidly at $80^{\circ}C$ and $100^{\circ}C$ for 20 and 10 min. There were no lipoxygenase-2 and -3 activities in Jinpum soybean with and without heating. Trypsin inhibitor was lost 91.9%, 78,1% and 58.6% of the activity after heat treatment at $100^{\circ}C$ for 50 min, at $80^{\circ}C$ for 100 min and at $60^{\circ}C$ for 100 min, respectively. The tannin content was increased by heat treatment. The content of and phytic acid was increase after heating at $60^{\circ}C$ for 100 min, unchanged at $80^{\circ}C$ for 100 min and decreased at $100^{\circ}C$ for 100 min.

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