사람唾液 Superoxide Dismutase의 精製 및 性狀

이봉재,이병래,전현우,양종대,이근배 朝鮮大學校 附設 醫學硏究所 1986 The Medical Journal of Chosun University Vol.11 No.1

The purification and some properties of superoxide dismutase(O_(2-)-O_(2-) oxidoreductase, ECI. 15.1.1) of human saliva have been studied. Human saliva was collected from medical students, male and female together with smoking male students. The activity of superoxide dismutase was measured by the method of McCord and Fridovich. The superoxide dismutase was purified by ammonium sulfate precifitation, DEAE-cellulose and sephadex C-100 column chromatography. The results are summerized as follows : 1. Human saliva contain measurable amounts of superoxide dismutase. 2. The activities of superoxide dismutase. were 0.72U/㎖ in male and 0.73U/㎖ in female saliva. The activity of this enzyme was 0.81U/㎖ in smoker. Accodingly, there was no recognizabIe differences in the enzyme activity between smoker and non-smoker. 4. The human salivary superoxide dismutase was purified approximately 90 folds, and the molecular weight of the purified enzyme was estimated to be approximately 32,000 by gel filtretion. 4. Purified salivary superoxide dismutase has contained 1.83 atoms of copper and 2.00 atoms of zinc per molecule and was strikingly similar to the superoxide dismutase previonsly isolated from other cukaryotes by other investigatury. 5. The ultraviolet absorption spectrum of purified salivan- superoxide dismutase was 268㎚ which was similar to the value previously seen with the cytosolic enzyme of eukaryotes.

간질(Fasciola hepatica)성층의 Superoxide Dismutase정제와 부분 특성 : Purification and Partial Characterization of Superoxide Dismutase from Fasciola hepatica Adult Worms

강은영,송철용,나동진 중앙대학교 기초과학연구소 1991 基礎科學硏究所 論文集 Vol.5 No.-

간질(F. hepatica)성충에서 superoxide dismutase(SOD)를 ammonium sulfate 침전, DEAE-Trisacryl Mcolumn chromatography와 UItrogel AcA 54 column chromatography를 실시하여 순수 정제하고, 이들 효소의 활성도를 Noritaka et al.(l982)의 방법으로 활성도를 측정하였다. 그리고 이들 효소의 특성을 관찰하여 다음과 같은 결과들을 얻었다. 1. 간질(F hepatica)의 세포질에는 구리와 아연을 함유한 superoxide dismutase가 존재하며, 이들의 비활성도는 3.6mg 이었다. 2. 순수 정제된 superoxide dismutase의 비활성도는 101.9mg 이었으며, 분자량은 Ultrogel AcA 54 gel filtrations으로 측정한 결과 34Kda. 이었고, SDS-PAGE에 의한 subunit의 분자량은 17Kda.으로서 dimer로 구성된 효소이었다. 3. 순수 정제된 효소는 3mM cyanide 농도에서 효소의 활성이 100%억제되었고, 20mM azide 농도에서는 23%억제되었다. 4.정제된 효소는 완충용액의 pH가 10.0에서 효소활성이 4.7배 증가하였다. The major superoxide dismutase of the Fasciola hepatica adult worms has been purified to homogeneity. The purification procedures included ammonium sulfate precipitation, DEAE-Trisacryl M column chromatography and AcA 54 gel filtration column chromatography. The activity of superoxide dismutase was measured by the method of Noritada et al.(1982). The superoxide dismutase with native molecular weight of approximately 34Kda. was purified from crude extract using AcA 54 gel filtration column chromatography. By SDS-polyacrylamide gel electrophoresis, the molecular weight of enzyme was estimated to be 17Kda. And so, it was composed of two subunits of equal size, which were not covalently joined. The activity of superoxide dismutase was inhibited 49.6% and 100% at 0.5mM and 3.0mM cyanide, respectively. But 10mM and 20mM azide caused 24.7% and 27% inhibition. The activity of this enzyme was appeared 4.7 times more active when the buffer pH raised to 10.0.

