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Sample records for catalase

  1. Recombinant Helicobacter pylori catalase

    Institute of Scientific and Technical Information of China (English)

    Yang Bai; Ya-Li Zhang; Jian-Feng Jin; Ji-De Wang; Zhao-Shan Zhang

    2003-01-01

    AIM: To construct a recombinant strain which highly expresses catalase of Helicobacter pylori(H.pylori) and assay the activity of H. pylori catalase.METHODS: The catalase DNA was amplified from H. pylori chromosomal DNA with PCR techniques and inserted into the prokaryotie expression vector pET-22b (+), and then was transformed into the BL21 (DE3) E. coli strain which expressed catalase recombinant protein. The activity of H.pylori catalase was assayed by the Beers & Sizers.RESULTS: DNA sequence analysis showed that the sequence of catalase DNA was the same as GenBank's research. The catalase recombinant protein amounted to 24.4 % of the total bacterial protein after induced with IPTG for 3 hours at 37 ℃ and the activity of H. pylori catalase was high in the BL21 (DE3) E. coli strain.CONCLUSION: A clone expressing high activity H. pylori catalase is obtained, laying a good foundation for further studies.

  2. Bentonite-supported catalase

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    H. CEYLAN

    2005-05-01

    Full Text Available The properties of the clay bentonite as a support for enzyme immobilization were studied using the enzyme catalase. Such an immobilization does not result in enzyme inactivation and constitutes a valuable method for immobilizing catalase at high ionic strength. The bentonite-supported catalase was characterized in terms of pH and ionic strength dependencies, thermal and storage stability and kinetic parameters. These studies indicate that bentonite is a valuable support for the simple adsorption of enzymes.

  3. In vitro assembly of catalase.

    Science.gov (United States)

    Baureder, Michael; Barane, Elisabeth; Hederstedt, Lars

    2014-10-10

    Most aerobic organisms contain catalase, which functions to decompose hydrogen peroxide. Typical catalases are structurally complex homo-tetrameric enzymes with one heme prosthetic group buried in each subunit. It is not known how catalase in the cell is assembled from its constituents. The bacterium Enterococcus faecalis cannot synthesize heme but can acquire it from the environment to form a cytoplasmic catalase. We have in E. faecalis monitored production of the enzyme polypeptide (KatA) depending on the availability of heme and used our findings to devise a procedure for the purification of preparative amounts of in vivo-synthesized apocatalase. We show that fully active catalase can be obtained in vitro by incubating isolated apoprotein with hemin. We have characterized features of the assembly process and describe a temperature-trapped hemylated intermediate of the enzyme maturation process. Hemylation of apocatalase does not require auxiliary cell components, but rapid assembly of active enzyme seemingly is assisted in the cell. Our findings provide insight about catalase assembly and offer new experimental possibilities for detailed studies of this process.

  4. 7 CFR 58.432 - Catalase.

    Science.gov (United States)

    2010-01-01

    ... Material § 58.432 Catalase. The catalase preparation shall be a stable, buffered solution, neutral in pH, having a potency of not less than 100 Keil units per milliliter. The source of the catalase, its... 7 Agriculture 3 2010-01-01 2010-01-01 false Catalase. 58.432 Section 58.432...

  5. Resuscitation effects of catalase on airborne bacteria.

    OpenAIRE

    Marthi, B; Shaffer, B. T.; Lighthart, B; Ganio, L

    1991-01-01

    Catalase incorporation into enumeration media caused a significant increase (greater than 63%) in the colony-forming abilities of airborne bacteria. Incubation for 30 to 60 min of airborne bacteria in collection fluid containing catalase caused a greater than 95% increase in colony-forming ability. However, catalase did not have any effects on enumeration at high relative humidities (80 to 90%).

  6. The Role of Catalase in Pulmonary Fibrosis

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    Takigawa Tomoko

    2010-12-01

    Full Text Available Abstract Background Catalase is preferentially expressed in bronchiolar and alveolar epithelial cells, and acts as an endogenous antioxidant enzyme in normal lungs. We thus postulated epithelial damage would be associated with a functional deficiency of catalase during the development of lung fibrosis. Methods The present study evaluates the expression of catalase mRNA and protein in human interstitial pneumonias and in mouse bleomycin-induced lung injury. We examined the degree of bleomycin-induced inflammation and fibrosis in the mice with lowered catalase activity. Results In humans, catalase was decreased at the levels of activity, protein content and mRNA expression in fibrotic lungs (n = 12 compared to control lungs (n = 10. Immunohistochemistry revealed a decrease in catalase in bronchiolar epithelium and abnormal re-epithelialization in fibrotic areas. In C57BL/6J mice, catalase activity was suppressed along with downregulation of catalase mRNA in whole lung homogenates after bleomycin administration. In acatalasemic mice, neutrophilic inflammation was prolonged until 14 days, and there was a higher degree of lung fibrosis in association with a higher level of transforming growth factor-β expression and total collagen content following bleomycin treatment compared to wild-type mice. Conclusions Taken together, these findings demonstrate diminished catalase expression and activity in human pulmonary fibrosis and suggest the protective role of catalase against bleomycin-induced inflammation and subsequent fibrosis.

  7. Characterization of Catalase from Psychrotolerant Psychrobacter piscatorii T-3 Exhibiting High Catalase Activity

    OpenAIRE

    Hidetoshi Matsuyma; Isao Yumoto; Hideyuki Kimoto; Kazuaki Yoshimune

    2012-01-01

    A psychrotolerant bacterium, strain T-3 (identified as Psychrobacter piscatorii), that exhibited an extraordinarily high catalase activity was isolated from the drain pool of a plant that uses H2O2 as a bleaching agent. Its cell extract exhibited a catalase activity (19,700 U·mg protein−1) that was higher than that of Micrococcus luteus used for industrial catalase production. Catalase was approximately 10% of the total proteins in the cell extract of the strain. The catalase (PktA) was purif...

  8. Growth of catalase A and catalase T deficient mutant strains of Saccharomyces cerevisiae on ethanol and oleic acid : Growth profiles and catalase activities in relation to microbody proliferation

    NARCIS (Netherlands)

    Klei, Ida J. van der; Rytka, Joanna; Kunau, Wolf H.; Veenhuis, Marten

    1990-01-01

    The parental strain (A+T+) of Saccharomyces cerevisiae and mutants, deficient in catalase T (A+T-), catalase A (A-T+) or both catalases (A-T-), grew on ethanol and oleic acid with comparable doubling times. Specific activities of catalase were low in glucose- and ethanol-grown cells. In the two olei

  9. The three catalases in Deinococcus radiodurans: Only two show catalase activity.

    Science.gov (United States)

    Jeong, Sun-Wook; Jung, Jong-Hyun; Kim, Min-Kyu; Seo, Ho Seong; Lim, Heon-Man; Lim, Sangyong

    2016-01-15

    Deinococcus radiodurans, which is extremely resistant to ionizing radiation and oxidative stress, is known to have three catalases (DR1998, DRA0146, and DRA0259). In this study, to investigate the role of each catalase, we constructed catalase mutants (Δdr1998, ΔdrA0146, and ΔdrA0259) of D. radiodurans. Of the three mutants, Δdr1998 exhibited the greatest decrease in hydrogen peroxide (H2O2) resistance and the highest increase in intracellular reactive oxygen species (ROS) levels following H2O2 treatments, whereas ΔdrA0146 showed no change in its H2O2 resistance or ROS level. Catalase activity was not attenuated in ΔdrA0146, and none of the three bands detected in an in-gel catalase activity assay disappeared in ΔdrA0146. The purified His-tagged recombinant DRA0146 did not show catalase activity. In addition, the phylogenetic analysis of the deinococcal catalases revealed that the DR1998-type catalase is common in the genus Deinococcus, but the DRA0146-type catalase was found in only 4 of 23 Deinococcus species. Taken together, these results indicate that DR1998 plays a critical role in the anti-oxidative system of D. radiodurans by detoxifying H2O2, but DRA0146 does not have catalase activity and is not involved in the resistance to H2O2 stress.

  10. Immobilization of bovine catalase onto magnetic nanoparticles.

    Science.gov (United States)

    Doğaç, Yasemin İspirli; Teke, Mustafa

    2013-01-01

    The scope of this study is to achieve carrier-bound immobilization of catalase onto magnetic particles (Fe₃O₄ and Fe₂O₃NiO₂ · H₂O) to specify the optimum conditions of immobilization. Removal of H2O2 and the properties of immobilized sets were also investigated. To that end, adsorption and then cross-linking methods onto magnetic particles were performed. The optimum immobilization conditions were found for catalase: immobilization time (15 min for Fe₃O₄; 10 min for Fe2O₃NiO₂ · H₂O), the initial enzyme concentration (1 mg/mL), amount of magnetic particles (25 mg), and glutaraldehyde concentration (3%). The activity reaction conditions (optimum temperature, optimum pH, pH stability, thermal stability, operational stability, and reusability) were characterized. Also kinetic parameters were calculated by Lineweaver-Burk plots. The optimum pH values were found to be 7.0, 7.0, and 8.0 for free enzyme, Fe₃O₄-immobilized catalases, and Fe₂O₃NiO₂ · H₂O-immobilized catalases, respectively. All immobilized catalase systems displayed the optimum temperature between 25 and 35°C. Reusability studies showed that Fe₃O₄-immobilized catalase can be used 11 times with 50% loss in original activity, while Fe2O₃NiO₂ · H₂O-immobilized catalase lost 67% of activity after the same number of uses. Furthermore, immobilized catalase systems exhibited improved thermal and pH stability. The results transparently indicate that it is possible to have binding between enzyme and magnetic nanoparticles.

  11. RESTORATION INDUCED BY CATALASE IN IRRADIATED MICROORGANISMS

    Science.gov (United States)

    Latarjet, Raymond; Caldas, Luis Renato

    1952-01-01

    1. E. coli, strain K-12, and B. megatherium 899, irradiated in strict but still undefined physiological conditions with certain heavy doses of ultraviolet light, are efficiently restored by catalase, which acts on or fixes itself upon the bacteria in a few minutes. This restoration (C. R.), different from photorestoration, is aided by a little visible light. 2. At 37° the restorability lasts for about 2 hours after UV irradiation; the restored cells begin to divide at the same time as the normal survivors. 3. C. R. is not produced after x-irradiation. 4. B. megatherium Mox and E. coli, strain B/r show little C. R.; E. coli strain B shows none. None of these three strains is lysogenic, whereas the two preceding catalase-restorable strains are. 5. Phage production in the system "K-12 infected with T2 phage" is restored by catalase after UV irradiation, whereas phage production in the system "infected B" is not. 6. With K-12, catalase does not prevent the growth of phage and the lysis induced by UV irradiation (Lwoff's phenomenon). 7. Hypotheses are discussed concerning: (a) the chemical nature of this action of catalase; (b) a possible relation between C. R. and lysogenicity of the sensitive bacteria; (c) the consequences of such chemical restorations on the general problem of cell radiosensitivity. PMID:14898028

  12. PURIFICATION OF CATALASE ENZYME FROM PLEUROTUS OSTREATUS

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    Susmitha.S

    2014-03-01

    Full Text Available The oyster mushroom Pleurotus ostreatus is the most commonly cultivated mushroom, and are effective for antitumor, antibacterial, anti viral and hematological agents and in immune modulating treatments. Several compounds from oyster mushrooms, potentially beneficial for human health have been isolated and studied. The aim of this research is to purify an enzyme catalase from Pleurotus ostreatus through Sephadox G-75 column, its molecular weight was determined by polyacrylamide gel electrophoresis and the catalase enzyme stability were observed at various temperature and different pH condition. Under denaturing conditions, polyacrylamide gel electrophoresis revealed dissociation of a major component of molecular weight 62,000 kDa, which constituted 90% of the total protein of the stained gel, suggesting that the native enzyme is tetrameric. The optimum temperature and pH for the purified enzyme catalase from Pleurotus ostreatus enzymatic reaction were 30°C and pH 7.5.

  13. Potential enzyme toxicity of oxytetracycline to catalase

    Energy Technology Data Exchange (ETDEWEB)

    Chi Zhenxing; Liu Rutao; Zhang Hao, E-mail: Trutaoliu@sdu.edu.cn [School of Environmental Science and Engineering, Shandong University, China-America CRC for Environment and Health, Shandong Province, 27 Shanda South Road, Jinan 250100 (China)

    2010-10-15

    Oxytetracycline (OTC) is a kind of widely used veterinary drugs. The residue of OTC in the environment is potentially harmful. In the present work, the non-covalent toxic interaction of OTC with catalase was investigated by the fluorescence spectroscopy, UV-vis absorption and circular dichroism (CD) spectroscopy at physiological pH 7.4. OTC can interact with catalase to form a complex mainly by van der Waals' interactions and hydrogen bonds with one binding site. The association constants K were determined to be K{sub 293K} = 7.09 x 10{sup 4} L mol{sup -1} and K{sub 311K} = 3.31 x 10{sup 4} L mol{sup -1}. The thermodynamic parameters ({Delta}H{sup o}, {Delta}G{sup o} and {Delta}S{sup o}) of the interaction were calculated. Based on the Foerster theory of non-radiative energy transfer, the distance between bound OTC and the tryptophan residues of catalase was determined to be 6.48 nm. The binding of OTC can result in change of the micro-environment of the tryptophan residues and the secondary structure of catalase. The activity of catalase was also inhibited for the bound OTC. This work establishes a new strategy to probe the enzyme toxicity of veterinary drug residues and is helpful for clarifying the molecular toxic mechanism of OTC in vivo. The established strategy can be used to investigate the potential enzyme toxicity of other small organic pollutants and drugs.

  14. 21 CFR 184.1034 - Catalase (bovine liver).

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Catalase (bovine liver). 184.1034 Section 184.1034... Listing of Specific Substances Affirmed as GRAS § 184.1034 Catalase (bovine liver). (a) Catalase (bovine liver) (CAS Reg. No. 81457-95-6) is an enzyme preparation obtained from extracts of bovine liver. It...

  15. Bacillus subtilis Vegetative Catalase Is an Extracellular Enzyme

    OpenAIRE

    Naclerio, G; Baccigalupi, L; Caruso, C; De Felice, M; Ricca, E

    1995-01-01

    Strong catalase activity was secreted by Bacillus subtilis cells during stationary growth phase in rich medium but not in sporulation-inducing medium. N-terminal sequencing indicated that the secreted activity was due to the vegetative catalase KatA, previously considered an endocellular enzyme. Extracellular catalase protected B. subtilis cells from oxidative assault.

  16. Preparation of highly efficient manganese catalase mimics.

    Science.gov (United States)

    Triller, Michael U; Hsieh, Wen-Yuan; Pecoraro, Vincent L; Rompel, Annette; Krebs, Bernt

    2002-10-21

    The series of compounds [Mn(bpia)(mu-OAc)](2)(ClO(4))(2) (1), [Mn(2)(bpia)(2)(muO)(mu-OAc)](ClO(4))(3).CH(3)CN (2), [Mn(bpia)(mu-O)](2)(ClO(4))(2)(PF(6)).2CH(3)CN (3), [Mn(bpia)(Cl)(2)](ClO)(4) (4), and [(Mn(bpia)(Cl))(2)(mu-O)](ClO(4))(2).2CH(3)CN (5) (bpia = bis(picolyl)(N-methylimidazol-2-yl)amine) represents a structural, spectroscopic, and functional model system for manganese catalases. Compounds 3 and 5 have been synthesized from 2 via bulk electrolysis and ligand exchange, respectively. All complexes have been structurally characterized by X-ray crystallography and by UV-vis and EPR spectroscopies. The different bridging ligands including the rare mono-mu-oxo and mono-mu-oxo-mono-mu-carboxylato motifs lead to a variation of the Mn-Mn separation across the four binuclear compounds of 1.50 A (Mn(2)(II,II) = 4.128 A, Mn(2)(III,III) = 3.5326 and 3.2533 A, Mn(2)(III,IV) = 2.624 A). Complexes 1, 2, and 3 are mimics for the Mn(2)(II,II), the Mn(2)(III,III), and the Mn(2)(III,IV) oxidation states of the native enzyme. UV-vis spectra of these compounds show similarities to those of the corresponding oxidation states of manganese catalase from Thermus thermophilus and Lactobacillus plantarum. Compound 2 exhibits a rare example of a Jahn-Teller compression. While complexes 1 and 3 are efficient catalysts for the disproportionation of hydrogen peroxide and contain an N(4)O(2) donor set, 4 and 5 show no catalase activity. These complexes have an N(4)Cl(2) and N(4)OCl donor set, respectively, and serve as mimics for halide inhibited manganese catalases. Cyclovoltammetric data show that the substitution of oxygen donor atoms with chloride causes a shift of redox potentials to more positive values. To our knowledge, complex 1 is the most efficient binuclear functional manganese catalase mimic exhibiting saturation kinetics to date.

  17. A Eukaryote without Catalase-Containing Microbodies : Neurospora crassa Exhibits a Unique Cellular Distribution of Its Four Catalases

    NARCIS (Netherlands)

    Schliebs, Wolfgang; Würtz, Christian; Kunau, Wolf-Hubert; Veenhuis, Marten; Rottensteiner, Hanspeter; Wuertz, Christian

    2006-01-01

    Microbodies usually house catalase to decompose hydrogen peroxide generated within the organelle by the action of various oxidases. Here we have analyzed whether peroxisomes (i.e., catalase-containing microbodies) exist in Neurospora crassa. Three distinct catalase isoforms were identified by native

  18. Methanol teratogenicity in mutant mice with deficient catalase activity and transgenic mice expressing human catalase.

    Science.gov (United States)

    Siu, Michelle T; Wiley, Michael J; Wells, Peter G

    2013-04-01

    The role of catalase in methanol (MeOH) teratogenesis is unclear. In rodents it both detoxifies reactive oxygen species (ROS) and metabolizes MeOH and its formic acid (FA) metabolite. We treated pregnant mice expressing either high (hCat) or low catalase activity (aCat), or their wild-type (WT) controls, with either MeOH (4g/kg ip) or saline. hCat mice and WTs were similarly susceptible to MeOH-initiated ophthalmic abnormalities and cleft palates. aCat and WT mice appeared resistant, precluding assessment of the developmental impact of catalase deficiency. Catalase activity was respectively increased at least 1.5-fold, and decreased by at least 35%, in hCat and aCat embryos and maternal livers. MeOH and FA pharmacokinetic profiles were similar among hCat, aCat and WT strains. Although the hCat results imply no ROS involvement, embryo culture studies suggest this may be confounded by maternal factors and/or a requirement for higher catalase activity in the hCat mice.

  19. Catalase and NO CATALASE ACTIVITY1 promote autophagy-dependent cell death in Arabidopsis

    DEFF Research Database (Denmark)

    Hackenberg, Thomas; Andersen, Trine Juul; Auzina, Aija

    2013-01-01

    Programmed cell death often depends on generation of reactive oxygen species, which can be detoxified by antioxidative enzymes, including catalases. We previously isolated catalase-deficient mutants (cat2) in a screen for resistance to hydroxyurea-induced cell death. Here, we identify an Arabidop......Programmed cell death often depends on generation of reactive oxygen species, which can be detoxified by antioxidative enzymes, including catalases. We previously isolated catalase-deficient mutants (cat2) in a screen for resistance to hydroxyurea-induced cell death. Here, we identify......-induced autophagy appeared normal in the nca1 and cat2 mutants. By contrast, autophagic degradation induced by avrRpm1 challenge was compromised, indicating that catalase acted upstream of immunity-triggered autophagy. The direct interaction of catalase with reactive oxygen species could allow catalase to act...

  20. Properties of erythrocyte catalase from heterozygotes for Japanese type acatalasemia.

    Directory of Open Access Journals (Sweden)

    Ogata,Masana

    1979-06-01

    Full Text Available The level of blood catalase activity in heterozygotes for Japanese type acatalasemia was demonstrated to be about half of normal levels by means of titration and spectrophotometric methods. A distribution plot of catalase activities in heterozygous blood was completely separate from that of normal blood. Comparative analysis of the partially purified erythrocyte catalase preparations obtained from normal and heterozygous individuals revealed no distinct differences between them regarding stability to heat, sodium dodecyl sulfate and some enzyme inhibitors or pH dependency. The erythrocyte catalase in heterozygotes for Japanese type acatalasemia contains about half the normal specific activity and as stable as that in normal individuals.

  1. Developmental expression of a catalase inhibitor in maize

    Energy Technology Data Exchange (ETDEWEB)

    Sorenson, J.C.; Scandalios, J.G.

