• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.854-860
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• 1999

Superoxide dismutase (SOD) was purified to homogeneity from fruiting bodies of edible mushroom, Lentinus edodes, by ammonium sulfate precipitation, diethylaminoethyl (DEAE)-Sepharose FF ion-exchange chromatography, Sephacryl S-200 gel filtration chromatography, and preparative PAGE. The molecular weight of the purified enzyme was estimated to be approximately 54 kDa by gel filtration chromatography, and the enzyme was shown to be consisted of two identical subunits of molecular weight 27 kDa by SDS-PAGE. The isoelectric point of the enzyme was 4.9 as determined by isoelectric focusing. The enzyme had optimal pH and temperature of pH 8.0 and $20^{\circ}C$, respectively. The activity of the enzyme was inhibited by hydrogen peroxide, but inhibited less by cyanide and azide. The native enzyme was found to contain 0.89g-atom of iron, 0.75g-atom of zinc, and 0.46g-atom of copper per mol of enzyme. Analysis of amino acids composition revealed that the SOD from L. edodes contained a relatively large amount of glutamic acid/glutamine, proline, cysteine, isoleucine, and leucine, but only a small amount of aspartic acid/asparagine, tyrosine, and tryptophan when compared to the other iron-containing SODs.

• Molecules and Cells
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• v.21 no.1
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• pp.161-165
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• 2006

Dichlorodihydrofluorescein ($DCFH_2$) is a widely used probe for intracellular $H_2O_2$. However, $H_2O_2$ can oxidize $DCFH_2$ only in the presence of a catalyst, whose identity in cells has not been clearly defined. We compared the peroxidase activity of Cu,Zn-superoxide dismutase (CuZnSOD), cytochrome c, horseradish peroxidase (HRP), $Cu^{2+}$, and $Fe^{3+}$ under various conditions to identify an intracellular catalyst. Enormous increase by bicarbonate in the rate of $DCFH_2$ oxidation distinguished CuZnSOD from cytochrome c and HRP. Cyanide inhibited the reaction catalyzed by CuZnSOD but accelerated that by $Cu^{2+}$ and $Fe^{3+}$. Oxidation of $DCFH_2$ by $H_2O_2$ in the presence of a cell lysate was also enhanced by bicarbonate and inhibited by cyanide. Confocal microscopy of $H_2O_2$-treated cells showed enhanced DCF fluorescence in the presence of bicarbonate and attenuated fluorescence for the cells pre-incubated with KCN. Moreover, DCF fluorescence was intensified in CuZnSOD-transfected HaCaT and RAW 264.7 cells. We propose that CuZnSOD is a potential intracellular catalyst for the $H_2O_2$-dependent oxidation of $DCFH_2$.

• The Korean Journal of Food And Nutrition
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• v.7 no.4
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• pp.373-382
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• 1994

This study was observed the effect of n6 and n3 polyunsaturated fatty acids of dietary corn oil and fish oil which was supplemented with similar levels of tocopherol in high fat diet on the levels of tocopherol, malondialdehyde ( MDA) productions of plasma and tissues of rats. Also RBC hemolysis, superoxide dismutase (SOD) and glutathione peroxidase (GSH Px) activities In liver were determined. Male Sprague Dawley rats were fed high fat (40%Cal) diet which was different only In fatty acid composition for 6 weeks. Dietary (arts were beef tallow(BT) as a source of saturated fatty acid, corn oil(CO) for n6 linoleic acid (LA) and fish oil(FO) for n3 eicosapentaenoic acid(EPA) + docosahexaenoic acid (DHA). Plasma and liver tocopherol levels were lowered by n3 PUy4 but there was no difference in malondialdehyde(MDA) level by different dietary PUFA. However, MDA content of RBC and hemolysis were increased by n3 PUFA. MDA content, superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in liver were increased in more unsaturated dietary fat groups. Especially, SOD activity was increased in proportion to the degree of fat unsaturation.

• BMB Reports
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• v.40 no.5
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• pp.684-689
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• 2007

The endogenous neurotoxin, 1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline (salsolinol), has been considered a potential causative factor for the pathogenesis of Parkinson's disease (PD). In the present study, we examined the pattern of human Cu,Zn-superoxide dismutase (SOD) modification elicited by salsolinol. When Cu,Zn-SOD was incubated with salsolinol, some protein fragmentation and some higher molecular weight aggregates were occurred. Salsolinol led to inactivation of Cu,Zn-SOD in a concentration-dependent manner. Free radical scavengers and catalase inhibited the salsolinol-mediated Cu,Zn-SOD modificaiton. Exposure of Cu,Zn-SOD to salsolinol led also to the generation of protein carbonyl compounds. The deoxyribose assay showed that hydroxyl radicals were generated during the oxidation of salsolinol in the presence of Cu,Zn-SOD. Therefore, the results indicate that free radical may play a role in the modification and inactivation of Cu,Zn-SOD by salsolinol.

• BMB Reports
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• v.32 no.5
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• pp.440-444
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• 1999

Membrane lipid peroxidation processes yield reactive aldehydes that may react with copper,zinc superoxide dismutase (Cu,Zn SOD), one of the key antioxidant enzymes against oxidative stress. We investigated this possibility and found that exposing Cu,Zn SOD to malondialdehyde (MDA) or 4-hydroxynonenal (HNE) caused the loss of dismutase activity, cross-linking of peptides, and an increase in protein oxidation, reflected by the increased level of carbonyl groups. When Cu,Zn SOD that had been exposed to MDA or HNE was subsequently analyzed by amino acid analysis, histidine content was found to be significantly lost. Both MDA-and HNE-treated Cu,Zn SOD were resistant to proteolysis, which may imply that damaged proteins exist in vivo for a longer period of time than the native enzyme. The lipid peroxidation-mediated damage to Cu,Zn SOD may result in the perturbation of cellular antioxidant defense mechanisms, and subsequently lead to a pro-oxidant condition.

