• Title/Summary/Keyword: Mg - dependent enzyme

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Effects of Butanol Extract of Aralia elata on Lipid Peroxidation (두릅나무 부탄올 추출물이 지질 과산화에 미치는 영향)

  • 서보권;정연봉;김용규;신옥진;이종철
    • YAKHAK HOEJI
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    • v.37 no.3
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    • pp.270-277
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    • 1993
  • It is well known that lipidperoxide, formed in vivo, induced the denaturation of enzyme and destruction of cell membrane to acute injury of tissue. Aralia elata have physiological activates, the improvement of lipid metabolism, antidiabetic activity etc., which was thought to have the relationship to lipid peroxidation. The anti-lipidperoxidative effect of Aralia elata have not yet established. In this study, we examined the anti-lipidperoxidative effects of Aralia elata (Butanol fraction) on CCI$_{4}$ induced lipidperoxidation in rats, and elucidated the anti-lipidperoxidative mechanism. In rat liver homogenate intoxicated with CCI$_{4}$ (0.5 ml/100g), BuOH fraction of Aralia elata (80 mg/Kg/day) exhibited 85.41% anti-lipidperoxidative effect but in serum 69.63% inhibitory effects, respectively. In mitochondrial and microsomal fraction showed inhibition of 55.85% and 69.30%, respectively. In order to elucidate the mechanism of anti-lipidperoxidation effects of Aralia elata, enzymatic (NADPH dependent) and non-enzymatic (Ascorbic acid catalyzed) reaction, in vitro, were performed. In enzymatic reation, Aralia elata exhibited 59.43% anti-lipidperoxidation effects, but in non-enzymatic reaction exhibited 43.27% inhibition. Therefore, it is noteworthy that antioxidative power of them may mainly results from the inhibition by enzymatic reaction.

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Effect of Artemisia anomala S. Moore on Antioxidant Activity and Melanogenesis (유기노의 항산화 활성 및 멜라닌 생성 억제 효과)

  • Lee, Bum-Chun;Kim, Jin-Hwa;Kim, Jin-Hui;Pyo, Hyeong-Bae;Zhang, Yong-He;Park, Hum-Dai;Cho, Young-Ho
    • Korean Journal of Pharmacognosy
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    • v.36 no.4 s.143
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    • pp.273-277
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    • 2005
  • In mammalian melanocytes, melanin synthesis is controlled by tyrosinase, the key enzyme in the pigment synthesis. In this study, to develop a new whitening agent, we have investigated the antioxidant and the inhibitory effect of Artemisia anomala extract on tyrosinase activity and melanigenesis in the B16/F1 melanoma cells. The inhibition ratio of tyrosinase activity of butanol fraction from A. anomala was higher than that of arbutin ($97.5{\pm}0.5%$ at the concentration of 2 mg/ml). The butanol fraction was shown scavenging activities of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals and superoxide anion radicals in a dose dependent manner. The highest inhibitory activity of melanogenesis was also butanol fraction ($25.0{\pm}3%$ at the concentration of $200\;{\mu}g/ml$). From these results, we suggest that the A. anomala extract might be used to be a potential agent for skin whitening.

Molecular Cloning and Characterization of Mannitol-1-Phosphate Dehydrogenase from Vibrio cholerae

  • Rambhatla, Prashanthi;Kumar, Sanath;Floyd, Jared T.;Varela, Manuel F.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.9
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    • pp.914-920
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    • 2011
  • Vibrio cholerae utilizes mannitol through an operon of the phosphoenolpyruvate-dependent phosphotransferase (PTS) type. A gene, mtlD, encoding mannitol-1-phosphate dehydrogenase was identified within the 3.9 kb mannitol operon of V. cholerae. The mtlD gene was cloned from V. cholerae O395, and the recombinant enzyme was functionally expressed in E. coli as a $6{\times}$His-tagged protein and purified to homogeneity. The recombinant protein is a monomer with a molecular mass of 42.35 kDa. The purified recombinant MtlD reduced fructose 6-phosphate (F6P) using NADH as a cofactor with a $K_m$ of $1.54{\pm}0.1$ mM and $V_{max}$ of $320.8{\pm}7.81\;{\mu}mol$/min/mg protein. The pH and temperature optima for F6P reduction were determined to be 7.5 and $37^{\circ}C$, respectively. Using quantitative real-time PCR analysis, mtlD was found to be constitutively expressed in V. cholerae, but the expression was up-regulated when grown in the presence of mannitol. The MtlD expression levels were not significantly different between V. cholerae O1 and non-O1 strains.

