• Journal of Microbiology and Biotechnology
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• v.10 no.5
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• pp.700-706
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• 2000

Iron- and zinc-containing superoxide dismutase (FeZnSOD) and nickel-containing superoxide dismutase (NiSOD) are cytoplamic enzymes in Streptomyces griseus. The sodF gene coding for FeZnSOD was cloned from genomic Southern hybridization analysis with a 0.5-kb DNA probe, which was PCR-amplified with facing primers corresponding to the N-terminal amino acid of the purified FeZnSOD of S. griseus and a C-terminal region which is conserved among bacterial FeSODs and MnSODs. The sodF open reading frame (ORF) was comprised of 213 amino acid (22,430 Da), and the deduced sequence of the protein was highly homologous (86% identity) to that of FeZnSOD of Streptomyces coelicolor. The FeZnSOD expression of exponentially growing S. griseus cell was approximately doubled as the cell growth reached the early stationary phase. The growth-associated expression of FeZnSOD was mainly controlled at the transcriptional level, and the regulation was exerted through the 110 bp regulatory DNA upstream from the ATG initiation codon of the sodF gene.

• Korean Journal of Environmental Agriculture
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• v.29 no.1
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• pp.77-85
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• 2010

Reactive oxygen species (ROS) are very harmful to living organisms due to the potential oxidation of membrane lipids, DNA, proteins, and carbohydrates. transformed E.coli strain QC 871, superoxide dismutase (SOD) double-mutant, with three sequence variant MnSOD1, MnSOD2, and MnSOD3 manganese superoxide dismutase (MnSOD) gene isolated from wheat. Although all QC 871 transformants grown at $37^{\circ}C$ expressed mRNA of MnSOD variants, only MnSOD2 transformant had functional SOD activity. MnSOD3 expressed active protein when grown at $22^{\circ}C$, however, MnSOD1 did not express functional protein at any growing and induction conditions. The sequence comparison of the wheat MnSOD variants revealed that the only amino acid difference between the sequence MnSOD2 and sequences MnSOD1 and 3 is phenylalanine/serine at position 58 amino acid. We made MnSOD2S58F gene, which was made by altering the phenylalaine to serine at position 58 in MnSOD2. The expressed MnSOD2S58F protein had functional SOD activity, even at higher levels than the original MnSOD2 at all observed temperatures. These data suggest that amino acid variation can result in highly active forms of MnSOD and the MnSOD2S58F gene can be an ideal target used for transforming crops to increase tolerance to environmental stresses.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.5
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• pp.875-880
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• 2000

Reactive oxygen species are produced under diabetic conditions and possibly cause various forms of tissue damage in patients with diabetes mellitus. The aim of this study was to examine the effects of high glucose on the alloxan-induced beta cell injury. The insulinoma (RINm5F) cells were clutured either with high glucose (22.2 mM) or normoglucose (5.6 mM) in RPMI 1460 media for 3 days. The SOD activities were determined by spectrophotometric assay and nitroblue tetrazolium (NBT) stain. The effects of high glucose on the cytotoxicity of alloxan were also investigated in RINm5F cells and the cells viability were determined by 3-(4,5-dimethylthiazolyl)-2,5-diphenyltetrazolium bromide (MTT) methods. Results showed that the CuZn-SOD activity was decreased but Mn-SOD activity was increased significantly in RINm5F cells cultured with high glucose (22.2 mM) media. The cytotoxicity of alloxan was increased by high glucose compared with normoglucose in RINm5F cells. Diethyl-dithiocarbarmate (DDC), as inhibitor of CuZn-SOC, also potentiate the alloxan-induced cytotoxocity in RINm5F cells. These results suggest that, in RINm5F cells, short term culture with high glucose media decreases Cu-Zn-SOD activity and the decreased activity of CuZn-SOD many one of the causative factors of beta-cell injury induced by high glucose.