Streptomyces subrutilus P5가 생산하는 철 함유 superoxide dismutase의 분비

박재승,김재헌,Park, Jae-seung,Kim, Jae-heon 한국미생물학회 2015 미생물학회지 Vol.51 No.2

본 연구에서는 Streptomyces subrutilus P5의 생장과 세포내 외 철 함유 superoxide dismutase 활성을 비교 분석하여 철함유 superoxide dismutase의 분비 시점을 확인하고 분자 수준에서 이 효소의 분비에 관여하는 유전정보를 확인하고자 하였다. Streptomyces subrutilus P5의 균체 생장은 건체 중량을 측정하여 결정하였다. Glucose는 log phase에서 급격히 소모되어 24시간 후에 이르러 완전히 고갈되었다. 세포내의 철 함유 superoxide dismutase는 배양 후 3시간에 나타나며 세포외 철 함유 superoxide dismutase는 배양 후 7.5시간부터 나타난다. 따라서 superoxide dismutase는 용균에 의해서가 아니라 능동적인 분비기작에 의해서 세포 외로 분비된 것으로 추측할 수 있다. Streptomyces subrutilus P5의 sodF에는 signal peptide 유전정보가 존재하지 않았다. 그러나 sodF의 상류지역에서 다른 세균의 type III 분비단백질 유전자와 유사한 type III 분비상자가 발견되었다. Streptomyces 균주에서 type III 분비단백질이 존재할 가능성이 있음을 처음으로 제시하였다. 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.

흰쥐 담즙울체간의 Superoxide Dismutase의 활성

박소경,곽춘식 啓明大學校 醫科大學 1999 계명의대학술지 Vol.18 No.4

The activity change of superoxide dismutase and its mechanism in rat cholestatic liver were evaluated. The activity of superoxide dismutase was measured in the liver which obtained from common bile duct ligated rats (CBDL) and sham operated rats, respectively. And mRNA expression of this enzyme was also measured by reverse transcription-polymerase chain reaction in each group. The activity of superoxide dismutase was higher in the CBDL group, but mRNA expression of superoxide dismutase was similar to the sham operated group. These results indicate that increased activity of superoxide dismutase in the cholestatic liver was due to the increased catalytic activity for this enzyme. The increase of superoxide dismutase activity may result in hepatic damage in the cholestasis by activating hydrogen peroxide release.

광화학적으로 제조된 Superoxide Radical을 이용한 Superoxide Dismutase를 생산하는 해양미생물의 탐색 및 효소정제

조기웅 한국미생물학회 2002 미생물학회지 Vol.38 No.2

해양세균으로부터 superoxide dismutase를 생산하는 균주를 광화학적 superoxide ion 형성 및 nitrite정량방법으로 검색하여 고활성 균주 B-446을 확보하고 본 균주로부터 35-75% ammonium sulfate precipitation, DEAE-Sephadex A-25 ion exchange chromatography, Sephadex C-200 gel filtration chromatography, High-Q anion exchange chromatography그리고 HPLC-GPC를 이용하여 수율 6%, 정제도 32.3 배의 정제된 SOD를 얻었으며 정제 과정중 효소활성 분석에 본 광화학적 superoxide ion생성 방법으로 사용하였다. A marine bacterium producing superoxide dismutase, strain number B446, was screened with nitrite quantitation method using hydroxy amine and photochemically generated superoxide ion, and the superoxide dismutase was purified through 35-75% ammonium sulfate precipitation, DEAE-Sephadex A-25 ion exchange chromatography, Sephadex G-200 gel filtration chromatography, and High-Q anion exchange chromatography to a yield of 6% and purification fold of 32.3.

