• Title/Summary/Keyword: Mn/FeSOD

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Purification and Characterization of Superoxide Dismutase in Sphingomonas sp. KS 301 (Sphingomonas sp. KS 301의 Superoxide Dismutase 정제 및 특성)

  • Kang, Hee-Jeong;Jeong, Jae-Hoon;Choi, Ji-Hye;Son, Seung-Yeol
    • 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.

Construction of Gene-Specific Primers for Various Antioxidant Isoenzyme Genes and Their Expressions in Rice (Oryza sativa L.) Seedlings Obtained from Gamma-irradiated Seeds

  • Kim, Jin-Hon;Chung, Byung-Yeoup;Kim, Jae-Sung;Wi, Seung-Gon;Yang, Dae-Hwa;Lee, Choon-Hwan;Lee, Myung-Chul
    • Journal of Photoscience
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    • v.11 no.3
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    • pp.115-120
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    • 2004
  • For the expression study of antioxidant isoenzyme genes in rice (Oryza sativa L.) plants, extensive searches for genes of superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT) isoforms were performed through the GenBank database. The genes for two cytosolic and one plastidic CuZn-SOD, one Fe-SOD, two Mn-SOD, two cytosolic and two chloroplastic (stromal and thylakoid) APX, and three CAT isoforms were available in japonica-type rice. These isoforms were named as cCuZn-SOD1, cCuZn-SOD2, pCuZn-SOD, Fe-SOD, Mn-SOD1, Mn-SOD2, cAPXa, cAPXb, Chl_sAPX, Chl_tAPX, CATa, CATb, and CATc, respectively. Since they shared a high degree of homology in the nucleotide and amino acid sequences, the gene-specific primers for the genes were designed directly from their full-length cDNAs found in the database except for the CATa gene. These primers were used in the RT-PCR analysis to investigate the differential expression of antioxidant isoenzyme genes in rice plants from the seeds irradiated with low doses (2, 4, 8, and 16 Gy) of gamma-radiation. The gammairradiation slightly increased the transcripts of pCuZn-SOD, while those of Fe-SOD, cAPXb, and CATb decreased. However, no substantial differences were observed in the expression of all the isoenzyme genes between the control and irradiated groups. In this study, gene specific primers for thirteen SOD, APX and CAT isoenzymes were constructed from the full-length cDNAs. The results of RT-PCR analysis obtained by using these primers suggests that the expression levels of SOD, APX, and CAT isoenzyme genes in rice seedlings were hardly affected by gamma-irradiation at the seed stage.

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A Study on SOD Activity and Serum Antioxidant Mineral Concentrations in Obese Adolescents (비만 청소년의 SOD 활성도 및 혈청 항산화무기질 농도에 관한 연구)

  • Lee, Da-Hong
    • Journal of Nutrition and Health
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    • v.40 no.1
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    • pp.41-48
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    • 2007
  • The purpose of this study was to investigate the relationship between obesity and, erythrocyte SOD (superoxide dismutase) activity and serum antioxidant mineral (Fe, Zn, Cu, Mn and Se) concentrations of adolescents. Subjects were assigned to one of two groups such as obese ($BMI{\geq}25$, 32 boys, 24 girls) and normal group (18.5 < BMI < 23, 27 boys, 30 girls) Subjects were evaluated based on anthropometric measurements, 24-hr dietary recalls and blood analysis. The mean age of the total subjects was 13.8 years. The mean weight (p < 0.001), BMI (p < 0.001) and body fat (p < 0.001) of obese were higher than those of normal group. There was no significant difference in nutrient intake between obese and normal groups. SOD activity of obese group was not significantly different from normal groups, in both males and females. However, in the males, serum Cu concentration of obese were significantly lower than those of normal group. In the females, Serum Mn concentration of obese were significantly lower then those of normal group. In the correlation analysis, BMI of the subjects had significantly negative correlations with serum Cu, Zn and Mn. To summarize the results, increase of obesity may lead to decrease of serum antioxidant minerals such as Cu, Zn and Mn.

An Efficient Method for the Expression and Reconstitution of Thermostable Mn/Fe Superoxide Dismutase from Aeropyrum pernix K1

  • Lee, Hee-Jin;Kwon, Hye-Won;Koh, Jong-Uk;Lee, Dong-Kuk;Moon, Ja-Young;Kong, Kwang-Hoon
    • 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.

Molecular Cloning of the Superoxide Dismutase Gene from Orientia tsutsugamushi, the Causative Agent of Scrub Typhus

  • Koh, Young-Sang;Yun, Ji-Hyun;Kim, Se-Jae
    • Journal of Microbiology
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    • v.40 no.2
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    • pp.151-155
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    • 2002
  • A Superoxide Dismutase (SOD) gene from the obligate intracellular bacterium Orientia tsutsugamushi has been cloned by using the polymerase chain reaction with degenerate oligonucleotide primers corresponding to conserved regions of known SODs. Nucleotide sequencing revealed that the predicted amino acid sequence was significantly more homologous to known iron-containing SODs (FeSOD) than to manganese-containing SODs (MnSOD). Conserved regions in bacterial FeSOD could also be seen. Isolation of the oriential SOD gene may provide an opportunity to examine its role in the intracellular survival of this bacterium.