Toxicogenomics Study on ${\alpha}-Naphthylisothiocyanate\;(ANIT)$ Induced Hepatotoxictiy in Mice

  • Hwang, Ji-Yoon;Lim, Jung-Sun;Jeong, Sun-Young;Park, Han-Jin;Cho, Jae-Woo;Yoon, Seok-Joo
    • Molecular & Cellular Toxicology
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    • v.2 no.1
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    • pp.48-53
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    • 2006
  • [ ${\alpha}-Naphthylisothiocyanate$ ] (ANIT) induces intrahepatic cholestasis, involving damage to biliary epitheial cells. This study investigates hepatic gene expression and histopathological alterations in response to ANIT treatment in order to elucidate early time response of ANIT-induced hepatotoxicity. ANIT was treated with single dose (3, 6, and 60 mg/kg) in corn oil by oral gavage. Serum biochemical and histopathological observation were performed for evaluation of hepatotoxicity level. Affymetrix oligo DNA chips were used for gene expression profile by ANIT-induced hetpatoxicity. Hepatic enzyme levels (ALT, AST, and ALP) were increased in 24 hr high dose group. In microscopic observations, moderate hepatocellular necrosis, were confirmed 24 hr high dose groups. We found that gene expression patterns were dependent on time and dose. Our selected genes were related inflammation and immunomodulation. In this study, ANIT-induced hepatotoxicity was involved in acute phase responses and provides evidence for role of neutrophil could be mechanism associated with ANIT-mediated hepatotoxicity.

Biological activity of functional foods and anti-microbial activity of phenolics from Sambucus sieboldiana var. pendula leaves

  • Park, Hye-Jin;Lee, Eun-Ho;Jeong, Da-Eun;Han, Chae-Won;Kim, Byung-Oh;Kang, In-Kyu;Cho, Young-Je
    • Korean Journal of Food Science and Technology
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    • v.54 no.4
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    • pp.386-390
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    • 2022
  • This study aimed to investigate the biological and antibacterial activities of phenolics from Sambucus sieboldiana var. pendula leaf extract. The phenolic contents of the S. sieboldiana var. pendula extracted with water and ethanol was 11.60 and 12.39 mg/g, respectively. The inhibitory effects of the water and ethanol extracts on the angiotensin converting enzyme (ACE) were 71.94-92.08 and 48.42-78.33%, respectively, and on xanthine oxidase (XOase) were 42.11-100 and 35.61-100%, respectively, at a phenolic concentration of 50-200 ㎍/mL. All these effects were found to be concentration-dependent. Additionally, the leaf extracts exhibited excellent antibacterial activities against Streptococcus mutans, Staphylococcus aureus, and Propionibacterium acnes. Hence, S. sieboldiana var. pendula was confirmed to have excellent antihypertensive, antigout, and antimicrobial properties.

Molecular Gene Cloning, Expression, and Characterization of Bovine Brain Glutamate Dehydrogenase

