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1.
Parallel increases in intracellular catalase activity and resistance to extracellular H2O2 and to hyperbaric O2 toxicity were observed when Bacteroides distasonis VPI 4243 (ATCC 8503, type strain) was grown in either complex or defined medium containing graded amounts of hemin. Virtually all of the cells with high catalase activity (greater than 200 U/mg) remained viable upon exposure at 37 degrees C to 100-lb/in2 O2 on agar surfaces for 1 h, whereas low-catalase cells (less than 10 U/mg) lost 1.2 log units of viable cells during that treatment. Upon exposure to 500 microM H2O2, high-catalase cells lost 0.4 log units of the initial viable colonies during the same period in which low-catalase cells lost 3 log units of viable cells. The superoxide dismutase activity was the same in each test culture. These data support the role of intracellular catalase in protecting B. distasonis from oxidative damage resulting from hyperbaric oxygenation or H2O2 exposure. Catalase activity elicited by adding hemin to cells grown previously in medium lacking hemin was inhibited only 40% by prior incubation of the cells with chloramphenicol (30 micrograms/ml) and only 22% with rifampin (5 micrograms/ml). A model which is consistent with these data involves the production of an apocatalase in cells grown in low-hemin medium. Addition of hemin to the cells would result in a rapid chloramphenicolor rifampin-insensitive stimulation of catalase activity followed by further de novo biosynthesis of catalase. 相似文献
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Eremin AN 《Prikladnaia biokhimiia i mikrobiologiia》2001,37(1):53-62
For preparationing the polyenzyme antioxidant complex, containing superoxide dismutase (SOD), catalase and horseradish peroxidase (HRP), the different successivities of those enzymes co-immobilization were compared. The optimum successivity is provided by simultaneous co-immobilization of covalently bound HRP with the SOD and catalase. The catalytic enzyme activity and the catalase operational stability was kinetically characterized in various samples. For one sample, the influence of ascorbate, glutathione and ethanol on the catalase kinetic parameters was studied. A possible scheme of different processes at the H2O2 decomposition in the presence of co-immobilized SOD, catalase, HRP and the substrates-reductans was discussed. 相似文献
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Characterization of the O2-induced manganese-containing superoxide dismutase from Bacteroides fragilis 总被引:6,自引:0,他引:6
E M Gregory 《Archives of biochemistry and biophysics》1985,238(1):83-89
A manganese-containing superoxide dismutase (MnSOD) has been isolated from extracts of O2-induced Bacteroides fragilis. The enzyme, Mr 43,000, was a dimer composed of noncovalently associated subunits of equal size. A preparation whose specific activity was 1760 U/mg had 1.1 g-atoms Mn, 0.3 g-atoms Fe, and 0.2 g-atoms Zn per mol dimer. Exposing the enzyme to 5 M guanidinium chloride, 20 mM 8-hydroxyquinoline abolished enzymatic activity. Dialysis of the denatured apoprotein in buffer containing either Fe (NH4)2(SO4)2 or MnCl2 restored O2-. scavenging activity. The iron-reconstituted enzyme was inhibited 89% by 2 mM NaN3, similar to other Fe-containing superoxide dismutases. The Mn-reconstituted and native MnSOD were inhibited approximately 50% by 20 mM NaN3. Addition of ZnSO4 to dialysis buffer containing either the iron or manganese salt inhibited restoration of enzymatic activity to the denatured apoprotein. MnSOD migrated as a single protein band coincident with a single superoxide dismutase activity band in 7.5 or 10% acrylamide gels. Isoelectric focusing resulted in a major isozymic form with pI 5.3 and a minor form at pI 5.0. Mixtures of the MnSOD and the iron-containing superoxide (FeSOD), isolated from anaerobically maintained B. fragilis [E. M. Gregory and C. H. Dapper (1983) Arch. Biochem. Biophys. 220, 293-300], migrated as a single band on acrylamide gels and isoelectrically focused to a major protein band (pI 5.3) and a minor band at pI 5.0. The amino acid composition of MnSOD was virtually identical to that of the FeSOD. The data are consistent with synthesis of a single superoxide dismutase apoprotein capable of accepting either Mn or Fe to form the holoenzyme. 相似文献
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Coisolation of glutathione peroxidase, catalase and superoxide dismutase from human erythrocytes 总被引:1,自引:0,他引:1