임신성 고혈압 임신부에서 지질과산화 및 항산화제 활성도

이종섭 ( Jong Seob Lee ),한도규 ( Do Gyu Han ),정우연 ( Woo Youn Jung ),이영기 ( Young Gi Lee ),이두진 ( Doo Jin Lee ),이승호 ( Sung Ho Lee ),박윤기 ( Yoon Ki Park ) 대한주산의학회 2002 Perinatology Vol.13 No.1

연구목적:임신 제3삼분기에서 임신성 고혈압의 병인론으로 제기되는 산화제와 항산화제의 활성도를 평가하기 위하여 정상 임신 및 임신성 고혈압 임신부의 혈액에서 산화계는 유리기에 의한 지질과산화의 정도로 malondialdehyde, 유리기의 제거에 관여하는 항산화계는 항산화효소인 superoxide dismutase와 총 항산화제의 활성을 측정, 비교 평가하고자 하였다. 방법:21명의 건강한 정상 임신부를 대조군으로, 22명의 건강한 비 임신부와 18명의 임신성고혈압 임신부를 대상군으로 하여 임신 제 3 삼분기에서 혈액을 채취하여 산화계는 지질과산화산물인 malondialdehyde, 항산화계는 superoxide dismutase와 총 항산화제를 측정하여 비교하였다. 고혈압이 지질과산화에 미치는 영향을 알기 위하여 임신부의 malondialdehyde 농도와 수축기 및 이완기 혈압과의 상관관계를 조사하였다. 결과:Malondialdehyde는 임신성 고혈압군에서 정상 임신군보다 유의하게 증가하였다(p<0.05). Superoxide dismutase는 임신성 고혈압군에서 정상 임신군보다 유의하게 감소하였으나(p<0.05), 총 항산화제는 양 군간에 유의한 차이가 없었다. 임신부에서 혈중 malondialdehyde와 수축기 및 이완기 혈압 사이에서 높은 양의 상관관계가 있었다(r=0.475, r=0.546, p<0.05). 결론:임신성 고혈압에서 지질과산화의 최종산물인 malondialdehyde 증가와 항산화효소 특히 superoxide dismutase의 감소가 임신성 고혈압의 병태생리에 관여한다고 생각된다. Objective:To assess maternal circulating levels of lipid peroxides, superoxide dismutase and total antioxidants of women with pregnancy-induced hypertension. Methods:Cross-sectional study consisting of 18 preeclamptic, 21 uncomplicated pregnant and 22 healthy non-pregnant women. Fasting venous blood samples were collected during the 3rd trimester of antepartum period and maternal circulating levels of malondialdehyde as a lipid peroxidation product, superoxide dismutase activity and total antioxidants were measured. Results:In the preeclamptic group, lipid peroxides were significantly increased, otherwise the activity of superoxide dismutase in the erythrocytes was significantly decreased compared to normal pregnant women. The value of serum total antioxidants was similar in both groups. Strong correlation was detected between malondialdehyde and blood pressure in the pregnant women. Conclusion:Preeclampsia is associated with decrease of antioxidant enzyme activity while lipid peroxidation was increased during the 3rd trimester of pregnancy. Therefore, a significant elevated lipid peroxidation and reduced superoxide dismutase activity may contribute to pathophysiology and pathogenesis of preeclampsia via vascular endothelial cell damage.

Biomimetic Superoxide Disproportionation Catalyst for Anti-Aging Lithium-Oxygen Batteries

Hwang, Chihyun,Yoo, JongTae,Jung, Gwan Yeong,Joo, Se Hun,Kim, Jonghak,Cha, Aming,Han, Jung-Gu,Choi, Nam-Soon,Kang, Seok Ju,Lee, Sang-Young,Kwak, Sang Kyu,Song, Hyun-Kon American Chemical Society 2019 ACS NANO Vol.13 No.8

