• Journal of Life Science
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• v.13 no.5
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• pp.658-667
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• 2003

Antioxidative enzymes (superoxide dismutase; SOD, ascorbate peroxidase; APX, glutathione reductase; GR) play major roles in scavenging mechanism of reactive oxygen species which were involved in various stress conditions including salt. In order to investigate the relation between their growth responses (dry weight) and the changes of antioxidative enzymes activity, salt-tolerant spinach beet having 15cm of shoot length were treated with various salt levels (0, 50, 200, 1000 mM NaCl) for 24 hours. Spinach beet exhibited an increase in the activity of antioxidative enzymes by salt, the maximal activity at 200 mM NaCl and the lowest activity at 50 mM NaCl in 2 hrs. after treatments. As a result of PAGE, it has been confirmed that spinach beet contained 3 isoforms (Fe-SOD, CuZn-SOD and Mn-SOD) of SOD and main isoform was CuZn- SOD form. In case of APX, isoforms of the low molecular weight(No. 7, 8) were showed strong expression especially at 200 and 400 mM NaCl treatment. Meanwhile, GR did not show specific pattern of isoforms among the salt treatments. Especially, in case of 50 mM treatment, plant showed the lowest activity of SOD with the best growth, a low enzyme activity was induced by inactivation of the Mn-SOD. Therefore, we suggested that the decrease of SOD activity at a low salt level (50 mM NaCl) or the increase of enzyme activity at a high salt level (200 mM NaCl) may be related to expression of the Mn-SOD isoform. These antioxidative enzymes showed the increase of activity in a short time by salt addition. So, it is considered that spinach beet copes effectively with a stressful condition such as salt by operating effective antioxidative defense mechanism rapidly under high salt level.

• Korean Journal of Microbiology
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• v.43 no.2
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• pp.83-90
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• 2007

Sphingomonas sp. KS 301, which was isolated from oil contaminated soil, was shown to have five different SODs (SODI, II, III, IV, V) which can be separated by DEAE-Sepharose chromatography, and SOD III was finally purified in this study by ammonium sulfate precipitation, DEAE-Sepharose chromatography, Superose 12 gel filtration and Uno-Q1 ion exchange chromatography. The molecular weight of SOD III was 23 kDa as determined by SDS-PAGE and the apparent molecular weight of the native enzyme was estimated to be approximately 71 kDa by Superose-12 gel filtration chromatography. These data suggest that the purified SOD consists of at least two subunits. The specific activity of the SOD III was higher than Mn type or Fe type SOD of Escherichia coli by 5 fold. To determine the type of SOD III, inhibitory effects of $NaN_{3},\;H_{2}O_{2},\;KCN$ were examined. 10 mM $NaN_{3}$ was able to inhibit 56% of the SOD III activity, which indicates that this SOD is Mn type. The optimum pH of the SOD III was 7.0 and the optimum temperature was $20^{\circ}C$. N-terminal amino acid sequence of purified SOD III was most similar to those of Psudomonase ovalis and Vibrio cholerae among bacteria.

• Korean Journal of Microbiology
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• v.40 no.2
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• pp.94-99
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• 2004

The activities of enzymes that act on oxygen derivatives in Rhodobacter sphaeroides D-230 were investigated under various culture conditions. Intracellular SOD activity from the cells grown in aerobic or anaerobic culture conditions was highest at pH 7.0 and pH 8.0, respectively. On the other hand, extracellular SOD activity was highest at pH 6.0. Catalase activity was highest at neutral pH in both cases. Growth of R. sphaeroides D-230 in aerobic or anaerobic culture conditions was inhibited by methyl viologen. As R. sphaeroides D-230 was cul-tured aerobically, SOD activity was increased about 2-fold by addition of iron ion. But $Mn^+2$ had little effect on the SOD activity of R. sphaeroides D-230 grown in aerobically. NaCN, the inhibitor of Cu$.$Zn-SOD, did not inhibit SOD activity. But, $NaN_3$, the inhibitor of Mn-SOD, inhibited SOD activity in anaerobic cultures con-dition. Therefore, R. sphaeroides D-230 produce Mn-SOD in anaerobic condition, although Fe-Sod is produced in aerobic condition. The activity of catalase was induced by methyl viologen, however, extremely inhibited by NaCN and $NaN_3$.

• Journal of Microbiology and Biotechnology
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• v.17 no.8
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• pp.1399-1402
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• 2007

The virulence of superoxide dismutase (SOD) mutants of Vibrio vulnificus, as tested by intraperitoneal injection into mice, decreases in the order of sodC mutant, sodA mutant, and sodB mutant lacking CuZnSOD, MnSOD, and FeSOD, respectively. The survival of SOD mutants under superoxide stress also decreases in the same order. The virulence of soxR mutant, which is unable to induce MnSOD in response to superoxide, is similar to that of the sodA mutant, as the survival of the soxR mutant under superoxide stress is similar to that of the sodA mutant. Consistently, the lowered survival of the soxR mutant is complemented not only with soxR but also with sodA. Thus, the virulence of V. vulnificus is significantly affected by the cellular level of SOD activity, and an increase in SOD level through MnSOD induction by SoxR under superoxide stress is essential for virulence.

