• Journal of Microbiology and Biotechnology
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• 제9권3호
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• pp.271-275
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• 1999

The intracellular superoxide dismutase (SOD) from Staphylococcus sciuri was isolated to homogeneity by continuous steps, including ammonium sulfate fractionation, DEAE-ion-exchange chromatography, gel filtration, and phenyl hydrophobic gel chromatography. Pure SOD had a specific activity of 4,625 U/mg and was purified 158-fold with a yield of 31 % from a cell free extract. The molecular weight of the purified SOD was determined to be approximately 35.5 kDa by gel filtration and the enzyme was also shown to be composed of dimeric subunits on denaturing SDS-PAGE. The enzyme activity remained stable at pH 5 to 11 and also to heat treatment of up to $50^{\circ}C$ at pH 7.8, with 80% relative activity. The enzyme was insensitive to cyanide, hydrogen peroxide, and azide, indicating that it is a manganese-containing SOD. The EPR spectrum showed the enzyme containing manganese as a cofactor.

• Fisheries and Aquatic Sciences
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• 제6권4호
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• pp.209-212
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• 2003

Lucigenin (Lg)- and luminol (Lm)-dependent chemiluminescence (CL) was used to compare the respiratory burst of rockfish (Sebastes schlegeli) phagocytes after stimulation with phorbol myristate acetate (PMA). To establish which reactive oxygen species (ROS) contributes to the observed CL, the modulators of ROS metabolism, such as superoxide dismutase (SOD), catalase, and sodium azide $(NaN_3)$ were used. Although LgCL responses were inhibited significantly by the addition of either SOD or catalase, in comparison to the control, significantly lower LgCL responses were recorded by SOD than catalase. LmCL also showed significantly decreased responses by the addition of SOD and catalase. However, there were no statistical differences in CL responses between SOD and catalase additions. More profound and significant decrease of LmCL responses were recorded by simultaneous addition of SOD and catalase. Sodium azide markedly enhanced LgCL responses, while it significantly inhibited LmCL responses. These results indicate that LgCL and LmCL can be used to measure extracellular $O_2$ production and myeloperoxidase (MPO)-mediated ROS production in fish phagocytes, respectively. Furthermore, LmCL can be used for analyzing intracellular ROS production by simultaneous addition of both SOD and catalase.

• 한국미생물·생명공학회지
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• 제15권6호
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• pp.381-387
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• 1987

생체 내에서 $O_2$를 특이적으로 제거시켜 줌으로써 의학적 응용가치가 매우 높은 것으로 알려진 superoxide dismutase(SOD)에 대하여 연구하였다. 먼저 이 효소의 활성을 여러 속의 세균을 대상으로 검토해된 결과 비교적 높은 활성을 나타낸 Bacillus circulans를 본 실험의 공시균주로 선택한 다음 SOD의 최적 생산 조건을 설정하였다. SOD의 생산은 각 세균에 따라서 양적인 차이는 있으나 종 및 속에 관계없이 다양하게 존재하는 것으로 나타났다. 효소의 생산 조건은 탄소원으로서 1% glucose, 질소원으로서 2% polypeptone, 무기염으로서 0.1% NaCl을 첨가하였을 때 가장 양호하였고 최적 pH는 6.0이었다. 이상의 조성을 가진 배지를 500$m\ell$용 진탕 플라스크에 100$m\ell$을 넣어 3$0^{\circ}C$ 에서 20시간 호기적 배양을 하였을 때 효소 생산 및 균체량이 최대에 도달하였다. 조효소액은 극히 불안정하여 안정제의 검토가 불가피하였다. 안정제로서는 최종농도가 5% 되도록 ethyl alcohol을 첨가한 경우가 효과적이었으며 pH 안정성은 pH5.0인 acetate buffer에서 가장 안정하였다. 이상의 조건 하에서 조효소액은 20일 이상 동안 약 80%의 잔존활성을 가지고 비교적 안정한 상태로 유지되었다.

• 생약학회지
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• 제46권1호
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• pp.17-22
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• 2015

Previous phytochemical studies of Vigna angularis (Ohwi) Ohwi & Ohashi (Leguminosae) have shown the presence of saponins and flavonoids. From the seed of V. angularis, genistein-7-O-${\beta}$-D-glucopyranoside (genistin) was isolated. Lifespan-extending effect of genistin was elucidated using Caenorhabditis elegans model system. Genistin showed potent lifespan extension of worms under normal culture condition. This compound also exhibited the protective effects against thermal and oxidative stress conditions. In the case of heat stress, genistin-treated worms exhibited enhanced survival rate, compared to control worms. In addition, genistin-fed worms lived longer than control worms under oxidative stress induced by paraquat. To verify the possible mechanism of genistin-mediated increased lifespan and stress resistance of worms, we investigated whether genistin might alter superoxide dismutase (SOD), catalase activities and intracellular ROS levels. Our results showed that genistin was able to elevate SOD and catalase activities of worms and reduce intracellular ROS accumulation in a dose-dependent manner.

