• Title/Summary/Keyword: SOD2

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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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Expression of Superoxide Dismutase Isoenzyme Genes and Enzyme Activities in Rice Irradiated with a High-Dose Gamma Ray (고선량 감마선을 조사한 벼에서 SOD isoenzyme들의 유전자 발현 및 효소활성)

  • Chae Hyo-Seok;Kim Jin-Hong;Chung Byung-Yeoup;Kim Jae-Sung;Wi Seung-Gon;Baek Myung-Hwa;Cho Jae-Young
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.180-185
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    • 2006
  • We investigated relations between physiological damages and gene expression and enzyme activities of superoxide dismutase(SOD) isoenzymes in leaves of rice (Oriza sativa L. cv. Ilpoombyeo) plants irradiated with a high-dose gamma-ray. Gamma-irradiation with 500 Gy caused significant decreases in the contents of protein, chlorophyll and carotenoid in the rice leaves by 24 h, especially reducing the chlorophyll contents up to 26% relative to the control. In contrast, gene expressions of SOD isoenzymes were kept higher in the irradiated leaves until 24 h after the irradiation than in the control and they started to noticeably decrease at 48 h, finally being lower in the irradiated leaves at 72 h than in the control. In the case of enzyme activities of SOD isoenzymes, some CuZn-SOD isoenzymes showed slightly increased activities until 48 h after the irradiation but at 72 h, all isoenzyme activities markedly decreased in the irradiated leaves below the control levels. In conclusion, 500 Gy gamma-irradiation used in the current study caused decreases in the contents of protein, chlorophyll and carotenoid as symptoms for physiological damages. Although such physiological damages were not directly related to the gene expressions and enzyme activities of SOD isoenzymes until 24 h after the irradiation, the damages at 72 h were reasonably attributable to their reduction.

Nitrite Scavenging Ability and SOD-like Activity of a Sterol Glucoside form Chrysanthemum coronarium L. var. spatiosum (쑥갓 스테롤배당체의 아질산염소거작용 및 SOD 유사활성)

  • Cho, Min-Jung;Park, Mi-Jung;Lee, Heum-Sook
    • Korean Journal of Food Science and Technology
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    • v.39 no.1
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    • pp.77-82
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    • 2007
  • From the total methanolic extract of Chrysanthemum coronarium L. var. spatiosum (Compositae), nitrite scavenging ability and superoxide dismutase (SOD)-like activity were analyzed as antioxidative characteristics. After successive partitioning with chloroform, n-butanol, and water, the chloroform fraction showed the most significant nitrite scavenging ability with an $IC_{50}$ value of 39 ppm compared with the values of vitamin C and chlorogenic acid, 15 ppm and 36 ppm, respectively. The active fraction was subjected to silica gel and Sephadex LH-20 column chromatography, and the compound was isolated and identified as ${\beta}-sitosterol-O-{\beta}-D-glucoside$ using $^{1}H-NMR$ and $^{13}C-NMR$ spectral data. The glucoside was further hydrolyzed and confirmed as a glycosylated ${\beta}-sitosterol$. The compound and its aglycone, ${\beta}-sitosterol$, showed different nitrite scavenging and SOD-like activity. The $IC_{50}$ value of nitrite scavenging ability of the compound was 335 ppm at pH 1.5, while that of its aglycone was 41 ppm. As for the SOD-like activity, the $EC_{50}$ values of the sterol and the glucoside were 1,291 ppm and >2,000 ppm, respectively, compared with those of vitamin C and chlorogenic acid, 38 ppm and 449 ppm, respectively.

Gamma Radiation-Induced Changes of Antioxidant Enzymes in Callus Cultures of Cassava(Manihot esculenta Crantz) (감마선에 의한 카사바 (Manihot esculenta Crantz) 배양세포의 항산화효소 활성 변화)

  • 이행순;유순희;권석윤;김재성;곽상수
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.1
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    • pp.53-58
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    • 1999
  • The gamma radiation-induced changes of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) in callus cultures of cassava (Manihot esculenta Crantz) selected as a high yield of cell line for SOD were investigated. In normal cultures, the cell growth reached a maximum at 30 days after subculture (DAS), followed by a rapid decrease with further cultures. The SOD and POD specific activities (units/mg protein) showed the highest at the immediately after subculture and subsequently decreased to 20 DAS, and then increased to 30 DAS, whereas the CAT activity showed the lowest at just after subculture, and it continuously increased from 15 DAS to 30 DAS, showing a good correlation with the cell growth. Irradiation of gamma-ray of 50 and 70 Gy on 7 DAS inhibited significantly the cell growth by 50% and 80% at 14 days after treatment (DAT), respectively. In the cells irradiated with 70 Gy, SOD and POD specific activities increased by 4 and 2.5 folds at 14 DAT, respectively, whereas CAT activity was not affected. The results indicate that SOD and POD may be involved in the antioxidative mechanism in relation to oxidative stresses induced by subcultures and by gamma radiation in callus cultures of cassava.