• Journal of Life Science
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• v.19 no.4
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• pp.442-448
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• 2009

We investigated photooxidation and responses of antioxidant enzymes involved in scavenging reactive oxygen species (ROS) after light-chilling ($4^{\circ}C$) for 2 days and post chilling ($25^{\circ}C$) in rice leaves. Chilling leaves indicated a 50% reduction in photosynthetic efficiency ($F_v/F_m$ ratio) and a 48% increase of $H_2O_2$, respectively, compared to the control group. In comparison with the control, activities of superoxide dismutase (SOD) and glutathione reductase (GR) increased at light-chilling and post-chilling. CuZn-SOD and Mn-SOD among SOD forms were detected in rice leaves, while Fe-SOD was not found. The increase of SOD and GR activity may serve as a basis for defense against chilling injury as it dismutase superoxide generated by light-chilling. Catalase (CAT) activity decreased during light-chilling, while activity of APX showed remarkable increase during light-chilling in rice leaves. Among CAT isoforms analyzed by 10% native PAGE, activities of isoform -2 and -3 were inhibited during light-chilling. From the elevated APX activity and decreased CAT activity, we suggest that these two enzymes show mutual supplementary relationships, indicating different tendency during light-chilling.

• Animal cells and systems
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• v.14 no.1
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• pp.1-10
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• 2010

Previously we showed that CHIP, a co-chaperone of Hsp70 and E3 ubiquitin ligase, can promote the degradation of mutant SOD1 linked to familial amyotrophic lateral sclerosis (fALS) via a mechanism not involving SOD1 ubiquitylation. Here we present evidence that CHIP functions in the interaction of mutant SOD1 with 26S proteasomes. Bag-1, a coupling factor between molecular chaperones and the proteasomes, formed a complex with SOD1 in an hsp70-dependent manner but had no direct effect on the degradation of mutant SOD1. Instead, Bag-1 stimulated interaction between CHIP and the proteasome-associated protein VCP (p97), which do not associate normally. Over-expressed CHIP interfered with the association between mutant SOD1 and VCP. Conversely, the binding of CHIP to mutant SOD1 was inhibited by VCP, implying that the chaperone complex and proteolytic machinery are competing for the common substrates. Finally we observed that mutant SOD1 strongly associated with the 19S complex of proteasomes and CHIP over-expression specifically reduced the interaction between S6/S6' ATPase subunits and mutant SOD1. These results suggest that CHIP, together with ubiquitin-binding proteins such as Bag-1 and VCP, promotes the degradation of mutant SOD1 by facilitating its translocation from ATPase subunits of 19S complex to the 20S core particle.

• YAKHAK HOEJI
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• v.49 no.1
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• pp.86-91
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• 2005

Flavonoids are class of polyphenolic compounds widely distributed in the plant kingdom, which display a variety of biological activities, including antiviral, antithrombotic, antiinflammatory, antihistaminic, antioxidant and free-radica 1 scavenging abilities. The antioxidant enzyme (AOE) system plays an important role in the defense against oxidative stress insults. To determine whether flavonoid, myricetin can exert antioxidative effects not only directly by modulating the AOE system but also scavenging free radical, we investigated the influence of the flavonoid myricetin on cell viability, different antioxidant enzyme activities, ROS level and the expression of different antioxidant emzyme in B16F10 murine melanoma cells. Myricetin in a concentration range from 6.25 to $50\;{\mu}M$ decreased superoxide dismutase (SOD) and glutathione peroxidase (GPx) enzyme activities, but catalase (CAT) activity was increased. In the myricetin-treated group, ROS levels were decreased dose-dependently. Antioxidant enzyme expression was measured by RT-PCR. Myricetin treatment of B16F10 cells increased catalase expression. Expression levels of copper zinc superoxide dismutase (CuZn SOD) were not affected by exposure of myricetin. Manganese superoxide dismutase (Mn SOD) and GPx expression levels decreased slightly after myricetin treatment. In conclusion, the antioxidant capacity of myricetin was due to CAT and free-radical scavenging.

• 한국체육학회지인문사회과학편
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• v.51 no.3
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• pp.437-445
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• 2012

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and manganese superoxide dismutase (MnSOD) has been hypothesized as a mediator in the activation of multiple pathways implicated in the pathogenesis of diabetic disease. The objective of this study was to understand the mechanism that aerobic exercise activate GAPDH and MnSOD in pancreatic cells. To achieve the purpose of this study, thirty male Sprague-Dawley rats were assigned to control group, diabetic group and diabetic exercise group. 10 rats were forced to exercise according to exercise protocol for 8weeks and 20 rats were untrained for control and diabetic group. Pancreatic tissue were extracted from the each. Expressions of GAPDH and MnSOD in diabetic pancreatic tissues were significantly decreased compare to control group. However, swimming (trained diabetic group) significantly increased expressions of GAPDH and MnSOD compare to diabetic group, respectively. In hyperglycemia, GAPDH and MnSOD in pancreatic cells is activated by aerobic exercise, and this inactivates multiple pathways implicated in the pathogenesis of diabetic disease. In conclusion, these findings suggest that increased activity of GAPDH and MnSOD by exercise have beneficial effects on mitochondrial dysfunction and arresting the progression of diabetic disease.