    1976-01-01

    The expression of an endogenous catalase inhibitor has been studied during development of Zea mays. In the 3-day seedling, the inhibitor is expressed primarily in the scutellum and in the aleurone layer of the endosperm. These tissues also show the highest catalase activity at this stage. Inhibitor expression has also been studied temporally in the scutellum, roots, and shoot over the first 12 days of germination. Inhibitor expression shows an inverse relationship with catalase activity in the scutellum and in the shoot. The relationship is less rigid in the root, due probably to the low levels of inhibitor found in that tissue. The role of the inhibitor in catalase regulation is discussed.

  2. Blood superoxiddismutase and catalase: enzymes activity under oxidative stress conditions

    Directory of Open Access Journals (Sweden)

    Каріна Леонідівна Шамелашвілі

    2015-05-01

    Full Text Available The activity of catalase and superoxide dismutase depends not only on the used compounds of rhenium, and also on their dimensional structure and form of applying. It is established that the cis- and trans-isomers of complex compounds of rhenium did countervailing effect on superoxide dismutase and catalase activities. Cis-isomers of Rhenium dycarboxylats agreed increased activity of superoxide dismutase and catalase. While under the action of trans-isomers, where increased activity of superoxide dismutase, catalase activity decreased

  3. Catalases are NAD(PH-dependent tellurite reductases.

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    Iván L Calderón

    Full Text Available Reactive oxygen species damage intracellular targets and are implicated in cancer, genetic disease, mutagenesis, and aging. Catalases are among the key enzymatic defenses against one of the most physiologically abundant reactive oxygen species, hydrogen peroxide. The well-studied, heme-dependent catalases accelerate the rate of the dismutation of peroxide to molecular oxygen and water with near kinetic perfection. Many catalases also bind the cofactors NADPH and NADH tenaciously, but, surprisingly, NAD(PH is not required for their dismutase activity. Although NAD(PH protects bovine catalase against oxidative damage by its peroxide substrate, the catalytic role of the nicotinamide cofactor in the function of this enzyme has remained a biochemical mystery to date. Anions formed by heavy metal oxides are among the most highly reactive, natural oxidizing agents. Here, we show that a natural isolate of Staphylococcus epidermidis resistant to tellurite detoxifies this anion thanks to a novel activity of its catalase, and that a subset of both bacterial and mammalian catalases carry out the NAD(PH-dependent reduction of soluble tellurite ion (TeO(3(2- to the less toxic, insoluble metal, tellurium (Te(o, in vitro. An Escherichia coli mutant defective in the KatG catalase/peroxidase is sensitive to tellurite, and expression of the S. epidermidis catalase gene in a heterologous E. coli host confers increased resistance to tellurite as well as to hydrogen peroxide in vivo, arguing that S. epidermidis catalase provides a physiological line of defense against both of these strong oxidizing agents. Kinetic studies reveal that bovine catalase reduces tellurite with a low Michaelis-Menten constant, a result suggesting that tellurite is among the natural substrates of this enzyme. The reduction of tellurite by bovine catalase occurs at the expense of producing the highly reactive superoxide radical.

  4. Immobilization and characterization of bovine liver catalase on eggshell

    Directory of Open Access Journals (Sweden)

    ÖZLEM ALPTEKİN

    2008-06-01

    Full Text Available Bovine liver catalase immobilized on eggshell particles was characterized and the reusability of the immobilized catalase was investigated in a batch type reactor. For immobilized catalase onto ground eggshell (ICATG, the optimum initial amount of catalase was 85 mg g-1 of eggshells, the optimum pH was 6.0 (75 mM citrate buffer and the temperature was 30 °C. The Vmax and Km values of ICATG were determined as 29.1±1.2 U/mg of protein and 41.9±2.7 mM, respectively. The reusability of ICATG was tested and the remaining activity of ICATG was found to be 73 % of the initial activity after 80 cycles of batch operation. The amount of catalase bound onto the carrier was estimated by using the results of induced coupled plasma measurements. The catalytic efficiencies (kcat/Km of free catalase and ICATG were found to be 1.4´106 and 2.8´103 dm3 s-1 mol-1, respectively. Catalase immobilization onto eggshell is economic and has good reusability. Hence, it can be concluded that eggshell is an efficient carrier for immobilizing catalase.

  5. Catalase Deficiency Accelerates Diabetic Renal Injury Through Peroxisomal Dysfunction

    Science.gov (United States)

    Hwang, Inah; Lee, Jiyoun; Huh, Joo Young; Park, Jehyun; Lee, Hi Bahl; Ho, Ye-Shih; Ha, Hunjoo

    2012-01-01

    Mitochondrial reactive oxygen species (ROS) play an important role in diabetes complications, including diabetic nephropathy (DN). Plasma free fatty acids (FFAs) as well as glucose are increased in diabetes, and peroxisomes and mitochondria participate in FFA oxidation in an interconnected fashion. Therefore, we investigated whether deficiency of catalase, a major peroxisomal antioxidant, accelerates DN through peroxisomal dysfunction and abnormal renal FFA metabolism. Diabetes was induced by multiple injections of low-dose streptozotocin into catalase knock-out (CKO) and wild-type (WT) C57BL/6 mice. Murine mesangial cells (MMCs) transfected with catalase small interfering RNA followed by catalase overexpression were used to further elucidate the role of endogenous catalase. Despite equivalent hyperglycemia, parameters of DN, along with markers of oxidative stress, were more accelerated in diabetic CKO mice than in diabetic WT mice up to 10 weeks of diabetes. CKO mice and MMCs showed impaired peroxisomal/mitochondrial biogenesis and FFA oxidation. Catalase deficiency increased mitochondrial ROS and fibronectin expression in response to FFAs, which were effectively restored by catalase overexpression or N-acetylcysteine. These data provide unprecedented evidence that FFA-induced peroxisomal dysfunction exacerbates DN and that endogenous catalase plays an important role in protecting the kidney from diabetic stress through maintaining peroxisomal and mitochondrial fitness. PMID:22315314

  6. Peroxisomal catalase deficiency modulates yeast lifespan depending on growth conditions

    NARCIS (Netherlands)

    Kawalek, Adam; Lefevre, Sophie D.; Veenhuis, Marten; van der Klei, Ida J.

    2013-01-01

    We studied the role of peroxisomal catalase in chronological aging of the yeast Hansenula polymorpha in relation to various growth substrates. Catalase-deficient (cat) cells showed a similar chronological life span (CLS) relative to the wild-type control upon growth on carbon and nitrogen sources th

  7. 21 CFR 173.135 - Catalase derived from Micrococcus lysodeikticus.

    Science.gov (United States)

    2010-04-01

    ... FOOD FOR HUMAN CONSUMPTION Enzyme Preparations and Microorganisms § 173.135 Catalase derived from... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Catalase derived from Micrococcus lysodeikticus. 173.135 Section 173.135 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND...

  8. Properties of catalase subfractions separated by chromatofocusing of acatalasemia hemolysates.

    Directory of Open Access Journals (Sweden)

    Ogata,Masana

    1982-02-01

    Full Text Available Erythrocyte catalase in normal and Japanese type acatalasemia hemolysates was separated by chromatofocusing into several fractions in the pH range of 6.1 to 5.7. Normal hemolysate gave a major peak of catalase activity with a pH of 6.1 to 5.5, while acatalasemia hemolysate gave several small peaks in this pH range and a main peak with a pH of 6.6 to 6.2. The main protein band in catalase active fractions separated from normal erythrocytes had a molecular weight of 60,000 by SDS polyacrylamide gel electrophoresis. A similar faint protein band having a molecular weight of 60,000 was also found in acatalasemia hemolysate in addition to a fairly intense band with a molecular weight of about 30,000. Catalase active fractions from normal erythrocytes reacted with antihuman erythrocyte catalase rabbit serum by double immunodiffusion.

  9. Regulation of catalase expression in healthy and cancerous cells.

    Science.gov (United States)

    Glorieux, Christophe; Zamocky, Marcel; Sandoval, Juan Marcelo; Verrax, Julien; Calderon, Pedro Buc

    2015-10-01

    Catalase is an important antioxidant enzyme that dismutates hydrogen peroxide into water and molecular oxygen. The catalase gene has all the characteristics of a housekeeping gene (no TATA box, no initiator element sequence, high GC content in promoter) and a core promoter that is highly conserved among species. We demonstrate in this review that within this core promoter, the presence of DNA binding sites for transcription factors, such as NF-Y and Sp1, plays an essential role in the positive regulation of catalase expression. Additional transcription factors, such as FoxO3a, are also involved in this regulatory process. There is strong evidence that the protein Akt/PKB in the PI3K signaling pathway plays a major role in the expression of catalase by modulating the activity of FoxO3a. Over the past decade, other transcription factors (PPARγ, Oct-1, etc.), as well as genetic, epigenetic, and posttranscriptional processes, have emerged as crucial contributors to the regulation of catalase expression. Altered expression levels of catalase have been reported in cancer tissues compared to their normal counterparts. Deciphering the molecular mechanisms that regulate catalase expression could, therefore, be of crucial importance for the future development of pro-oxidant cancer chemotherapy.

  10. Properties of catalase subfractions separated by chromatofocusing of acatalasemia hemolysates.

    OpenAIRE

    Ogata, Masana; Mizugaki,Junko

    1982-01-01

    Erythrocyte catalase in normal and Japanese type acatalasemia hemolysates was separated by chromatofocusing into several fractions in the pH range of 6.1 to 5.7. Normal hemolysate gave a major peak of catalase activity with a pH of 6.1 to 5.5, while acatalasemia hemolysate gave several small peaks in this pH range and a main peak with a pH of 6.6 to 6.2. The main protein band in catalase active fractions separated from normal erythrocytes had a molecular weight of 60,000 by SDS polyacrylamide...

  11. Multiple Catalase Genes Are Differentially Regulated in Aspergillus nidulans

    OpenAIRE

    Kawasaki, Laura; Aguirre, Jesús

    2001-01-01

    Detoxification of hydrogen peroxide is a fundamental aspect of the cellular antioxidant responses in which catalases play a major role. Two differentially regulated catalase genes, catA and catB, have been studied in Aspergillus nidulans. Here we have characterized a third catalase gene, designated catC, which predicts a 475-amino-acid polypeptide containing a peroxisome-targeting signal. With a molecular mass of 54 kDa, CatC shows high similarity to other small-subunit monofunctional catalas...

  12. Catalase Enhances Growth and Biofilm Production of Mycoplasma pneumoniae.

    Science.gov (United States)

    Simmons, Warren L; Dybvig, Kevin

    2015-08-01

    Mycoplasma pneumoniae causes chronic respiratory disease in humans. Factors thought to be important for colonization include the ability of the mycoplasma to form a biofilm on epithelial surfaces and the production of hydrogen peroxide to damage host tissue. Almost all of the mycoplasmas, including M. pneumoniae, lack superoxide dismutase and catalase and a balance should exist between peroxide production and growth. We show here that the addition of catalase to cultures enhanced the formation of biofilms and altered the structure. The incorporation of catalase in agar increased the number of colony-forming units detected and hence could improve the clinical diagnosis of mycoplasmal diseases.

  13. Isolation of the catalase T structural gene of Saccharomyces cerevisiae by functional complementation.

    OpenAIRE

    Spevak, W; Fessl, F; Rytka, J; Traczyk, A; Skoneczny, M; Ruis, H

    1983-01-01

    The catalase T structural gene of Saccharomyces cerevisiae was cloned by functional complementation of a mutation causing specific lack of the enzyme (cttl). Catalase T-deficient mutants were obtained by UV mutagenesis of an S. cerevisiae strain bearing the cas1 mutation, which causes insensitivity of catalase T to glucose repression. Since the second catalase protein of S. cerevisiae, catalase A, is completely repressed on 10% glucose, catalase T-deficient mutant colonies could be detected u...

  14. The searching of active catalase producers among the microscopic fungi

    Directory of Open Access Journals (Sweden)

    Tamara SIRBU

    2011-11-01

    Full Text Available The screening among 158 fungal strains from different taxonomic groups - 128 new soil strains, from plant roots and 30 NCNM strains was carried out on the criterion of catalase synthesis. It was demonstrated that some new strains were more active than those from NCNM. All tested strains from the Penicillium and Aspergillus (108 genus had catalase activity and 26 representatives of other strains did not. Penicillium funiculosum strain, isolated from selected soils of Moldova, was selected as a potential catalase producer. This strain had a catalase activity of 244 units / ml. The optimal parameters for crop cultivation were: temperature of 28 gr.C, the cultivation period - 6 days, the pH value of the nutritional medium – 6.6.

  15. A Laboratory Experiment of the Purification of Catalase.

    Science.gov (United States)

    Busquets, Montserrat; Franco, Rafael

    1986-01-01

    Describes a simple method for purifying catalase for the study of proteins. Procedures are systematically and diagramatically presented. Also identifies polyacrylamide gel electrophoresis, kinetic studies, and apparent molecular weight determination as possible techniques to be used in studying proteins. (ML)

  16. The catalase activity of diiron adenine deaminase

    Energy Technology Data Exchange (ETDEWEB)

    Kamat S. S.; Swaminathan S.; Holmes-Hampton, G. P.; Bagaria, A.; Kumaran, D.; Tichy, S. E.; Gheyi, T.; Zheng, X.; Bain, K.; Groshong, C.; Emtage, S.; Sauder, J. M.; Burley, S. K.; Lindahl, P. A.; Raushel, F. M.

    2011-12-01

    Adenine deaminase (ADE) from the amidohydrolase superfamily (AHS) of enzymes catalyzes the conversion of adenine to hypoxanthine and ammonia. Enzyme isolated from Escherichia coli was largely inactive toward the deamination of adenine. Molecular weight determinations by mass spectrometry provided evidence that multiple histidine and methionine residues were oxygenated. When iron was sequestered with a metal chelator and the growth medium supplemented with Mn{sup 2+} before induction, the post-translational modifications disappeared. Enzyme expressed and purified under these conditions was substantially more active for adenine deamination. Apo-enzyme was prepared and reconstituted with two equivalents of FeSO{sub 4}. Inductively coupled plasma mass spectrometry and Moessbauer spectroscopy demonstrated that this protein contained two high-spin ferrous ions per monomer of ADE. In addition to the adenine deaminase activity, [Fe{sup II}/Fe{sup II}]-ADE catalyzed the conversion of H{sub 2}O{sub 2} to O{sub 2} and H{sub 2}O. The values of k{sub cat} and k{sub cat}/K{sub m} for the catalase activity are 200 s{sup -1} and 2.4 x 10{sup 4} M{sup -1} s{sup -1}, respectively. [Fe{sup II}/Fe{sup II}]-ADE underwent more than 100 turnovers with H{sub 2}O{sub 2} before the enzyme was inactivated due to oxygenation of histidine residues critical for metal binding. The iron in the inactive enzyme was high-spin ferric with g{sub ave} = 4.3 EPR signal and no evidence of anti-ferromagnetic spin-coupling. A model is proposed for the disproportionation of H{sub 2}O{sub 2} by [Fe{sup II}/Fe{sup II}]-ADE that involves the cycling of the binuclear metal center between the di-ferric and di-ferrous oxidation states. Oxygenation of active site residues occurs via release of hydroxyl radicals. These findings represent the first report of redox reaction catalysis by any member of the AHS.

  17. Ionic adsorption of catalase on bioskin: kinetic and ultrastructural studies.

    Science.gov (United States)

    Solas, M T; Vicente, C; Xavier, L; Legaz, M E

    1994-03-15

    Bioskin is a natural polymer produced by Acetobacter xylinum and several yeasts in culture. It contains glucosamine and N-acetyl galactosamine which promote ionic adsorption of catalase at the adequate pH value. High values of ionic strength are required to enzyme desorption. Adsorption of catalase on bioskin fibers has been visualized by scanning electron microscopy associated to a dispersion X-ray analyzer. At low enzyme density, the affinity of the immobilized catalase for hydrogen peroxide was 30% lower than that of the free enzyme. This affinity decreased dramatically at higher density of immobilized enzyme and could not be increased by agitation of the enzyme reaction mixture. Immobilized catalase retains about 70% of its initial activity after 16 d storage, whereas soluble enzyme is completely inactivated after 3 d at room temperature. The haeme group of catalase is not protected after immobilization since it is accessible to both EDTA and phloroglucinol, chelating agents which inactivate catalase by removing the iron atom from the haeme group.

  18. Growth-Dependent Catalase Localization in Exiguobacterium oxidotolerans T-2-2T Reflected by Catalase Activity of Cells

    OpenAIRE

    Yoshiko Hanaoka; Fumihiko Takebe; Yoshinobu Nodasaka; Isao Hara; Hidetoshi Matsuyama; Isao Yumoto

    2013-01-01

    A psychrotolerant and H2O2-resistant bacterium, Exiguobacterium oxidotolerans T-2-2(T), exhibits extraordinary H2O2 resistance and produces catalase not only intracellularly but also extracellularly. The intracellular and extracellular catalases exhibited the same enzymatic characteristics, that is, they exhibited the temperature-dependent activity characteristic of a cold-adapted enzyme, their heat stabilities were similar to those of mesophilic enzymes and very high catalytic intensity. In ...

  19. Embryonic catalase protects against ethanol-initiated DNA oxidation and teratogenesis in acatalasemic and transgenic human catalase-expressing mice.

    Science.gov (United States)

    Miller, Lutfiya; Shapiro, Aaron M; Wells, Peter G

    2013-08-01

    Reactive oxygen species (ROS) are implicated in fetal alcohol spectrum disorders (FASD) caused by alcohol (ethanol, EtOH). Although catalase detoxifies hydrogen peroxide, embryonic catalase activity is only about 5% of maternal levels. To determine the roles of ROS and embryonic catalase in FASD, pregnant mice with enhanced (expressing human catalase, hCat) or deficient (acatalasemic, aCat) catalase activity, or their respective wild-type (WT) controls, were treated ip on gestational day 9 with 4 or 6g/kg EtOH or its saline vehicle, and embryos and fetuses were, respectively, evaluated for oxidatively damaged DNA and structural anomalies. Untreated hCat and aCat dams had, respectively, more and less offspring than their WT controls. hCat progenies were protected from all EtOH fetal anomalies at the low dose (p fetal anomalies (p fetal anomalies (p teratogenesis. Endogenous embryonic catalase, despite its low level, is an important embryoprotective enzyme for EtOH teratogenesis and a likely determinant of individual risk.

  20. Increased microglial catalase activity in multiple sclerosis grey matter.

    Science.gov (United States)

    Gray, Elizabeth; Kemp, Kevin; Hares, Kelly; Redondo, Julianna; Rice, Claire; Scolding, Neil; Wilkins, Alastair

    2014-04-22

    Chronic demyelination, on-going inflammation, axonal loss and grey matter neuronal injury are likely pathological processes that contribute to disease progression in multiple sclerosis (MS). Although the precise contribution of each process and their aetiological substrates is not fully known, recent evidence has implicated oxidative damage as a major cause of tissue injury in MS. The degree of tissue injury caused by oxidative molecules, such as reactive oxygen species (ROS), is balanced by endogenous anti-oxidant enzymes which detoxify ROS. Understanding endogenous mechanisms which protect the brain against oxidative injury in MS is important, since enhancing anti-oxidant responses is a major therapeutic strategy for preventing irreversible tissue injury in the disease. Our aims were to determine expression and activity levels of the hydrogen peroxide-reducing enzyme catalase in MS grey matter (GM). In MS GM, a catalase enzyme activity was elevated compared to control GM. We measured catalase protein expression by immune dot-blotting and catalase mRNA by a real-time polymerase chain reaction (RT-PCR). Protein analysis studies showed a strong positive correlation between catalase and microglial marker IBA-1 in MS GM. In addition, calibration of catalase mRNA level with reference to the microglial-specific transcript AIF-1 revealed an increase in this transcript in MS. This was reflected by the extent of HLA-DR immunolabeling in MS GM which was significantly elevated compared to control GM. Collectively, these observations provide evidence that microglial catalase activity is elevated in MS grey matter and may be an important endogenous anti-oxidant defence mechanism in MS.

