• Bulletin of the Korean Chemical Society
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• 제32권7호
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• pp.2405-2409
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• 2011

The psychrophilic bacteria Psychromonas arctica survives at subzero temperatures by having adapted several protective mechanisms against freezing and oxidative stresses. Many reactive oxygen species are likely generated in P. arctica as a result of reduced metabolic turnover rates. A previous study identified the pasod gene for superoxide dismutase from P. arctica using a series of PCR amplifications. Here, upon cloning into a His-tag fused plasmid, the sod gene from P. arctica (pasod) was successfully expressed by IPTG induction. His-tagged PaSOD was subsequently purified by $Ni^{2+}$-NTA affinity chromatography. The purified PaSOD exhibited a higher SOD activity than that of Escherichia coli (EcSOD) at all temperatures. The difference in activity between PaSOD and EcSOD becomes even more significant at 4$^{\circ}C$, indicating that PaSOD plays a functional role in the cold adaptation of P. arctica in the Arctic.

• Journal of Microbiology
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• 제39권1호
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• pp.37-41
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• 2001

The fission yeast Schizosaccharomyces pombe contains two distinct superoxide dismutase (SOD) activities, one in the cytosol encoded by the $sod2^{+}$ gene and the other in mitochondria. The $sod2^{+}$ gene encoding putative mitochondrial manganese superoxide dismutase (MnSOD) was isolated from the S. pombe genomic library using a PCR fragment as the probe. The nucleotide sequence of the $sod2^{+}$ gene and its flanking region (4051 bp HindIII fragment) was determined. An intron of 123 nt in size was predicted and confirmed by sequencing the cDNA following reverse transcription PCR. The predicted Sod2p consists of 218 amino acid residues with a molecular mass of 24,346 Da. The deduced amino acid sequence showed a high degree of homology with other MnSODs, especially in the metal binding residues at the active site and their relative positions. The transcriptional start site was mapped by primer extension at 231 at upstream from the ATG codon. A putative TATA box(TATAAAA) was located 58 nt upstream from the transcriptional start site and putative polyadenylation sites were located at 1000, 1062, and 1074 nt downstream from the ATG start codon.

• Archives of Plastic Surgery
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• 제40권5호
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• pp.517-521
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• 2013

Background Diabetes is characterized by chronic hyperglycemia, which can increase reactive oxygen species (ROS) production by the mitochondrial electron transport chain. The formation of ROS induces oxidative stress and activates oxidative damage-inducing genes in cells. No research has been published on oxidative damage-related extracellular superoxide dismutase (EC-SOD) protein levels in human diabetic skin. We investigated the expression of EC-SOD in diabetic skin compared with normal skin tissue in vivo. Methods The expression of EC-SOD protein was evaluated by western blotting in 6 diabetic skin tissue samples and 6 normal skin samples. Immunohistochemical staining was also carried out to confirm the EC-SOD expression level in the 6 diabetic skin tissue samples. Results The western blotting showed significantly lower EC-SOD protein expression in the diabetic skin tissue than in the normal tissue. Immunohistochemical examination of EC-SOD protein expression supported the western blotting analysis. Conclusions Diabetic skin tissues express a relatively small amount of EC-SOD protein and may not be protected against oxidative stress. We believe that EC-SOD is related to the altered metabolic state in diabetic skin, which elevates ROS production.

• BMB Reports
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• 제37권3호
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• pp.325-329
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• 2004

Oxidation of catecholamines may contribute to the pathogenesis of Parkinson's disease (PD). The effect of the oxidized products of catecholamines on the modification of Cu,Zn-superoxide dismutase (SOD) was investigated. When Cu,Zn-SOD was incubated with the oxidized 3,4-dihydroxyphenylalanine (DOPA) or dopamine, the protein was induced to be aggregated. The deoxyribose assay showed that hydroxyl radicals were generated during the oxidation of catecholamines in the presence of copper ion. Radical scavengers, azide, N-acetylcysteine, and catalase inhibited the oxidized catecholamine-mediated Cu,Zn-SOD aggregation. Therefore, the results indicate that free radicals may play a role in the aggregation of Cu,Zn-SOD. When Cu,Zn-SOD that had been exposed to catecholamines was subsequently analyzed by an amino acid analysis, the glycine and histidine residues were particularly sensitive. These results suggest that the modification of Cu,Zn-SOD by oxidized catecholamines might induce the perturbation of cellular antioxidant systems and led to a deleterious cell condition.