• Korean Journal of Microbiology
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• v.51 no.2
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• pp.108-114
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• 2015

We tried to analyze the growth time for secretion of the iron containing superoxide dismutase by comparing the intra-and extracellular enzyme activity from Streptomyces subrutilus P5 and analyze possible genetic information for this enzyme secretion. The mycelial dry weights and glucose concentrations in culture filtrates were determined during growth. Glucose was consumed rapidly during logarithmic growth phase and almost exhausted at 24 h of cultivation. While the intracellular activity of iron containing superoxide dismutase was first appeared at three hours, the extracellular activity of this enzyme appeared from 7.5 h of cultivation, early logarithmic growth phase. This early presence of the superoxide dismutase might not be the result of cell lysis but active secretion pathway. There was no information for signal peptide responsible for the enzyme secretion in sodF. However, we found a type three secretion box in the promoter region of sodF that has been known for the genes of type III secreted proteins in other bacteria. This is the first report on the possible existence of type III secretion in Streptomyces.

• Journal of Physiology & Pathology in Korean Medicine
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• v.33 no.5
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• pp.267-274
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• 2019

This study aims to evaluate the effects of Shengmaisan (SMS) and three types of ShengmaisanJiaweifang on the inhibitory effect of melanin synthesis in B16F10 cells, the mechanism of action through tyrosinase, and the antioxidant effect through superoxide dismutase (SOD) activity. In this study, we used ShengmaisanJiaweifangs (SMS, SMSRR, SMSAD, SMSAR) to research the whitening effects in B16F10 cell lines. Shengmaisan (SMS) was a herbal medicine composed of Ginseng Radix, Liriopis Tuber, and Schisandrae Fructus. ShengmaisanJiaweifangs included SMSRR (SMS added with Rehmanniae Radix), SMSAD (SMS added with Asparagi Radix) and SMSAR (SMS added with Astragali Radix). We measured the cell viability, the inhibition rate of the melanin biosynthesis, and the activity of tyrosinase and SOD in malignant melanoma, B16F10 cells, to survey the whitening effect and the mechanism of the impact on the sample. As a result, SMSRR significantly suppressed the cell viability of B16F10 at more than $500{\mu}g/m{\ell}$ and significantly inhibited the generation of melanin induced by ${\alpha}$-MSH at more than $250{\mu}g/m{\ell}$. SMSRR ($500{\mu}g/m{\ell}$) decreased the activity of tyrosinase while increased the activity of SOD. Therefore, we considered that the SMSRR would be able to produce high value-added products more SMS if used as a commercial.

• The Journal of Korean Medicine
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• v.31 no.4
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• pp.20-37
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• 2010

Objective: This study was performed to investigate the effect of Yagwan-cheunghyeoltang (YCT) on insulin secretion in RIN-m5F cells. Methods: After treatment with various concentrations of YCT to RIN-m5F cells, cell viability, free radical-scavenging activity, SOD activity, and insulin secretion were measured. Additionally, insulin-related gene expressions were measured using real-time RT-PCR. Results: 1. YCT didn't show any influence on RIN-m5F cells viability. 2. YCT showed free radical-scavenging activity by 16% at $100{\mu}g/m{\ell}$ of concentration. 3. YCT showed enhancement of SOD activity by 60% at $100{\mu}g/m{\ell}$ of concentration. 4. YCT significantly increased insulin secretion in RIN-m5F cells in a dose-dependent manner. 5. YCT up-regulated INS-1, INS-2, IRS-1, IRS-2 and IRS-3 mRNA expressions compared to the control group. 6. YCT down-regulated INS-R, GCK, GLP-1R and GLP-2R mRNA expressions compared to the control group. Conclusion: YCT has pharmaceutical properties enhancing insulin production and controlling glucose-associated metabolism, and could be a candidate for drug development after further research.