  1. Response of soil catalase activity to chromium contamination

    Institute of Scientific and Technical Information of China (English)

    Zofia St(e)pniewska; Agnieszka Woli(n)ska; Joanna Ziomek

    2009-01-01

    The impact of chromium (III) and (VI) forms on soil catalase activity is presented.The Orthic Podzol, Haplic Phaeozem and Mollic Gleysol from different depths were used in the experiment.The soil samples were amended with solution of Cr(III) using CrCl3, and with Cr(VI) using K2Cr2O7 in the concentration range from 0 to 20 mg/kg, whereas the samples without the addition of chromium served as control.Catalase activity was assayed by one of the commonly used spectrophotometric methods.As it is demonstrated in the experiment, both Cr(III) and Cr(VI) forms have ability to reduce soil catalase activity.A chromium dose of 20 mg/kg caused the inhibition of catalase activity and the corresponding contamination levels ranged from 75% to 92% for Cr(III) and 68% to 76% for Cr(VI), with relation to the control.Catalase activity reached maximum in the soil material from surface layers (0-25 cm), typically characterized by the highest content of organic matter creating favorable conditions for microorganisms.

  2. HIGHLY SENSITIVE CATALASE ELECTRODE BASED ON POLYPYRROLE FILMS WITH MICROCONTAINERS

    Institute of Scientific and Technical Information of China (English)

    Yu-ying Gao; Gao-quan Shi

    2006-01-01

    Highly sensitive catalase electrodes for sensing hydrogen peroxide have been fabricated based on polypyrrole films with microcontainers. The microcontainers have a cup-like morphology and are arranged in a density of 4000 units cm-2.Catalase was immobilized into the polypyrrole films with microcontainers (Ppy-mc), which were coated on a Pt substrate electrode. The catalase/Ppy-mc/Pt electrode showed linear response to hydrogen peroxide in the range of 0-18 mmol/L at a potential of -0.3 V (versus SCE). Its sensitivity was measured to be approximately 3.64 μA (mmol/L)-1 cm-2, which is about two times that of the electrode fabricated from a flat Ppy film (catalase/Ppy-flat/Pt electrode). The electrode is highly selective for hydrogen peroxide and its sensitivity is interfered by potential interferents such as ascorbic acid, urea and fructose. Furthermore, such catalase electrodes showed long-term storage stability of 15 days under dry conditions at 4℃.

  3. Fused-expression and characterization of catalase-TAT%Catalase-TAT的融合表达与表征

    Institute of Scientific and Technical Information of China (English)

    李仁宽; 孙立国; 吕元辉; 刘树滔; 饶平凡

    2012-01-01

    利用RT-PCR技术从人的肝细胞中克隆出成熟蛋白过氧化氢酶(catalase)的基因,通过PCR将其与TAT基因融合形成融合基因(catalase-TAT);将catalase-TAT基因构建到表达质粒pET21a(+)中,转入大肠杆菌Rosetta2 (DE3);通过IPTG诱导,转化子成功表达出catalase-TAT融合蛋白.重组菌发酵菌体破碎液经过硫酸铵沉降和Sp-5pw阳离子交换色谱,分离得到电泳纯的目的蛋白,其纯化倍数为18.51倍;经理化性质分析确定了此融合蛋白的最适pH和温度稳定性;细胞实验表明融合蛋白catalase-TAT能够明显提高胞内的过氧化氢酶活性.%Human catalase gene was amplified from liver cells via RT - PCR and fused to TAT. The catalase - TAT was cloned into the expression vector pET21a ( + ) and transformed into Rosettal ( DE3). After induction with IPTG, the recombinant catalase - TAT fusion protein was successfully expressed. The lecombinant bacteria were sonicated and the targeted protein was purified to a single band on SDS - PAGE after ammonium sulfate precipitation and Sp -5pw cation exchange chromatogra-phy, with the purification factor of 18. 51. With physical and chemical analyses, the optimum pH and thermal stability of the fusion protein was determined.

  4. Metallic mercury uptake by catalase Part 1 In Vitro metallic mercury uptake by various kind of animals' erythrocytes and purified human erythrocyte catalase

    OpenAIRE

    劒持,堅志

    1980-01-01

    The uptake of metallic mercury was studied using erythrocytes with different catalase activities taken from various kind of animals. The results were: 1) The uptake of metallic mercury by erythrocytes paralleled the activity of catalase in the erythrocytes with and without hydrogen peroxide, suggesting that the erythrocyte catalase activity is related to the uptake of metallic mercury. 2) The uptake of metallic mercury occurred not only with purified human erythrocyte catalase but also with h...

  5. Embryonic catalase protects against ethanol embryopathies in acatalasemic mice and transgenic human catalase-expressing mice in embryo culture

    Energy Technology Data Exchange (ETDEWEB)

    Miller-Pinsler, Lutfiya [Department of Pharmacology and Toxicology, Faculty of Medicine, University of Toronto, Toronto, Ontario (Canada); Wells, Peter G., E-mail: pg.wells@utoronto.ca [Division of Biomolecular Sciences, Faculty of Pharmacy, University of Toronto, Toronto, Ontario (Canada); Department of Pharmacology and Toxicology, Faculty of Medicine, University of Toronto, Toronto, Ontario (Canada)

    2015-09-15

    Reactive oxygen species (ROS) have been implicated in the mechanism of ethanol (EtOH) teratogenicity, but the protective role of the embryonic antioxidative enzyme catalase is unclear, as embryonic activity is only about 5% of maternal levels. We addressed this question in a whole embryo culture model. C57BL/6 mouse embryos expressing human catalase (hCat) or their wild-type (C57BL/6 WT) controls, and C3Ga.Cg-Cat{sup b}/J catalase-deficient, acatalasemic (aCat) mouse embryos or their wild-type C3HeB/FeJ (C3H WT) controls, were explanted on gestational day (GD) 9 (plug = GD 1), exposed for 24 h to 2 or 4 mg/mL EtOH or vehicle, and evaluated for functional and morphological changes. hCat and C57BL/6 WT vehicle-exposed embryos developed normally, while EtOH was embryopathic in C57BL/6 WT embryos, evidenced by decreases in anterior neuropore closure, somites developed, turning and head length, whereas hCat embryos were protected (p < 0.001). Maternal pretreatment of C57BL/6 WT dams with 50 kU/kg PEG-catalase (PEG-cat) 8 h prior to embryo culture, which increases embryonic catalase activity, blocked all EtOH embryopathies (p < 0.001). Vehicle-exposed aCat mouse embryos had lower yolk sac diameters compared to WT controls, suggesting that endogenous ROS are embryopathic. EtOH was more embryopathic in aCat embryos than WT controls, evidenced by reduced head length and somite development (p < 0.01), and trends for reduced anterior neuropore closure, turning and crown–rump length. Maternal pretreatment of aCat dams with PEG-Cat blocked all EtOH embryopathies (p < 0.05). These data suggest that embryonic catalase is a determinant of risk for EtOH embryopathies. - Highlights: • Ethanol (EtOH) exposure causes structural embryopathies in embryo culture. • Genetically enhanced catalase (hCat) protects against EtOH embryopathies. • Genetically deficient catalase (aCat) exacerbates EtOH embryopathies. • Embryonic catalase is developmentally important. • Et

  6. Recycling of textile bleaching effluents for dyeing using immobilized catalase

    OpenAIRE

    Costa, Silgia; Tzanov,Tzanko; Carneiro, Ana Filipa Gonçalves da Costa; Gübitz, Georg M.; Paulo, Artur Cavaco

    2002-01-01

    Catalase was immobilized on alumina carrier and crosslinked with glutaraldehyde. Storing stability, temperature and pH profiles of enzyme activity were studied in a column reactor with recirculation and in a batch stirred-tank reactor. The immobilized enzyme retained 44% of its activity at pH 11, 30 °C and 90% at 80 °C, pH 7. The half-life time of the immobilized catalase was increased to 2 h at pH 12, and 60 °C. Acceptable results were achieved when the residual water from the washing proces...

  7. Improving catalase-based propelled motor endurance by enzyme encapsulation

    Science.gov (United States)

    Simmchen, Juliane; Baeza, Alejandro; Ruiz-Molina, Daniel; Vallet-Regí, Maria

    2014-07-01

    Biocatalytic propulsion is expected to play an important role in the future of micromotors as it might drastically increase the number of available fuelling reactions. However, most of the enzyme-propelled micromotors so far reported still rely on the degradation of peroxide by catalase, in spite of being vulnerable to relatively high peroxide concentrations. To overcome this limitation, herein we present a strategy to encapsulate the catalase and to graft the resulting enzyme capsules on motor particles. Significant improvement of the stability in the presence of peroxide and other aggressive agents has been observed.Biocatalytic propulsion is expected to play an important role in the future of micromotors as it might drastically increase the number of available fuelling reactions. However, most of the enzyme-propelled micromotors so far reported still rely on the degradation of peroxide by catalase, in spite of being vulnerable to relatively high peroxide concentrations. To overcome this limitation, herein we present a strategy to encapsulate the catalase and to graft the resulting enzyme capsules on motor particles. Significant improvement of the stability in the presence of peroxide and other aggressive agents has been observed. Electronic supplementary information (ESI) available. See DOI: 10.1039/c4nr02459a

  8. Catalases as biocatalysts in technical applications : current state and perspectives

    NARCIS (Netherlands)

    Lončar, Nikola; Fraaije, Marco W

    2015-01-01

    Catalases represent a class of enzymes which has found its place among industrially relevant biocatalysts due to their exceptional catalytic rate and high stability. Textile bleaching prior to the dyeing process is the main application and has been performed on a large scale for the past few decades

  9. Embryonic catalase protects against ethanol embryopathies in acatalasemic mice and transgenic human catalase-expressing mice in embryo culture.

    Science.gov (United States)

    Miller-Pinsler, Lutfiya; Wells, Peter G

    2015-09-15

    Reactive oxygen species (ROS) have been implicated in the mechanism of ethanol (EtOH) teratogenicity, but the protective role of the embryonic antioxidative enzyme catalase is unclear, as embryonic activity is only about 5% of maternal levels. We addressed this question in a whole embryo culture model. C57BL/6 mouse embryos expressing human catalase (hCat) or their wild-type (C57BL/6 WT) controls, and C3Ga.Cg-Cat(b)/J catalase-deficient, acatalasemic (aCat) mouse embryos or their wild-type C3HeB/FeJ (C3H WT) controls, were explanted on gestational day (GD) 9 (plug=GD 1), exposed for 24h to 2 or 4mg/mL EtOH or vehicle, and evaluated for functional and morphological changes. hCat and C57BL/6 WT vehicle-exposed embryos developed normally, while EtOH was embryopathic in C57BL/6 WT embryos, evidenced by decreases in anterior neuropore closure, somites developed, turning and head length, whereas hCat embryos were protected (pcatalase (PEG-cat) 8h prior to embryo culture, which increases embryonic catalase activity, blocked all EtOH embryopathies (pcatalase is a determinant of risk for EtOH embryopathies.

  10. Development of lyophilization cycle and effect of excipients on the stability of catalase during lyophilization

    OpenAIRE

    Lale, Shantanu V; Goyal, Monu; Bansal, Arvind K.

    2011-01-01

    Introduction: The purpose of the present study was to screen excipients such as amino acids and non-aqueous solvents for their stabilizing effect on catalase, a model protein, for lyophilization. The present study also includes optimization of lyophilization cycle for catalase formulations, which is essential from the commercial point of view, since lyophilization is an extremely costly process. Materials and Methods: Activity of catalase was determined using catalase activity assay. Differen...

  11. Direct electrochemistry of Penicillium chrysogenum catalase adsorbed on spectroscopic graphite.

    Science.gov (United States)

    Dimcheva, Nina; Horozova, Elena

    2013-04-01

    The voltammetric studies of Penicillium chrysogenum catalase (PcCAT) adsorbed on spectroscopic graphite, showed direct electron transfer (DET) between its active site and the electrode surface. Analogous tests performed with the commercially available bovine catalase revealed that mammalian enzyme is much less efficient in the DET process. Both catalases were found capable to catalyse the electrooxidation of phenol, but differed in the specifics of catalytic action. At an applied potential of 0.45V the non-linear regression showed the kinetics of the bioelectrochemical oxidation catalysed by the PcCAT obeyed the Hill equation with a binding constant K=0.034±0.002 M(2) (Hill's coefficient n=2.097±0.083, R(2)=0.997), whilst the catalytic action of the bovine catalase was described by the Michaelis-Menten kinetic model with the following parameters: V(max,app)=7.780±0.509 μA, and K(M,app)=0.068±0.070 mol L(-1). The performance of the electrode reaction was affected by the electrode potential, the pH, and temperature. Based on the effect of pH and temperature on the electrode response in presence of phenol a tentative reaction pathway of its bioelectrocatalytic oxidation has been hypothesised. The possible application of these findings in biosensing phenol up to concentration 30 mM at pHs below 7 and in absence of oxidising agents (oxygen or H(2)O(2)) was considered.

  12. Catalase-only nanoparticles prepared by shear alone: Characteristics, activity and stability evaluation.

    Science.gov (United States)

    Huang, Xiao-Nan; Du, Xin-Ying; Xing, Jin-Feng; Ge, Zhi-Qiang

    2016-09-01

    Catalase is a promising therapeutic enzyme; however, it carries risks of inactivation and rapid degradation when it is used in practical bioprocess, such as delivery in vivo. To overcome the issue, we made catalase-only nanoparticles using shear stress alone at a moderate shear rate of 217s(-1) in a coaxial cylinder flow cell. Properties of nanoparticles, including particle size, polydispersity index and zeta potential, were characterized. The conformational changes of pre- and post-sheared catalase were determined using spectroscopy techniques. The results indicated that the conformational changes of catalase and reduction in α-helical content caused by shear alone were less significant than that by desolvation method. Catalase-only nanoparticles prepared by single shear retained over 90% of its initial activity when compared with the native catalase. Catalase nanoparticles lost only 20% of the activity when stored in phosphate buffer solution for 72h at 4°C, whereas native catalase lost 53% under the same condition. Especially, the activity of nanogranulated catalase was decreased only slightly in the simulated intestinal fluid containing α-chymotrypsin during 4h incubation at 37°C, implying that the catalase nanoparticle was more resistant to the degradation of proteases than native catalase molecules. Overall, catalase-only nanoparticles offered a great potential to stabilize enzymes for various pharmaceutical applications.

  13. The relevance of the non-canonical PTS1 of peroxisomal catalase

    NARCIS (Netherlands)

    Williams, Chris; Aksam, Eda Bener; Gunkel, Katja; Veenhuis, Marten; van der Klei, Ida J.

    2012-01-01

    Catalase is sorted to peroxisomes via a C-terminal peroxisomal targeting signal 1 (PTS1), which binds to the receptor protein Pex5. Analysis of the C-terminal sequences of peroxisomal catalases from various species indicated that catalase never contains the typical C-terminal PTS1 tripeptide-SKL, bu

  14. Overexpression of Catalase Diminishes Oxidative Cysteine Modifications of Cardiac Proteins.

    Directory of Open Access Journals (Sweden)

    Chunxiang Yao

    Full Text Available Reactive protein cysteine thiolates are instrumental in redox regulation. Oxidants, such as hydrogen peroxide (H2O2, react with thiolates to form oxidative post-translational modifications, enabling physiological redox signaling. Cardiac disease and aging are associated with oxidative stress which can impair redox signaling by altering essential cysteine thiolates. We previously found that cardiac-specific overexpression of catalase (Cat, an enzyme that detoxifies excess H2O2, protected from oxidative stress and delayed cardiac aging in mice. Using redox proteomics and systems biology, we sought to identify the cysteines that could play a key role in cardiac disease and aging. With a 'Tandem Mass Tag' (TMT labeling strategy and mass spectrometry, we investigated differential reversible cysteine oxidation in the cardiac proteome of wild type and Cat transgenic (Tg mice. Reversible cysteine oxidation was measured as thiol occupancy, the ratio of total available versus reversibly oxidized cysteine thiols. Catalase overexpression globally decreased thiol occupancy by ≥1.3 fold in 82 proteins, including numerous mitochondrial and contractile proteins. Systems biology analysis assigned the majority of proteins with differentially modified thiols in Cat Tg mice to pathways of aging and cardiac disease, including cellular stress response, proteostasis, and apoptosis. In addition, Cat Tg mice exhibited diminished protein glutathione adducts and decreased H2O2 production from mitochondrial complex I and II, suggesting improved function of cardiac mitochondria. In conclusion, our data suggest that catalase may alleviate cardiac disease and aging by moderating global protein cysteine thiol oxidation.

  15. Structure–Function Relationships in Fungal Large-Subunit Catalases

    Energy Technology Data Exchange (ETDEWEB)

    Diaz, A.; Valdez, V; Rudino-Pinera, E; Horjales, E; Hansberg, W

    2009-01-01

    Neurospora crassa has two large-subunit catalases, CAT-1 and CAT-3. CAT-1 is associated with non-growing cells and accumulates particularly in asexual spores; CAT-3 is associated with growing cells and is induced under different stress conditions. It is our interest to elucidate the structure-function relationships in large-subunit catalases. Here we have determined the CAT-3 crystal structure and compared it with the previously determined CAT-1 structure. Similar to CAT-1, CAT-3 hydrogen peroxide (H{sub 2}O{sub 2}) saturation kinetics exhibited two components, consistent with the existence of two active sites: one saturated in the millimolar range and the other in the molar range. In the CAT-1 structure, we found three interesting features related to its unusual kinetics: (a) a constriction in the channel that conveys H{sub 2}O{sub 2} to the active site; (b) a covalent bond between the tyrosine, which forms the fifth coordination bound to the iron of the heme, and a vicinal cysteine; (c) oxidation of the pyrrole ring III to form a cis-hydroxyl group in C5 and a cis-{gamma}-spirolactone in C6. The site of heme oxidation marks the starts of the central channel that communicates to the central cavity and the shortest way products can exit the active site. CAT-3 has a similar constriction in its major channel, which could function as a gating system regulated by the H{sub 2}O{sub 2} concentration before the gate. CAT-3 functional tyrosine is not covalently bonded, but has instead the electron relay mechanism described for the human catalase to divert electrons from it. Pyrrole ring III in CAT-3 is not oxidized as it is in other large-subunit catalases whose structure has been determined. Different in CAT-3 from these enzymes is an occupied central cavity. Results presented here indicate that CAT-3 and CAT-1 enzymes represent a functional group of catalases with distinctive structural characteristics that determine similar kinetics.

  16. Antigenic role of stress-induced catalase of Salmonella typhimurium in cell-mediated immunity.

    OpenAIRE

    Kagaya, K; Miyakawa, Y; Watanabe, K; Fukazawa, Y.

    1992-01-01

    The ability of the H2O2-induced catalase of Salmonella typhimurium to induce cell-mediated immunity against S. typhimurium infection in mice was examined. When exponentially growing cells of S. typhimurium were treated with 20 microM H2O2, the cells resisted killing by 1 mM H2O2 and showed the induction of a new species of catalase in addition to the constitutively produced one. Two molecules of catalases in S. typhimurium were isolated from mutant strains: H2O2-induced catalase (catalase II,...

  17. Catalytic Properties and Immobilization Studies of Catalase from Malva sylvestris L.

    Directory of Open Access Journals (Sweden)

    G. Arabaci

    2013-01-01

    Full Text Available Catalase was partially purified from Malva sylvestris L. and immobilized onto chitosan. Then, its catalytic properties were investigated. (NH42SO4 precipitation and dialysis were performed in the extracted enzyme. Further purification was performed with sephadex G-200 column. Kinetic studies of the purified enzyme activity were measured and characterized. The inhibitory effects of KCN, NaN3, CuSO4, and EDTA on M. sylvestris L. catalase activity were observed except NaCl. Furthermore, M. sylvestris L. catalase was immobilized covalently with glutaraldehyde onto chitosan particles. The pH and temperature optima as well as the changes in the kinetics (Km, Vmax of the immobilized and free M. sylvestris L. catalase were determined. The Km value for immobilized catalase (23.4 mM was higher than that of free enzyme (17.6 mM. Optimum temperature was observed higher than that of the free enzyme. The optimum pH was the same for both free and immobilized catalases (pH 7.50. Immobilized catalase showed higher storage and thermal stabilities than free catalases. Free catalase lost all its activity within 60 days whereas immobilized catalase lost 45% of its activity during the same incubation period at 4°C. The remaining immobilized catalase activity was about 70% after 8 cycles of batch operations.

