• Title/Summary/Keyword: $P_{sod}$

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Photosynthetic Pigment Concentrations and Changes of SOD Activities on Liana, Equisetum Arvense and Artemisia Princeps Exposured to Ozone (오존에 노출된 덩굴식물류, 쇠뜨기, 쑥의 광색소 함량과 SOD 활성 변화)

  • 박은희;김종갑;이재천;한심희
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.4 no.3
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    • pp.159-163
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    • 2002
  • This study was analyzed to photosynthetic pigment concentrations and changes of SOD activities on seven species of liana of A. heterophylla, P. scandens, V. thunbergii, P. tricuspidata, C. trilobus, L. japonica and T. kirilowii, and two species of E. arvense and A. princeps of non climbing plants. Concentrations of chlorophyll a and b, total chlorophylls and total carotenoids of P. tricuspidata in 100 ppb ozone site were the most increased. It was the most increased to P. scandens in ratio of chlorophylls and carotenoids, and E. arvense in ratio of chlorophyll a and b. There was difference to ratio of chlorophyll a and b of liana and non liana. At ratio of chlorophyll a and b of 100 ppb ozone site and the control it was more sensitive to chlorophyll a than chlorophyll b, and P. tricuspidata was the most sensitive at comparing with species, and it was more sensitive to liana than non liana. In SOD activities A. princeps was the most increased to 3535.7 unit/g, and P. scandens was the fewest increased to 109.3 unit/g, and A. heterophylla was only decreased to 131.7 unit/g in comparing to 100 ppb ozone sites and the control.

Effects on Lipid Peroxidation and Antioxidants of Paraquat in the Liver of Senescence-Accelerated Mouse(SAM) (제초제인 Paraquat가 SAM의 간조직에서 항산화효소의 활성 및 지질과산화에 미치는 영향)

  • 양미경;박문숙
    • Journal of environmental and Sanitary engineering
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    • v.14 no.2
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    • pp.8-17
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    • 1999
  • This research employed a senescence-accelerated mouse(SAM) to explore the possibility that differences exist among the major antioxidatns, lipid peroxidation in terms of ability to protect such animal treatment PQ, SAM-R/1 and SAM-P/8 were administered with PQ(200ppm/Kg) orally. The toxicity of PQ on SAM was determined as a bioassays of SOD, catalase and lipid peroxidation in the mouse liver. The data show that the SOD activity was induced by paraqwuat terement in both SAM-R/1 and SAM-P/8. The degree of lipid peroxidation was increased with PQ treatment. This means that SOD rather than catalase may protect against oxygen radical toxicity. Finally, over data lead to the toxicity of PQ and its function may efect to the antioxidants including SOD, catalase and lipid peroxidation in both SAM-R/1 and SAM-P/8 .

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Molecular Cloning and High-Level Expression of Human Cytoplasmic Superoxide Dismutase Gene in Escherichia coli (사람의 세포질 Superoxide Dismutase 유전자의 클로닝과 대장균내에서의 대량발현에 관한 연구)

  • 이우길;김영호;양중익;노현모
    • Korean Journal of Microbiology
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    • v.28 no.2
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    • pp.91-97
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    • 1990
  • Complementary DNA (cDNA) coding for human cytoplasmic superoxide dismutase (SOD1) (superoxide: superoxide oxidoreductase E.C.1.15.1.1) was isolated from human liver cDNA library of $\lambda$gt11 by in situ plaque hybridization. The insery cDNA gas the 5' untranslational region (UTR) and 3'UTR of SOD1 gene. Polymerase Chain Reaction (PCR) method was used fro subcloning of SOD1 structural gene. Using synthetic sense strand primer (24mer) containing a start codon and antisense strand primer (24mer), SOD1 structural gene was selectively amplified. Amplified DNA was directly cloned into the HincII site of pUC19 plasmid. Insery cDNA was subcloned into M13 mp19 and sequenced by dideowy chain termination method with Sequenase. The nucleotide sequence of insert cDNA had an open reading frame (ORF) coding for 153 amino acid residues. The structural gene of cytoplasmic SOD was placed under the control of bacteriophage $\lambda P_{L}$ regulatory sequences, generating a highly efficient expression plasmid. The production of human SOD1 in E. coli cells was about 7% of total cellular proteins and recombinant human SOD1 possessed its own enzymatic acitivity.