Glutathione peroxidase (GSH-Px; glutathione: hydrogen peroxide oxidoreductase; EC 1.11.1.9), catalase (H2O2: H2O2 oxidoreductase; EC 1.11.1.6) and superoxide dismutase (superoxide: superoxide oxidoreductase; EC 1.15.1.1) were coisolated from human erythrocyte lysate by chromatography on DEAE-cellulose. Glutathione peroxidase was separated from superoxide dismutase and catalase by thiol-disulfide exchange chromatography and then purified to approximately 90% homogeneity by gel permeation chromatography and dye-ligand affinity chromatography. Catalase and superoxide dismutase were separated from each other and purified further by gel permeation chromatography. Catalase was then purified to approximately 90% homogeneity by ammonium sulfate precipitation and superoxide dismutase was purified to apparent homogeneity by hydrophobic interaction chromatography. The results for glutathione peroxidase represent an improvement of approximately 10-fold in yield and 3-fold in specific activity compared with the established method for the purification of this enzyme. The yields for superoxide dismutase and catalase were high (45 mg and 232 mg, respectively, from 820 ml of washed packed cells), and the specific activities of both enzymes were comparable to values found in the literature. 相似文献
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The effects of superoxide dismutase (SOD) and catalase on the autoxidation rate of L-ascorbic acid (ASA) in the absence of metal ion catalysts were examined. The stabilization of ASA by SOD was confirmed, and the enzyme activity of SOD, which scavenges the superoxide anion formed during the autoxidation of ASA, contributed strongly to this stabilization. The stabilization of ASA by catalase was observed for the first time; however, the specific enzyme ability of catalase would not have been involved in the stabilization of ASA. Such proteins as bovine serum albumin (BSA) and ovalbumin also inhibited the autoxidation of ASA, therefore it seems that non-specific interaction between ASA and such proteins as catalase and BSA might stabilize ASA and that the non-enzymatic superoxide anion scavenging ability of proteins might be involved. 相似文献
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Fimbriae from the oral anaerobe Bacteroides gingivalis: physical, chemical, and immunological properties. 总被引:15,自引:3,他引:12
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F Yoshimura T Takasawa M Yoneyama T Yamaguchi H Shiokawa T Suzuki 《Journal of bacteriology》1985,163(2):730-734
Circular dichroism spectra indicated the predominance of beta-sheet structure in Bacteroides gingivalis fimbriae regardless of the presence of sodium dodecyl sulfate. By using a computer program, the alpha-helix, beta-sheet, and beta-turn contents and the remainder were estimated to be 0, 55, 18, and 27%, respectively, judging from the circular dichroism spectra of the fimbriae. Heating for 5 min at 100 degrees C in sodium dodecyl sulfate was necessary to denature the fimbriae into their constituent protein (fimbrilin) monomers with a reduced content of beta-sheet structure. The amino-terminal amino acid sequence of the fimbrilin was different from partial or complete amino acid sequences of fimbrilins so far determined from Bacteroides nodosus, which falls into the same nonfermentative species of the genus Bacteroides as B. gingivalis, and from various other bacteria. Fimbrilin monomers had an isoelectric point of 6.0. Examination of antibodies against fimbriae and sodium dodecyl sulfate-denatured fimbrilin by enzyme-linked immunosorbent assay reinforced a previous notion (F. Yoshimura, K. Takahashi, Y. Nodasaka, and T. Suzuki, J. Bacteriol. 160:949-957, 1984) that different sets of antigenic determinants seemed to be exposed on their surfaces. 相似文献
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CuZn superoxide dismutase, Mn superoxide dismutase, catalase and glutathione peroxidase in glutathione-deficient human fibroblasts 总被引:1,自引:0,他引:1
The effect of genetically determined glutathione deficiency on the fibroblast content of CuZn superoxide dismutase, Mn superoxide dismutase, catalase and glutathione peroxidase was investigated. No significant differences between glutathione-deficient and -proficient human fibroblasts were revealed. There was a large variation in the content of the investigated enzymes in fibroblasts grown and analysed on different occasions. Whereas the contents of CuZn superoxide dismutase, catalase and glutathione peroxidase did not deviate much from what has been found in other human cell-lines and tissues, the fibroblasts were found to contain exceptional amounts of Mn superoxide dismutase. 相似文献
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The primary structure of superoxide dismutase purified from anaerobically maintained Bacteroides gingivalis 总被引:5,自引:0,他引:5
