• 제목/요약/키워드: oxygen mutant

검색결과 85건 처리시간 0.024초

Control of Singlet Oxygen-induced Oxidative Damage in Escherichia coli

  • Kim, Sun-Yee;Kim, Eun-Ju;Park, Jeen-Woo
    • BMB Reports
    • /
    • 제35권4호
    • /
    • pp.353-357
    • /
    • 2002
  • Singlet oxygen ($^1O_2$) is highly reactive form of molecular oxygen that may harm living systems by oxidizing critical cellular macromolecules. The oxyR gene product regulates the expression of the enzymes and proteins that are needed for cellular protection against oxidative stress. In this study, the role of oxyR in cellular defense against a singlet oxygen was investigated using Escherichia coli oxyR mutant strains. Upon exposure to methylene blue and visible light, which generates singlet oxygen, the oxyR overexpression mutant was much more resistant to singlet oxygen-mediated cellular damage when compared to the oxyR deletion mutant in regard to growth kinetics, viability and protein oxidation. Induction and inactivation of major antioxidant enzymes, such as superoxide desmutase and catalase, were observed after their exposure to a singlet oxygen generating system in both oxyR strains. However, the oxyR overexpression mutant maintained significantly higher activities of anticxidant enzymes than did the oxyR deletion mutant. These results suggest that the oxyR regulon plays an important protective role in singlet oxygen-mediated cellular damage, presumably through the protection of antioxidant enzymes.

Streptomyces coelicolor의 연속 배양시 산소 분압에 따른 방어 효소의 활성 변화 (Effect of Partial Oxygen Pressure on the Growth and Defense Enzyme Activities of Streptomyces coelicolor in continuous culture system)

  • 박용두;이계준;노정혜
    • 한국미생물·생명공학회지
    • /
    • 제22권5호
    • /
    • pp.538-543
    • /
    • 1994
  • Effect of partial oxygen pressure on the cell growth and the activities of oxidative defense enzymes were measured in the continuous culture of Streptomyces coelicolor. Both the wild type and the mutant strain resistant to hydrogen peroxide were cultured and the dry cell weight of the two cultures were measured at different oxygen tensions. Growth of the wild type was inhibited by oxygen at above 0.5 vvm. Growth of the hydrogen peroxide resistant mutant was stimulated by pure oxygen at 0.5 vvm but was inhibited by oxygen at 1.0 vvm. Therefore, growth of the hydrogen peroxide resistant mutant was less affected by the deleterious oxidative stress of oxygen. Activities of the several defense enzymes were also measured at different oxygen tensions. Activities of catalase and glucose-6-phosphate dehydrogenase increased significantly as oxygen pressure increased in the wild type culture. In the mutant, however, increase in those enzyme activities was not obvious whereas the uninduced levels of the above enzymes were higher than those of wild type. As judged by Western blotting, the amount of the major catalase increased as the oxygen pressure increased. This indicates that the induction of the catalase activity by oxygen pressure is mostly due to the increase in the expression level for the major catalase.

  • PDF

Bifidobacterium longum 산소변이주의 분리와 변이주의 산소내성 (Isolation and Characterization of Oxygen-tolerant Mutant of Bifidobacterium longum.)

  • 안준배;김광엽;박종현
    • 한국미생물·생명공학회지
    • /
    • 제26권6호
    • /
    • pp.476-482
    • /
    • 1998
  • Bifidobacterium의 효과적인 산업적 이용을 위해서는 산소에 내성이 강한 균주를 선발하는 연구뿐만 아니라, 산소 존재하에서 비피도박테리아의 내성기작을 규명하는 기초적인 연구도 필요하다. 산소에 민감한 B. adolescentis 균주와 산소내성 B. longum JI-1균주의 용존산소 제거능을 비교한 결과산소 민감성 균주는 총존산소를 전혀 제거하지 못하였으나 산소내성 균주는 10분 이내에 3%이상의 용존산소를 제거하여 총존산소 제거능과 Bifidobacterium의 산소내성과는 밀접한 관계가 있음을 알 수 있었다. 이 B. longum JI-1 균주를 미호기성 조건에서 배양하여 내산소성 돌연변이 균주인 B. longum ADJ-1 균주를 분리하였다. B. longum ADJ-1과 모균주의 특성을 당자화능, NADH 산화효소 및 세포지방산등으로 비교하였을 때 차이점을 발견하지 못했으며, 균생육은 B. longum ADJ-1이 모균주의 80% 정도를 보여 주었다. 그러나 B. longum ADJ-1는 매우 두꺼운 slime layer를 형성하였는데 confocal scanning laser microscopy에 의한 분석 결과 돌연변이 균주는직경이 약 6 $\mu\textrm{m}$에 이르는 층을 형성한 반면 모균주는 약 3 $\mu\textrm{m}$인 층을 형성하였다. 그리고 돌연변이 균주는 산소에 24시간 이상 노출되었을 경우 모균주에 비하여 더 큰 산소내성을 보여주었다. 그러므로 돌연변이 균주의 산소내성의 증가는 slime layer 차이에서 유래된 것으로 생각되며 이는 Bifidobacterium의 내산소성 기작중의 하나인 것으로 보인다.