  18. Cloning, Expression, and Characterization of a Novel Thermophilic Monofunctional Catalase from Geobacillus sp. CHB1

    Directory of Open Access Journals (Sweden)

    Xianbo Jia

    2016-01-01

    Full Text Available Catalases are widely used in many scientific areas. A catalase gene (Kat from Geobacillus sp. CHB1 encoding a monofunctional catalase was cloned and recombinant expressed in Escherichia coli (E. coli, which was the first time to clone and express this type of catalase of genus Geobacillus strains as far as we know. This Kat gene was 1,467 bp in length and encoded a catalase with 488 amino acid residuals, which is only 81% similar to the previously studied Bacillus sp. catalase in terms of amino acid sequence. Recombinant catalase was highly soluble in E. coli and made up 30% of the total E. coli protein. Fermentation broth of the recombinant E. coli showed a high catalase activity level up to 35,831 U/mL which was only lower than recombinant Bacillus sp. WSHDZ-01 among the reported catalase production strains. The purified recombinant catalase had a specific activity of 40,526 U/mg and Km of 51.1 mM. The optimal reaction temperature of this recombinant enzyme was 60°C to 70°C, and it exhibited high activity over a wide range of reaction temperatures, ranging from 10°C to 90°C. The enzyme retained 94.7% of its residual activity after incubation at 60°C for 1 hour. High yield and excellent thermophilic properties are valuable features for this catalase in industrial applications.

  19. Cloning, Expression, and Characterization of a Novel Thermophilic Monofunctional Catalase from Geobacillus sp. CHB1

    Science.gov (United States)

    2016-01-01

    Catalases are widely used in many scientific areas. A catalase gene (Kat) from Geobacillus sp. CHB1 encoding a monofunctional catalase was cloned and recombinant expressed in Escherichia coli (E. coli), which was the first time to clone and express this type of catalase of genus Geobacillus strains as far as we know. This Kat gene was 1,467 bp in length and encoded a catalase with 488 amino acid residuals, which is only 81% similar to the previously studied Bacillus sp. catalase in terms of amino acid sequence. Recombinant catalase was highly soluble in E. coli and made up 30% of the total E. coli protein. Fermentation broth of the recombinant E. coli showed a high catalase activity level up to 35,831 U/mL which was only lower than recombinant Bacillus sp. WSHDZ-01 among the reported catalase production strains. The purified recombinant catalase had a specific activity of 40,526 U/mg and Km of 51.1 mM. The optimal reaction temperature of this recombinant enzyme was 60°C to 70°C, and it exhibited high activity over a wide range of reaction temperatures, ranging from 10°C to 90°C. The enzyme retained 94.7% of its residual activity after incubation at 60°C for 1 hour. High yield and excellent thermophilic properties are valuable features for this catalase in industrial applications. PMID:27579320

  20. Novel insights in mammalian catalase heme maturation: effect of NO and thioredoxin-1.

    Science.gov (United States)

    Chakravarti, Ritu; Gupta, Karishma; Majors, Alana; Ruple, Lisa; Aronica, Mark; Stuehr, Dennis J

    2015-05-01

    Catalase is a tetrameric heme-containing enzyme with essential antioxidant functions in biology. Multiple factors including nitric oxide (NO) have been shown to attenuate its activity. However, the possible impact of NO in relation to the maturation of active catalase, including its heme acquisition and tetramer formation, has not been investigated. We found that NO attenuates heme insertion into catalase in both short-term and long-term incubations. The NO inhibition in catalase heme incorporation was associated with defective oligomerization of catalase, such that inactive catalase monomers and dimers accumulated in place of the mature tetrameric enzyme. We also found that GAPDH plays a key role in mediating these NO effects on the structure and activity of catalase. Moreover, the NO sensitivity of catalase maturation could be altered up or down by manipulating the cellular expression level or activity of thioredoxin-1, a known protein-SNO denitrosylase enzyme. In a mouse model of allergic inflammatory asthma, we found that lungs from allergen-challenged mice contained a greater percentage of dimeric catalase relative to tetrameric catalase in the unchallenged control, suggesting that the mechanisms described here are in play in the allergic asthma model. Together, our study shows how maturation of active catalase can be influenced by NO, S-nitrosylated GAPDH, and thioredoxin-1, and how maturation may become compromised in inflammatory conditions such as asthma.

  1. Cloning, Expression, and Characterization of a Novel Thermophilic Monofunctional Catalase from Geobacillus sp. CHB1.

    Science.gov (United States)

    Jia, Xianbo; Chen, Jichen; Lin, Chenqiang; Lin, Xinjian

    2016-01-01

    Catalases are widely used in many scientific areas. A catalase gene (Kat) from Geobacillus sp. CHB1 encoding a monofunctional catalase was cloned and recombinant expressed in Escherichia coli (E. coli), which was the first time to clone and express this type of catalase of genus Geobacillus strains as far as we know. This Kat gene was 1,467 bp in length and encoded a catalase with 488 amino acid residuals, which is only 81% similar to the previously studied Bacillus sp. catalase in terms of amino acid sequence. Recombinant catalase was highly soluble in E. coli and made up 30% of the total E. coli protein. Fermentation broth of the recombinant E. coli showed a high catalase activity level up to 35,831 U/mL which was only lower than recombinant Bacillus sp. WSHDZ-01 among the reported catalase production strains. The purified recombinant catalase had a specific activity of 40,526 U/mg and K m of 51.1 mM. The optimal reaction temperature of this recombinant enzyme was 60°C to 70°C, and it exhibited high activity over a wide range of reaction temperatures, ranging from 10°C to 90°C. The enzyme retained 94.7% of its residual activity after incubation at 60°C for 1 hour. High yield and excellent thermophilic properties are valuable features for this catalase in industrial applications.

  2. Effect of TiO₂ nanoparticles on the structure and activity of catalase.

    Science.gov (United States)

    Zhang, Hong-Mei; Cao, Jian; Tang, Bo-Ping; Wang, Yan-Qing

    2014-08-05

    TiO₂ nanoparticles are the most widely used metal oxide nanoparticles and have oxidative toxicity. Catalase is an important antioxidant enzyme. Here the understanding of an effect of TiO₂ nanoparticles on the activity and structure of catalase is crucial to characterize the toxicity of TiO₂ nanoparticles. These experimental data revealed that TiO₂ nanoparticles could bind to catalase by the electrostatic and hydrogen bonding forces. On binding TiO₂ nanoparticles, catalase got destabilized with the decrease of α-helices content, the solvent polarity of environment around the fluorescence chromophores on catalase were also affected. In addition, TiO₂ nanoparticles also affected the activity of catalase. TiO₂ nanoparticles acted as an activator of catalase activity at a low molar concentration and as an inhibitor at a higher molar concentration. With regard to human health, the present study could provide a better understanding of the potential nanotoxicity of TiO₂ nanoparticles.

  3. [Fermentation production of microbial catalase and its application in textile industry].

    Science.gov (United States)

    Zhang, Dongxu; Du, Guocheng; Chen, Jian

    2010-11-01

    Microbial catalase is an important industrial enzyme that catalyzes the decomposition of hydrogen peroxide to water and oxygen. This enzyme has great potential of application in food, textile and pharmaceutical industries. The production of microbial catalase has been significantly improved thanks to advances in bioprocess engineering and genetic engineering. In this paper, we review the progresses in fermentation production of microbial catalase and its application in textile industry. Among these progresses, we will highlight strain isolation, substrate and environment optimization, enzyme induction, construction of engineering strains and application process optimization. Meanwhile, we also address future research trends for microbial catalase production and its application in textile industry. Molecular modification (site-directed mutagenesis and directed revolution) will endue catalase with high pH and temperature stabilities. Improvement of catalase production, based on the understanding of induction mechanism and the process control of recombinant stain fermentation, will further accelerate the application of catalase in textile industry.

  4. A new PANI biosensor based on catalase for cyanide determination.

    Science.gov (United States)

    Özcan, Hakkı Mevlüt; Aydin, Tuba

    2016-01-01

    Cyanide is one of the most widespread of compounds measured in environmental analysis due to their toxic effects on environment and health. We report a highly sensitive, reliable, selective amperometric sensor for determination of cyanide, using a polyaniline conductive polymer. The enzyme catalase was immobilized by electropolymerization. The steps during the immobilization were controlled by electrochemical impedance spectroscopy. Optimum pH, temperature, aniline concentration, enzyme concentration, and the number of scans obtained during electropolymerization, were investigated. In addition, the cyanide present in artificial waste water samples was determined. In the characterization studies of the biosensor, some parameters such as reproducibility and storage stability, were analyzed.

  5. Atypical refsum disease with pipecolic acidemia and abnormal catalase distribution.

    Science.gov (United States)

    Baumgartner, M R; Jansen, G A; Verhoeven, N M; Mooyer, P A; Jakobs, C; Roels, F; Espeel, M; Fourmaintraux, A; Bellet, H; Wanders, R J; Saudubray, J M

    2000-01-01

    We describe an 18-year-old patient with psychomotor retardation and abnormally short metatarsi and metacarpals but no other signs of classic Refsum disease. Molecular analysis of the phytanoyl-coenzyme A hydroxylase gene revealed a homozygous deletion causing a frameshift. Surprisingly, L-pipecolic acid was elevated in plasma, and microscopy of the liver showed a reduced number of peroxisomes per cell and a larger average peroxisome size. These abnormal peroxisomes lacked catalase as did peroxisomes in fibroblasts of this patient. Such generalized peroxisomal abnormalities are not present in classic Refsum disease.

  6. Plating isolation of various catalase-negative microorganisms from soil

    Science.gov (United States)

    Labeda, D. P.; Hunt, C. M.; Casida, L. E., Jr.

    1974-01-01

    A unique plating procedure was developed that allows isolation, but not enumeration, of representatives of the catalase-negative soil microflora. The numbers recovered, however, are low as compared to the numbers recovered when the modified dilution-to-extinction isolation procedure is used. The latter procedure provides prolonged inoculation in sealed tubes containing a nutritionally rich broth medium over small submerged agar slants. In contrast, the plating procedure utilizes nutritionally minimal media and the shorter incubations mandated by the inherent problems associated with plating.

  7. Identification of a Catalase-Phenol Oxidase in Betalain Biosynthesis in Red Amaranth (Amaranthus cruentus).

    Science.gov (United States)

    Teng, Xiao-Lu; Chen, Ning; Xiao, Xing-Guo

    2015-01-01

    Betalains are a group of nitrogen-containing pigments that color plants in most families of Caryophyllales. Their biosynthesis has long been proposed to begin with hydroxylation of L-tyrosine to L-DOPA through monophenolase activity of tyrosinase, but biochemical evidence in vivo remains lacking. Here we report that a Group 4 catalase, catalase-phenol oxidase (named as AcCATPO), was identified, purified and characterized from leaves of Amaranthus cruentus, a betalain plant. The purified enzyme appeared to be a homotrimeric protein composed of subunits of about 58 kDa, and demonstrated not only the catalase activity toward H2O2, but also the monophenolase activity toward L-tyrosine and diphenolase activity toward L-DOPA. Its catalase and phenol oxidase activities were inhibited by common classic catalase and tyrosinase inhibitors, respectively. All its peptide fragments identified by nano-LC-MS/MS were targeted to catalases, and matched with a cDNA-encoded polypeptide which contains both classic catalase and phenol oxidase active sites. These sites were also present in catalases of non-betalain plants analyzed. AcCATPO transcript abundance was positively correlated with the ratio of betaxanthin to betacyanin in both green and red leaf sectors of A. tricolor. These data shows that the fourth group catalase, catalase-phenol oxidase, is present in plant, and might be involved in betaxanthin biosynthesis.

  8. Identification of a catalase-phenol oxidase in betalain biosynthesis in red amaranth (Amaranthus cruentus

    Directory of Open Access Journals (Sweden)

    Xiao-Lu eTeng

    2016-01-01

    Full Text Available Betalains are a group of nitrogen-containing pigments that color plants in most families of Caryophyllales. Their biosynthesis has long been proposed to begin with hydroxylation of L-tyrosine to L-DOPA through monophenolase activity of tyrosinase, but biochemical evidence in vivo remains lacking. Here we report that a Group 4 catalase, catalase-phenol oxidase (named as AcCATPO, was identified, purified and characterized from leaves of Amaranthus cruentus, a betalain plant. The purified enzyme appeared to be a homotrimeric protein composed of subunits of about 58 kDa, and demonstrated not only the catalase activity toward H2O2, but also the monophenolase activity toward L-tyrosine and diphenolase activity toward L-DOPA. Its catalase and phenol oxidase activities were inhibited by common classic catalase and tyrosinase inhibitors, respectively. All its peptide fragments identified by nano-LC-MS/MS were targeted to catalases, and matched with a cDNA-encoded polypeptide which contains both classic catalase and phenol oxidase active sites. These sites were also present in catalases of non-betalain plants analyzed. AcCATPO transcript abundance was positively correlated with the ratio of betaxanthin to betacyanin in both green and red leaf sectors of A. tricolor. These data shows that the fourth group catalase, catalase-phenol oxidase, is present in plant, and might be involved in betaxanthin biosynthesis.

  9. Wood utilization is dependent on catalase activities in the filamentous fungus Podospora anserina.

    Science.gov (United States)

    Bourdais, Anne; Bidard, Frederique; Zickler, Denise; Berteaux-Lecellier, Veronique; Silar, Philippe; Espagne, Eric

    2012-01-01

    Catalases are enzymes that play critical roles in protecting cells against the toxic effects of hydrogen peroxide. They are implicated in various physiological and pathological conditions but some of their functions remain unclear. In order to decipher the role(s) of catalases during the life cycle of Podospora anserina, we analyzed the role of the four monofunctional catalases and one bifunctional catalase-peroxidase genes present in its genome. The five genes were deleted and the phenotypes of each single and all multiple mutants were investigated. Intriguingly, although the genes are differently expressed during the life cycle, catalase activity is dispensable during both vegetative growth and sexual reproduction in laboratory conditions. Catalases are also not essential for cellulose or fatty acid assimilation. In contrast, they are strictly required for efficient utilization of more complex biomass like wood shavings by allowing growth in the presence of lignin. The secreted CATB and cytosolic CAT2 are the major catalases implicated in peroxide resistance, while CAT2 is the major player during complex biomass assimilation. Our results suggest that P. anserina produces external H(2)O(2) to assimilate complex biomass and that catalases are necessary to protect the cells during this process. In addition, the phenotypes of strains lacking only one catalase gene suggest that a decrease of catalase activity improves the capacity of the fungus to degrade complex biomass.

  10. Wood utilization is dependent on catalase activities in the filamentous fungus Podospora anserina.

    Directory of Open Access Journals (Sweden)

    Anne Bourdais

    Full Text Available Catalases are enzymes that play critical roles in protecting cells against the toxic effects of hydrogen peroxide. They are implicated in various physiological and pathological conditions but some of their functions remain unclear. In order to decipher the role(s of catalases during the life cycle of Podospora anserina, we analyzed the role of the four monofunctional catalases and one bifunctional catalase-peroxidase genes present in its genome. The five genes were deleted and the phenotypes of each single and all multiple mutants were investigated. Intriguingly, although the genes are differently expressed during the life cycle, catalase activity is dispensable during both vegetative growth and sexual reproduction in laboratory conditions. Catalases are also not essential for cellulose or fatty acid assimilation. In contrast, they are strictly required for efficient utilization of more complex biomass like wood shavings by allowing growth in the presence of lignin. The secreted CATB and cytosolic CAT2 are the major catalases implicated in peroxide resistance, while CAT2 is the major player during complex biomass assimilation. Our results suggest that P. anserina produces external H(2O(2 to assimilate complex biomass and that catalases are necessary to protect the cells during this process. In addition, the phenotypes of strains lacking only one catalase gene suggest that a decrease of catalase activity improves the capacity of the fungus to degrade complex biomass.

  11. LESION SIMULATING DISEASE1 interacts with catalases to regulate hypersensitive cell death in Arabidopsis.

    Science.gov (United States)

    Li, Yansha; Chen, Lichao; Mu, Jinye; Zuo, Jianru

    2013-10-01

    LESION SIMULATING DISEASE1 (lsd1) is an important negative regulator of programmed cell death (PCD) in Arabidopsis (Arabidopsis thaliana). The loss-of-function mutations in lsd1 cause runaway cell death triggered by reactive oxygen species. lsd1 encodes a novel zinc finger protein with unknown biochemical activities. Here, we report the identification of CATALASE3 (CAT3) as an lsd1-interacting protein by affinity purification and mass spectrometry-based proteomic analysis. The Arabidopsis genome contains three homologous catalase genes (CAT1, CAT2, and CAT3). Yeast two-hybrid and coimmunoprecipitation analyses demonstrated that lsd1 interacted with all three catalases both in vitro and in vivo, and the interaction required the zinc fingers of lsd1. We found that the catalase enzymatic activity was reduced in the lsd1 mutant, indicating that the catalase enzyme activity was partially dependent on lsd1. Consistently, the lsd1 mutant was more sensitive to the catalase inhibitor 3-amino-1,2,4-triazole than the wild type, suggesting that the interaction between lsd1 and catalases is involved in the regulation of the reactive oxygen species generated in the peroxisome. Genetic studies revealed that lsd1 interacted with CATALASE genes to regulate light-dependent runaway cell death and hypersensitive-type cell death. Moreover, the accumulation of salicylic acid was required for PCD regulated by the interaction between lsd1 and catalases. These results suggest that the lsd1-catalase interaction plays an important role in regulating PCD in Arabidopsis.

  12. Purification and Characterization of Catalase from Marine Bacterium Acinetobacter sp. YS0810

    Directory of Open Access Journals (Sweden)

    Xinhua Fu

    2014-01-01

    Full Text Available The catalase from marine bacterium Acinetobacter sp. YS0810 (YS0810CAT was purified and characterized. Consecutive steps were used to achieve the purified enzyme as follows: ethanol precipitation, DEAE Sepharose ion exchange, Superdex 200 gel filtration, and Resource Q ion exchange. The active enzyme consisted of four identical subunits of 57.256 kDa. It showed a Soret peak at 405 nm, indicating the presence of iron protoporphyrin IX. The catalase was not apparently reduced by sodium dithionite but was inhibited by 3-amino-1,2,4-triazole, hydroxylamine hydrochloride, and sodium azide. Peroxidase-like activity was not found with the substrate o-phenylenediamine. So the catalase was determined to be a monofunctional catalase. N-terminal amino acid of the catalase analysis gave the sequence SQDPKKCPVTHLTTE, which showed high degree of homology with those of known catalases from bacteria. The analysis of amino acid sequence of the purified catalase by matrix-assisted laser desorption ionization time-of-flight mass spectrometry showed that it was a new catalase, in spite of its high homology with those of known catalases from other bacteria. The catalase showed high alkali stability and thermostability.

  13. Endothelin-1 stimulates catalase activity through the PKCδ-mediated phosphorylation of serine 167.

    Science.gov (United States)

    Rafikov, Ruslan; Kumar, Sanjiv; Aggarwal, Saurabh; Hou, Yali; Kangath, Archana; Pardo, Daniel; Fineman, Jeffrey R; Black, Stephen M

    2014-02-01

    Our previous studies have shown that endothelin-1 (ET-1) stimulates catalase activity in endothelial cells and in lambs with acute increases in pulmonary blood flow (PBF), without altering gene expression. The purpose of this study was to investigate the molecular mechanism by which this occurs. Exposing pulmonary arterial endothelial cells to ET-1 increased catalase activity and decreased cellular hydrogen peroxide (H2O2) levels. These changes correlated with an increase in serine-phosphorylated catalase. Using the inhibitory peptide δV1.1, this phosphorylation was shown to be protein kinase Cδ (PKCδ) dependent. Mass spectrometry identified serine 167 as the phosphorylation site. Site-directed mutagenesis was used to generate a phospho-mimic (S167D) catalase. Activity assays using recombinant protein purified from Escherichia coli or transiently transfected COS-7 cells demonstrated that S167D catalase had an increased ability to degrade H2O2 compared to the wild-type enzyme. Using a phospho-specific antibody, we were able to verify that pS167 catalase levels are modulated in lambs with acute increases in PBF in the presence and absence of the ET receptor antagonist tezosentan. S167 is located on the dimeric interface, suggesting it could be involved in regulating the formation of catalase tetramers. To evaluate this possibility we utilized analytical gel filtration to examine the multimeric structure of recombinant wild-type and S167D catalase. We found that recombinant wild-type catalase was present as a mixture of monomers and dimers, whereas S167D catalase was primarily tetrameric. Further, the incubation of wild-type catalase with PKCδ was sufficient to convert wild-type catalase into a tetrameric structure. In conclusion, this is the first report indicating that the phosphorylation of catalase regulates its multimeric structure and activity.