  • Kim, Dae-Won;Eum, Won-Sik;Jang, Sang-Ho;Yoon, Chang-Sik;Kim, Young-Hoon;Choi, Soo-Hyun;Choi, Hee-Soon;Kim, So-Young;Kwon, Hyeok-Yil;Kang, Jung-Hoon;Kwon, Oh-Shin;Cho, Sung-Woo;Park, Jin-Seu;Choi, Soo-Young
    • BMB Reports
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    • v.36 no.6
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    • pp.545-551
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    • 2003
  • A cDNA of bovine brain glutamate dehydrogenase (GDH) was isolated from a cDNA library by recombinant PCR. The isolated cDNA has an open-reading frame of 1677 nucleotides, which codes for 559 amino acids. The expression of the recombinant bovine brain GDH enzyme was achieved in E. coli. BL21 (DE3) by using the pET-15b expression vector containing a T7 promoter. The recombinant GDH protein was also purified and characterized. The amino acid sequence was found 90% homologous to the human GDH. The molecular mass of the expressed GDH enzyme was estimated as 50 kDa by SDS-PAGE and Western blot using monoclonal antibodies against bovine brain GDH. The kinetic parameters of the expressed recombinant GDH enzymes were quite similar to those of the purified bovine brain GDH. The $K_m$ and $V_{max}$ values for $NAD^+$ were 0.1 mM and $1.08\;{\mu}mol/min/mg$, respectively. The catalytic activities of the recombinant GDH enzymes were inhibited by ATP in a concentration-dependent manner over the range of 10 - $100\;{\mu}M$, whereas, ADP increased the enzyme activity up to 2.3-fold. These results indicate that the recombinant-expressed bovine brain GDH that is produced has biochemical properties that are very similar to those of the purified GDH enzyme.

Screening of Natural Products for Endothelial and Renal Nitric Oxide Production

  • Kim, Hyeyoung;Han, Sang-Won
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1997.04a
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    • pp.90-90
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    • 1997
  • Natural products, which have been used for the treatment of hypertension, diuresis and nephritis in traditional oriental medicine, were selected for the screening of nitric oxide (NO) production in endothelial cells and kidney tissues in vitro as well as in vivo by measuring the conversion of [$\^$14/C]-L-arginine to [$\^$14/C]-L-citrulline, a coproduct of the enzyme reaction with NO. Confluent monolayer of endothelial cells were used for the screening of 16 natural products. Among the natural products, Zizyphus jujuba and Codonopsis pilosula stimulated endothelial NO synthase activity. Thus, both confluent monolayer of endothelial cells and kidney homogenates (glomeruli, cortical tubules, meudllae) were treated with Zizyphus jujuba and Codonopsis pilosula (final concentration 10 $\mu\textrm{g}$/$m\ell$) and NO releases were compared with those by receptor - dependent agonists, bradykinin and ADP and receptor - independent calcium ionophore A23187 in vitro. In rat experiment, NO releases in glomeruli, cortical tubules and medullae and plasma renin activity were assessed after intraperitoneal injection of Zizyphus jujuba and Codonopsis pilosula (10 mg/kg/day for 4 days). As a result, both Zizyphus jujuba and Codonopsis pilosula significantly increased NO releases in cultured endothelial cells, kidney tissues in vitro as well as in vivo. Stimulation of NO releases by Zizyphus jujuba and Codonopsis pilosula was similar to those by receptor - dependent agonists, bradykinin and ADP and receptor - independent calcium ionophore A23187 in cultured endothelial cells. However, plasma renin activity was not influenced by these two natural products. In conclusion, stimulatory effects of Zizyphus jujuba and Codonopsis pilosula on NO release in kidney may contribute their hypotensive effects and antinephritic action possibly by increasing renal blood flow.

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Ginsenosides attenuate the 3-nitropropioic acid-induced rat striatal degeneration in an age-dependent manner