• Journal of environmental and Sanitary engineering
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• v.15 no.2
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• pp.1-9
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• 2000

This research employed a senescence-accelerated mouse(SAM) to explore the possibility that differences exits among the major antioxidants, superoxid dismutase(SOD), in terms of ability to protect such animal treated with Cu, Fe and Mn. To assess the antioxidants function of metal ions on SAM-R/1 and SAM-P/8 were administered with Cu, Fe and Mn orally. The effect of metal ions on SAM towards reversing oxygen sensitivity was determined as a bioassays of SOD in the mouse liver. The data show that the SOD activity was induced by each metal ions in both SAM-R/1 and SAM-P/8. It suggested that induced SOD by each metal ions may protect against oxidative mediated stress. Finally, overall data lead to the possibility of metal ions as an antioxidants or each metal ions act producer of oxygen radicals in the liver of SAM-R/1 and SAM-P/8.

• Journal of Nutrition and Health
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• v.20 no.1
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• pp.15-24
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• 1987

To study effects of heated oil on lipid peroxidation in rat liver, rats were fed 3 and 6 weeks by intubating oils heated for l1(HA group) and 24 hours (HB group) at 18$0^{\circ}C$. The contents of lipid peroxides and vitamin E, and the activities of super oxide dismutase and glutathione peroxidase in liver were measured. Histological changes of the liver tissue were observed. In both HA and HE groups, the contents of lipid peroxides and the activities of superoxide dismutase were increased, but the activities of glutathione peroxidase and the contents of vitamin E in liver were decreased when compared to the control group which was fed fresh cora oils. During the oil feeding period, the activities of superoxide dismutase and the contents of vitamin E were not significantly changed, but the activities of glutathione peroxidase were decreased, and lipid peroxides were increased in the 3 weeks than in 6 weeks. In HB liver, the heterochromatin of nucleus increased, mitochondria swellen, cristae in mitochondria disappeared, fat droplet and secondary lysosome increased and lumen of rough endoplasmic reticulum enlarged, compared with that of the control group. These phenomena in HA group were less pronounced.

• Journal of the Korean Chemical Society
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• v.38 no.1
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• pp.74-79
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• 1994

The participation of superoxide in initiating tissue damage derived from xenobiotics is best illustrated by paraquat intoxication. In the present study, the roles of superoxide dismutase and catalase on paraquat-induced cell injury were investigated using primary cultured rat skin fibroblast. The degree of cell injury was assassed by the conversion of reduced MTT to a blue formazan. Paraquat produced concentration-and time-related cell injury in cultured rat skin fibroblast. Paraquat induced-cell injury was aggravated by pretreatment of aminotriazol (AT: 10 mM), an catalase inhibitor, and attenuated by addition of catalase (100∼500 unit/ml). However, the effects of diethyldithiocarbamate (DDC : 10 mM), copper- and zinc-containing superoxide dismutase (Cu, Zn-SOD) inhibitor, and Cu, Zn-SOD on paraquat-induced injury were not significant. These results suggest that hydrogen peroxide might be more responsible factor than superoxide in the pathogenesis of paraquat-induced cell injury.

• Molecules and Cells
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• v.39 no.3
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• pp.242-249
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• 2016

A balance between production and degradation of reactive oxygen species (ROS) is critical for maintaining cellular homeostasis. Increased levels of ROS during oxidative stress are associated with disease conditions. Antioxidant enzymes, such as extracellular superoxide dismutase (EC-SOD), in the extracellular matrix (ECM) neutralize the toxicity of superoxide. Recent studies have emphasized the importance of EC-SOD in protecting the brain, lungs, and other tissues from oxidative stress. Therefore, EC-SOD would be an excellent therapeutic drug for treatment of diseases caused by oxidative stress. We cloned both the full length (residues 1-240) and truncated (residues 19-240) forms of human EC-SOD (hEC-SOD) into the donor plasmid pFastBacHTb. After transposition, the bacmid was transfected into the Sf9-baculovirus expression system and the expressed hEC-SOD purified using FLAG-tag. Western blot analysis revealed that hEC-SOD is present both as a monomer (33 kDa) and a dimer (66 kDa), as detected by the FLAG antibody. A water-soluble tetrazolium (WST-1) assay showed that both full length and truncated hEC-SOD proteins were enzymatically active. We showed that a potent superoxide dismutase inhibitor, diethyldithiocarbamate (DDC), inhibits hEC-SOD activity.

• Korean Journal of Food Science and Technology
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• v.28 no.5
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• pp.865-869
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• 1996

Biological effect of aqueous extracts of 12 plants which showed superoxide dismutase (SOD)-like activity in vitro was evaluated using Drosophila melanogaster. Survival percentage of the flies was a criterion of effect when the flies were exported to paraquat, which generated superoxide anion radical in vivo. When flies were co-administered with paraquat and aqueous extracts of garlic, lettuce, kiwi, and nameko, they showed no defensive effect against of oxygen toxicity. If flies were exposed to 60 mM paraquat after adaptation to feed containing plant extracts with SOD-like activity for 10 days, however, survival percentage of flies fed with phytochemicals was $35{\sim}63%$ while that percentage of flies fed without phytochemical was only 11%. This result indicated that adaptation of flies to plant extracts with SOD-like activity could prevent the flies from oxidative injury. On the other hand, lettuce, kiwi, nameko, onion, persimmon, fern brake and cauliflower showed a reparative effect on an oxidative stress. Dropwort, shiitake, agaric mushroom and broccoli did not show such an effect.