• Title/Summary/Keyword: Mg - dependent enzyme

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Physiological Activities of Gymnopilus spectabilis Mycelium Extract and Supernatant of its Broth (갈황색 미치광이버섯 균사체 추출물 및 배양액의 생리활성)

  • Son, Jung-A;Seok, Soon-Ja;Lee, Kyoung-Jin;Lee, Kang-Hyo;Park, Jeong-Sik;Park, Ki-Moon
    • The Korean Journal of Mycology
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    • v.35 no.2
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    • pp.85-95
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    • 2007
  • This study was carried out to investigate the physiological activities of the ethanol extract from Gymnopilus spectabilis mycelium (EGM) and of the supernatant obtained from fermentation broth (SGB). The contents of polysaccharides, phenol compounds and total ${\beta}-glucans$ of EGM were found to be 80.14%, 3.5 mg/ml and 5.91%, respectively and those for SGB were 78.68%, 3.32 mg/ml and 3.28%, respectively. Both EGM and SGB exhibited dose-dependent nitrate-scavenging abilities at pH 1.2. In addition, both EGM and SGB on the autoxidation rate of the linoleic acid demonstrated powerful antioxidant activities at 1 mg/ml level. With respect to fibrolytic activity, EGM showed 1,180 unit/g, which was the same activity as streptokinase, while SGB was 1,011 unit/g. The angiotensin converting enzyme inhibition activity of EMG determined by both the normal and pretreatment methods were estimated to be 8.2% and 10.2%, respectively. However, SGB showed no corresponding activity. The growth inhibitory effects of EGM on AGS, A549, HeLa and NCTC cells were over 58.88%, respectively. And the growth inhibitory effects of the SGB on HeLa and NCTC cells were 44.92 and 76.76%, respectively. Also, EGM and SGB activated the components of the alternative complement pathway from 51 and 62% at the concentration of 100 mg/ml, The xanthine oxidase inhibition activities of EGM and SGB (1 mg/ml) were 9.53 and 16.92%, respectively.

Anti-asthmatic Activities of the Extract of Lonicera japonica (인동 추출물의 항천식 효과)

  • Ryu, Keun-Ho;Han, Chang-Kyun;Rhee, Hae-In;Kim, Taek-Soo;Jung, In-Ho;Lee, Sung-Jae;Im, Guang-Jin;Lee, Kang-Jin;Jeong, Ki-Won;Kim, Dae-Kee;Kim, Key-H.;Cho, Yong-Baik
    • Korean Journal of Pharmacognosy
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    • v.30 no.4
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    • pp.377-383
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    • 1999
  • The anti-asthmatic activities of the extract of Lonicera japonica (BuOH fraction) and its mode of action were investigated using several in vitro and in vivo models. Lonicera japonica was extracted with 30% ethanol (v/v) and successively partitioned into BuOH. The BuOH fraction reduced antigen-induced contraction of isolated trachea from sensitized guinea pigs in a concentration-dependent manner. The BuOH fraction also inhibited histamine release from rat peritoneal mast cells induced by antigen or calcium ionophore A23187 ($IC_{50}=0.26$ and 0.32mg/ml, respectively). Eosinophil infiltration into bronchoalveolar lavage fluids induced by aeroallergen challenge in passively sensitized guinea pigs was inhibited by the BuOH fraction at a dose of 800mg/kg (51.7%). In addition, the BuOH fraction inhibited leukotriene $B_4$ prodution in rat basophilic leukemia cells ($IC_{50}=0.42\;mg/ml$) as well as phosphodiesterase 4 (PDE4) isolated from rat brain ($IC_{50}=0.015\;mg/ml$). All results from this study strongly suggest that the BuOH fraction of Lonicera japonica may be useful in the treatment of asthma and its mode of action may be related with inhibition of both 5-lipoxygenase and PDE4 enzyme.