  • Kim, Jong-Hoon;Nah, Seung-Yeol
    • Journal of Ginseng Research
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    • v.29 no.2
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    • pp.100-106
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    • 2005
  • The number of reporting the effects on ginseng's physiological, pharmacological, and behavioral effects has been increased every year. Major active components of Panax ginseng, are the ginsenosides, which are mainly triterpenoid dammarane derivatives. 3-Nitropropionic acid (3-NP) is blown to induce cellular energy deficit and oxidative stress related neurotoxicity via an irreversible inhibition of the mitochondrial enzyme succinate dehydrogenase (SDH). Intraperitoneal injection of 3-NP produces striatal degeneration. Aged animals was more vulnerable to 3-NP than young animal. We used three different ages of 5-, 8-, and 26-week-old rats. 3-NP alone treatment induced striatal lesion and increased lesion volume with age-dependent manner in 5-, 8-, and 26-week-old rats by $30.2{\pm}5.8$, $v$, and $51.3{\pm}8.4mm^3$, respectively. However, pretreatment of GTS (100 mg/kg/day) before 3-NP reduced striatal lesion in 5-,8-, and 26-week-old rats by $3.15{\pm}6.1$, $8.89{\pm}1.9$, and $27.3{\pm}5.6mm^3$, respectively. Pretreatment of GTS also significantly increased survival rate in 5-week-old rats (3-NP alone: GTS +3-NP = $40.4{\pm}6.3$: $72.5{\pm}9.5\%$) than 8-week-old rats (3-NP alone: GTS + 3-NP : $13.5{\pm}5.2\%$ : $45.1{\pm}3.1\%$). In 26-week-old rats, 3-NP alone treated group died on day 18, whereas GTS +3-NP-treated group prolonged lifespan to 30 days. Thus, pretreatment of GTS before administration of 3-NP extended lifespan in all ages. The present results indicate that aged animals are more vulnerable to 3-NP and GTS pretreatment protected 3-NP-induced striatal damage in different ages of animals.

Effect of Glycyrrhizae Radix on the Immune Responses(I) - Immuno-regulatory Action of 50% Methanol Extract - (감초가 면역반응에 미치는 영향 (I) - 50% 메탄올 엑스의 면역조절작용 -)

  • 한종현;오찬호;은재순
    • YAKHAK HOEJI
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    • v.35 no.3
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    • pp.154-164
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    • 1991
  • These experiments were conducted to investigate the effects of Glycyrrhizae Radix extract(GR) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [$^{3}$H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}Ca^{2+}$ to P388D$_{1}$ cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GR(10$^{-3}$g/ml). Histamine production in mouse spleen cell culture was significantly increased by 48 hour incubation added 0.25$\mu\textrm{g}$/ml of Con A. Con A-dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 $\mu\textrm{g}$/ml of Con A. GR depressed histamine contents at 10$^{-9}$~10$^{-4}$g/ml. and Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-5}$~10$^{-4}$g/ml. IL-1 activity was significantly decreased by 10$^{-8}$~10$^{-4}$g/ml of GR. $Ca^{2+}$ uptake was not changed by GR, but antibody production markedly increased at 10.0~50.0 mg/kg of GR. From the above results, it is suggested that GR have immuno-regulatory action; GR decreased cell-mediated immune response and increased antibody production by B lymphocyte at high doses.

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Alterations in Growth and Morphology of Ganoderma lucidum and Volvariella volvaceae in Response to Nanoparticle Supplementation

  • Singh, Swarnjeet;Kuca, Kamil;Kalia, Anu
    • Mycobiology
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    • v.48 no.5
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    • pp.383-391
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    • 2020
  • Use of nanoparticles (NPs) in several commercial products has led to emergence of novel contaminants of air, soil and water bodies. The NPs may exhibit greater ecotoxicity due to nano-scale dependent properties over their bulk counterparts. The present investigation explores the effect of in vitro supplementation of TiO2, silica and silver NPs on radial growth and ultrastructural changes in the hyphae and spores of two mushroom genera, Ganoderma lucidum and Volvariella volvaceae. A concentration dependent decrease in radial growth on NP amended potato dextrose agar medium was recorded. However, in comparison to control, there was decrease in radial diameter on supplementation with TiO2 NPs while an increase was recorded for silica and silver NPs amendments as compared to their bulk salts at same concentrations after 48 h of incubation. Optical microscopy studies showed decrease in the number of spores while increase in spore diameter and thinning of hyphal diameter on NPs supplementation. Scanning electron microscopy analysis of fungal growth showed presence of deflated and oblong spores in two fruiting strains of Ganoderma while Volvariella exhibited decreased sporulation. Further, hyphal thinning and branching was recorded in response to NP amendments in both the test mushrooms. Enhancement of protein content was observed on NP compared to bulk supplementation for all cultures, concentrations and hours of incubation except for TiO2 NPs. Likewise, bulk and NP supplementations (at 100 mg L-1) resulted in enhanced laccase activity with occurrence of laccase specific protein bands on SDS-PAGE analysis.