  14. A gasometric method to determine erythrocyte catalase activity

    Directory of Open Access Journals (Sweden)

    A.J.S. Siqueira

    1999-09-01

    Full Text Available We describe a new gasometric method to determine erythrocyte catalase activity by the measurement of the volume of oxygen produced as a result of hydrogen peroxide decomposition in a system where enzyme and substrate are separated in a special reaction test tube connected to a manometer and the reagents are mixed with a motor-driven stirrer. The position of the reagents in the test tube permits the continuous measurement of oxygen evolution from the time of mixing, without the need to stop the reaction by the addition of acid after each incubation time. The enzyme activity is reported as KHb, i.e., mg hydrogen peroxide decomposed per second per gram of hemoglobin (s-1 g Hb-1. The value obtained for catalase activity in 28 samples of hemolyzed human blood was 94.4 ± 6.17 mg H2O2 s-1 g Hb-1. The results obtained were precise and consistent, indicating that this rapid, simple and inexpensive method could be useful for research and routine work.

  15. A chaperone function of NO CATALASE ACTIVITY1 is required to maintain catalase activity and for multiple stress responses in Arabidopsis.

    Science.gov (United States)

    Li, Jing; Liu, Juntao; Wang, Guoqiang; Cha, Joon-Yung; Li, Guannan; Chen, She; Li, Zhen; Guo, Jinghua; Zhang, Caiguo; Yang, Yongqing; Kim, Woe-Yeon; Yun, Dae-Jin; Schumaker, Karen S; Chen, Zhongzhou; Guo, Yan

    2015-03-01

    Catalases are key regulators of reactive oxygen species homeostasis in plant cells. However, the regulation of catalase activity is not well understood. In this study, we isolated an Arabidopsis thaliana mutant, no catalase activity1-3 (nca1-3) that is hypersensitive to many abiotic stress treatments. The mutated gene was identified by map-based cloning as NCA1, which encodes a protein containing an N-terminal RING-finger domain and a C-terminal tetratricopeptide repeat-like helical domain. NCA1 interacts with and increases catalase activity maximally in a 240-kD complex in planta. In vitro, NCA1 interacts with CATALASE2 (CAT2) in a 1:1 molar ratio, and the NCA1 C terminus is essential for this interaction. CAT2 activity increased 10-fold in the presence of NCA1, and zinc ion binding of the NCA1 N terminus is required for this increase. NCA1 has chaperone protein activity that may maintain the folding of catalase in a functional state. NCA1 is a cytosol-located protein. Expression of NCA1 in the mitochondrion of the nca1-3 mutant does not rescue the abiotic stress phenotypes of the mutant, while expression in the cytosol or peroxisome does. Our results suggest that NCA1 is essential for catalase activity.

  16. Comperative study of catalase immobilization on chitosan, magnetic chitosan and chitosan-clay composite beads.

    Science.gov (United States)

    Başak, Esra; Aydemir, Tülin; Dinçer, Ayşe; Becerik, Seda Çınar

    2013-12-01

    Catalase was immobilized on chitosan and modified chitosan. Studies were carried out on free-immobilized catalase concerning the determination of optimum temperature, pH, thermal, storage stability, reusability, and kinetic parameters. Optimum temperature and pH for free catalase and catalase immobilized were found as 35°C and 7.0, respectively. After 100 times of repeated tests, the immobilized catalases on chitosan-clay and magnetic chitosan maintain over 50% and 60% of the original activity, respectively. The ease of catalase immobilization on low-cost matrices and good stability upon immobilization in the present study make it a suitable product for further use in the food industry.

  17. Roles of Catalase and Trehalose in the Protection from Hydrogen Peroxide Toxicity in Saccharomyces cerevisiae.

    Science.gov (United States)

    Nishimoto, Takuto; Watanabe, Takeru; Furuta, Masakazu; Kataoka, Michihiko; Kishida, Masao

    2016-01-01

    The roles of catalase and trehalose in Saccharomyces cerevisiae subject to hydrogen peroxide (H2O2) treatment were examined by measuring the catalase activity and intracellular trehalose levels in mutants lacking catalase or trehalose synthetase. Intracellular trehalose was elevated but the survival rate after H2O2 treatment remained low in mutants with deletion of the Catalase T gene. On the other hand, deletion of the trehalose synthetase gene increased the catalase activity in mutated yeast to levels higher than those in the wild-type strain, and these mutants exhibited some degree of tolerance to H2O2 treatment. These results suggest that Catalase T is critical in the yeast response to oxidative damage caused by H2O2 treatment, but trehalose also plays a role in protection against H2O2 treatment.

  18. Stimulation of KatG catalase activity by peroxidatic electron donors.

    Science.gov (United States)

    Ndontsa, Elizabeth N; Moore, Robert L; Goodwin, Douglas C

    2012-09-15

    Catalase-peroxidases (KatGs) use a peroxidase scaffold to support robust catalase activity, an ability no other member of its superfamily possesses. Because catalase turnover requires H(2)O(2) oxidation, whereas peroxidase turnover requires oxidation of an exogenous electron donor, it has been anticipated that the latter should inhibit catalase activity. To the contrary, we report peroxidatic electron donors stimulated catalase activity up to 14-fold, particularly under conditions favorable to peroxidase activity (i.e., acidic pH and low H(2)O(2) concentrations). We observed a "low-" and "high-K(M)" component for catalase activity at pH 5.0. Electron donors increased the apparent k(cat) for the "low-K(M)" component. During stimulated catalase activity, less than 0.008 equivalents of oxidized donor accumulated for every H(2)O(2) consumed. Several classical peroxidatic electron donors were effective stimulators of catalase activity, but pyrogallol and ascorbate showed little effect. Stopped-flow evaluation showed that a Fe(III)-O(2)(·-)-like intermediate dominated during donor-stimulated catalatic turnover, and this intermediate converted directly to the ferric state upon depletion of H(2)O(2). In this respect, the Fe(III)-O(2)(·-) -like species was more prominent and persistent than in the absence of the donor. These results point toward a much more central role for peroxidase substrates in the unusual catalase mechanism of KatG.

  19. Cloning and Sequencing of a Candida albicans Catalase Gene and Effects of Disruption of This Gene†

    OpenAIRE

    Wysong, Deborah R.; Christin, Laurent; Sugar, Alan M.; Robbins, Phillips W.; Diamond, Richard D.

    1998-01-01

    Catalase plays a key role as an antioxidant, protecting aerobic organisms from the toxic effects of hydrogen peroxide, and in some cases has been postulated to be a virulence factor. To help elucidate the function of catalase in Candida albicans, a single C. albicans-derived catalase gene, designated CAT1, was isolated and cloned. Degenerate PCR primers based on highly conserved areas of other fungal catalase genes were used to amplify a 411-bp product from genomic DNA of C. albicans ATCC 102...

  20. Extracellular localization of catalase is associated with the transformed state of malignant cells.

    Science.gov (United States)

    Böhm, Britta; Heinzelmann, Sonja; Motz, Manfred; Bauer, Georg

    2015-12-01

    Oncogenic transformation is dependent on activated membrane-associated NADPH oxidase (NOX). However, the resultant extracellular superoxide anions are also driving the NO/peroxynitrite and the HOCl pathway, which eliminates NOX-expressing transformed cells through selective apoptosis induction. Tumor progression is dependent on dominant interference with intercellular apoptosis-inducing ROS signaling through membrane-associated catalase, which decomposes H2O2 and peroxynitrite and oxidizes NO. Particularly, the decomposition of extracellular peroxynitrite strictly requires membrane-associated catalase. We utilized small interfering RNA (siRNA)-mediated knockdown of catalase and neutralizing antibodies directed against the enzyme in combination with challenging H2O2 or peroxynitrite to determine activity and localization of catalase in cells from three distinct steps of multistage oncogenesis. Nontransformed cells did not generate extracellular superoxide anions and only showed intracellular catalase activity. Transformed cells showed superoxide anion-dependent intercellular apoptosis-inducing ROS signaling in the presence of suboptimal catalase activity in their membrane. Tumor cells exhibited tight control of intercellular apoptosis-inducing ROS signaling through a high local concentration of membrane-associated catalase. These data demonstrate that translocation of catalase to the outside of the cell membrane is already associated with the transformation step. A strong local increase in the concentration of membrane-associated catalase is achieved during tumor progression and is controlled by tumor cell-derived H2O2 and by transglutaminase.

  1. Study of catalase electrode for organic peroxides assays.

    Science.gov (United States)

    Horozova, Elena; Dimcheva, Nina; Jordanova, Zinaida

    2002-12-01

    The catalytic activity of immobilized catalase (EC 1.11.1.6) for two model peroxide compounds (dibenzoyl peroxide and 3-chloroperoxibenzoic acid) in a non-aqueous medium was used to prepare an organic-phase enzyme electrode (OPEE). The enzyme was immobilized within a polymeric film on spectrographic graphite. The amperometric signal of the enzyme electrode in substrate solutions was found to be due to the reduction of oxygen generated in the enzyme layer. The electrode response is proportional to peroxide concentrations up to about 40 microM within the potential range from -450 to -650 mV (vs. Ag/AgCl), and the response time is at most 90 s. The enzyme electrode retains about 35% of its initial activity after a 3-week storage at room temperature.

  2. Superoxide Dismutase and Catalase Genotypes in Pediatric Migraine Patients.

    Science.gov (United States)

    Saygi, Semra; Erol, İlknur; Alehan, Füsun; Yalçın, Yaprak Yılmaz; Kubat, Gözde; Ataç, Fatma Belgin

    2015-10-01

    This study compared superoxide dismutase (SOD) and catalase (CAT) alleles in 97 consecutive children and adolescents with migraine to 96 healthy children and adolescents. Isolated genomic DNA was used as a template for SOD1 (35 A/C), SOD2 16 C/T, and CAT2 [(-262 C/T) and (-21 A/T)] allele genotyping. The SOD2 16 C/T genotype and C allele frequency differed significantly between controls and migraine (P = .047; P = .038). CAT -21 AA genotype and A allele frequency were significantly higher in both migraine with aura patients (P = .013; P = .004) and migraine without aura patients (P = .003; P = .001) compared to controls. To our knowledge, this is the first demonstration of differences in SOD and CAT genotypes between pediatric migraine patients and age-matched controls. Further studies on the functional implications of these genetic variants on neural antioxidant capacity and the use of antioxidant modulators for migraine treatment are warranted.

  3. Characterization of a catalase-deficient strain of Neisseria gonorrhoeae: evidence for the significance of catalase in the biology of N. gonorrhoeae.

    Science.gov (United States)

    Johnson, S R; Steiner, B M; Cruce, D D; Perkins, G H; Arko, R J

    1993-01-01

    We obtained a catalase-deficient (Kat-) strain of Neisseria gonorrhoeae isolated from a patient who had been unsuccessfully treated with penicillin. Quantitative enzyme assays and electrophoresis of cell extracts on native polyacrylamide gels subsequently stained for catalase and peroxidase activities failed to detect both enzymes. The strain exhibited no growth anomalies or unusual requirements when grown under ordinary laboratory conditions. However, the Kat- strain proved extremely sensitive to exogenous hydrogen peroxide, and analysis of the bacterial DNA after such exposure showed extensive single-strand breakage in both chromosomal and plasmid DNAs. Partial characterization of the gonococcal catalase from a Kat+ laboratory strain revealed that the enzyme had the physical and chemical properties of both catalase and peroxidase. Images PMID:8454325

  4. Study of catalase adsorption on two mixed-mode ligands and the mechanism involved therein.

    Science.gov (United States)

    Shiva Ranjini, S; Vijayalakshmi, M A

    2012-11-01

    Mixed-mode chromatography sorbents n-hexylamine HyperCel™ (HEA) and phenylpropylamine HyperCel™ (PPA) were evaluated for the study of adsorption of catalase from two different sources. Various parameters such as buffer composition, ionic strength and pH were investigated to study the mechanism of interaction of commercially available pre-purified catalase from Bovine liver, purified catalase from black gram (Vigna mungo) and crude extract of black gram containing catalase with these mixed-mode ligands. A simple and economical screening protocol for identifying optimal buffer conditions for adsorption and desorption of catalase was established with micro volumes of the sorbent in batch mode. With HEA HyperCel, it was observed that pre-purified catalase from both bovine liver and black gram was completely retained at pH 7.0, irrespective of the presence or absence of NaCl in the adsorption buffer, whereas the catalase from crude extract of black gram was completely retained only in the presence of 0.2 M salt in the adsorption buffer. The elution of catalase from both the sources was accomplished by lowering the pH to 4.5 in absence of salt. In case of PPA HyperCel, catalase from both the sources was very strongly adsorbed under different buffer conditions studied, and elution did not yield a significant catalase activity. From the screening experiments, it could be concluded that the interaction of catalase with HEA HyperCel could be dominated by hydrophobic forces with minor contributions from ionic interaction and with PPA HyperCel, it could be a combination of different non-covalent interactions acting on different loci on the surface of the protein.

  5. Inhibition of catalase by tea catechins in free and cellular state: a biophysical approach.

    Science.gov (United States)

    Pal, Sandip; Dey, Subrata Kumar; Saha, Chabita

    2014-01-01

    Tea flavonoids bind to variety of enzymes and inhibit their activities. In the present study, binding and inhibition of catalase activity by catechins with respect to their structure-affinity relationship has been elucidated. Fluorimetrically determined binding constants for (-)-epigallocatechin gallate (EGCG) and (-)-epicatechin gallate (ECG) with catalase were observed to be 2.27×106 M(-1) and 1.66×106 M(-1), respectively. Thermodynamic parameters evidence exothermic and spontaneous interaction between catechins and catalase. Major forces of interaction are suggested to be through hydrogen bonding along with electrostatic contributions and conformational changes. Distinct loss of α-helical structure of catalase by interaction with EGCG was captured in circular dichroism (CD) spectra. Gallated catechins demonstrated higher binding constants and inhibition efficacy than non-gallated catechins. EGCG exhibited maximum inhibition of pure catalase. It also inhibited cellular catalase in K562 cancer cells with significant increase in cellular ROS and suppression of cell viability (IC50 54.5 µM). These results decipher the molecular mechanism by which tea catechins interact with catalase and highlight the potential of gallated catechin like EGCG as an anticancer drug. EGCG may have other non-specific targets in the cell, but its anticancer property is mainly defined by ROS accumulation due to catalase inhibition.

  6. Inhibition of catalase by tea catechins in free and cellular state: a biophysical approach.

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    Sandip Pal

    Full Text Available Tea flavonoids bind to variety of enzymes and inhibit their activities. In the present study, binding and inhibition of catalase activity by catechins with respect to their structure-affinity relationship has been elucidated. Fluorimetrically determined binding constants for (--epigallocatechin gallate (EGCG and (--epicatechin gallate (ECG with catalase were observed to be 2.27×106 M(-1 and 1.66×106 M(-1, respectively. Thermodynamic parameters evidence exothermic and spontaneous interaction between catechins and catalase. Major forces of interaction are suggested to be through hydrogen bonding along with electrostatic contributions and conformational changes. Distinct loss of α-helical structure of catalase by interaction with EGCG was captured in circular dichroism (CD spectra. Gallated catechins demonstrated higher binding constants and inhibition efficacy than non-gallated catechins. EGCG exhibited maximum inhibition of pure catalase. It also inhibited cellular catalase in K562 cancer cells with significant increase in cellular ROS and suppression of cell viability (IC50 54.5 µM. These results decipher the molecular mechanism by which tea catechins interact with catalase and highlight the potential of gallated catechin like EGCG as an anticancer drug. EGCG may have other non-specific targets in the cell, but its anticancer property is mainly defined by ROS accumulation due to catalase inhibition.

  7. [Effect of protonofore 2,4-dinitrophenol on catalase activity of intact Escherichia coli bacteria].

    Science.gov (United States)

    Semchyshyn, H M; Lushchak, V I

    2004-01-01

    Catalase activity of intact E. coli bacteria had a broad pH-optimum (4.0-7.5). Addition of protonofore 2,4-dinitrophenol (200 microM) caused pH-dependency modification (6.5-7.0). Both the catalase activity and curves mode of native cells in the presence of dinitrophenol and cell-free extracts are almost identical. The loss of catalase activity at acid pH was caused by cells destruction and dinitrophenol addition, that makes it possible to suppose that this activity is connected with some membrane component functioning. The induction of "acid" catalase by oxidative stress was blocked with addition of chloramphenicol--protein synthesis inhibitor in prokaryotes. Probably this membrane complex is a part of OxyR regulon, because it is activated by hydrogen peroxide. The activation was not detected in the strain E. coli UM202, which is lacked of catalase HPI (H2O2 response). The catalase activity at acid pH was not observed in the strain E. coli AB1157, that produces both catalase forms and probably has the membrane defect. However, known genetic characteristics of AB1157 stain not let to identify a gene responsible for the "acid" catalase activity.

  8. Covalent Immobilization of Catalase onto Regenerated Silk Fibroins via Tyrosinase-Catalyzed Cross-Linking.

    Science.gov (United States)

    Wang, Ping; Qi, Chenglong; Yu, Yuanyuan; Yuan, Jiugang; Cui, Li; Tang, Gengtie; Wang, Qiang; Fan, Xuerong

    2015-09-01

    Regenerated silk fibroins could be used as medical scaffolds and carrier materials for enzyme immobilization. In the present work, tyrosinase enzyme was used for enzymatic oxidation of silk fibroins, followed by immobilization of catalase onto the fibroin surfaces through physical adsorption and covalent cross-linking as well. Spectrophotometry, SDS-PAGE, and Fourier transform infrared spectroscopy (FTIR) were used to examine the efficiency of enzymatic oxidation and catalase immobilization, respectively. The results indicate that tyrosine residues in silk fibroins could be oxidized and converted to the active o-quinones. Incubating silk fibroins with catalase and tyrosinase led to a noticeable change of molecular weight distribution, indicating the occurrence of the cross-links between silk fibroins and catalase molecules. Two different pathways were proposed for the catalase immobilizations, and the method based on grafting of catalase onto the freeze-dried fibroin membrane is more acceptable. The residual enzyme activity for the immobilized catalase exhibited higher than that of the control after repeated washing cycles. Meanwhile, the thermal stability and alkali resistance were also slightly improved as compared to free catalase. The mechanisms of enzymatic immobilization are also concerned.

  9. Purification and characterization of an intracellular catalase-peroxidase from Penicillium simplicissimum

    NARCIS (Netherlands)

    Fraaije, Marco W.; Roubroeks, Hanno P.; Hagen, Wilfred R.; Berkel, Willem J.H. van

    1996-01-01

    The first dimeric catalase-peroxidase of eucaryotic origin, an intracellular hydroperoxidase from Penicillium simplicissimum which exhibited both catalase and peroxidase activities, has been isolated. The enzyme has an apparent molecular mass of about 170 kDa and is composed of two identical subunit

  10. Effect of Catalase on Biocatalytic Synthesis of Pyruvate by Enzymes from Pseudomonas sp.

    Institute of Scientific and Technical Information of China (English)

    Jing Song GU; Yuan Xiu WANG; Qiang JIAO

    2004-01-01

    Pyruvate was produced from DL-lactate by a kind of green-chemical biocatalyst - cell-free extract from bacterial strain Pseudomonas sp. SM-6. Catalase in cell-free extract, which could stabilize the pyruvate formed by lactate oxidase, played an important role in pyruvate preparation. The effect of catalase in conversion process was evaluated.

  11. Location of catalase in crystalline peroxisomes of methanol-grown Hansenula polymorpha

    NARCIS (Netherlands)

    Keizer, Ineke; Roggenkamp, Rainer; Harder, Willem; Veenhuis, Marten

    1992-01-01

    We have studied the intraperoxisomal location of catalase in peroxisomes of methanol-grown Hansenula polymorpha by (immuno)cytochemical means. In completely crystalline peroxisomes, in which the crystalline matrix is composed of octameric alcohol oxidase (AO) molecules, most of the catalase protein

  12. Catalase activity in the soil of the wood biogeocenoses in the Samara-river region

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    A. F. Kulik

    2009-11-01

    Full Text Available Research of catalase activity changes in connection with the free-radical oxidation is soil of natural and artificial ecosystems is conducted. The catalase is a plants’ enzyme of antioxidative protection. The catalase activity is a marker of variety and improvement of soils. It is important for the problems solutions in applied soil science. The aberrations of catalase, peroxidase and polyphenol oxidase activities characterise not only the metabolizing of plants, microorganisms and soils, but the level of the environmental pollution. The general activity of enzymatic systems allows ascertaining their role in the forming of biota components’ resistance to the exogenous influence. The seasonal dynamics of the soils’ catalase activity subject to the type of a biogeocenosis is disclosed.