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Reduction of Oxidative Stress by Chondroitin Sulfate in the Ovariectomy-Induced Aging Rat (난소절제로 유도한 노화쥐에서 chondroitin sulfate에 의한 산화 스트레스의 감소효과)

  • 이진영;하배진
    • Journal of Life Science
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    • v.14 no.2
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    • pp.280-285
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    • 2004
  • The ovarian hormone-deficiency induced ovariectomy rat is widely used as an aging model due to its practicality, convenience, and cost effectiveness. The surgically ovariectomized rat induces reactive oxygen species (ROS) generation like aging phenomena. Free oxygen radicals have been proposed as important causative agents of aging. The purpose of this study was to investigate the effect of chondroitin sulfate (CS) to prevent ovariectomy (OVX)-induced oxidative stress. The OVX rats were given intraperitoneally CS at doses of 100 mg/kg and 200 mg/kg daily for fifteen weeks. Malondialdehyde (MDA) levels were determined as well as the activities of superoxide dismutase (SOD), catalase (CAT), reduced-glutathione (GSH), oxidized-glutathione (GSSG), glutathione peroxidase (GPx) in the liver. The liver antioxidative enzyme activity was elevated while MDA concentration decreased in all CS treated animals. The results demonstrated that CS reduced oxidative stress in a dose dependent manner. These results suggest that CS might be a useful candidate for antioxidative reagent.

Physiological Activity of Sarcodon aspratus Extracts (능이버섯(Sarcodon aspratus) 추출물의 생리활성)

  • 송재환;이현숙;황진국;한정환;노정근;금동혁;박기문
    • Food Science of Animal Resources
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    • v.23 no.2
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    • pp.172-179
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    • 2003
  • This study was carried out to find the preventive medical and therapeutic effects of Sarcodon asparatus on adult disease by employing several biological and biochemical assays. Nitrate scavenging ability(NSA) of Sarcodan asparatus extracts was displayed up to 99.9% at pH 1.2 in a dose-dependent manner. They also had 90.4% electron donating ability(EDA) at the concentration of 0.1 mg/mL. Extracts of Sarcodon asparatus were also able to function as a powerful antioxidant at all concentrations(0.01∼l.0 mg/mL). Furthermore, we observed that 1 mg/mL concentration of the extracts was more powerful than BHT, With respect to fibrolytic activity, Sarcodon asparatus showed 1,843.8 unit/g, which was higher than streptokinase(1,189 unit/g). The inhibitory effects of the extracts on angiotensin converting enzyme, measured by the normal and pretreatment methods, were 53 and 58%, respectively. We also performed cytotoxicity effect of Sarcodon asparatus extracts on a various cancer cell lines. The growth inhibitory effects of the extracts(5.0 mg/mL) on A549, HeLa, AGS, and SK-Hep-1 cells were 78.9, 55.3, 69.0, and 42.5 %, respectively. Interestingly, Sarcodon asparatusextracts induced mutation on Salmonella typhimurium TA98 and TA100 when Ames test was done.

Inhibitory Effects of Ethanol Extracts from Polygoni multiflori radix and Cynanchi wilfordii radix on Melanogenesis in Melanoma Cells (하수오와 백하수오의 에탄올 추출물에 의한 B16/F10 Melanoma 세포주의 멜라닌 생성 억제효과)

  • Seo, Hee;Seo, Geun-Young;Ko, Su-Zie;Park, Young-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.8
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    • pp.1086-1091
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    • 2011
  • Anti-oxidative activity and tyrosinase inhibitory activity of various ethanol extracts of Polygoni multiflori radix (PMR) and Cynanchi wilfordii radix (CWR) were compared to identify an anti-oxidant and whitening agent source from nature. We conducted an investigation into the anti-oxidant activities of PMR and CWR ethanol extracts by measuring total polyphenol content, total flavonoid content, and ABTS radical capacity. The total polyphenol contents of PMR and CWR were 17.31${\pm}$0.54 mg GA/eq g, and 2.75${\pm}$0.22 mg GA/eq g, respectively. The total flavonoid contents of PMR and CWR were 6.38${\pm}$0.39 mg naringine/eq g, and 1.34${\pm}$0.09 mg naringine/eq g, respectively. The 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) radical decolorization of PMR and CWR were 96.89${\pm}$0.21% at 1 mg/mL and 93.49${\pm}$0.76% at 50 mg/mL. Melanoma cells were cultured with the PMR and CWR ethanol extracts for 48 hr, and total melanin content as a final product and the activity of tyrosinase, a key enzyme, in melanogenesis, were estimated. The PMR and CWR ethanol extracts increased melanin content and tyrosinase activity in a dose-dependent manner. These results suggest that PMR and CWR ethanol extracts could be useful as a skin whitening agent.

