• Title/Summary/Keyword: Mn-peroxidase

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Biodegradation of Phenanthrene by Transformant Trametes versicolor MrP1 (구름버섯의 형질전환체 Trametes versicolor MrP1에 의한 Phenanthrene의 생분해)

  • Choi, Yun-Seong;Choi, Hyoung-Tae;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.273-278
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    • 2007
  • As a model compound of PAHs (polycyclic aromatic hydrocarbons) phenanthrene has been regarded as a toxic material, mutagen and carcinogen in various animals. Biodegradation conditions of phenanthrene such as pH, temperature, shaking speed, stabilizer and cofactor of degrading enzymes were investigated with Trametes versicolor and its transformant T. versicolor MrP1 in YMG medium, minimal medium and soil microcosm. T. versicolor MrP1 can overexpress mrp gene encoding Mn-repressed peroxidase that is involved in fungal degradation. Biodegradations of phenanthrene by T. versicolor and T. versicolor MrP1 were optimally performed in conditions of weak-acid (pH 6.0), $30^{\circ}C$, shaken culture and medium containing 5 mM veratryl alcohol or tryptophan. In these optimal conditions, biodegradation of phenanthrene by T. versicolor MrP1 is 31% higher than that of wild type strain in a minimal medium for 20 days. Biodegradation of phenanthrene by T. versicolor MrP1 was also higher than that of wild type in soil microcosm. T. versicolor MrP1 can be a excellent candidate for the bioremediation of PAHs contaminated environments.

Oxidative Stress Inhibitory Effects of Low Temperature-Aged Garlic (Allium sativum L.) Extracts through Free Radical Scavenging Activity (저온숙성마늘의 라디칼 소거 활성을 통한 산화스트레스 억제 효과)

  • Hwang, Kyung-A;Kim, Ga Ram;Hwang, Yu-Jin;Hwang, In-Guk;Song, Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.1
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    • pp.27-34
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    • 2016
  • Garlic has drawn attention as a food material for its anti-oxidative and anti-inflammatory properties as well as for prevention and treatment of cancer. In order to increase efficiency, various aging methods for garlic have been attempted. In particular, thermally processed garlic is known to have higher biological activities due to its various chemical changes during heat treatment. Therefore, in this study, we investigated the anti-oxidative effects of garlic extracts aged at low temperature ($60{\sim}70^{\circ}C$). In the results, 2,2-diphenyl-1-picrylhydrazyl and 2,2-azino-bis (3-ethylbenzo-thiazoline-6-sulfonate) radical scavenging activities and ferric reducing ability of low temperature-aged garlic (LTAG) were similar to those of raw garlic. LTAG also showed decreased lipopolysaccharide (LPS)-induced production of reactive oxygen species, although there were not significant differences among samples. In addition, xanthine oxidase activity was inhibited by LTAG; the 15 days and $60^{\circ}C$ extract showed outstanding inhibition compared with the others. To understand the molecular mechanisms behind the anti-oxidative activity of LTAG, we performed quantitative real-time PCR analysis. The 30 days and $70^{\circ}C$ extract upregulated mRNA expression of antioxidant enzymes such as Cu/Zn-superoxide dismutase (SOD), Mn-SOD, glutathione peroxidase, and catalase in LPS-stimulated RAW 264.7 cells. This result indicates that LTAG can be a functional food as a nature antioxidant and antioxidant substance.

Isolation and Characterization of a Novel Calcium/Calmodulin-Dependent Protein Kinase, AtCK, from Arabidopsis