  13. Effect of cimetidine on catalase activity of Pseudomonas aeruginosa: a suggested mechanism of action.

    Science.gov (United States)

    Masoud, Masoudeh; Ebrahimi, Farnoosh; Minai-Tehrani, Dariush

    2014-01-01

    Catalase is an important enzyme for the degradation of hydrogen peroxide in cells. Bacteria have potent catalase to deal with H2O2 in their medium culture. Any chemicals that inhibit catalase activity can be harmful for cells. Histamine H2 antagonist drugs such as cimetidine and ranitidine are used for the treatment of gastrointestinal tract disorders. The present results showed that cimetidine could inhibit the catalase activity of Pseudomonas aeruginosa in a competitive inhibition. The determination of IC50 value and Ki (6.5 μM) of cimetidine demonstrated that the enzyme binds to the drug with high affinity. Binding of the drug to the enzyme was pH-dependent and no binding was observed at basic pH (>9) and acidic pH (catalase activity.

  14. A simple method of catalase purification for the undergraduate experimental course.

    Science.gov (United States)

    Chen, Qian; Cheng, Meng; Wang, Yinnan; Yao, Ming; Chen, Yongchun; Gao, Yuan; Ding, Wenyuan

    2015-02-01

    Catalase is a characteristic enzyme of peroxisomes, of which it is the most abundant protein. This enzyme serves as a typical example of a peroxisomal enzyme and is important in the teaching of biochemistry and molecular biology. Although there is substantial information regarding catalase purification, purifying catalase for the junior‑grade undergraduate experimental course face challenges in obtaining materials and increasingly expensive purification equipment. This study presents a simple method for the purification of mouse liver catalase using ethanol‑chloroform treatment, sodium sulfate fractionation, dialysis and Sephadex G‑200 gel filtration chromatography. Catalase was purified 31.8‑fold with an 18.3% yield. The advantages of this method were its low operating environment requirements, simple procedure and reduced cost. Furthermore, the method was designed to improve students' comprehensive ability and manipulative ability and to introduce a sense of innovation in the fields of biochemistry and molecular biology during their junior year.

  15. Forchlorfenuron detection based on its inhibitory effect towards catalase immobilized on boron nitride substrate.

    Science.gov (United States)

    Xu, Qin; Cai, Lijuan; Zhao, Huijie; Tang, Jiaqian; Shen, Yuanyuan; Hu, Xiaoya; Zeng, Haibo

    2015-01-15

    An enzymatic procedure based on a catalase biosensor for the detection of forchlorfenuron (CPPU) has been reported in this work. Catalase was immobilized on boron nitride (BN) sheets dispersed in chitosan by adsorption. The immobilized catalase exhibited direct electron transfer character and excellent electrocatalytic activity towards H2O2 reduction. After introducing CPPU into the H2O2 containing phosphate buffer solution, the catalase-catalyzed H2O2 reduction current decreased. By measuring the current decrease, CPPU can be determined in the range of 0.5-10.0 µM with the detection limit of 0.07 μM. The non-competitive inhibition behavior of CPPU towards catalase was verified by the Lineweaver-Burk plots. Long stability character has been ascribed to this biosensor. Possible use of this biosensor in flow systems is illustrated. The proposed biosensor has been successfully applied to CPPU determination in fruits samples with satisfactory results.

  16. Natural resistance to ascorbic acid induced oxidative stress is mainly mediated by catalase activity in human cancer cells and catalase-silencing sensitizes to oxidative stress

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    Klingelhoeffer Christoph

    2012-05-01

    Full Text Available Abstract Background Ascorbic acid demonstrates a cytotoxic effect by generating hydrogen peroxide, a reactive oxygen species (ROS involved in oxidative cell stress. A panel of eleven human cancer cell lines, glioblastoma and carcinoma, were exposed to serial dilutions of ascorbic acid (5-100 mmol/L. The purpose of this study was to analyse the impact of catalase, an important hydrogen peroxide-detoxifying enzyme, on the resistance of cancer cells to ascorbic acid mediated oxidative stress. Methods Effective concentration (EC50 values, which indicate the concentration of ascorbic acid that reduced the number of viable cells by 50%, were detected with the crystal violet assay. The level of intracellular catalase protein and enzyme activity was determined. Expression of catalase was silenced by catalase-specific short hairpin RNA (sh-RNA in BT-20 breast carcinoma cells. Oxidative cell stress induced apoptosis was measured by a caspase luminescent assay. Results The tested human cancer cell lines demonstrated obvious differences in their resistance to ascorbic acid mediated oxidative cell stress. Forty-five percent of the cell lines had an EC50 > 20 mmol/L and fifty-five percent had an EC50 50 of 2.6–5.5 mmol/L, glioblastoma cells were the most susceptible cancer cell lines analysed in this study. A correlation between catalase activity and the susceptibility to ascorbic acid was observed. To study the possible protective role of catalase on the resistance of cancer cells to oxidative cell stress, the expression of catalase in the breast carcinoma cell line BT-20, which cells were highly resistant to the exposure to ascorbic acid (EC50: 94,9 mmol/L, was silenced with specific sh-RNA. The effect was that catalase-silenced BT-20 cells (BT-20 KD-CAT became more susceptible to high concentrations of ascorbic acid (50 and 100 mmol/L. Conclusions Fifty-five percent of the human cancer cell lines tested were unable to protect themselves

  17. Inhibition of the catalase activity from Phaseolus vulgaris and Medicago sativa by sodium chloride.

    Science.gov (United States)

    Tejera García, Noel A; Iribarne, Carmen; Palma, Francisco; Lluch, Carmen

    2007-08-01

    Changes in catalase activity during the development of the Rhizobium-legume symbiosis as well as its response in salinized plants of Phaseolus vulgaris and Medicago sativa, was studied. Besides, it was examined the behavior of the enzyme, isolated from leaves and root nodules, during in vitro incubation with NaCl doses. Nodule catalase activities of both legumes were assayed with several enzyme inhibitors and also purified. Leaf catalase activity of Phaseolus vulgaris and Medicago sativa decreased and increased respectively throughout the ontogeny, but root nodule catalase kept a high and stable value. This last result suggests that both legumes require the maintenance of high nodule catalase in nitrogen-fixing nodules. Under salt stress conditions leaf and nodule catalase activity decreased in both, grain and pasture legumes. Because catalase from leaf of Medicago sativa and nodules of Phaseolus vulgaris were relatively sensitive to NaCl during in vitro experiments, the detoxifying role of this enzyme for H(2)O(2) should be limited in such conditions. Both catalases, from determinate and indeterminate nodules, were affected neither by oxygen nor superoxide radicals but showed a strong (Phaseolus vulgaris) or partial (Medicago sativa) inhibition with dithiothreitol, dithionite and beta-mercaptoethanol. Besides, cyanide was the most potent inhibitor of nodule catalases. Finally, catalases partially purified by immobilized metal ion affinity chromatography migrated at 42 (Phaseolus vulgaris) and 46kDa (Medicago sativa) on SDS-PAGE, whereas native forms on sephacryl S-300 columns exhibited a molecular mass of 59 and 48kDa (Phaseolus vulgaris) and 88 and 53kDa (Medicago sativa).

  18. Human Islet Amyloid Polypeptide Fibril Binding to Catalase: A Transmission Electron Microscopy and Microplate Study

    Directory of Open Access Journals (Sweden)

    Nathaniel G. N. Milton

    2010-01-01

    Full Text Available The diabetes-associated human islet amyloid polypeptide (IAPP is a 37-amino-acid peptide that forms fibrils in vitro and in vivo. Human IAPP fibrils are toxic in a similar manner to Alzheimer's amyloid-β (Aβ and prion protein (PrP fibrils. Previous studies have shown that catalase binds to Aβ fibrils and appears to recognize a region containing the Gly-Ala-Ile-Ile sequence that is similar to the Gly-Ala-Ile-Leu sequence found in human IAPP residues 24-27. This study presents a transmission electron microscopy (TEM—based analysis of fibril formation and the binding of human erythrocyte catalase to IAPP fibrils. The results show that human IAPP 1-37, 8-37, and 20-29 peptides form fibrils with diverse and polymorphic structures. All three forms of IAPP bound catalase, and complexes of IAPP 1-37 or 8-37 with catalase were identified by immunoassay. The binding of biotinylated IAPP to catalase was high affinity with a KD of 0.77nM, and could be inhibited by either human or rat IAPP 1-37 and 8-37 forms. Fibrils formed by the PrP 118-135 peptide with a Gly-Ala-Val-Val sequence also bound catalase. These results suggest that catalase recognizes a Gly-Ala-Ile-Leu—like sequence in amyloid fibril-forming peptides. For IAPP 1-37 and 8-37, the catalase binding was primarily directed towards fibrillar rather than ribbon-like structures, suggesting differences in the accessibility of the human IAPP 24-27 Gly-Ala-Ile-Leu region. This suggests that catalase may be able to discriminate between different structural forms of IAPP fibrils. The ability of catalase to bind IAPP, Aβ, and PrP fibrils demonstrates the presence of similar accessible structural motifs that may be targets for antiamyloid therapeutic development.

  19. Catalase inhibits ionizing radiation-induced apoptosis in hematopoietic stem and progenitor cells.

    Science.gov (United States)

    Xiao, Xia; Luo, Hongmei; Vanek, Kenneth N; LaRue, Amanda C; Schulte, Bradley A; Wang, Gavin Y

    2015-06-01

    Hematologic toxicity is a major cause of mortality in radiation emergency scenarios and a primary side effect concern in patients undergoing chemo-radiotherapy. Therefore, there is a critical need for the development of novel and more effective approaches to manage this side effect. Catalase is a potent antioxidant enzyme that coverts hydrogen peroxide into hydrogen and water. In this study, we evaluated the efficacy of catalase as a protectant against ionizing radiation (IR)-induced toxicity in hematopoietic stem and progenitor cells (HSPCs). The results revealed that catalase treatment markedly inhibits IR-induced apoptosis in murine hematopoietic stem cells and hematopoietic progenitor cells. Subsequent colony-forming cell and cobble-stone area-forming cell assays showed that catalase-treated HSPCs can not only survive irradiation-induced apoptosis but also have higher clonogenic capacity, compared with vehicle-treated cells. Moreover, transplantation of catalase-treated irradiated HSPCs results in high levels of multi-lineage and long-term engraftments, whereas vehicle-treated irradiated HSPCs exhibit very limited hematopoiesis reconstituting capacity. Mechanistically, catalase treatment attenuates IR-induced DNA double-strand breaks and inhibits reactive oxygen species. Unexpectedly, we found that the radioprotective effect of catalase is associated with activation of the signal transducer and activator of transcription 3 (STAT3) signaling pathway and pharmacological inhibition of STAT3 abolishes the protective activity of catalase, suggesting that catalase may protect HSPCs against IR-induced toxicity via promoting STAT3 activation. Collectively, these results demonstrate a previously unrecognized mechanism by which catalase inhibits IR-induced DNA damage and apoptosis in HSPCs.

  20. A Review on Direct Electrochemistry of Catalase for Electrochemical Sensors

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    Periasamy Arun Prakash

    2009-03-01

    Full Text Available Catalase (CAT is a heme enzyme with a Fe(III/II prosthetic group at its redox centre. CAT is present in almost all aerobic living organisms, where it catalyzes the disproportionation of H2O2 into oxygen and water without forming free radicals. In order to study this catalytic mechanism in detail, the direct electrochemistry of CAT has been investigated at various modified electrode surfaces with and without nanomaterials. The results show that CAT immobilized on nanomaterial modified electrodes shows excellent catalytic activity, high sensitivity and the lowest detection limit for H2O2 determination. In the presence of nanomaterials, the direct electron transfer between the heme group of the enzyme and the electrode surface improved significantly. Moreover, the immobilized CAT is highly biocompatible and remains extremely stable within the nanomaterial matrices. This review discusses about the versatile approaches carried out in CAT immobilization for direct electrochemistry and electrochemical sensor development aimed as efficient H2O2 determination. The benefits of immobilizing CAT in nanomaterial matrices have also been highlighted.

  1. Catalase is a key enzyme in seed recovery from ageing during priming.

    Science.gov (United States)

    Kibinza, Serge; Bazin, Jérémie; Bailly, Christophe; Farrant, Jill M; Corbineau, Françoise; El-Maarouf-Bouteau, Hayat

    2011-09-01

    Ageing induces seed deterioration expressed as the loss of seed vigour and/or viability. Priming treatment, which consists in soaking of seeds in a solution of low water potential, has been shown to reinvigorate aged seeds. We investigate the importance of catalase in oxidation protection during accelerated ageing and repair during subsequent priming treatment of sunflower (Helianthus annuus L.) seeds. Seeds equilibrated to 0.29g H2Og(-1) dry matter (DM) were aged at 35°C for different durations and then primed by incubation for 7 days at 15°C in a solution of polyethylene glycol 8000 at -2MPa. Accelerated ageing affected seed germination and priming treatment reversed partially the ageing effect. The inhibition of catalase by the addition of aminotriazol during priming treatment reduced seed repair indicating that catalase plays a key role in protection and repair systems during ageing. Ageing was associated with H2O2 accumulation as showed by biochemical quantification and CeCl3 staining. Catalase was reduced at the level of gene expression, protein content and affinity. Interestingly, priming induced catalase synthesis by activating expression and translation of the enzyme. Immunocytolocalization of catalase showed that the enzyme co-localized with H2O2 in the cytosol. These results clearly indicate that priming induce the synthesis of catalase which is involved in seed recovery during priming.

  2. Monofunctional catalase P of Paracoccidioides brasiliensis: identification, characterization, molecular cloning and expression analysis.

    Science.gov (United States)

    Moreira, Sabrina F I; Bailão, Alexandre M; Barbosa, Mônica S; Jesuino, Rosalia S A; Felipe, M Sueli Soares; Pereira, Maristela; de Almeida Soares, Célia Maria

    2004-01-30

    Within the context of studies on genes from Paracoccidioides brasiliensis (Pb) potentially associated with fungus-host interaction, we isolated a 61 kDa protein, pI 6.2, that was reactive with sera of patients with paracoccidioidomycosis. This protein was identified as a peroxisomal catalase. A complete cDNA encoding this catalase was isolated from a Pb cDNA library and was designated PbcatP. The cDNA contained a 1509 bp ORF containing 502 amino acids, whose molecular mass was 57 kDa, with a pI of 6.5. The translated protein PbCATP revealed canonical motifs of monofunctional typical small subunit catalases and the peroxisome-PTS-1-targeting signal. The deduced and the native PbCATP demonstrated amino acid sequence homology to known monofunctional catalases and was most closely related to catalases from other fungi. The protein and mRNA were diminished in the mycelial saprobic phase compared to the yeast phase of infection. Protein synthesis and mRNA levels increased during the transition from mycelium to yeast. In addition, the catalase protein was induced when cells were exposed to hydrogen peroxide. The identification and characterization of the PbCATP and cloning and characterization of the cDNA are essential steps for investigating the role of catalase as a defence of P. brasiliensis against oxygen-dependent killing mechanisms. These results suggest that this protein exerts an influence in the virulence of P. brasiliensis.

  3. Do pH and flavonoids influence hypochlorous acid-induced catalase inhibition and heme modification?

    Science.gov (United States)

    Krych-Madej, Justyna; Gebicka, Lidia

    2015-09-01

    Hypochlorous acid (HOCl), highly reactive oxidizing and chlorinating species, is formed in the immune response to invading pathogens by the reaction of hydrogen peroxide with chloride catalyzed by the enzyme myeloperoxidase. Catalase, an important antioxidant enzyme, catalyzing decomposition of hydrogen peroxide to water and molecular oxygen, hampers in vitro HOCl formation, but is also one of the main targets for HOCl. In this work we have investigated HOCl-induced catalase inhibition at different pH, and the influence of flavonoids (catechin, epigallocatechin gallate and quercetin) on this process. It has been shown that HOCl-induced catalase inhibition is independent on pH in the range 6.0-7.4. Preincubation of catalase with epigallocatechin gallate and quercetin before HOCl treatment enhances the degree of catalase inhibition, whereas catechin does not affect this process. Our rapid kinetic measurements of absorption changes around the heme group have revealed that heme modification by HOCl is mainly due to secondary, intramolecular processes. The presence of flavonoids, which reduce active catalase intermediate, Compound I to inactive Compound II have not influenced the kinetics of HOCl-induced heme modification. Possible mechanisms of the reaction of hypochlorous acid with catalase are proposed and the biological consequences are discussed.

  4. The kinetic properties producing the perfunctory pH profiles of catalase-peroxidases.

    Science.gov (United States)

    Moore, Robert L; Powell, Luke J; Goodwin, Douglas C

    2008-06-01

    Many structure-function relationship studies performed on the catalase-peroxidase enzymes are based on limited kinetic data. To provide a more substantive understanding of catalase-peroxidase function, we undertook a more exhaustive evaluation of catalase-peroxidase catalysis as a function of pH. Kinetic parameters across a broad pH range for the catalase and peroxidase activities of E. coli catalase peroxidase (KatG) were obtained, including the separate analysis of the oxidizing and reducing substrates of the peroxidase catalytic cycle. This investigation identified ABTS-dependent inhibition of peroxidase activity, particularly at low pH, unveiling that previously reported pH optima are clearly skewed. We show that turnover and efficiency of peroxidase activity increases with decreasing pH until the protein unfolds. The data also suggest that the catalase pH optimum is more complex than it is often assumed to be. The apparent optimum is in fact the intersection of the optimum for binding (7.00) and the optimum for activity (5.75). We also report the apparent pK(a)s for binding and catalysis of catalase activity as well as approximate values for certain peroxidatic and catalatic steps.

  5. Over-Expression of Catalase in Myeloid Cells Confers Acute Protection Following Myocardial Infarction

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    E. Bernadette Cabigas

    2014-05-01

    Full Text Available Cardiovascular disease is the leading cause of death in the United States and new treatment options are greatly needed. Oxidative stress is increased following myocardial infarction and levels of antioxidants decrease, causing imbalance that leads to dysfunction. Therapy involving catalase, the endogenous scavenger of hydrogen peroxide (H2O2, has been met with mixed results. When over-expressed in cardiomyocytes from birth, catalase improves function following injury. When expressed in the same cells in an inducible manner, catalase showed a time-dependent response with no acute benefit, but a chronic benefit due to altered remodeling. In myeloid cells, catalase over-expression reduced angiogenesis during hindlimb ischemia and prevented monocyte migration. In the present study, due to the large inflammatory response following infarction, we examined myeloid-specific catalase over-expression on post-infarct healing. We found a significant increase in catalase levels following infarction that led to a decrease in H2O2 levels, leading to improved acute function. This increase in function could be attributed to reduced infarct size and improved angiogenesis. Despite these initial improvements, there was no improvement in chronic function, likely due to increased fibrosis. These data combined with what has been previously shown underscore the need for temporal, cell-specific catalase delivery as a potential therapeutic option.

  6. Effects of pergolide mesylate on transduction efficiency of PEP-1-catalase protein

    Energy Technology Data Exchange (ETDEWEB)

    Sohn, Eun Jeong; Kim, Dae Won; Kim, Young Nam; Kim, So Mi [Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702 (Korea, Republic of); Lim, Soon Sung [Department of Food Science and Nutrition and RIC Center, Hallym University, Chunchon 200-702 (Korea, Republic of); Kang, Tae-Cheon [Department of Anatomy and Neurobiology, College of Medicine, Hallym University, Chunchon 200-702 (Korea, Republic of); Kwon, Hyeok Yil [Department of Physiology, College of Medicine, Hallym University, Chunchon 200-702 (Korea, Republic of); Kim, Duk-Soo [Department of Anatomy, College of Medicine, Soonchunhyang University, Cheonan-Si 330-090 (Korea, Republic of); Cho, Sung-Woo [Department of Biochemistry and Molecular Biology, University of Ulsan College of Medicine, Seoul 138-736 (Korea, Republic of); Han, Kyu Hyung; Park, Jinseu; Eum, Won Sik [Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702 (Korea, Republic of); Hwang, Hyun Sook, E-mail: wazzup@hallym.ac.kr [Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702 (Korea, Republic of); Choi, Soo Young, E-mail: sychoi@hallym.ac.kr [Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702 (Korea, Republic of)

    2011-03-18

    Research highlights: {yields} We studied effects of pergolide mesylate (PM) on in vitro and in vivo transduction of PEP-1-catalase. {yields} PEP-1-catatase inhibited 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation. {yields} PM enhanced the transduction of PEP-1-catalase into HaCaT cells and skin tissue. {yields} PM increased anti-inflammatory activity of PEP-1-catalase. {yields} PM stimulated therapeutic action of anti-oxidant enzyme catalase in oxidative-related diseases. -- Abstract: The low transduction efficiency of various proteins is an obstacle to their therapeutic application. However, protein transduction domains (PTDs) are well-known for a highly effective tool for exogenous protein delivery to cells. We examined the effects of pergolide mesylate (PM) on the transduction of PEP-1-catalase into HaCaT human keratinocytes and mice skin and on the anti-inflammatory activity of PEP-1-catatase against 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation using Western blot and histological analysis. PM enhanced the time- and dose-dependent transduction of PEP-1-catalase into HaCaT cells without affecting the cellular toxicity. In a mouse edema model, PEP-1-catalase inhibited the increased expressions of inflammatory mediators and cytokines such as cyclooxygenase-2, inducible nitric oxide synthase, interleukin-6 and -1{beta}, and tumor necrosis factor-{alpha} induced by TPA. On the other hand, PM alone failed to exert any significant anti-inflammatory effects. However, the anti-inflammatory effect of co-treatment with PEP-1-catalase and PM was more potent than that of PEP-1-catalase alone. Our results indicate that PM may enhance the delivery of PTDs fusion therapeutic proteins to target cells and tissues and has potential to increase their therapeutic effects of such drugs against various diseases.