The Effects of Onion Extracts on Mercury-Induced Toxicity and Lipid Peroxidation in Rat Hepatocyte Primary Culture (랫드 간세포 일차배양에서 양파 추출물이 수은에 의해 유도된 독성 및 지질과산화에 미치는 영향)

  • Rhim, Tae-Jin;Lim, Sang-Cheol
    • Korean Journal of Environmental Agriculture
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    • v.24 no.2
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    • pp.146-152
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    • 2005
  • The objective of present study was to investigate the effect of onion extracts on mercuryinduced cytotoxicity, lipid peroxidation and antioxidant enzyme activities in primary monolayer cultures of rat hepatocytes. Primary cultures of rat hepatocytes were incubated for 6 hr in the presence of various concentrations (0, 1, 5, 10, 30 or 50 ppm) of $HgCl_2$. Cytotoxicity and cell viability were determined by measuring glutamic oxaloacetic transaminase (GOT) activity, lactate dehydrogenase (LDH) activity and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) value. Lipid peroxidation w as evaluated using thiobarbituric acid reactive substances (TBARS) assay. Effects of onion extract on antioxidant system were determined by measuring catalase, glutathione peroxidase (GSH-Px), glutathione reductase (GSH-Rd) activities as well as DPPH free radical scavenging activity. $HgCl_2$ at the concentration of 10 ppm increased GOT activity and TBARS concentration but decreased %MTT reduction, whereas $HgCl_2$ at the concentration of 30 ppm increased LDH activity, representing that $HgCl_2$ caused cytotoxicity and lipid peroxidation in dose-dependent manner, $HgCl_2$ at the concentration of 30 ppm significantly decreased catalase, GSH-Px and GSH-Rd activities. When primary cultures of rat hepatocytes were incubated with various concentrations (0, 0.01, 0.05, 0.1 or 0.3 mg/ml) of onion extract for 6 hr in the presence of 30 ppm of $HgCl_2$, onion extracts at the concentration of 0.05 mg/ml decreased GOT activity, but increased %MTT reduction by 30 ppm of $HgCl_2$. $HgCl_2-induced$ LDH activity and TBARS concentration were decreased by onion extract at the concentration of 0.01 mg/ml. Taken together, onion extract prevented H$HgCl_2-induced$ hepatocyte injury and lipid peroxidation. Onion extracts at the concentration of 0.1 mg/ml almost or completely inhibited $HgCl_2-induced$ catalase and GSB-Px activities. GSH-Rd activity, however, was not affected by onion extract. Free radical scavengjing activity was increased as concentration of onion extract increased. Onion extract at the concentrion of 5 mg/ml possesed mote than 93% scavenging activity comparing to 100% radical scavenging activity by pyrogallol solution as a reference. These results demonstrate that onion extracts suppressed mercury-induced cytoctoxicity and lipid peroxidation by scavenging free radical and increasing catalase and GSH-Px activities.

Expression and Characterization of Thiol-Specific Antioxidant Protein, DirA of Corynebacterium diphtheriae (코리네박테리움 디프테리아 티올 특이성 항산화단백 DirA의 발현 및 특성)