• 생약학회지
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• 제45권3호
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• pp.227-232
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• 2014

The seed of Vigna angularis (Ohwi) Ohwi & Ohashi (= Phaseolus angularis W. F. Wight, Leguminosae) is one of well-known folk foodstuffs in Korea, China and Japan. In the course of screening for antioxidants from natural plants in Korea by measuring the radical scavenging effect on 1,1-diphenyl-2-picrylhydrazyl (DPPH) a methanol extract of the seeds of V. angularis were found to show a potent antioxidant activity. And the ethyl acetate soluble fraction of V. angularis (VAEA) showed the most potent DPPH radical scavenging and superoxide quenching activities. To reveal the effect of antioxidant activities of VAEA, we tested the thermal and oxidative stress tolerances, activities of SOD and catalase, and intracellular ROS level in Caenorhabditis elegans. Consequently, VAEA-fed worms lived longer than control worms under the thermal and oxidative stress conditions. And VAEA elevated SOD and catalase activities of worms, and reduced intracellular ROS accumulation in a dose-dependent manner.

• Tuberculosis and Respiratory Diseases
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• 제50권5호
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• pp.579-590
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• 2001

연구배경 : 노화란 반응성산소종에 의한 산화적 손상이 축적되어 세포의 기능의 저하로 초래된다고 주장되고 있다. 본 연구에서 1, 4, 8, 12개월된 정상 생쥐의 폐에서 반응성 산소종 생성의 초기에 중요한 방어기전인 superoxide dismutases(SODs) mRNA와 hydroxy radical($OH{\cdot}$)에 의한 산화적 손상의 마지막 방어기전인 metallothionein(MT) mRNA의 함량과 paraquat에 의한 유도능을 조사하였다. 방 법 : 산화적손상은 세포내에서 superoxide을 생성하는 paraquat를 투여한 후 SODs와 MT mRNAs을 RT-PCR 방법으로 측정하였다. 결 과 : 정상생쥐의 폐의 노화과정에서 Mn-SOD와 Cu/Zn-SOD, 그리고 MT mRNA의 항정상태 함량을 조사한 결과, Mn-SOD mRNA는 8개월까지는 증가하다가 12개월에서 감소하였으나, Cu/Zn-SOD mRNA는 별 변화가 없었고 12개월에 감소하였다. 그러나 MT mRNA는 어느 연령에도 변화가 없었다. 산화성 스트레스를 유발하는 paraquat에 의하여 Mn-SOD mRNA는 4개월 부터 유도되지 않았으며, Cu/Zn-SOD mRNA는 전혀 유도되지 않았다. MT mRNA는 관찰한 전 연령에서 유도되었다. 결 론 : 노화과정에서 SODs유도능의 이상이 노화의 원인 인자로 작용하고, 반면에 MT의 정상적인 유도능이 산화적 스트레스에 대하여 방어 역할을 할 가능성을 시사해주고 있다. 이와 같은 결과는 연령이 증가함에 따라 Mn-SOD, Cu/Zn-SOD, 및 MT의 발현과 유도능의 차이로 인하여 반응성 산소종에 의한 산화적 손상의 정도의 차이가 노화의 속도를 결정하는 하나의 인자로 작용할 가능성 이 있을 것으로 생각된다.

• Parasites, Hosts and Diseases
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• 제53권3호
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• pp.253-258
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• 2015

Toxoplasma gondii, an obligate intracellular protozoan parasite of the phylum Apicomplexa, can infect all warm-blooded vertebrates, including humans, livestock, and marine mammals. The aim of this study was to investigate whether superoxide dismutase (SOD) of T. gondii can be used as a new marker for genetic study or a potential vaccine candidate. The partial genome region of the SOD gene was amplified and sequenced from 10 different T. gondii isolates from different parts of the world, and all the sequences were examined by PCR-RFLP, sequence analysis, and phylogenetic reconstruction. The results showed that partial SOD gene sequences ranged from 1,702 bp to 1,712 bp and A + T contents varied from 50.1% to 51.1% among all examined isolates. Sequence alignment analysis identified total 43 variable nucleotide positions, and these results showed that 97.5% sequence similarity of SOD gene among all examined isolates. Phylogenetic analysis revealed that these SOD sequences were not an effective molecular marker for differential identification of T. gondii strains. The research demonstrated existence of low sequence variation in the SOD gene among T. gondii strains of different genotypes from different hosts and geographical regions.