  7. The Molecular Mechanism of the Catalase-like Activity in Horseradish Peroxidase.

    Science.gov (United States)

    Campomanes, Pablo; Rothlisberger, Ursula; Alfonso-Prieto, Mercedes; Rovira, Carme

    2015-09-02

    Horseradish peroxidase (HRP) is one of the most relevant peroxidase enzymes, used extensively in immunochemistry and biocatalysis applications. Unlike the closely related catalase enzymes, it exhibits a low activity to disproportionate hydrogen peroxide (H2O2). The origin of this disparity remains unknown due to the lack of atomistic information on the catalase-like reaction in HRP. Using QM(DFT)/MM metadynamics simulations, we uncover the mechanism for reduction of the HRP Compound I intermediate by H2O2 at atomic detail. The reaction begins with a hydrogen atom transfer, forming a peroxyl radical and a Compound II-like species. Reorientation of the peroxyl radical in the active site, concomitant with the transfer of the second hydrogen atom, is the rate-limiting step, with a computed free energy barrier (18.7 kcal/mol, ∼ 6 kcal/mol higher than the one obtained for catalase) in good agreement with experiments. Our simulations reveal the crucial role played by the distal pocket residues in accommodating H2O2, enabling formation of a Compound II-like intermediate, similar to catalases. However, out of the two pathways for Compound II reduction found in catalases, only one is operative in HRP. Moreover, the hydrogen bond network in the distal side of HRP compensates less efficiently than in catalases for the energetic cost required to reorient the peroxyl radical at the rate-determining step. The distal Arg and a water molecule in the "wet" active site of HRP have a substantial impact on the reaction barrier, compared to the "dry" active site in catalase. Therefore, the lower catalase-like efficiency of heme peroxidases compared to catalases can be directly attributed to the different distal pocket architecture, providing hints to engineer peroxidases with a higher rate of H2O2 disproportionation.

  8. Catalase in testes and epididymidis of wistar rats fed zinc deficient diet.

    Science.gov (United States)

    Bedwal, S; Prasad, S; Nair, N; Saini, M R; Bedwal, R S

    2009-01-01

    Catalase activities have been evaluated in testes and caput and cauda epididymis of Wistar rats fed on zinc deficient diet for 2 and 4 weeks. The enzyme activity has been measured as chromic acetate formed by heating of dichromate (in acetic acid) in presence of H(2) O(2) with perchromic acid as an unstable intermediate. Observed non-significant increase in catalase activity in testes as well as in caput and cauda epididymis of 2 weeks experiments has been related to low levels of H(2) O(2) produced in two organs whereas significant (Pspermatozoa in epididymis. Thus, zinc deficiency increases catalase activity in testes and epididymis.

  9. Direct Electrochemistry of Catalase on Single Wall Carbon Nanotubes Modified Glassy Carbon Electrode

    Institute of Scientific and Technical Information of China (English)

    Qiang ZHAO; Lun Hui GUAN; Zhen Nan GU; Qian Kun ZHUANG

    2005-01-01

    Direct electrochemistry of catalase (Ct) has been studied on single wall carbon nanotubes (SWNTs) modified glassy carbon (GC) electrode. A pair of well-defined nearly reversible redox peaks is given at --0.48 V (vs. SCE) in 0.1 mol/L phosphate solution (pH 7.0).The peak current in cyclic voltammogram is proportional to the scan rate. The peak potential of catalase is shifted to more negative value when the pH increases. Catalase can adsorb on the SWNTs modified electrode.

  10. Catalase-negative, methicillin-resistant Staphylococcus aureus as a cause of septicemia Staphylococcus aureus catalase-negativo resistente a meticilina como causa de septicemia

    Directory of Open Access Journals (Sweden)

    Ana Lúcia Innaco de Carvalho

    2003-01-01

    Full Text Available A catalase-negative methicillin-resistant Staphylococcus aureus (MRSA was isolated from blood, venous catheter spike and bone marrow collected from an HIV-positive man with lobar pneumonia and sepsis after ten days of hospitalization. The isolate was resistant to oxacillin (positive for penicillin-binding protein 2', ceftriaxone, clindamycin and clarithromycin, and susceptible to vancomycin. This is the first case of septicemia due to a catalase-negative S. aureus reported in Brazil, and, to our knowledge, it is the first case of catalase-negative MRSA reported in the literature. We believe that the patient acquired the S. aureus infection within the hospital environment since it was isolated ten days after hospitalization, it was isolated in a venous catheter spike, and the antibiotic resistance profile is similar to other S. aureus isolates recovered from infections in our hospital.Em um paciente HIV-positivo, com pneumonia lobar e septicemia, foi isolada, após dez dias de internação, uma cepa de Staphylococcus aureus catalase-negativa, resistente a meticilina/oxacilina (MRSA, de culturas de sangue, cateter venoso central e medula óssea. A cepa era resistente a oxacilina (PBP 2' positivo, ceftriaxona, clindamicina e claritromicina, e sensível a vancomicina. Este é o primeiro caso, reportado no Brasil, de uma septicemia por S. aureus catalase-negativo e, em nosso conhecimento, o primeiro caso de um S. aureus catalase-negativo resistente a meticilina. Nós acreditamos que o paciente tenha adquirido a infecção no ambiente hospitalar, uma vez que esta cepa foi isolada aos dez dias de internação, foi isolada em cateter venoso central e o perfil de sensibilidade aos antimicrobianos é semelhante ao dos S. aureus de infecções nosocomiais que ocorrem em nosso hospital.

  11. An oxyferrous heme/protein-based radical intermediate is catalytically competent in the catalase reaction of Mycobacterium tuberculosis catalase-peroxidase (KatG).

    Science.gov (United States)

    Suarez, Javier; Ranguelova, Kalina; Jarzecki, Andrzej A; Manzerova, Julia; Krymov, Vladimir; Zhao, Xiangbo; Yu, Shengwei; Metlitsky, Leonid; Gerfen, Gary J; Magliozzo, Richard S

    2009-03-13

    A mechanism accounting for the robust catalase activity in catalase-peroxidases (KatG) presents a new challenge in heme protein enzymology. In Mycobacterium tuberculosis, KatG is the sole catalase and is also responsible for peroxidative activation of isoniazid, an anti-tuberculosis pro-drug. Here, optical stopped-flow spectrophotometry, rapid freeze-quench EPR spectroscopy both at the X-band and at the D-band, and mutagenesis are used to identify catalase reaction intermediates in M. tuberculosis KatG. In the presence of millimolar H2O2 at neutral pH, oxyferrous heme is formed within milliseconds from ferric (resting) KatG, whereas at pH 8.5, low spin ferric heme is formed. Using rapid freeze-quench EPR at X-band under both of these conditions, a narrow doublet radical signal with an 11 G principal hyperfine splitting was detected within the first milliseconds of turnover. The radical and the unique heme intermediates persist in wild-type KatG only during the time course of turnover of excess H2O2 (1000-fold or more). Mutation of Met255, Tyr229, or Trp107, which have covalently linked side chains in a unique distal side adduct (MYW) in wild-type KatG, abolishes this radical and the catalase activity. The D-band EPR spectrum of the radical exhibits a rhombic g tensor with dual gx values (2.00550 and 2.00606) and unique gy (2.00344) and gz values (2.00186) similar to but not typical of native tyrosyl radicals. Density functional theory calculations based on a model of an MYW adduct radical built from x-ray coordinates predict experimentally observed hyperfine interactions and a shift in g values away from the native tyrosyl radical. A catalytic role for an MYW adduct radical in the catalase mechanism of KatG is proposed.

  12. Simultaneous production of catalase, glucose oxidase and gluconic acid by Aspergillus niger mutant.

    Science.gov (United States)

    Fiedurek, J; Gromada, A; Pielecki, J

    1998-01-01

    The production of gluconic acid, extracellular glucose oxidase and catalase in submerged culture by a number of biochemical mutants has been evaluated. Optimization of stirrer speed, time cultivation and buffering action of some chemicals on glucose oxidase, catalase and gluconic acid production by the most active mutant, AM-11, grown in a 3-L glass bioreactor was investigated. Three hundred rpm appeared to be optimum to ensure good growth and best glucose oxidase production, but gluconic acid or catalase activity obtained maximal value at 500 or 900 rpm, respectively. Significant increase of dissolved oxygen concentration in culture (16-21%) and extracellular catalase activity were obtained when the traditional aeration was employed together with automatic dosed hydrogen peroxide.

  13. Immobilization of catalase on chitosan and amino acid- modified chitosan beads.

    Science.gov (United States)

    Başak, Esra; Aydemir, Tülin

    2013-08-01

    Bovine liver catalase was covalently immobilized onto amino acid-modified chitosan beads. The beads were characterized with SEM, FTIR, TGA and the effects of immobilization on optimum pH and temperature, thermostability, reusability were evaluated. Immobilized catalase showed the maximal enzyme activity at pH 7.0 at 30°C. The kinetic parameters, Km and Vmax, for immobilized catalase on alanine-chitosan beads and lysine-chitosan beads were estimated to be 25.67 mM, 27 mM and 201.39 μmol H2O2/min, 197.50 μmol H2O2/min, respectively. The activity of the immobilized catalase on Ala-CB and Lys-CB retained 40% of its high initial activity after 100 times of reuse.

  14. Occurrence of High Catalase-containing Acinetobacter in Spacecraft Assembly Facilities

    Science.gov (United States)

    McCoy, K. B.; Derecho, I.; La Duc, M. T.; Vaishampayan, P.; Venkateswaran, K. J.; Mogul, R.

    2010-04-01

    In summary, the measurement of high catalase specific activity values for spacecraft-associated Acinetobacter strains is potentially the result of adaptation towards the harsh conditions of the clean rooms and assembly process.

  15. Catalase in testes and epididymidis of wistar rats fed zinc deficient diet

    Directory of Open Access Journals (Sweden)

    Bedwal S

    2009-01-01

    Full Text Available Catalase activities have been evaluated in testes and caput and cauda epididymis of Wistar rats fed on zinc deficient diet for 2 and 4 weeks. The enzyme activity has been measured as chromic acetate formed by heating of dichromate (in acetic acid in presence of H 2 O 2 with perchromic acid as an unstable intermediate. Observed non-significant increase in catalase activity in testes as well as in caput and cauda epididymis of 2 weeks experiments has been related to low levels of H 2 O 2 produced in two organs whereas significant (P< 0.01/0.001 increase in catalase activity in 4-weeks experiments indicate for increased oxidative stress due to phagocytotic activity of Sertoli cells in testes and damaged spermatozoa in epididymis. Thus, zinc deficiency increases catalase activity in testes and epididymis.

  16. Interference of a commercial catalase preparation in laccase and peroxidase activities

    Directory of Open Access Journals (Sweden)

    Nara Ballaminut

    2009-10-01

    Full Text Available The influence of commercial catalase preparations (fungal and bovine origin on laccase and peroxidase activity assays was evaluated using enzymatic extracts obtained from several basidiomycetes grown under different culture conditions. No hydrogen peroxide was detected in the extracts. Inhibition of laccase activity by 40 to 80% was related to the catalase source. In addition, oxidation of the substrate (ABTS by fungal catalase in the absence of the enzymatic extract from basidiomycetes was observed. The results demonstrated the need for the evaluation of interference of the commercial catalase preparation when its use was required in the reaction mixture.A influência da preparação comercial de catalase (origem fúngica e bovina nos ensaios de atividade absence and presence of a fungal or bovine de lacase e de peroxidases foi avaliada empregando-se extratos enzimáticos obtidos do crescimento de diversos basidiomicetos em diferentes condições de cultivo. Não foi detectado H2O2 nos extratos. Inibição de 40 a 80% da atividade de lacase foi relacionada à fonte de catalase. Além disso, foi observada oxidação do substrato (ABTS pela catalase fúngica, na ausência de extrato enzimático do basidiomiceto. Os resultados evidenciaram a necessidade de se proceder a uma avaliação da interferência da preparação comercial de catalase, quando o seu uso se fizer necessário na mistura reacional.

  17. Immobilization of catalases from Bacillus SF on alumina for the treatment of textile bleaching effluents

    OpenAIRE

    Costa, Silgia; Tzanov,Tzanko; Paar, Andreas; Gudelj, Marinka; Gübitz, Georg M.; Paulo, Artur Cavaco

    2001-01-01

    A catalase preparation from a newly isolated Bacillus sp. was covalently immobilized on silanized alumina using glutaraldehyde as crosslinking agent. The effect of the coupling time of the enzyme-support reaction was determined in terms of protein recovery and immobilization yield and a certain balance point was found after which the activity recovery decreased. The activity profile of the immobilized catalase at high pH and temperature was investigated. The immobilized enzyme showed...

  18. Identification of a Catalase-Phenol Oxidase in Betalain Biosynthesis in Red Amaranth (Amaranthus cruentus)

    OpenAIRE

    Teng, Xiao-Lu; Chen, Ning; Xiao, Xing-Guo

    2016-01-01

    Betalains are a group of nitrogen-containing pigments that color plants in most families of Caryophyllales. Their biosynthesis has long been proposed to begin with hydroxylation of L-tyrosine to L-DOPA through monophenolase activity of tyrosinase, but biochemical evidence in vivo remains lacking. Here we report that a Group 4 catalase, catalase-phenol oxidase (named as AcCATPO), was identified, purified and characterized from leaves of Amaranthus cruentus, a betalain plant. The purified enzym...

  19. Identification of a catalase-phenol oxidase in betalain biosynthesis in red amaranth (Amaranthus cruentus)

    OpenAIRE

    Xiao-Lu eTeng; Ning eChen; Xing-Guo eXiao

    2016-01-01

    Betalains are a group of nitrogen-containing pigments that color plants in most families of Caryophyllales. Their biosynthesis has long been proposed to begin with hydroxylation of L-tyrosine to L-DOPA through monophenolase activity of tyrosinase, but biochemical evidence in vivo remains lacking. Here we report that a Group 4 catalase, catalase-phenol oxidase (named as AcCATPO), was identified, purified and characterized from leaves of Amaranthus cruentus, a betalain plant. The purified enzym...

  20. Catalases play differentiated roles in the adaptation of a fungal entomopathogen to environmental stresses.

    Science.gov (United States)

    Wang, Zheng-Liang; Zhang, Long-Bin; Ying, Sheng-Hua; Feng, Ming-Guang

    2013-02-01

    The catalase family of Beauveria bassiana (fungal entomopathogen) consists of catA (spore-specific), catB (secreted), catP (peroxisomal), catC (cytoplasmic) and catD (secreted peroxidase/catalase), which were distinguished in phylogeny and structure and functionally characterized by constructing single-gene disrupted and rescued mutants for enzymatic and multi-phenotypic analyses. Total catalase activity decreased 89% and 56% in ΔcatB and ΔcatP, corresponding to the losses of upper and lower active bands gel-profiled for all catalases respectively, but only 9-12% in other knockout mutants. Compared with wild type and complement mutants sharing similar enzymatic and phenotypic parameters, all knockout mutants showed significant (9-56%) decreases in the antioxidant capability of their conidia (active ingredients of mycoinsecticides), followed by remarkable phenotypic defects associated with the fungal biocontrol potential. These defects included mainly the losses of 40% thermotolerance (45°C) in ΔcatA, 46-48% UV-B resistance in ΔcatA and ΔcatD, and 33-47% virulence to Spodoptera litura larvae in ΔcatA, ΔcatP and ΔcatD respectively. Moreover, the drastic transcript upregulation of some other catalase genes observed in the normal culture of each knockout mutant revealed functionally complimentary effects among some of the catalase genes, particularly between catB and catC whose knockout mutants displayed little or minor phenotypic changes. However, the five catalase genes functioned redundantly in mediating the fungal tolerance to either hyperosmotic or fungicidal stress. The differentiated roles of five catalases in regulating the B. bassiana virulence and tolerances to oxidative stress, high temperature and UV-B irradiation provide new insights into fungal adaptation to stressful environment and host invasion.

  1. Fluorescence Spectrometry of the Interaction of Multi-Walled Carbon Nanotubes with Catalase

    Science.gov (United States)

    Fan, Y.; Li, Y.; Cai, H.; Li, J.; Miao, J.; Fu, D.; Yang, Q.

    2014-11-01

    The interaction of multi-walled carbon nanotubes (MWCNTs) with catalase is investigated using fluorescence and circular dichroism spectroscopic techniques. The results of the fluorescence experiments suggest that MWCNTs quench the intrinsic fluorescence of catalase via a static quenching mechanism. The circular dichroism spectral results reveal the unfolding of catalase with a significant decrease in the α-helix content in the presence of MWCNTs, which indicates that the conformation of catalase is changed in the binding process, thereby remarkably decreasing its activity. The binding constants and the number of binding sites of the MWCNT to the catalase are calculated at different temperatures. The thermodynamic parameters, such as the changes in free energy (ΔG), enthalpy (ΔH), and entropy (ΔS), are calculated using thermodynamic equations. The fact that all negative values of ΔG, ΔH, and ΔS are obtained suggests that the interaction of the MWCNTs with catalase is spontaneous, and that hydrogen bonding and van der Waals interactions play an important role in the binding process.

  2. Generation 9 polyamidoamine dendrimer encapsulated platinum nanoparticle mimics catalase size, shape, and catalytic activity.

    Science.gov (United States)

    Wang, Xinyu; Zhang, Yincong; Li, Tianfu; Tian, Wende; Zhang, Qiang; Cheng, Yiyun

    2013-04-30

    Poly(amidoamine) (PAMAM) encapsulated platinum nanoparticles were synthesized and used as catalase mimics. Acetylated generation 9 (Ac-G9) PAMAM dendrimer with a molecular size around 10 nm was used as a template to synthesize platinum nanoparticles. The feeding molar ratio of Pt(4+) and Ac-G9 is 2048, and the synthesized platinum nanoparticle (Ac-G9/Pt NP) has an average size of 3.3 nm. Ac-G9/Pt NP has a similar molecular size and globular shape with catalase (~11 nm). The catalytic activity of Ac-G9/Pt NP on the decomposition of H2O2 is approaching that of catalase at 37 °C. Ac-G9/Pt NP shows differential response to the changes of pH and temperature compared with catalase, which can be explained by different catalytic mechanisms of Ac-G9/Pt NP and catalase. Ac-G9/Pt NP also shows horseradish peroxidase activity and is able to scavenge free radicals such as di(phenyl)-(2,4,6-trinitrophenyl)iminoazanium (DPPH). Furthermore, Ac-G9/Pt NP shows excellent biocompatibility on different cell lines and can down-regulate H2O2-induced intracellular reactive oxygen species (ROS) in these cells. These results suggest that dendrimers are promising mimics of proteins with different sizes and Ac-G9/Pt NP can be used as an alternative candidate of catalase to decrease oxidation stress in cells.