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Intragastrically Applicated CCl4-Thiopental Sodium Enhanced Lipid Peroxidation and Liver Fibrosis (Cirrhosis) in Rat: Malonedialdehyde as a Parameter of Lipid Peroxidation Correlated with Hydroxyproline as a Parameter of Collagen Synthesis (Deposition)

  • Kim, Ki-Young;Cho, Syung-Eun;Yu, Byung-Soo
    • Toxicological Research
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    • v.25 no.2
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    • pp.71-78
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    • 2009
  • We investigated the pathogenesis of liver tissue damage during the lipid peroxidation and fibrogenesis with the observation of correlations between the parameters of collagen synthesis (and deposition) and lipid peroxidation in liver fibrosis (cirrhosis) rats. Rats were randomly divided into two groups, normal and $CCl_4$-thiopental sod. intoxicated group. And the one group was treated intragastrically with the mixture of $CCl_4$-thiopental sod. 3 times per week for 3 weeks. The liver tissue and sera were used for the measurement of hydroxyproline (HYP), malonedialdehyde (MDA) and superoxide dismutase (SOD). Biochemical parameters such as aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total-bilirubin and blood urea nitrogen (BUN) were measured. Additionally, the expression of collagen ${\alpha}1$(III) and $\beta$-actin mRNA was observed by RTPCR. The histological change in liver tissue was also observed by Masson's trichrome and H&E staining. Correlation analysis was carried by Spearman's rho method. All biochemical parameters except total-bilirubin were significantly higher in the $CCl_4$-thiopental sod. treated group than that of the normal group (p < 0.01). In the $CCl_4$-thiopental sod. treated group, Hyp as a parameter of collagen synthesis (deposition) and MDA as a metabolite of lipid peroxidation, were significantly elevated by 1.98 and 2.11 times higher than that of the normal group (p < 0.001) respectively. The activity of SOD in the $CCl_4$-thiopental sod. treated group is decreased significantly by 44.8% (p < 0.001). And collagen ${\alpha}1$(III) mRNA was more expressed in the $CCl_4$-thiopental sod. treated group than that of the normal group. However, the expression of $\beta$-actin mRNA is showed similar in both of groups. A good correlation was observed between the content of hyp and MDA concentration (r = 0.70, n = 40) in the two groups. And the correlation between the levels of hyp and SOD (r = -0.71, n = 25) is also reliable. However, no correlation were observed between MDA concentration and SOD (r = -0.40, n = 25) in the two groups. Elevated levels of MDA in $CCl_4$-thiopental sod. treated rats indicated enhancement of lipid peroxidation, which is accompanied by a decrease in SOD activity. Moreover, we could confirm that the parameters of collagen synthesis (and deposition) is in good correlation with the metabolite of lipid peroxidation (MDA) and the lipid peroxidation antagonizing enzyme (SOD). Hence, we propose that (1) lipid peroxidation and collagen synthesis (and deposition) could be enhanced by intragastrically application of $CCl_4$-thiopental sod. during a short terms. And (2) the intoxication of $CCl_4$-thiopental sod. could be used for monitoring of lipid peroxidation and collagen synthesis (and deposition) for test of antioxidant and antifibrotic agent.

Purification and Characterization of Superoxide Dismutase from Pseudomonas polycolor

  • LEE SANG-OK;IL-CHUN SEO;SOOK-HYUN CHUNG;TAE-HO LEE
    • Journal of Microbiology and Biotechnology
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    • v.3 no.3
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    • pp.188-193
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    • 1993
  • Superoxide dismutase (SOD) was purified from Pseudomonas polycolor to an electrophoretically homogeneous state and partially characterized. SOD was purified by ammonium sulfate fractionation, column chromatography on DEAE-Sephadex A-50, phenyl-Toyopearl 650 M, and gel filtration on Sephadex G-100. The molecular weight and subunit molecular weight of the purified enzyme were estimated to be 40, 000 and 20, 000, respectively. The purified enzyme remained stable at pH 9.0~11.0, $25^{\circ}C$ for 40 hr, but rapidly became inactive below 9.0. SOD was stable up to $45^{\circ}C$ at pH 9.0 with about 80% relative activity, but rapidly became inactive at temperature above that. The enzyme was insensitive to cyanide and fluoride, and sensitive to hydrogen peroxide and azide. The results suggest that the enzyme be an iron-containing SOD.