• Toxicological Research
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• 제17권4호
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• pp.249-253
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• 2001

Fumonisin B1, a mycotoxin, is thought to induce esophageal cancer in humans and apoptosis in animal cells by inhibiting ceramide synthase. Dumonisin Bl may also generate reactive oxygen species directly or indirectly, leading to DNA damage and lipid peroxidation. In this study, a DNA fragmentation assay, dichlorofluorescein (DCF) analysis, and single cell gel electrophoresis (SCGE) were used to investigate the involvement of cellular free radicals, specifically hydrogen peroxide, in the DNA damaging activity of fumonisin B1. From an in vitro DNA fragmentation assay, E. coli DNA, damage by fumonisin Bl was increased by the addition of superxide dismutase (SOD) and decreased by catalase. SCGE and DCF analysis in vivo showed that the nuclear DNA damage and intracellular free radicals in cultured rat hepatocytes treated with fumonisin B1 were increased with the concentration of fumonisin Bl . DNA damage and free radical generation were inhibited by the addition of catalase. Fumonisin Bl , in the presence of SOD, produces hydrogen peroxide causing oxidative DNA damage and protein malfunction, leading to genotoxicity and cytotoxicity of the toxin.

• Natural Product Sciences
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• 제20권2호
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• pp.80-85
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• 2014

In the course of screening for antioxidant compounds from natural plants in Korea by measuring the radical scavenging effect, a methanol extract of the flower buds of Daphne genkwa S. et Z. (Thymelaeaceae) was found to show a potent antioxidant activity. Subsequent activity-guided fractionation of methanol extract of D. genkwa led to the isolation of four compounds from the ethyl acetate soluble fraction. The chemical structures were elucidated as genkwanin (1), 3'-hydroxygenkwanin (2), apigenin (3), and tiliroside (4) by spectroscopic techniques. Among them, compound 2 showed the significant anti-oxidative effect on DPPH. And compound 2 showed the significant riboflavin-and xanthine-originated superoxide quenching activities. To verify the antioxidant enzymatic activities of compound 2, the SOD enzymatic activity was measured spectrophtometrically using prepared Caenorhabditis elegans homogenates. The results showed that compound 2 was able to elevate SOD activity of C. elegans in a dose dependent manner. Moreover, compound 2 decreased the intracellular ROS accumulation of worms.

• Tuberculosis and Respiratory Diseases
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• 제41권3호
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• pp.215-221
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• 1994

연구배경 : 산소기가 여러 종류의 급성 폐손상에 중요한 역할을 한다는 것은 이미 잘 알려진 사실이다. 생체내에는 여러 항산화 방어기전이 존재하는데, SOD는 두 개의 superoxide radical이 과산화수소와 산소로 dismutation되는 과정을 $10^4$배 촉진시키는 효소로서 산소기에 대한 일차적인 방어기전으로 작용한다. Eukaryotic 세포내에는 두 가지 종류의 SOD가 존재하는데, 하나는 세포질에 위치하고 이중체(dimeric)의 구조를 가지며 구리와 아연을 포함하는 효소(CuZnSOD)이고, 또 하나는 미토콘드리아에 있고 사중체(tetrameric)의 구조를 갖는 망간을 포함하는 효소(MnSOD)이다. 내독소에 의한 백서의 급성 폐손상 모델에서 내독소 투여 후 시간 경과에 따른 백서 폐장의 MnSOD와 CuZnSOD의 유전자 발현을 관찰하여 이를 급성 폐손상의 양상과 비교하고자 본 연구를 시행하였다. 방법 : 백서에 E. coli의 내독소를 투여한 후 0, 1, 2, 4, 6, 12, 18, 그리고 24시간 후에 백서를 희생시켜 폐장을 얻은 후 폐장의 총 RNA를 single step phenol extraction 방법으로 추출하였다(n=3, respectively). 총 RNA를 formaldehyde를 함유한 1.2% agarose gel 에 전기영동하고, gel의 RNA를 nylon membrane으로 transfer시켰다. Nylon membrane을 $^{32}P$로 labeling시킨 MnSOD와 CuZnSOD를 probe로 하여 hybridization하고 autoradiography를 시행하였다. 결과 : 내독소률 투여하고 4시간후부터 MnSOD mRNA가 발현되기 시작하여 6시간에 최고치를 보였고, 약 12시간까지 지속되었으며, 24시간이 경과한 후에는 대조군의 수준으로 감소되었다. CuZnSOD 유전자는 내독소를 투여하고, 1 시간후부터 발현되기 시작하여 24시간까지 지속되었는데, 18시간에 최고치에 도달하였다. 결론 : 이상의 결과는 SOD가 백서에서 내독소에 의해 유발된 급성 폐손상에 대한 방어기전에 관여 할 가능성을 시사하는 것으로 생각된다.