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Study on the Antioxidant Activity of the Extracts from the Lepista nuda (민자주방망이버섯(Lepista nuda)의 항산화성에 관한 연구)

  • Lee, Yang-Suk;Park, Dong-Cheol;Joo, Eun-Yong;Shin, Seung-Ryeul;Kim, Nam-Woo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.7
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    • pp.942-947
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    • 2005
  • This study was investigated to analyze the effect of extracts from the Lepista nuda, on the antioxidant activity to form a part of studies on the functional materials of L. nuda. Antioxidant activity of L. nuda extracts was evaluated by measuring the electron-donating ability (EDA), the superoxide dismutase (SOD)-like activity, and the nitrite-scavenging ability. The EDAs of water and ethanol extracts at the concentration of 1,000 ppm by the rotary heating method from L. nuda were $60.47\%$ and $60.13\%$, respectively and those of water and ethanol extracts by microwave-assisted method were $87.73\%$ and $84.84\%$, respectively. The measurements of SOD-like activity were in the range of $24.58\%\~42.03\%$ at 1,000 ppm. EDA and SOD were increased with the concentrations of extracts. The nitrite-scavenging ability at the concentration of 1,000 ppm was the highest and $29.77\%$ at pH 1.2, and was decreased with an increment of pH value. These results indicated that microwave­assisted water extract from L. nuda showed the highest activities on the EDA and nitrate-scavenging ability, while the rotary heating ethanol extract had the highest effect on the SOD-like activity.

A Mechanistic Study on the Early Stage-Events Involved in Low Temperature Stress in Clamydomonas reinhardtii (Clamydomonas reinhardtii의 냉해 초기과정에 관한 기작론적 연구)

  • Cho, Hyun-Soon;Kim, Chang-Sook;Jung, Jin
    • Applied Biological Chemistry
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    • v.37 no.6
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    • pp.433-440
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    • 1994
  • The exposure of Clamydomonas reinhardtii to low temperatures resulted in an accumulation of cellular pyruvate that dissipated when the chilled cells returned to ambient temperature. The dissipation of pyruvate accumulation was accompanied by an increase in the production level of superoxide radicals $(O_2^-)$ in cells. The formation of $O_2^-$ at an excessive level during the post-chilling period was apparently countered by a substantial activation of superoxide dismutase (SOD). All these results are similar to those observed previously in rice seedlings subjected to the cold-treatment, implicating that a common mechanism is probably underlying for the primary processes of chilling injury both in higher plants and in algae. It was also observed that the activation of Mn-containing SOD contributes the major share in the increase of SOD activity of whole algal cells. Because Mn-SOD is present only in mitochondria, the observation corroborates the concept that the $O_2^-$ scavenging enzyme would be induced to cope with the cold treatment-caused adverse situation in mitochondria where the toxic active oxygen is produced at rates far exceeding the normal rate.

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The Influence of Excercise in a Forest Environment on Blood Pressure, Heart Rate, MDA and SOD (산림 운동이 혈압, 심박수, 과산화지질, 항산화효소에 미치는 영향)

  • Choi, Jonghwan;Shin, Won Sop;Seo, Kyeong-Won;Cha, Wook-Seong;Yeon, Poung Sik;Yoo, Ri Wha
    • Journal of Korean Society of Forest Science
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    • v.97 no.4
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    • pp.417-422
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    • 2008
  • The purpose of this study was to investigate the effect of forest exercise on blood pressure, heart rate, MDA, and SOD. Twelve volunteers were placed in 2 groups: Forest Exercise Group (FEG, n=6; $age=44.2{\pm}2.68$) and Indoor Exercise Group (IEG, n=6; $age=45.6{\pm}2.97$). The subjects who joined this investigation were asked to participate in forest exercise and/or indoor exercise. Blood pressure, heart rate, MDA, and SOD were collected at rest, just after the submaximal exercise, and 30 minutes after each exercise. For data analysis, mean and standard deviation scores were calculated, and ANCOVA were used. The results were as followings : 1) There was no difference between FEG and IEG on systolic blood pressure after submaximal exercise and at 30 minutes after each exercise., however FEG showed better stability than IEG on diastolic blood pressure at 30 minutes after forest exercise. 2) Both groups showed similar change of heart rate and MDA after submaximal exercise and at 30 minutes after each exercise. 3) Both groups similarly activated on the SOD level after submaximal exercise. However, compared to IEG, FEG showed significant decrease on SOD level at 30 minutes after forest exercise. Therefore, this study suggests that forest exercise may have good effects on blood pressure and MDA, and cardiovascular disease prevention.