  • Myung-Jai Choi;Kanghwa Kim;Won-Ki Choi
    • Biomedical Science Letters
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    • v.4 no.1
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    • pp.1-9
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    • 1998
  • A Corynebacterium diphtheriae iron-repressible gene dirA, that was homologous to TSA of Saccharomyces cerevisiae and AhpC subunit of Salmonella typhimurium alkyl hydroperoxide reductase, was amplified with PCR and expressed in E. coli. The DirA purified from the transformed E. coli crude extracts prevented the inactivation of enzyme caused by metal-catalyzed oxidation (MCO) system containing thiols but not by ascorbate/Fe$^{3+}$/$O_2$ MCO system. The DirA concentration, which inhibited the inactivation of glutamine synthetase by 50% (IC$_{50}$) against MCO system, was 0.12 mg/ml. The multimeric forms of DirA were converted to the monomeric form in SDS-PAGE under the thioredoxin system comprised of NADPH, Saccharomyces cerevisiae thioredoxin reductase, and thioredoxin. Also, DirA showed thioredoxin dependent peroxidase activity. All of these results were consistent with the characteristics of a thiol specific antioxidant (TSA) protein having two conserved cysteine residues.

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Catalytic properties of wheat phytase that favorably degrades long-chain inorganic polyphosphate

  • An, Jeongmin;Cho, Jaiesoon
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.1
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    • pp.127-131
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    • 2020
  • Objective: This study was conducted to determine catalytic properties of wheat phytase with exopolyphosphatase activity toward medium-chain and long-chain inorganic polyphosphate (polyP) substrates for comparative purpose. Methods: Exopolyphosphatase assay of wheat phytase toward polyP75 (medium-chain polyP with average 75 phosphate residues) and polyP1150 (long-chain polyP with average 1150 phosphate residues) was performed at pH 5.2 and pH 7.5. Its activity toward these substrates was investigated in the presence of Mg2+, Ni2+, Co2+, Mn2+, or ethylenediaminetetraacetic acid (EDTA). Michaelis constant (Km) and maximum reaction velocity (Vmax) were determined from Lineweaver-Burk plot with polyP75 or polyP1150. Monophosphate esterase activity toward p-nitrophenyl phosphate (pNPP) was assayed in the presence of polyP75 or polyP1150. Results: Wheat phytase dephosphorylated polyP75 and polyP1150 at pH 7.5 more effectively than that at pH 5.2. Its exopolyphosphatase activity toward polyP75 at pH 5.2 was 1.4-fold higher than that toward polyP1150 whereas its activity toward polyP75 at pH 7.5 was 1.4-fold lower than that toward polyP1150. Regarding enzyme kinetics, Km for polyP75 was 1.4-fold lower than that for polyP1150 while Vmax for polyP1150 was 2-fold higher than that for polyP75. The presence of Mg2+, Ni2+, Co2+, Mn2+, or EDTA (1 or 5 mM) exhibited no inhibitory effect on its activity toward polyP75. Its activity toward polyP1150 was inhibited by 1 mM of Ni2+ or Co2+ and 5 mM of Ni2+, Co2+, or Mg2+. Ni2+ inhibited its activity toward polyP1150 the most strongly among tested additives. Both polyP75 and polyP1150 inhibited the monophosphate esterase activity of wheat phytase toward pNPP in a dose-dependent manner. Conclusion: Wheat phytase with an unexpected exopolyphosphatase activity has potential as a therapeutic tool and a next-generational feed additive for controlling long-chain polyP-induced inappropriate inflammation from Campylobacter jejuni and Salmonella typhimurium infection in public health and animal husbandry.

The Hepatotprotective and Antioxidative Effects of Onion (Allium cepa) Extracts in Rat Hepatocyte Primary Culture (양파(Allium cepa) 추출물의 간보호 및 항산화 효과)