  • Jeong, Jae Cheol;Shin, Dongjin;Lee, Jiyoung;Kang, Chang Ho;Baek, Dongwon;Cho, Moo Je;Kim, Min Chul;Yun, Dae-Jin
    • Molecules and Cells
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    • v.24 no.2
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    • pp.276-282
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    • 2007
  • Protein phosphorylation is one of the major mechanisms by which eukaryotic cells transduce extracellular signals into intracellular responses. Calcium/calmodulin ($Ca^{2+}/CaM$)-dependent protein phosphorylation has been implicated in various cellular processes, yet little is known about $Ca^{2+}/CaM$-dependent protein kinases (CaMKs) in plants. From an Arabidopsis expression library screen using a horseradish peroxidase-conjugated soybean calmodulin isoform (SCaM-1) as a probe, we isolated a full-length cDNA clone that encodes AtCK (Arabidopsis thaliana calcium/calmodulin-dependent protein kinase). The predicted structure of AtCK contains a serine/threonine protein kinase catalytic domain followed by a putative calmodulin-binding domain and a putative $Ca^{2+}$-binding domain. Recombinant AtCK was expressed in E. coli and bound to calmodulin in a $Ca^{2+}$-dependent manner. The ability of CaM to bind to AtCK was confirmed by gel mobility shift and competition assays. AtCK exhibited its highest levels of autophosphorylation in the presence of 3 mM $Mn^{2+}$. The phosphorylation of myelin basic protein (MBP) by AtCK was enhanced when AtCK was under the control of calcium-bound CaM, as previously observed for other $Ca^{2+}/CaM$-dependent protein kinases. In contrast to maize and tobacco CCaMKs (calcium and $Ca^{2+}/CaM$-dependent protein kinase), increasing the concentration of calmodulin to more than $3{\mu}M$ suppressed the phosphorylation activity of AtCK. Taken together our results indicate that AtCK is a novel Arabidopsis $Ca^{2+}/CaM$-dependent protein kinase which is presumably involved in CaM-mediated signaling.

Studies on Immunomodulatory and Antioxidant Activities of Astragali membranacei Radix according to the Cultivated Years (황기(黃芪)의 재배 년수에 따른 면역 및 항산화 활성 연구)

  • Jung, Chul
    • Korean Journal of Korean Medical Institute of Dermatology and Aesthetics
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    • v.1 no.1
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    • pp.53-90
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    • 2005
  • Purpose: Contents of astragaloside I, II and IV, cytotoxicity, anticancer activity, immunomodulatory activity and antioxidant capacity were to be compared as a function of the cultivated years as one, three, five and seven years. Method: Major components of Astragali membranacei Radix were separated as astragaloside I, astragaloside II, astragaloside IV by HPLC analysis. Cytotoxicity and anticancer activities were measured by MTT and SRB assay. For immunomodulatory activity, the secretion of IL -6 and $TNF-{\alpha}$, NK cell activation and macrophage activation were observed as well as kinetics of responding to human T cells by a microphysiometer. In vitro antioxidant activities were measured by several radical scavenging activities of superoxide anion radican, DPPH, LDL and linoleic acid. For in vivo activity, the activation of SOD, GSH-px, catalase, ALDH and ADH was measured as well the relative weight of liver. Result : 1. For HPLC analysis, the contents of all of astragaloside I, astragaloside II, astragaloside IV were in order of three, five, one and seven years. 2. The cytotoxicity of normal human lung cell line, HEL299 showed lower than 18% in adding 0.25 mg/ml, and 28.9% in adding 1.0 mg/ml of water extract of seven year root. For methanol extracts, three year root showed highest cytotoxicity as 35.2 % and there was no difference between the cultivated years. 3. For anticancer activities, methanol extracts of one and three year roots showed relatively high inhibition of human stomach cancer cells, AGS, breast cancer cells, MCF-7, lung cancer cells, A549 and liver cancer cell, Hep3B as well as high selectivities. 4. The water extract of seven year root could yield high secretion of IL-6 from both human Band T cells while the methanol extracts of three and five year roots secreted high amounts of IL-6 and $TNF-{\alpha}$ from both Band T cells. 5. As a result of in vitro antioxidant activities, both water and methanol extracts from five and seven year roots showed high activities for superoxide anion radical scavenging activity, inhibiting linoleic acid peroxide and contents of total phenols. 6. For in vivo tests, Mn-SOD and GSH-px activities and weight of liver were better in adding seven year root. For ALDH activity one year root was better and for ADH activity five year root. Overall speaking, seven year root showed relatively better antioxidant activities. Conclusion:There was difference of the contents of astragaloside I, astragaloside II, astragaloside IV according to cultivation year. Methanol extract showed better activities of anticancer and immune activation rather than water extract Interestingly enough, for methanol extracts, overall activities were improved as the cultivation year increased. There might be further investigation required for the clinical uses of the results as several biological activities varied according to the cultivated year of Astragali membranacei Radix.