<P>Reactive oxygen species or superoxide (O<SUB>2</SUB><SUP>-</SUP>), which damages or ages biological cells, is generated during metabolic pathways using oxygen as an electron acceptor in biological systems. Superoxide dismutase (SOD) protects cells from superoxide-triggered apoptosis by converting superoxide to oxygen and peroxide. Lithium-oxygen battery (LOB) cells have the same aging problems caused by superoxide-triggered side reactions. We transplanted the function of SOD of biological systems into LOB cells. Malonic acid-decorated fullerene (MA-C<SUB>60</SUB>) was used as a superoxide disproportionation chemocatalyst mimicking the function of SOD. As expected, MA-C<SUB>60</SUB> as the superoxide scavenger improved capacity retention along charge/discharge cycles successfully. A LOB cell that failed to provide a meaningful capacity just after several cycles at high current (0.5 mA cm<SUP>-2</SUP>) with 0.5 mAh cm<SUP>-2</SUP> cutoff survived up to 50 cycles after MA-C<SUB>60</SUB> was introduced to the electrolyte. Moreover, the SOD-mimetic catalyst increased capacity, <I>e</I>.<I>g</I>., more than a 6-fold increase at 0.2 mA cm<SUP>-2</SUP>. The experimentally observed toroidal morphology of the final discharge product of oxygen reduction (Li<SUB>2</SUB>O<SUB>2</SUB>) and density functional theory calculation confirmed that the solution mechanism of Li<SUB>2</SUB>O<SUB>2</SUB> formation, more beneficial than the surface mechanism from the capacity-gain standpoint, was preferred in the presence of MA-C<SUB>60</SUB>.</P> [FIG OMISSION]</BR>

홍모기(Cufexpipienspoiffens)의 난성숙 과정중 Superoxide dismutase의 활성

임태진,이승훈 한국통합생물학회 1994 동물학회지 Vol.37 No.3

Culexpfpfens pollens에 존재하는 superoxide dismutase의 연구를 위한 적정 분석 조건과 우화 전후. 시간 경과에 따른 superoxide dismutase 활성 경향에 대해서 연구하였다. C. pipiens에 존재하는 superoxide dismutase의 활성은 부화 직후부터 지속적으로 감소하다가. 우화 후 흡혈 자극에 의해서 급격하게 증가한다 흡혈 후 36시간이 경과했을 때. 최대의 논성도를 보인 후 급격히 감소하였다. 기관별 분석에서 흡혈 후 가장 높은 활성 증가는 중장에서 일어나고, 머리 흥부. 지꼴체 및 난소에서도 활성의 증가가 나타났다. 흡혈 전에는 2개의 동위효소(SOD-1, soD-2)가 존재하였으나. 흡혈후 36시간된 성체에서는 3개의 동위효소 밴드가 보이는데, 머리에서는 SOD-1만이 나타나고, 가슴에서는 SOD-2. 지방체와 난소에서는 SOD-1과 SOD-2이 존재하였고. 중장에서는 SOD-3가 나타났다. 그러므로 흡혈 후 나타나는 새로운 동위효소는 중장에서 합성됨을 알 수 있다.

Localization of Superoxide dismutase (SOD) as a Possible Exogeneous Function

Kim, Young Gen 한국노화학회 1991 한국노화학회지 Vol.1 No.1

Generation of Neuroblastoma Cell Lines Overexpressing hCu,Zn-Superoxide Dismutase Genomic DNA

kwak,Hahn-Shik 培材大學校 1996 培材論叢 Vol.1 No.-

Cu,Zn-superoxide dismutase의 과잉생산이 다운 증후군의 병인론과 관계가 있음을 조사하기 위하여 hCu,Zn-SOD 유전자를 overexpress하는 transfected cell을 만들고자 하였다. pHG165 vector에 hCu,Zn-SOD genomic DNA를 집어넣어 만든 재조합 DNA를 NCB-20 neuroblastoma cell에 transfection 시키고 G418에 저항성이 있는 세포군을 골라 Cu,Zn-SOD의 활성을 측정하였고, polyacrylamide gel electrophoresis 후에 activity staining을 실시하였으며, 항체를 사용한 immunoblot을 실시하여 hCu,Zn-SOD를 overexpress하는 세포종을 얻었다. To investigate the possible involvement of hCu,Zn-superoxide dismutase overproduction in the etiology of the Down's Syndrome, we tried to generate the transfected cells overexpressing the hCu,Zn-superoxide dismutase(hCu,Zn-SOD) gene. The recombinant DNA which contains hCu,Zn-SOD genomic DNA in the pHG165 vector was constructed and transfected in the NCB-20 neuroblastoma cells. The transfected cells were selected for G418 resistance and analyzed by the Cu,Zn-SOD assay, the activity staining after polyacrylamide gel electrophoresis and the immunoblot detection with antibody. We could therefore generate several cell lines which overexpress hCu,Zn-SOD gene.