  • Rhim, Tae-Jin;Lim, Sang-Cheol
    • Proceedings of the Plant Resources Society of Korea Conference
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    • v.18 no.1
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    • pp.52-60
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    • 2005
  • The objectives of present study were to investigate the hepatoprotective and antioxidative effects of onion extracts. Primary cultures of rat hepatocytes were incubated with 1.5 mM tert-butyl hydroperoxide(t-BHP), potent oxidizing agent for liver injury for 1 hr in the presence or absence of various concentrations (0, 0.01, 0.05, 0.1 or 0.3 mg/ml) of onion extract. Cytotoxicity and cell viability were determined by measuring glutamic oxaloacetic transaminase(GOT) activity, lactate dehydrogenase(LDH) activity and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) value. Lipid peroxidation was evaluated using thiobarbituric acid reactive substances(TBARS) assay. Effects on antioxidant system were determined by measuring catalase, glutathione peroxidase(GSH-Px), glutathione reductase(GSH-Rd) activities as well as DNA strand breaking assay. Incubation with t-BHP alone increased GOT and LDH activities and TBARS concentration but decreased MTT reduction. Onion extracts at the concentration of 0.05 mg/ml began to decrease GOT and LDH activities induced by 1.5 mM t-BHP. Decreased MTT reduction began to be increased by onion extract at the concentration of 0.01 mg/ml. Onion extracts at the concentration of 0.01 mg/ml began to decrease TBARS concentration induced by t-BHP. Taken together, onion extracts prevented t-BHP-induced hepatocyte injury and lipid peroxidation. Catalase, GSH-Px and GSH-Rd activities of hepatocytes were significantly decreased by 1.5 mM t-BHP for 1 hr incubation. Onion extracts, on the other hand, at the concentration of 0.1 mg/ml began to prevent t-BHP-induced decrease in catalase, GSH-Px and GSH-Rd activities. Onion extracts prevented hydroxyl radical-induced single-strand breakage in dose-dependent manner when plasmid DNA was incubated with various concentrations of onion extracts in the presence of Fenton regents producing hydroxyl radical. These results demonstrate that onion extracts suppressed t-BHP-induced cytoctoxicity, decreased viability and lipid peroxidation and increased GSH-Px, GSH-Rd and catalase activities. Thus hepatoprotective and antioxidant effects of onion extract seem to be due to, at least in part, the increase in antioxidant enzyme activities as well as prevention from hydroxyl radical-induced oxidation, followed by inhibition in lipid peroxidation.

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The Hepatotprotective and Antioxidative Effects of Onion (Allium cepa) Extracts in Rat Hepatocyte Primary Culture (양파(Allium cepa) 추출물의 간보호 및 항산화 효과)

  • Lim Sang-Cheol;Rhim Tae-Jin
    • Korean Journal of Plant Resources
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    • v.18 no.3
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    • pp.470-478
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    • 2005
  • The objective of present study was to investigate the hepatoprotective and antioxidative effects of onion extracts. Primary cultures of rat hepatocytes were incubated with 1.5 mM tort-butyl hydroperoxide(t-BHP), potent oxidizing agent to liver, for 1 hr in the presence or absence of various concentrations (0, 0.01, 0.05, 0.1 or 0.3 mg/ml) of onion extract. Incubation with t-BHP increased glutamic oxaloacetic transaminase(GOT) and lactate dehydrogenase(LDH) acitivities and thiobarbituric acid reactive substances(TBARS) concentration but decreased 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) reduction. Onion extracts at the concentration of 0.05 mg/ml decreased t-BHP-induced GOT and LDH activities. Onion extract at the concentration of 0.1 mg/ml increased t-BHP-induced MTT reduction. Onion extract at the concentration of 0.01 mg/ml decreased t-BHP-induced TBARS concentration. Taken together, onion extracts prevented t-BHP-induced hepatocyte injury and lipid peroxidation. Catalase, glutathione peroxidase(GSH-Px) and glutathione reductase(GSH-Rd) activities of hepatocytes were significantly decreased by t-BHP. Onion extracts at the concentration of 0.1 mg/ml prevented t-BHP-induced decrease in catalase, GSH-Px and GSH-Rd activities. Onion extracts prevented hydroxyl radical-induced single-strand breakage in dose-dependent manner when plasmid DNA was incubated with various concentrations of onion extracts in the presence of Fenton reagents producing hydroxyl radical. These results demonstrate that onion extracts suppressed t-BHP-induced cytoctoxicity, decreased viability and lipid peroxidation and increased GSH-Px, GSH-Rd and catalase activities. Thus hepatoprotective and antioxidant effects of onion extract seem to be due to, at least in part, the increase in antioxidant enzyme activities as well as prevention from hydroxyl radical-induced oxidation, followed by inhibition of lipid peroxidation.