• Journal of Microbiology and Biotechnology
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• v.8 no.4
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• pp.305-309
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• 1998

The superoxide dismutase (SOD) activity of seven Bifidobacterium spp. strains was examined by an indirect SOD assay method. Some Bifidobacterium spp. showed significant levels of SOD activity. However, we could not observe any significant differences between anaerobic and aerobic cultures. Furthermore, although several Bifidobacterium spp. exhibited some degree of tolerance to paraquat which produces superoxide radicals, the apparent SOD activity of these strains was not correlated with their resistance to paraquat. In addition, when we added increasing amounts of manganese or iron to MRS medium which had been prepared without either of the metal ions, the apparent SOD activity of cell free extracts (CFEs) was increased with increasing concentration of both metal ions. To our surprise, the heat-denatured CFEs also showed nearly identical correlative patterns. Based on these results, the apparent SOD activity was likely due to a nonenzymatic dismutation. These results strongly suggest that high concentration of divalent metal ions ($Mn^{2+}$, $Fe^{2+}$) in MRS medium result in nonenzymatic dismutation which can lead to false positive SOD activities in Bifidobacerium spp.

• Journal of Microbiology and Biotechnology
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• v.20 no.4
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• pp.727-731
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• 2010

The gene APE0743 encoding the superoxide dismutase (ApSOD) of a hyperthermophilic archaeon Aeropyrum pernix K1 was cloned and overexpressed as a GST fusion protein at a high level in Escherichia coli. The expressed protein was simply purified by the process of glutathione affinity chromatography and thrombin treatment. The ApSOD was a homodimer of 25 kDa subunits and a cambialistic SOD, which was active with either Fe(II) or Mn(II) as a cofactor. The ApSOD was highly stable against high temperature. This thermostable ApSOD is expected to be applicable as a useful biocatalyst for medicine and bioindustrial processes.

• The Korean Journal of Ecology
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• v.26 no.3
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• pp.115-121
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• 2003

Leaves of two-week old seedlings of tomato (Lycopersicon esculentum) were treated with various concentrations (0∼100 M) of $CdCl_2$ for up to 9 days and subsequent growth of seedlings, symptoms of oxidative stress and isozyme activities of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and peroxidase (POX) were investigated. Compared with the non-treated control, Cd exposure decreased biomass but increased Cd accumulation, hydrogen peroxide production and lipid peroxidation as malondialdehyde (MDA) formation in leaves and roots. Further studies on the developmental changes of isozyme activities showed that Fe-SOD, Cu/Zn-SOD and one of three APX isozymes decreased and CAT and one of four POX isozymes increased in leaves, whereas Fe-SOD, one of three POX isozymes and two of four APX isozymes decreased and CAT increased in roots, showing different expression of isozymes in leaves and roots with Cd exposure level and time. Based on our results, we suggest that the reduction of seedling growth by Cd exposure is the oxidative stress resulting from the over production of $H_2O_2$ and the insufficient activities of antioxidant enzymes particularly involved in the scavenging of $H_2O_2$. Further, the decreased activities of SOD and APX isozymes of chloroplast origin, the increased activities of CAT and POX and high $H_2O_2$ contents with Cd exposure might indicate that Cd-induced oxidative stress starts outside chloroplast.

• 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 $DCFH_2$.

• 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.

• BMB Reports
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• v.28 no.3
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• pp.249-253
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• 1995

The nonenzymatic glycation of copper, zinc-superoxide dismutase (Cu,Zn-SOD) led to inactivation and fragmentation of the enzyme. The glycated Cu,zn-SOD was isolated by boronate affinity chromatography. The formation of 8-hydroxy-2'-deoxyguanosine (8-OH-dG) in calf thymus DNA and the generation of strand breaks in pBhiescript plasmid DNA by a metal-catalyzed oxidation (MCO) system composed of $Fe^{3+}$, $O_2$, and glutathione (GSH) as an electron donor was enhanced more effectively by the glycated CU,Zn-SOD than by the nonglycated enzyme. The capacity of glycated Cu,Zn-SOD to enhance damage to DNA was inhibited by diethylenetriaminepentaacetic acid (DETAPAC), azide, mannitol, and catalase. These results indicated that incubation of glycated CU,Zn-SOD with GSH-MCO may result in a release of $Cu^{2+}$ from the enzyme. The released $Cu^{2+}$ then likely participated in a Fenton-type reaction to produce hydroxyl radicals, which may cause the enhancement of DNA damage.