  • PDF

Survival Factor Gene FgSvf1 Is Required for Normal Growth and Stress Resistance in Fusarium graminearum

  • Li, Taiying;Jung, Boknam;Park, Sook-Young;Lee, Jungkwan
    • The Plant Pathology Journal
    • /
    • 제35권5호
    • /
    • pp.393-405
    • /
    • 2019
  • Survival factor 1 (Svf1) is a protein involved in cell survival pathways. In Saccharomyces cerevisiae, Svf1 is required for the diauxic growth shift and survival under stress conditions. In this study, we characterized the role of FgSvf1, the Svf1 homolog in the homothallic ascomycete fungus Fusarium graminearum. In the FgSvf1 deletion mutant, conidial germination was delayed, vegetative growth was reduced, and pathogenicity was completely abolished. Although the FgSvf1 deletion mutant produced perithecia, the normal maturation of ascospore was dismissed in deletion mutant. The FgSvf1 deletion mutant also showed reduced resistance to osmotic, fungicide, and cold stress and reduced sensitivity to oxidative stress when compared to the wild-type strain. In addition, we showed that FgSvf1 affects glycolysis, which results in the abnormal vegetative growth in the FgSvf1 deletion mutant. Further, intracellular reactive oxygen species (ROS) accumulated in the FgSvf1 deletion mutant, and this accumulated ROS might be related to the reduced sensitivity to oxidative stress and the reduced resistance to cold stress and fungicide stress. Overall, understanding the role of FgSvf1 in F. graminearum provides a new target to control F. graminearum infections in fields.

Major Fe-Superoxide Dismutase (FeSOD) Activity in Pseudomonas putida is Essential for Survival Under Conditions of Oxidative Stress During Microbial Challenge and Nutrient Limitation

  • Kim, Young-Cheol;Kim, Cheol-Soo;Cho, Baik-Ho;Anderson, Anne-J.
    • Journal of Microbiology and Biotechnology
    • /
    • 제14권4호
    • /
    • pp.859-862
    • /
    • 2004
  • An isolate of Pseudomonas putida has been found to aggressively colonize root tips and induce plant resistance to Fusarium wilt. However, P. putida mutants lacking Fe-superoxide dismutase (SOD) or both FeSOD and MnSOD activities are less competitive in root tip colonization. In the current study, the growth of an FeSOD mutant was found to be more sensitive than that of the wild-type or a MnSOD mutant to oxidative stress imposed by paraquat treatment and culturing with the soil fungus Talaromyces flavus, which generates reactive oxygen species. Also, the loss of culturability with an aging stationary-phase culture was greater for a double SOD mutant than an FeSOD mutant, while no reduction in culturability was observed with the wild-type and a MnSOD mutant under the same protracted stationary-phase conditions. Accordingly, it was concluded that FeSOD activity is the major form of SOD in P. putida and plays an essential role in survival under stress conditions when increased oxidative stress is encountered.

고용존산소 농도에서의 산소내성 Pseudomonas aeruginosa 돌연변이주 M-10 생장거동 (Characterization of Pseudomonas aeruginosa Growth under High Dissolved Oxygen Concentration : Selection of the High Dissolved Oxygen Resistant Mutant pseudomonas aeruginosa M-10)

  • 이항우
    • 미생물학회지
    • /
    • 제35권4호
    • /
    • pp.283-288
    • /
    • 1999
  • 본 연구를 통하여 활성슬러지중의 주요 세균인 Pseudomonas aeruginosa를 이용한 생장 특성과 세포 생장에 있어서의 고DO 농도의 영향에 대한 특성을 이용, 페수처리계의 응용성을 조사하였다. 그 결과, 본 균은 DO 농도의 증가에 따라서 세포 수율은 14배 정도 감소되는 것으로 나타났다. 또한 세포 수율의 감소는 압력의 증가에 의한 것이 아니라 DO 농도의 증가에 의한 것으로 나타났다. 이러한 사실을 바탕으로 낮은 DO 농도에서도 세포 수율의 감소효과를 가질 수 있는 변이주를 변이원 NTG 농도 100 ${\mu}$g/㎖, 30${\circ}C$, 1시간 처리하여 얻은 다음 D-cycloserine에 의한 농축배양을 통해 목적 변이주를 얻었다. 이 목적 변이주에 대한 세포 수율은 DO 농도의 증가에 따라 감소하는 경향을 나타내었으며 그 수율은 DO 농도 36 ppm에서 야생주에 비교할 때, 약 55%의 감소율을 나타내는 것으로 보아 종래의 폭기방식을 채용할 경우에도 슬러지량을 충분히 감소시킬 수 있을 것으로 기대된다.