• BMB Reports
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• 제46권11호
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• pp.555-560
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• 2013

Acrolein is the most reactive aldehydic product of lipid peroxidation and is found to be elevated in the brain when oxidative stress is high. The effects of acrolein on the structure and function of human Cu,Zn-superoxide dismutase (SOD) were examined. When Cu,Zn-SOD was incubated with acrolein, the covalent crosslinking of the protein was increased, and the loss of enzymatic activity was increased in a dose-dependent manner. Reactive oxygen species (ROS) scavengers and copper chelators inhibited the acrolein-mediated Cu,Zn-SOD modification and the formation of carbonyl compound. The present study shows that ROS may play a critical role in acrolein-induced Cu,Zn-SOD modification and inactivation. When Cu,Zn-SOD that has been exposed to acrolein was subsequently analyzed by amino acid analysis, serine, histidine, arginine, threonine and lysine residues were particularly sensitive. It is suggested that the modification and inactivation of Cu,Zn-SOD by acrolein could be produced by more oxidative cell environments.

• Journal of Microbiology and Biotechnology
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• 제3권3호
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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.

• Biotechnology and Bioprocess Engineering:BBE
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• 제3권1호
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• pp.15-18
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• 1998

The entanol productivity of superoxide dismutase (SOD)-deficient mutants of Saccharo-Myces cerevisiae was examined under the oxidative stress by Paraquat. It was observed that MnSOD-deficient mutant of S. cerevisiae had higher ethanol productivity than wild type or CuZnSOD-deficient yeast both in aerobic and in anaerobic culture condition. Pyruvated dehydrogenase activity decreased by 35% and alcohol dehydrogenase activity increased by 32% were observed in MnSOD-deficient yeast grown aerobically. When generating oxygen radicals by Paraquat, the ehanol productivity was increased by 40% in CuZnSOD-deficient or wild strain, resulting from increased activity of alcohol dehydrogenase and decreased a activity of pyruvate dehydrogenase. However, the addition of ascorbic acid with Paraquat returned the enzyme activities at the level of control. These results imply that SOD-deficiency in yeast strains may cause the metabolic flux to shift into anaerobic ethanol fermentation in order to avoid their oxidative damages by Paraquat.

• Journal of Ginseng Research
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• 제11권1호
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• pp.24-31
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• 1987

We studied a assay method on the measurement of superoxide dismutase (SOD Superoxide : superoxide oxidoreductase, EC. 1. 15. 1. 1) activity with photoreduced flavin and nitroblue tetrazolium (NBT) as superoxide (${O_2}^{-}$) source and detector, respectively. The $\Delta$E (1000 ng SOD$.$$min.)^{-1}$ of photoreduced flavin-NBT system was 0.08, whereas that of xanthine-xanthine-cytochrome system used broadly in experiments was 0.014. Therefore, the new method was regarded more simple and utilizable than xanthine-xanthine cytochrome system method. In the present paper, we also carried out to investigate the SOD activity and isozyme pattern for the parpose of study of leaf-burning disease in ginseng (Panax ginseng C.A. Meyer) leaves.

• KSBB Journal
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• 제18권6호
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• pp.517-521
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• 2003

본 연구에서는 절대적 호기성인 Vitreoscilla가 지닌 FeSOD의 특성을 밝히고자 paraquat와 iron을 사용하여 실험하였다. Vitreoscilla에서 FeSOD의 활성도는 초산화 음이온을 생성하는 paraquat에 대해 커다란 영향 없이 일정하게 발현되었다. 이 결과는 Vitreoscilla는 호기적인 물질대사 동안 생성되어지는 초산화 음이온에 대해 방어할 수 있을 만큼 충분한 FeSOD를 평상시 지니고 있음을 의미한다. 또한 다른 요소에 의한 FeSOD의 발현 가능성이 고려되어졌다. 보조 인자인 Fe 를 처리한 결과 활성도가 증가됨을 알 수 있었다. 이는 CN-resistant respiration을 증가시키는 초산화 음이온에 대한 조절 기작과는 달리 apoenzyme 상태가 보조 인자인 금속 이온 (Fe)에 의해 holoenzyme 상태로 전환되었음을 의미한다. PQ와 Fe를 함께 처리하였을 경우, FeSOD의 증가는 PQ에 의해서 생성된 초산화 음이온에 의해 apo 상태에서 hole 상태로 FeSOD의 활성이 증진되는 synergism effect로 설명되어진다. 결론적으로 metalloenzyme인 SOD의 경우는 이런 전환이 활성 속도를 제한하거나 조절할 수도 있는 posttranslational 수준의 조절 기작이 존재함을 알 수 있었다.

• Journal of Ginseng Research
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• 제21권3호
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• pp.153-159
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• 1997

Living organisms have antioxidant enzymes, such as superoxide dismutase, catalase SE glutathione peroxidase, that protect themselves from the toxic effect of superoxide free radicals. Some report says that intracellular oxidation stress is involved in pathogenesis of chronic complications of diabetes mellitus. This study was performed to evaluate the effect of red ginseng on lipid peroxidation of red blood cell and antioxidant SOD activity of serum in NIDDM patients. As a result, there were trends for decrease of lipid peroxidases of RBC and Increase of SOD activity of serum in ginseng group but that were not statistically significant. Therefore, we suggest long term and large sized control study is necessary to confirm the protective effects of red ginseng on oxidative damage in NIDDM patients.