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Collaborative Effect of CuZnSOD and Human AP Endonuclease against Oxidative Stress

  • Kim Young Gon
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2004.05a
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    • pp.47-50
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    • 2004
  • The defenses against free radical damage include specialized repair enzymes that correct oxidative damages in DNA, and detoxification systems such as superoxide dismutases. These defenses may be coordinated genetically as global responses. We hypothesized that the expression of the SOD and the DNA repair genes would inhibit DNA damage under oxidative stress. Therefore, the protection of E. coli mutants deficient in SOD and DNA repair genes $(sod^-\;xth^-\;and\;nfo^-)$ was demonstrated by transforming the mutant strain with a plasmid pYK9 which encoded Photobacterium leiognathi CuZnSOD and human AP endonuclease. The results show that survival rates were increased in $sod^+\;xth^-\;nfo^+$ cells compared to $sod^-\;xth^-\;ap^+,\;sod^-\;xth^-\;ap^-,\;and\;sod^+\;xth^-\;ap^-$ cells under oxidative stress generated from 0.1 mM Paraquat or 3 mM $H_2O_2$. The data suggested that, at least, SOD and DNA repair enzymes may have collaborate protection and repair of the damaged DNA. Additionally, both enzymes are required for protection against free radicals.

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Expression of Human SOD1 and Mutant SOD1 (G93A) in E. coli and Identification of SOD1 as a Substrate of HtrA2 Serine Protease (대장균에서의 human SOD1과 mutant SOD1 (G93A) 단백질의 발현과 HtrA2의 기질 여부 확인에 관한 연구)

  • Kim, Goo-Young;Kim, Sang-Soo;Park, Hyo-Jin;Rhim, Hyang-Shuk
    • Journal of Life Science
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    • v.16 no.5
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    • pp.716-722
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    • 2006
  • Superoxide dismutase (SOD) is physiologically important in regulating cellular homeostasis and apoptotic cell death, and its mutations are the cause of familial amyotrophic lateral sclerosis (FALS). Mitochondrial serine protease HtrA2 has a pro-apoptotic function and has known to be associated with neurodegenerative disorders. To investigate the relationship between genes associated with apoptotic cell death, such as HtrA2 and SOD1, we utilized the pGEX expression system to develop a simple and rapid method for purifying wild-type and ALS-associated mutant SOD1 proteins in a suitable form for biochemical studies. We purified SOD1 and SOD1 (G93A) proteins to approximately 90% purity with relatively high yields (3 mg per liter of culture). Consistent with the result in mammalian cells, SOD1 (G93A) was more insoluble than wild-type SOD1 in E. coli, indicating that research on the aggregate formation of SOD1 may be possible using this pGEX expression system in E. coli. We investigated the HtrA2 serine protease activity on SOD1 to assess the relationship between two proteins. Not only wild-type SOD1 but also ALS-associated mutant SOD1 (G93A) were cleaved by HtrA2, resulting in the production of the 19 kDa and 21 kDa fragments that were specific for anti-SOD1 antibody. Using protein gel electrophoresis and immunoblot assay, we compared the relative molecular masses of thrombin-cleaved GST-SOD1 and HtrA2-cleaved SOD1 fragments and can predict that the HtrA2-cleavage sites within SOD1 are the peptide bonds between leucine 9-lysine 10 (L9-K10) and glutamine 23-lysine 24 (Q23-K24). Our study indicates that SOD1 is one of the substrate for HtrA2, suggesting that both HtrA2 and SOD1 may be important for modulating the HtrA2-SOD1-mediated apopotic cell death that is associated with the pathogenesis of neurodegenerative disorder.