The superoxide dismutase (SOD) of Bacteroides gingivalis can use either iron or manganese as a cofactor in its catalytic activity. In this study, the complete amino acid sequence of this SOD purified from anaerobically maintained B. gingivalis cells was determined. The proteins consisted of 191 amino acid residues and had a molecular mass of 21,500. The sequence of B. gingivalis SOD showed 44-51% homology with those for iron-specific SODs (Fe-SODs) and 40-45% homology with manganese-specific SODs (Mn-SODs) from several bacteria. However, this sequence homology was considerably less than that seen among the Fe-SOD (65-74%) or Mn-SOD family (42-60%). This indicates that B. gingivalis SOD, which accepts either iron or manganese as metal cofactor, is a structural intermediate between the Fe-SOD and Mn-SOD families. 相似文献
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Both superoxide dismutase (SOD) and catalase are key enzymes in the antioxidant system of the cells that work to maintain low steady-state concentrations of the reactive oxygen species. When exposed to a singlet oxygen-producing system composed of dye, such as methylene blue or rose bengal, and visible light both SOD and catalase were susceptible to oxidative modification and damage as indicated by the loss of activity, fragmentation and aggregation of peptide as well as by the formation of carbonyl groups. Histidine, a powerful quenching agent for singlet oxygen, and the polyamines, such as spermine and spermidine, were effective at protecting the activity loss mediated by illuminated dye, whereas spin traps were only mildly effective. The structural alterations of modified enzymes were indicated by the increase in susceptibility to proteases, the change in absorption spectra and in fluorescence spectra. The singlet oxygen-mediated damage to SOD and catalase may result in the perturbation of cellular antioxidant defense mechanisms and subsequently lead to a pro-oxidant condition. 相似文献
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1. Activities of superoxide dismutase (superoxide: superoxide oxidoreductase, EC 1.15.1.1) have been estimated in eye tissues. In rabbit eye, superoxide dismutase is present in corneal epithelium, corneal endothelium, lens, iris, ciliary body and retina. In lens the activity is in capsule epithelium. 2. Copper chelator diethyldithiocarbamate inhibited lens superoxide dismutase in vitro and in vivo in rabbit. 3. H2O2 caused inhibition of superoxide dismutase activity of lens extract, and this inhibition was potentiated by the catalase inhibitor 3-amino-1H-1,2,4-triazole (3-aminotriazole) or NaN3. 3-Aminotriazole or NaN3 had no effect on lens superoxide dismutase. Thus endogenous catalase of lens affords protection to the lens superoxide dismutase from inactivation by H2O2. 4. In rabbit having early cataract (vacuolar stage) induced by feeding-3-aminotriazole, there was a decrease in superoxide dismutase of lens, a fall in ascorbic acid of ocular humors and lens, and a 2--3-Fold increase in H2O2 of aqueous humor and vitreous humor. We conclude that catalase of eye affords protection to the lens from H2O2 and it also protects superoxide dismutase of lens from inactivation by H2O2. Superoxide dismutase, in turn, protects the lens from the superoxide radical, O2.-. It is likely that inhibition of these enzymes may lead to production of the highly reactive oxidant, the hydroxyl radical, under pathological conditions when H2O2 concentration in vivo exceeds physiological limits as in cataract induced by 3-aminotriazole. A scheme of reaction mechanism has been proposed to explain the relative functions of ocular catalase and superoxide dismutase. Such a mechanism may be involved in cataractogenic process in the human. 相似文献
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Methods have been developed for the measurements of catalase and superoxide dismutase (SOD) in single, isolated muscle fibers. These fibers are also classified according to fiber type. Catalase is determined using a fluorescent method for the measurement of hydrogen peroxide consumed. SOD measurements are carried out using a modification of established techniques whereby the inhibition of oxidation of epinephrine by SOD is assayed fluorometrically. Both enzymes may be determined in submicrogram samples of dried muscle. This approach avoids the complication of the inclusion of nonmuscle tissue with varying enzymatic activities which is frequently experienced when using homogenates of muscle, particularly diseased muscle. In addition, these techniques can be used to determine the inherent variation in SOD and catalase activities within individual fibers of the same fiber type. The Km and Vmax for catalase, determined using homogenates of human muscle, were found to be 12 mM and 1.45 mumol/min/mg dry wt, respectively. Catalase of muscle was inhibited 50% by 2 microM sodium azide. Mn-SOD contributes less than one-fifth of the total SOD activity. Therefore the activity is largely due to the Cu-Zn form of SOD. These methods are applicable to a wide variety of tissues. 相似文献