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Biodegradation of 4-t-Octylphenol by Basidioradulum molare and Schizopora paradoxa and Estrogenecity Reduction of its Metabolites (옥틸페놀(4-t-Octylphenol)의 Basidioradulum molare와 Schizopora paradoxa에 의한 분해 및 에스트로겐성 저감효과)

  • Lee, Soo-Min;Ku, Bon-Wook;Lee, Jae-Won;Choi, Don-Ha;Jeung, Eui-Bae;Choi, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.32 no.4
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    • pp.27-35
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    • 2004
  • Recalcitrant 4-t-Octylphenol used as a surfactant was subjected to the biodegradation with wood rot fungi, Basidioradulum molare and Schizopora paradoxa. Two fungi were grown in the culture medium containing various concentrations of 4-t-Octylphenol in order to investigate their resistance against 4-t-octylphenol Schizopora paradoxa was reached to the full growth within 14 incubation days in the concentration of more than 200 ppm of 4-t-Octylphenol, while Basidioradulum molare showed the inhibitory mycelium growth as the concentration was increased Schizopora paradoxa and Basidioradulum molare biodegraded 95% and 36% of initial concentration of 4-t-Octylphenol at first incubation day, respectively. However, the biodegradation capability reached to more than 95% after 3 incubation days. During the biodegradation of 4-t-Octylphenol, the activity of manganese dependent peroxidase was induced by the addition of 4-t-Octylphenol in the culture medium of Schizopora paradoxa, but that of laccase was maximal before the addition. The reduction of estrogenecity was assayed by MCF-7 cell proliferation test and measurement of pS2 mRNA expression. The level of pS2 mRNA was decreased down to the level of baseline at first incubation day. Also, estrogenecity of 4-t-Ocrylphenol completely disappeared after treatment with supernatant by Schizopora paradoxa and Basidioradulum molare from first incubation day of culture down to the levels of vehicle.

Effect of Pyroligneous Liquor on Oxygen Radicals and Their Scavenger Enzymes in Liver of CD Rats (흰쥐 간조직의 활성산소 및 제거효소에 미치는 목초액의 영향)

  • Choi, Jin-Ho;Cho, Weon-Ki
    • Journal of Nutrition and Health
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    • v.40 no.2
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    • pp.111-117
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    • 2007
  • This study was designed to investigate the effects of pyroligneous liquor on oxygen radicals and their scavenger enzymes in the liver of Cri/Bgi CD rats (7 rats per group). Male rats were fed a basic diet prepared in our Lab., PL-0 (Control), PL-1, PL-25, PL-50 and PL-75 groups were Prepared to be 0%, 1%, 25%, 50% and 75%with distilled water using pyroligneous liquor (35% of Choa Co. Ltd.), and were administrated orally for 8 weeks. Superoxide radical contents in liver mitochondria and microsomes were significantly decreased to 12-14%, 11-15%, respectively, in these PL-25 and PL-50groups compared with the control group. Hydroxyl radical content in mitochondria and microsomes were markedly decreased to 12-20% and 17%, respectively, in these PL-25 and PL-50% groups compared with the control group. Hydrogen peroxide content in mitochondria and microsomes were significantly decreased about 15-12% and 22-20% in liver of PL-25 and PL-50 groups compared with the control group. Mn-SOD and Cu/Zn-SOD activities in liver of PL-25 and PL-50 groups were remarkably increased to 15-25%, 11-16%, respectively, compared with the control group. GPx activities in mitochondria and microsomes were significantly increased in the liver of PL-25 and PL-50 groups compared with the control group. CAT activities in mitochondria and cytosol were significantly increased to 12-14%, 15-27%, respectively, in the liver of PL-25 and PL-50 groups compared with the control group. These results suggest that long term administration orally of 25 and 50% pyroligneous liquor may effectively inhibit the formation of oxygen free radicals, and also scavenger enzyme activities significantly increase through the administration orally.