Characterization of Superoxide Dismutase in Lactococcus lactis

  • Chang, Woo-Suk;So, Jae-Seong
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.732-736
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    • 1999
  • The superoxide dismutase (SOD) in Lactococcus lactis was measured quantitatively and qualitatively under various culture conditions. The L. lactis SOD was induced by oxidative stress. As the concentration of paraquat to produce superoxide radicals increased, the growth of L. lactis decreased with concomitant increase of SOD activity. The SOD activity was found to be growth-phase dependent: when aerobically grown cells entered to the stationary phase, the activity increased gradually until the late stationary phase. From inhibition studies, L. lactis SOD was found to be insensitive to KCN and $H_2O_2$ which are known to inhibit Cu/ZnSOD and FeSOD, respectively. Moreover, as the concentration of manganese in the medium increased, the activity of SOD also increased. These data strongly suggested that L. lactis possessed a single manganese-containing SOD (MnSOD). Finally, a putative sod gene fragment of 510 bp was identified in L. lactis using a polymerase chain reaction (PCR) with degenerate primers designed from the deduced DNA sequences of known SOD genes.

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False Positive SOD Activity of Bifidobacterium spp. Grown in MRS Medium

  • Chang, Woo-Suk;So, Jae-Seong
    • 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.

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Photochemical Damage and Responses of Antioxidant Enzymes in Rice Leaves Induced to Light-Chilling (Light-chilling에 의해 유도된 벼 잎에서의 광합성 변화와 항산화 효소의 반응)

  • Koo, Jeung-Suk;Choo, Yeon-Sik;Lee, Chin-Bum
    • 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.

Cloning, Expression, and Characterization of Superoxide dismutase from Aquifex Pyophilus, a Hyperthermophilic Bacteria

  • Rhim, Jae-Hwan;Yesun Han;Kim, Sung-Hou;Yunje Cho
    • Proceedings of the Korean Biophysical Society Conference
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    • 1996.07a
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    • pp.30-30
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    • 1996
  • A suproxide dismutase gene of Aquifex pyroprolus, a novel marine hypenhermophilic bacterium, was cloned, expressed, and characterized. The SOD of A pyrophilus (ApSOD) is an iron-containing homo-oligomeric protein with a monomeric molecular weight of 24.2 kDa. the amino acid sequence is similar to those of known Mn- and Fe-SODs from thermophilic archaea, and metal binding residues in all SOD sequences from different species are also conserved in A. pyrophilus SOD. (omitted)

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Changes in the Activities of Antioxidant Enzymes during Chilling Stress in Chilling-Tolerant and Chilling-Sensitive Cultivars of Cucurbita spp. (내저온성과 민감성 호박 품종의 저온 스트레스에 대한 항산화효소의 활성 차이)

  • Kang, Nam-Jun;Kwon, Joon-Kook;Cho, Yong-Seop;Choi, Young-Hah
    • Journal of Bio-Environment Control
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    • v.16 no.1
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    • pp.54-61
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    • 2007
  • To determine whether antioxidant enzyme systems are related to chilling tolerance, changes of antioxidant enzyme activities during the chilling stress were determined in the leaves of a chilling-tolerant cultivar (Cucurbita ficifolia, cv. Heukjong) and a chilling-sensitive cultivar (Cucurbita moschata, cv. Jaerae 13). Leaves of chilling-tolerant plant have two major isoforms, Fe-SOD and Mn-SOD, at the Rm values of 0.20 and 0.52, respectively. In leaves of chilling-sensitive plant, two major isozymes of SOD was observed, one isoform is Mn-SOD at the Rm value of 0.20, and the other isoform is Cu/zn-SOD at the nm value of 0.58. When plants were treated with chilling stress, Cu/zn-SOD at the Rm value of 0.58 was newly expressed at 10 days after chilling stress in the chilling-tolerant plants, and density of this band increased at five days after chilling stress in the chilling-sensitive plants. One APX isozyme band was observed in unstressed plants of both cultivars. Under the chilling stress one APX isozyme band was newly expressed at 10 days after chilling stress in the chilling-tolerant cultivar. Significant genotype differences were observed fnr POD isozyme banding patterns such as few main isozyme bands in chilling-tolerant plants, and one band in chilling-sensitive plants. Densities of three POD isozyme bands at the Rm of 0.36, 0.40 and 0.54 increased at 10 days after chilling stress in the chilling-tolerant plants, while two bands at the nm of 0.36 and 0.54 increased at 10 days and 20 days after chilling stress in the chilling-sensitive plants, respectively. Activities of SOD, APX and POD significantly increased during five days after chilling stress in both cultivars. In the chilling-tolerant cultivar, activities of these enzymes were higher in chilling-stressed plant than in unstressed plants. However, activities of these enzymes in the chilling-sensitive cultivar decreased rapidly after five days of chilling stress, and were lower in chilling stressed plants than in unstressed plants.