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Superoxide anion production and superoxide dismutase and catalase activities in Coxiella burnetii. 总被引:6,自引:0,他引:6
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Coxiella burnetii was examined for superoxide anion (O2-) production and superoxide dismutase and catalase activities. The organism generated O2- at pH 4.5 but not at pH 7.4. The rickettsia displayed superoxide dismutase activity distinguishable from that of the host cell (L-929 mouse fibroblast). Catalase activity was maximal at pH 7.0 and diminished at pH 4.5. These enzymes may account, in part, for the ability of this obligate intracellular parasite to survive within phagocytes. 相似文献
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The polarographic method of catalytic currents applied to a wave of oxygen permits the simultaneous assay of superoxide dismutase and catalase in biological materials with high speed and reproducibility and minimal manipulation of tissues. Washed red blood cells and tissue homogenates give rise to a strong polarographic maximum, apparently due to heme proteins, which interferes with the measurement. This maximum is suppressed by addition of approximately 0.2% plasma. Therefore, the determination of the two enzymes in red blood cells can be carried out by direct addition of whole blood to the polarographic solution. Thirty microliters of blood are enough for optimal determination of both enzymes. The method can determine superoxide dismutase and catalase at concentrations as low as 2 × 10−11m and 5 × 10−10m, respectively, and shows a linear correlation between measured activity and enzyme levels. The average values of the two enzymes in human red blood cells was found by this method to be 2.6 × 10−6m for catalase and 1.8 × 10−6m for superoxide dismutase, which agree with previously reported values. 相似文献
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Superoxide dismutase (EC 1.15.1.1) has been isolated and characterised from the blue-green alga Spirulina platensis and from aerobically-grown Rhodopseudomonas spheroides, a purple, non-sulphur bacterium. The former enzyme contains 1 gatom of iron and the latter 1 gatom of manganese per mol; both enzymes have a molecular weight of 37 000-38 000, being composed of two non-covalently joined subunits of equal size. Various spectral studies have been carried out including absorbance, circular dichroism and electron spin resonance. Catalytic activity has been studied as a function of pH and shows a decrease at alkaline pH values. The manganoenzyme is generally more stable to various potentially denaturing conditions and is resistant to inactivation by hydrogen peroxide. Amino acid compositions and N-terminal residue determinations are presented. 相似文献
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Rosalind A. Rosenthal Karl D. Huffman Leslie W. Fisette Christy A. Damphousse Wyeth B. Callaway Bernard Malfroy Susan R. Doctrow 《Journal of biological inorganic chemistry》2009,14(6):979-991
Superoxide dismutase/catalase mimetics, such as salen Mn complexes and certain metalloporphyrins, catalytically neutralize
reactive oxygen and nitrogen species, which have been implicated in the pathogenesis of many serious diseases. Both classes
of mimetic are protective in animal models of oxidative stress. However, only AEOL11207 and EUK-418, two uncharged Mn porphyrins,
have been shown to be orally bioavailable. In this study, EUK-418 and several new analogs (the EUK-400 series) were synthesized
and shown to exhibit superoxide dismutase, catalase, and peroxidase activities in vitro. Some also protected PC12 cells against
staurosporine-induced cell death. All EUK-400 compounds were stable in simulated gastric fluid, and most were substantially
more lipophilic than the salen Mn complexes EUK-189 and EUK-207, which lack oral activity. Pharmacokinetics studies demonstrate
the presence of all EUK-400 series compounds in the plasma of rats after oral administration. These EUK-400 series compounds
are potential oral therapeutic agents for cellular damage caused by oxidative stress.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献