Antioxidant Effect of Hot water and Ethanol extracts from Cheonnyuncho (Opuntia humifusa) on Reactive Oxygen Species (ROS) Production in 3T3-L1 Adipocytes (3T3-L1 지방세포내 ROS 생성에 대한 천년초 열수 및 에탄올 추출물의 항산화 효과)

  • Yoon, Bo-Ra;Lee, Young-Jun;Kim, Sun-Gu;Jang, Jung-Young;Lee, Hyo-Ku;Rhee, Seong-Kap;Hong, Hee-Do;Choi, Hyeon-Son;Lee, Boo-Yong;Lee, Ok-Hwan
    • Food Science and Preservation
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    • v.19 no.3
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    • pp.443-450
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    • 2012
  • Recently, NADPH oxidase 4 (NOX4)-mediated generation of intracellular reactive oxygen species (ROS) was proposed to accelerate adipogenesis of 3T3-L1 cell. We have previously shown that Cheonnyuncho (Opuntia humifusa) extract significantly inhibited adipocyte differentiation via downregulation of $PPAR{\gamma}$ (peroxisome proliferator-activated receptor gamma) gene expression. In this study, we focused on the molecular mechanism(s) of NOX4, G6PDH (glucose-6-phosphate dehydrogenase) and antioxidant enzymes in anti-oxidative activities of 3T3-L1 adipocytes. Our results indicate that Cheonnyuncho extracts markedly inhibits ROS production during adipogenesis in 3T3-L1 cells. Cheonnyuncho extracts suppressed the mRNA expression of the pro-oxidant enzyme such as NOX4 and the NADPH-producing G6PDH enzyme. In addition, treatment with Cheonnyuncho extract was found to upregulate mRNA levels of antioxidant enzymes such as Mn-SOD (manganese-superoxide dismutase), Cu/Zn-SOD (copper/zinc-SOD), glutathione peroxidase (GPx), glutathion reductase (GR), and catalase, all of which are important for endogenous antioxidant responses. These data suggest that Cheonnyuncho extract may be effective in preventing the rise of oxidative stress during adipocyte differentiation through mechanism(s) that involves direct down regulation of NOX4 and G6PDH gene expression or via upregulation of endogenous antioxidant responses.

Effect of buchu (Allium tuberosum) on lipid peroxidation and antioxidative defense system in streptozotocin-induced diabetic rats (부추가 Streptozotocin 유발 당뇨쥐의 지질과산화와 항산화방어체계에 미치는 영향)

  • 송영선;정현실;노경희;조혜연;박지영;최춘연;권태완
    • Journal of Life Science
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    • v.13 no.3
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    • pp.333-342
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    • 2003
  • The pathogenic effort of high glucose, possibly in concert with fatty acids, is mediated to vascular complications of diabetes via increased production of reactive oxygen species(ROS), reactive nitrogen species(RNS), and subsequent oxidative stress. This study was carried out to investigate the suppressive effect of buchu(Allium tuberosum) on oxidative stress in streptozotocin(STZ)-induced diabetes in Sprague Dawley male rats. The effect of buchu supplementation (10%) on lipid peroxidation, and antioxidative defense system in blood and liver was compared among normal rats fed basal diet(normal) and diabetic rats fed basal diet(DM-control) or 10% buchu-supplemented diet(DM-buchu). Diabetes was experimentally induced by the femoral muscle injection of 50 mg STZ per kg of body weight. Animals were sacrificed after 4 wks of experimental diets feeding. The induction of diabetes by STZ elevated the level of lipid peroxidation represented by thiobarbituric acid-reactive substances(TBARS) and conjugated dienes in plasma, LDL, liver, and erythrocytes. 10% buchu-supplemented diet significantly reduced the levels of conjugated dienes in erythrocytes(p<0.05) and lowered TBARS in liver and LDL to the levels of control. Induction of diabetes by STZ elevated Mn-superoxide dismutase(Mn-SOD) activity and lowered activities of glutathionine reductase(GSH-red) and glutathionine peroxidase(GSH-px). Catalase activity was not affected by the induction of diabetes by STZ. However, buchu supplementation to diabetic rats significantly elevated catalase activity(p<0.05) and slightly elevated GSH-px and GSH-red activities in liver. GSH levels of blood and liver were lowered or not changed by induction of diabetes by STZ, respectively, while buchu supplementation to diabetic rats significantly elevated hepatic GSH level (p<0.05). In conclusion, it can be concluded that buchu might be a food source to attenuate oxidative stress in diabetic patients by inhibiting lipid peroxidation, by increasing hepatic GSH level, and by inducing anti-oxidative enzyme systems.