The Effects of the Makgeolri on the Antioxidative Activity in the Endotoxin LPS-treated Rats (내독소인 LPS로 처치된 흰쥐에 대한 막걸리 항산화 활성효과)

  • Kwon, Ryun-Hee;Chae, Go-Yeon;Ho, Boe-Jin
    • Journal of Food Hygiene and Safety
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    • v.26 no.2
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    • pp.166-170
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    • 2011
  • Modem people have begun to have the nationwide interest in the rice wine called Makgeolri which is one of the traditional Korean alcoholic liquors. This study was performed to investigate the effects of San sung Makgeolri extract (SM) on antioxidation together with the determination of pH and dissolved oxygen (DO) in the progress of fermentation in the lipopolysaccharide(LPS)-treated rats. We examined the levels of SOD (superoxide dismutase), CAT (catalase), GPx (glutathione peroxidase) in liver homogenates and the histopathological observations in liver tissue. LPS-treated group markedly decreased the levels of SOD, CAT and GPx. But SM + LPS-treated group significantly increased the levels of them. Furthermore, the antioxidative effects of SM were supported by the histopathological observations in liver tissue which showed severe inflammation and necrosis in LPS-treated group, compared to the attenuated inflammation and necrosis in SM + LPS-treated group. This results suggested that SM could be a candidate of antioxidative material in spite of alcoholic liquors.

EFFECTS OF GINSENOSIDE $Rb_2$ ON THE ANTIOXIDANTS IN SENESCENCE - ACCELERATED MICE(SAM - R/1)

  • Chung H.Y.;Kim K.W.;Oura H.;Yokozawa T.
    • Proceedings of the Ginseng society Conference
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    • 1993.09a
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    • pp.30-32
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    • 1993
  • In biological system, there are enzymes such as superoxide dismutase(SOD), catalase and glutathione(GSH) peroxidase which scavenge reactive oxygen species as well as antioxidants such as ceruloplasmin, albumin and nonprotein-bound SH including GSH related to defense mechanism. In the present study, the protective effects of ginsenoside $Rb_2$ against oxidative stress were investigated in the SAM-R/1 mice. Treatment with ginsenoside $Rb_2$ significantly increased Cu, Zn-SOD and Mn-SOD in the liver. Ginsenoside $Rb_2$ tended to increase hepatic catalase activity and significantly increased serum albumin and nonprotein-bound SH levels in the liver. But treatment with ginsenoside $Rb_2$ showed a significant decrease in hepatic malondialdehyde(MDA) levels compared to control group. Furthermore, we compared the effects in the hepatic SOD, MDA and serum albumin. These findings suggest that the increase of antioxidants by ginsenoside $Rb_2$ results in the protective effects against reactive oxygen species.

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The Effects of Onion and Garlic on Copper-Phenanthroline Complex Induced DNA Degradation (Copper-Phenanthroline 복합체에 의해 유도되는 DNA 손상에 대한 양파와 마늘의 억제효과)

  • 박평심;이명렬
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.4
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    • pp.367-371
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    • 1992
  • In the present study, the effects of onion and garlic on copper and 1, 10-phenanthroline com plex induced DNA degradation were investigated by the decreased level of thiobarbiturin acid (TBA) reactive materials. Phenanthroline is specific for copper and the reaction releases TBA reactive material from DNA which can be measured by absorbance at 535nm. The levels of TBA reactive materials were decreased by adding onion or garlic ghomogenate into reaction mixture but the onion had more strong potency and the effect of onion was not changed by boiliing. Superoxide dismutase (SOD) and catalase have no inhibitory effects on copper induced DNa damage but reduced glutathone was more effective. The activities of antioxidant enzymes and the contents of sulfhydryl groups in onion and garlic were also investigated. The activity of SOD was more higher in garlic, but catalase and glutathione peroxidase activities were higher in onion. The contents of induced DNA damage were not by antioxidant enzymes such as SOD, catalase and glutathione peroxidase or sulfhydryl groups, but a substance which is more stable in high temperature.

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