  • PDF

Isolation and characterization of Vitreoscilla mutant defective in catalase-peroxidase hydroperoxidase I

  • Kim, Hee-Jung;Moon, Ja-Young;Lee, John-Hwa;Park, Kie-In
    • 한국동물위생학회지
    • /
    • 제30권3호
    • /
    • pp.291-304
    • /
    • 2007
  • Mutants of an obligate aerobic bacterium, Vitreoscilla, that have deficiency in heat-labile catalase-peroxidase hydroperoxidase I (HPI) were created by EMS treatment. The catalase-peroxidase HPI-deficient mutant showed substantially lower peroxidase activity in exponential and mid-stationary phase compared with the wild type strain. In late stationary phase, the mutant exhibited no peroxidase activity. Peroxidase deficiency in the mutant was revealed by polyacrylamide gels stained for peroxidase activity. Characteristically, catalase levels in the mutant increased about 14- and 8-fold during growth in the exponential and stationary phases, respectively, compared to those in the wild type, suggesting a compensatory effect for protection from $H_2O_2$ toxicity. The mutant showed differences in physiology from the wild type: retardation in growth rate and decrease in oxygen consumption. Both the wild type and the catalase-peroxidase HPI-deficient mutant of Vitreoscilla had lower growth rates in media containing increasing $H_2O_2$ concentrations. However, the mutant exhibited an additionally decreased growth rate after 6 to 8 h of growth compared to the wild type. The wild type was resistent up to 20 mM $H_2O_2$, whereas the mutant was very sensitive to high concentrations of exogenous $H_2O_2$. Although elevated catalase levels would provide protection of the bacteria from the deleterious effect of $H_2O_2$, it did not appear to be complete. Cell-free extracts of the mutant showed decreased NADH oxidation rates and higher accumulation of $H_2O_2$ during this oxidation. These results may account for the impaired growth and earlier onset of death phase by the catalase-peroxidase HPI-deficient mutant of Vitreoscilla.

Protective Role of Thioredoxin Peroxidase Against Ionizing Radiation

  • Lee, Su-Min;Kim, Sun-Yee;Park, Jeen-Woo
    • BMB Reports
    • /
    • 제31권6호
    • /
    • pp.572-577
    • /
    • 1998
  • A soluble protein from Saccharomyces cerevisiae provides protection against a thiol-containing oxidation system but not against an oxidation system without thiol. This 25-kDa protein acts as a peroxidase but requires the NADPH-dependent thioredoxin system or a thiol-containing intermediate, and was thus named thioredoxin peroxidase. The protective role of thioredoxin peroxidase against ionizing radiation, which generates reactive oxygen species harmful tocellular function, was investigated in wild-type and mutant yeast strains in which the tsa gene encoding thioredoxin peroxidase was disrupted by homologous recombination. Upon exposure to ionizing radiation, there was a distinct difference between these two strains in regard to viability and the level of protein carbonyl content, which is the indicative marker of oxidative damage to protein. Activities of other antioxidant enzymes, such as catalase, superoxide dismutase, glucose-6-phosphate dehydrogenase, and glutathione reductase were increased at 200-600 Gy of irradiation in wild-type cells. However, the activities of antioxidant enzymes were not significantly changed by ionizing radiation in thioredoxin peroxidase-deficient mutant cells. These results suggest that thioredoxin peroxidase acts as an antioxidant enzyme in cellular defense against ionizing radiation through the removal of reactive oxygen species as well as in the protection of antioxidant enzymes.

  • PDF

A NOVEL PHOTOHETEROTROPHIC MUTANT FOR psaB GENE OF Synechocystis sp. PCC 6803 GENERATED FROM TARGETED MUTAGENESIS

  • Kim, Soohyun;Kim, Seung-Il;Choi, Jong-Soon;Chung, Young-Ho;Chun, Soon-Bai;Park, Young-Mok
    • Journal of Photoscience
    • /
    • 제3권1호
    • /
    • pp.23-28
    • /
    • 1996
  • To investigate the structure and function of photosystem I, cartridge mutagenesis technique was used to inactivate the psaB gene of photosystem I. From the screen, many strains which have potential defects in photosystem I were generated. Biochemical analysis revealed that B2, one of the mutant, had a reduced amount of chlorophyll. Electron transfer activitx from photosystem II to photosystem I as oxygen uptake was the rate of 64 % of wild type. Also B2 showed a decreased photosystem I activity when measured by 77 K fluorescence emission spectrum. Particularly, immunodetection analysis showed that the B2 had reduced amount of PsaA/PsaB, but a normal range of PsaC and PsaD. Here we present a photoheterotrophic mutant for psaB gene as a unique model strain for future study of structural/functional relationship and biogenesis of photosystem I.

  • PDF