Anti-atherogenic Effect of Isoflavone through Hypolipidemic, Anti-oxidative and Anti-inflammatory Actions in C57BL/6 Mice (C57BL/6 Mice에서 이소플라본의 지질강하, 항산화, 항염증효과를 통한 항동맥경화 효과)

  • Cho, Hye-Yeon;Yang, Jeong-Lye;Noh, Kyung-Hee;Kim, Jin-Ju;Kim, Young-Hwa;Huh, Kyung-Hye;Song, Young-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.3
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    • pp.276-283
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    • 2007
  • This study was carried out to investigate the effect of isoflavone on the atherogenic effect in C57BL/6 mice. C57BL/6 female mice, 5 weeks of age, were fed on chow diets for 2 weeks during adjustment period. Mice weighing approximately $17.9{\pm}0.9\;g$ were divided into 4 groups and were fed on the experimental diets containing isoflavone for 8 weeks. Experimental groups were control (atherogenic diet), IF-10 (atherogenic diet with isoflavone 10 mg/100 g diet), IF-40 (atherogenic diet with isoflavone 40 mg/100 g diet) and IF-100 (atherogenic diet with isoflavone 100 mg/100 g diet). Food efficiency ratio was not different among the experimental groups. Plasma triglyceride (TG) concentrations were lower after 4 weeks in isoflavone supplementation groups than in control group, whereas monocyte chemoattractant protein-1 and thiobarbituric acid reactive substances (TBARS) levels of plasma were significantly (p<0.05) decreased in the isoflavone supplementation groups in a dose dependent manner. Both hepatic TG and cholesterol levels were significantly lowered in IF-100 than control. Hepatic glutathione concentrations were higher in the IF-100 group than in the other groups. Hepatic antioxidant enzyme activities including glutathione-reductase, glutathione-peroxidase, catalase, and Mn-superoxide dismutase were significantly higher in the isoflavone supplemen-tation groups in a dose dependent manner. From the above results, it is concluded that isoflavone may reduce the risk of atherosclerosis via hypolipidemic, anti-oxidative and anti-inflammatory effects.

Attenuation of Oxidative Stress-Induced HepG2 Cellular Damage by Cirsiumjaponicum Root Extract (HepG2 세포에서 대계 추출물에 의한 산화적 스트레스 유발 세포 손상의 억제)

  • Da Jung Ha;Seohwi Kim;Byunwoo Son;Myungho Jin;Sungwoo Cho;Sang Hoon Hong;Yung Hyun Choi;Sang Eun Park
    • Journal of Life Science
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    • v.33 no.12
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    • pp.1002-1014
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    • 2023
  • The root of Cirsium japonicum var. maackii (Maxim.) has long been used in traditional medicine to prevent the onset and progression of various diseases and has been reported to exert a wide range of physiological effects, including antioxidant activity. However, research on its effects on hepatocytes remains scarce. This study used the human hepatocellular carcinoma HepG2 cell line to investigate the antioxidant activity of ethanol extract of C. japonicum root (EECJ) on hepatocytes. Hydrogen peroxide (H2O2) was used to mimic oxidative stress. The results showed that EECJ significantly reverted the decrease in cell viability and suppressed the release of lactate dehydrogenase in HepG2 cells treated with H2O2. Moreover, an analysis of changes in cell morphology, flow cytometry, and microtubule-associated protein light chain 3 (LC3) expression showed that EECJ significantly inhibited HepG2 cell autophagy induced by H2O2. Furthermore, it attenuated H2O2-induced apoptosis and cell cycle disruption by blocking intracellular reactive oxygen species and mitochondrial superoxide production, indicating strong antioxidant activity. EECJ also restored the decreased levels of intracellular glutathione (GSH) and enhanced the expression and activity of superoxide dismutase and GSH peroxidase in H2O2-treated HepG2 cells. Although an analysis of the components contained in EECJ and in vivo validation using animal models are needed, these findings indicate that EECJ is a promising candidate for the prevention and treatment of oxidative stress-induced liver cell damage.