Study on the Intracellular Superoxide Dismutase Produced by Bacillus circulans (Bacillus circulans가 생산하는 Superoxide Dismutase에 관한 연구)

  • Lee, Sang-Ok;Tae-Ho Lee
    • Microbiology and Biotechnology Letters
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    • v.15 no.6
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    • pp.381-387
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    • 1987
  • Distribution of superoxide dismutase (SOD) which catalyzes the dismutation of superoxide radicals to hydrogen peroxide and oxygen has been examined in various genera of bacteria. SOD was produced by various bacteria independent of genus and species with variation in superoxide dismutase activity of each bacteria. Bacillus circulans which produced relatively large amount of SOD was selected and used to investigate the optimum culture conditions and further studies. The compositions of optimum culture medium for the enzyme production were 1% glucose, 2% polypeptone, 0.l% NaCl, and 0.2mM of methyl viologen and initial pH was 6.0. The highest enzyme production was observed after 20 hours of cultivation at 3$0^{\circ}C$ on a reciprocal shaker. The enzyme activity was maintained stably for a relatively long period by the addition of 5% ethanol in pH 5.0, 0.01M acetate buffer.

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Effects of Opuntia humifusa Supplementation on Lipid Peroxidation and SOD Protein Expression in the Liver, Kidney, and Skeletal Muscle of Rats Fed a High-fat Diet (손바닥 선인장 보충이 고지방식이 흰쥐의 간장, 신장 골격근에 지질괴산화와 SOD단백질 발현에 미치는 영향)

  • Kwon, Dae-Keun;Kang, Jun-Yong;Park, Jin-Ho;Ryu, Sung-Pil;Song, Young-Ju
    • Journal of Life Science
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    • v.22 no.7
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    • pp.857-862
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    • 2012
  • This study was conducted to investigate the effects of Opuntia humifusa supplementation on lipid peroxidation and superoxide dismutase (SOD) protein expression at resting state in various organs of rats fed a high-fat diet. Sixteen Sprague-Dawley male rats, 6 weeks of age, were randomly divided into two groups: a control diet group (CG, n=8) and an experimental diet group (EG, n=8). They were given a high-fat diet (CG) or a diet supplemented with 5% of O. humifusa (EG) for 8 weeks. The results showed that the malondialdehyde (MDA) levels of the kidney and the liver were significantly lower in the EG group than in the CG group (p<0.01). In addition, the MDA levels in the skeletal muscle of the EG group tended to be lower than those in the CG group, but this difference was not significant. The Cu, Zn-SOD protein expression in the kidney of the EG group was significantly increased compared with that of the CG group (p<0.01). The Mn-SOD protein expression in the skeletal muscle of the EG group was significantly increased compared with that of the CG group (p<0.01). These results suggest that O. humifusa supplementation has antioxidative properties, which are exerted in a specific organ manner, and that it inhibits the action of lipid peroxidation and the expression of SOD in rats fed a high-fat diet.

Iron Containing Superoxide Dismutase of Streptomyces subrutilus P5 Increases Bacterial Heavy Metal Resistance by Sequestration (Streptomyces subrutilus P5의 철 함유 Superoxide Dismutase의 중금속 격리에 의한 세균의 중금속 저항성 증가)

  • Kim, Jae-Heon;Han, Kwang Yong;Jung, Ho Jin;Lee, Jungnam
    • Korean Journal of Microbiology
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    • v.50 no.3
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    • pp.179-184
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    • 2014
  • Mitigation of heavy metal toxicity by iron containing superoxide dismutase (FeSOD) of Streptomyces subrutilus P5 was investigated. For E. coli $DH5{\alpha}$, the survival rate in the presence of 0.1 mM lead ions was only 7% after 120 min; however, with the addition of $0.1{\mu}M$ of purified native FeSOD the survival rate increased to 39%. This detoxification effect was also shown with 0.01 mM copper ions (survival increased from 6% to 50%), and the effect was stronger than with the use of EDTA. E. coli M15[pREP4] producing 6xHis-tagged FeSOD was constructed, and this showed an increase in survival rates throughout the incubation time; in the presence of 0.1 mM lead ions,the final increase at 60 min was from 3% to 19%. The FeSOD absorbed about 123 g-atom lead per subunit; therefore, we suggest that FeSOD could sequestrate toxic heavy metals to enhance bacterial survival against heavy metal contamination.