  3. Reversible adsorption of catalase onto Fe(3+) chelated poly(AAm-GMA)-IDA cryogels.

    Science.gov (United States)

    Aktaş Uygun, Deniz; Uygun, Murat; Akgöl, Sinan; Denizli, Adil

    2015-05-01

    In this presented study, poly(acrylamide-glycidyl methacrylate) [poly(AAm-GMA)] cryogels were synthesized by cryopolymerization technique at sub-zero temperature. Prepared cryogels were then functionalized with iminodiacetic acid (IDA) and chelated with Fe(3+) ions in order produce the metal chelate affinity matrix. Synthesized cryogels were characterized with FTIR, ESEM and EDX analysis, and it was found that the cryogel had sponge like structure with interconnected pores and their pore diameter was about 200 μm. Fe(3+) chelated poly(AAm-GMA)-IDA cryogels were used for the adsorption of catalase and optimum adsorption conditions were determined by varying the medium pH, initial catalase concentration, temperature and ionic strength. Maximum catalase adsorption onto Fe(3+) chelated poly(AAm-GMA)-IDA cryogel was found to be 12.99 mg/g cryogel at 25 °C, by using pH 5.0 acetate buffer. Adsorbed catalase was removed from the cryogel by using 1.0M of NaCl solution and desorption yield was found to be 96%. Additionally, reusability profile of the Fe(3+) chelated poly(AAm-GMA)-IDA cryogel was also investigated and it was found that, adsorption capacity of the cryogels didn't decrease significantly at the end of the 40 reuses. Catalase activity studies were also tested and it was demonstrated that desorbed catalase retained 70% of its initial activity.

  4. Inhibitory effects of a novel Val to Thr mutation on the distal heme of human catalase.

    Science.gov (United States)

    Mashhadi, Zahra; Boeglin, William E; Brash, Alan R

    2014-11-01

    True catalases efficiently breakdown hydrogen peroxide, whereas the catalase-related enzyme allene oxide synthase (cAOS) is completely unreactive and instead metabolizes a fatty acid hydroperoxide. In cAOS a Thr residue adjacent to the distal His restrains reaction with H2O2 (Tosha et al. (2006) J. Biol. Chem. 281:12610; De Luna et al. (2013) J. Phys. Chem. B 117: 14635) and its mutation to the consensus Val of true catalases permits the interaction. Here we investigated the effects of the reciprocal experiment in which the Val74 of human catalase is mutated to Thr, Ser, Met, Pro, or Ala. The Val74Thr substitution decreased catalatic activity by 3.5-fold and peroxidatic activity by 3-fold. Substitution with Ser had similar negative effects (5- and 3-fold decreases). Met decreased catalatic activity 2-fold and eliminated peroxidatic activity altogether, whereas the Val74Ala substitution was well tolerated. (The Val74Pro protein lacked heme). We conclude that the conserved Val74 of true catalases helps optimize catalysis. There are rare substitutions of Val74 with Ala, Met, or Pro, but not with Ser of Thr, possibly due their hydrogen bonding affecting the conformation of His75, the essential distal heme residue for activity in catalases.

  5. Binding of Cimetidine to Balb/C Mouse Liver Catalase; Kinetics and Conformational Studies.

    Science.gov (United States)

    Jahangirvand, Mahboubeh; Minai-Tehrani, Dariush; Yazdi, Fatemeh; Minai-Tehrani, Arash; Razmi, Nematollah

    2016-01-01

    Catalase is responsible for converting hydrogen peroxide (H2O2) into water and oxygen in cells. This enzyme has high affinity for hydrogen peroxide and can protect the cells from oxidative stress damage. Catalase is a tetramer protein and each monomer contains a heme group. Cimetidine is a histamine H2 receptor blocker which inhibits acid release from stomach and is used for gasterointestinal diseases. In this research, effect of cimetidine on the activity of liver catalase was studied and the kinetic parameters of this enzyme and its conformational changes were investigated. Cell free extract of mouse liver was used for the catalase assay. The activity of the catalase was detected in the absence and presence of cimetidine by monitoring hydrogen peroxide reduction absorbance at 240 nm. The purified enzyme was used for conformational studies by Fluorescence spectrophotometry. The data showed that cimetidine could inhibit the enzyme in a non-competitive manner. Ki and IC50 values of the drug were determined to be about 0.75 and 0.85 uM, respectively. The Arrhenius plot showed that activation energy was 6.68 and 4.77 kJ/mol in the presence and absence of the drug, respectively. Fluorescence spectrophotometry revealed that the binding of cimetidine to the purified enzyme induced hyperchromicity and red shift which determined the conformational change on the enzyme. Cimetidine could non-competitively inhibit the liver catalase with high affinity. Binding of cimetidine to the enzyme induced conformational alteration in the enzyme.

  6. Not so monofunctional--a case of thermostable Thermobifida fusca catalase with peroxidase activity.

    Science.gov (United States)

    Lončar, Nikola; Fraaije, Marco W

    2015-03-01

    Thermobifida fusca is a mesothermophilic organism known for its ability to degrade plant biomass and other organics, and it was demonstrated that it represents a rich resource of genes encoding for potent enzymes for biocatalysis. The thermostable catalase from T. fusca has been cloned and overexpressed in Escherichia coli with a yield of 400 mg/L. Heat treatment of disrupted cells at 60 °C for 1 h resulted in enzyme preparation of high purity; hence, no chromatography steps are needed for large-scale production. Except for catalyzing the dismutation of hydrogen peroxide, TfuCat was also found to catalyze oxidations of phenolic compounds. The catalase activity was comparable to other described catalases while peroxidase activity was quite remarkable with a k obs of nearly 1000 s(-1) for catechol. Site directed mutagenesis was used to alter the ratio of peroxidase/catalase activity. Resistance to inhibition by classic catalase inhibitors and an apparent melting temperature of 74 °C classifies this enzyme as a robust biocatalyst. As such, it could compete with other commercially available catalases while the relatively high peroxidase activity also offers new biocatalytic possibilities.

  7. Milk catalase activity as an indicator of thermization treatments used in the manufacture of cheddar cheese.

    Science.gov (United States)

    Hirvi, Y; Griffiths, M W

    1998-02-01

    Pilot-scale studies were carried out to determine the effect of different heat treatments on catalase activity during the manufacture and maturation of Cheddar cheese. Three trials were conducted to monitor catalase activity using disk flotation and polarographic methods. Cheese was manufactured from raw milk and from milk that had been treated at 60, 65 and 72 degrees C for 16 s using a high temperature, short time heat exchanger. Catalase activity was also determined in samples of commercial milk and in samples of mild, medium, sharp, and extra sharp Cheddar cheeses obtained from different manufacturers in order to verify that the enzyme could be used as an indicator of the type of heat treatment applied to cheese milk. Catalase activity was present in cheese made from raw milk but was only present at low concentrations in cheese manufactured from thermized milk. However, high catalase activity was observed in commercial samples of sharp and extra sharp Cheddar cheese that was apparently due to the growth of catalase-producing yeasts in the cheese during maturation.

  8. Chromatin remodeling regulates catalase expression during cancer cells adaptation to chronic oxidative stress.

    Science.gov (United States)

    Glorieux, Christophe; Sandoval, Juan Marcelo; Fattaccioli, Antoine; Dejeans, Nicolas; Garbe, James C; Dieu, Marc; Verrax, Julien; Renard, Patricia; Huang, Peng; Calderon, Pedro Buc

    2016-10-01

    Regulation of ROS metabolism plays a major role in cellular adaptation to oxidative stress in cancer cells, but the molecular mechanism that regulates catalase, a key antioxidant enzyme responsible for conversion of hydrogen peroxide to water and oxygen, remains to be elucidated. Therefore, we investigated the transcriptional regulatory mechanism controlling catalase expression in three human mammary cell lines: the normal mammary epithelial 250MK primary cells, the breast adenocarcinoma MCF-7 cells and an experimental model of MCF-7 cells resistant against oxidative stress resulting from chronic exposure to H2O2 (Resox), in which catalase was overexpressed. Here we identify a novel promoter region responsible for the regulation of catalase expression at -1518/-1226 locus and the key molecules that interact with this promoter and affect catalase transcription. We show that the AP-1 family member JunB and retinoic acid receptor alpha (RARα) mediate catalase transcriptional activation and repression, respectively, by controlling chromatin remodeling through a histone deacetylases-dependent mechanism. This regulatory mechanism plays an important role in redox adaptation to chronic exposure to H2O2 in breast cancer cells. Our study suggests that cancer adaptation to oxidative stress may be regulated by transcriptional factors through chromatin remodeling, and reveals a potential new mechanism to target cancer cells.

  9. Catalase overexpression reduces the germination time and increases the pathogenicity of the fungus Metarhizium anisopliae.

    Science.gov (United States)

    Morales Hernandez, Claudia Erika; Padilla Guerrero, Israel Enrique; Gonzalez Hernandez, Gloria Angelica; Salazar Solis, Eduardo; Torres Guzman, Juan Carlos

    2010-07-01

    Catalases and peroxidases are the most important enzymes that degrade hydrogen peroxide into water and oxygen. These enzymes and superoxide dismutase are the first lines of cell defense against reactive oxygen species. Metarhizium anisopliae displays an increase in catalase-peroxidase activity during germination and growth. To determine the importance of catalase during the invasion process of M. anisopliae, we isolated the cat1 gene. cat1 cDNA expression in Escherichia coli and the subsequent purification of the protein confirmed that the cat1 gene codes for a monofunctional catalase. Expression analysis of this gene by RT-PCR from RNA isolated from fungus grown in liquid cultures showed a decrease in the expression level of the cat1 gene during germination and an increase during mycelium growth. The expression of this gene in the fungus during the infection process of the larvae of Plutella xylostella also showed a significant increase during invasive growth. Transgenic strains overexpressing the cat1 gene had twice the catalase activity of the wild-type strain. This increase in catalase activity was accompanied by a higher level of resistance to exogenous hydrogen peroxide and a reduction in the germination time. This improvement was also observed during the infection of P. xylostella larvae. M. anisopliae transgenic strains overexpressing the cat1 gene grew and spread faster in the soft tissue of the insect, reducing the time to death of the insect by 25% and the dose required to kill 50% of the population 14-fold.

  10. Adeno-Associated Viral-Mediated Catalase Expression Suppresses Optic Neuritis in Experimental Allergic Encephalomyelitis

    Science.gov (United States)

    Guy, John; Qi, Xiaoping; Hauswirth, William W.

    1998-11-01

    Suppression of oxidative injury by viral-mediated transfer of the human catalase gene was tested in the optic nerves of animals with experimental allergic encephalomyelitis (EAE). EAE is an inflammatory autoimmune disorder of primary central nervous system demyelination that has been frequently used as an animal model for the human disease multiple sclerosis (MS). The optic nerve is a frequent site of involvement common to both EAE and MS. Recombinant adeno-associated virus containing the human gene for catalase was injected over the right optic nerve heads of SJL/J mice that were simultaneously sensitized for EAE. After 1 month, cell-specific catalase activity, evaluated by quantitation of catalase immunogold, was increased approximately 2-fold each in endothelia, oligodendroglia, astrocytes, and axons of the optic nerve. Effects of catalase on the histologic lesions of EAE were measured by computerized analysis of the myelin sheath area (for demyelination), optic disc area (for optic nerve head swelling), extent of the cellular infiltrate, extravasated serum albumin labeled by immunogold (for blood-brain barrier disruption), and in vivo H2O2 reaction product. Relative to control, contralateral optic nerves injected with the recombinant virus without a therapeutic gene, catalase gene inoculation reduced demyelination by 38%, optic nerve head swelling by 29%, cellular infiltration by 34%, disruption of the blood-brain barrier by 64%, and in vivo levels of H2O2 by 61%. Because the efficacy of potential treatments for MS are usually initially tested in the EAE animal model, this study suggests that catalase gene delivery by using viral vectors may be a therapeutic strategy for suppression of MS.

  11. Adventitial gene transfer of catalase attenuates angiotensin II-induced vascular remodeling.

    Science.gov (United States)

    Liu, Cun-Fei; Zhang, Jia; Shen, Kai; Gao, Ping-Jin; Wang, Hai-Ya; Jin, Xin; Meng, Chao; Fang, Ning-Yuan

    2015-04-01

    Vascular adventitia and adventitia‑derived reactive oxygen species (ROS) contribute to vascular remodeling following vascular injury. A previous ex vivo study in adventitial fibroblasts showed that catalase, one of most important anti‑oxide enzymes, was downregulated by angiotensin II (AngII). The aim of the present study was to investigate whether adventitial gene transfer of catalase affects AngII‑induced vascular remodeling in vivo. Adenoviruses co‑expressing catalase and enhanced green fluorescent protein (eGFP) or expressing eGFP only were applied to the adventitial surface of common carotid arteries of Sprague‑Dawley rats. Alzet minipumps administering AngII (0.75 mg/kg/day) were then implanted subcutaneously for 14 days. Systolic blood pressure and biological parameters of vascular remodeling were measured in each group. Adventitial fibroblasts were cultured and p38 mitogen‑activated protein kinase (MAPK) phosphorylation was measured using western blot analysis. The results showed that adventitial gene transfer of catalase had no effect on AngII‑induced systolic blood pressure elevation. However, catalase adenovirus transfection significantly inhibited AngII‑induced media hypertrophy compared with that of the control virus (Pcatalase transfection significantly attenuated AngII‑induced ROS generation, macrophage infiltration, collagen deposition and adventitial α‑smooth muscle actin expression. Furthermore, catalase transfection significantly inhibited the AngII‑induced increase in p38MAPK phosphorylation. In conclusion, the results of the present study demonstrated that adventitial gene transfer of catalase significantly attenuated AngII‑induced vascular remodeling in rats via inhibition of adventitial p38MAPK phosphorylation.

  12. The role and regulation of catalase in respiratory tract opportunistic bacterial pathogens.

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    Eason, Mia M; Fan, Xin

    2014-09-01

    Respiratory tract bacterial pathogens are the etiologic agents of a variety of illnesses. The ability of these bacteria to cause disease is imparted through survival within the host and avoidance of pathogen clearance by the immune system. Respiratory tract pathogens are continually bombarded by reactive oxygen species (ROS), which may be produced by competing bacteria, normal metabolic function, or host immunological responses. In order to survive and proliferate, bacteria have adapted defense mechanisms to circumvent the effects of ROS. Bacteria employ the use of anti-oxidant enzymes, catalases and catalase-peroxidases, to relieve the effects of the oxidative stressors to which they are continually exposed. The decomposition of ROS has been shown to provide favorable conditions in which respiratory tract opportunistic bacterial pathogens such as Haemophilus influenzae, Mycobacterium tuberculosis, Legionella pneumophila, and Neisseria meningitidis are able to withstand exposure to highly reactive molecules and yet survive. Bacteria possessing mutations in the catalase gene have a decreased survival rate, yet may be able to compensate for the lack of catalatic activity if peroxidatic activity is present. An incomplete knowledge of the mechanisms by which catalase and catalase-peroxidases are regulated still persists, however, in some bacterial species, a regulatory factor known as OxyR has been shown to either up-regulate or down-regulate catalase gene expression. Yet, more research is still needed to increase the knowledge base in relation to this enzyme class. As with this review, we focus on major respiratory tract opportunistic bacterial pathogens in order to elucidate the function and regulation of catalases. The importance of the research could lead to the development of novel treatments against respiratory bacterial infections.

  13. Evaluation on the Toxic Effects of NanoAg to Catalase.

    Science.gov (United States)

    Zhang, Bin; Zhai, Wenxin; Liu, Rutao; Yu, Zehua; Shen, Hengmei; Hu, Xinxin

    2015-02-01

    Protein is the functional actor of life. Research on protein damage induced by nanomaterials may give insight into the toxicity mechanisms of nanoparticles. Studying nano silver over the impact of the structure and function of catalase (CAT) at the molecular level, is of great significance for a comprehensive evaluation of their toxic effects. The toxic effects of nanoAg on catalase were thoroughly investigated using steady state and time resolved fluorescence quenching measurements, ultraviolet-visible absorption spectroscopy, resonance light scattering spectroscopy (RLS), circular dichroism spectroscopy (CD) and transmission electron microscopy (TEM). NanoAg could decrease the amount of alpha-helix and increase the beta sheet structure, leading to loose the skeleton structure of catalase. The characteristic fluorescence of catalase was obviously quenched, which showed the exposal of internal hydrophobic amino acids enhanced, and its quenching type is dynamic quenching. The result of RLS and TEM showed that the distribution and size of nanoAg become more uniform and smaller after their interaction, resulting in a decrease of RLS intensity. NanoAg could make the activity of catalase rise. By changing the structure of catalase, nanoAg increases its enzymatic activity to a certain extent, breaking down its balance in vivo, thereby affecting the normal physiological activities. NanoAg has obvious toxic effects on catalase. This paper provided a new perspective and method for the toxic effects of nanoAg to biological macromolecules; provided basic data and reference gist for the hygienics and toxicology studies of nanoAg. It is conducive to the toxicity prevention and control work of nanoAg, promoting nano-technology applied to human production and living better.

  14. Reduction of hydrogen peroxide accumulation and toxicity by a catalase from Mycoplasma iowae.

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    Rachel E Pritchard

    Full Text Available Mycoplasma iowae is a well-established avian pathogen that can infect and damage many sites throughout the body. One potential mediator of cellular damage by mycoplasmas is the production of H2O2 via a glycerol catabolic pathway whose genes are widespread amongst many mycoplasma species. Previous sequencing of M. iowae serovar I strain 695 revealed the presence of not only genes for H2O2 production through glycerol catabolism but also the first documented mycoplasma gene for catalase, which degrades H2O2. To test the activity of M. iowae catalase in degrading H2O2, we studied catalase activity and H2O2 accumulation by both M. iowae serovar K strain DK-CPA, whose genome we sequenced, and strains of the H2O2-producing species Mycoplasma gallisepticum engineered to produce M. iowae catalase by transformation with the M. iowae putative catalase gene, katE. H2O2-mediated virulence by M. iowae serovar K and catalase-producing M. gallisepticum transformants were also analyzed using a Caenorhabditis elegans toxicity assay, which has never previously been used in conjunction with mycoplasmas. We found that M. iowae katE encodes an active catalase that, when expressed in M. gallisepticum, reduces both the amount of H2O2 produced and the amount of damage to C. elegans in the presence of glycerol. Therefore, the correlation between the presence of glycerol catabolism genes and the use of H2O2 as a virulence factor by mycoplasmas might not be absolute.

  15. Purification and characterization of catalase from sprouted black gram (Vigna mungo) seeds.

    Science.gov (United States)

    Kandukuri, Sai Srikar; Noor, Ayesha; Ranjini, S Shiva; Vijayalakshmi, M A

    2012-03-15

    Black gram (Vigna mungo) is a legume which belongs to Fabaceae family. It is a rich source of protein. It has been known to have interesting small molecule antioxidant activity. However, its enzymatic antioxidant properties have not been explored much. In the present work we studied catalase, a principal antioxidant enzyme from black gram seeds. Day four sprouted black gram seeds were found to have a significant catalase content approximately of 15,240 U/g seeds. IMAC (Seph 4B-IDA-Zn(II)) was used for purifying this catalase, a purification fold of 106 and a high specific activity of 25,704 U/mg was obtained. The K(m) and V(max) of the purified catalase were found to be 16.2 mM and 2.5 μmol/min. The effect of inhibitors like Sodium azide (NaN(3)) and EDTA and different metal ions on catalase activity were studied. NaN(3), Fe(3+)and Cu(2+) were found to have profound inhibitory effects on the enzyme activity. Other metal ions like Ni(2+), Ca(2+), Mg(2+) and Mn(2+) had both enhancing and inhibitory effects. The enzyme showed optimal activity at a temperature of 40°C and pH 7.0. It was stable over a broad range of pH 6.0-10.0 and had a half life of 7h 30 min at 50°C.

  16. Arrhenius activation energy of damage to catalase during spray-drying.

    Science.gov (United States)

    Schaefer, Joachim; Lee, Geoffrey

    2015-07-15

    The inactivation of catalase during spray-drying over a range of outlet gas temperatures could be closely represented by the Arrhenius equation. From this an activation energy for damage to the catalase could be calculated. The close fit to Arrhenius suggests that the thermally-induced part of inactivation of the catalase during the complex drying and particle-formation processes takes place at constant temperature. These processes are rapid compared with the residence time of the powder in the collecting vessel of the cyclone where dried catalase is exposed to a constant temperature equal to approximately the drying gas outlet temperature. A lower activation energy after spray drying with the ultrasonic nozzle was found than with the 2-fluid nozzle under otherwise identical spray drying conditions. It is feasible that the ultrasonic nozzle when mounted in the lid of the spray dryer heats up toward the drying gas inlet temperature much more that the air-cooled 2-fluid nozzle. Calculation of the Arrhenius activation energy also showed how the stabilizing efficacy of trehalose and mannitol on the catalase varies in strength across the range of drying gas inlet and outlet temperatures examined.

  17. Ectopic catalase expression in mitochondria by adeno-associated virus enhances exercise performance in mice.

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    Dejia Li

    Full Text Available Oxidative stress is thought to compromise muscle contractility. However, administration of generic antioxidants has failed to convincingly improve performance during exhaustive exercise. One possible explanation may relate to the inability of the supplemented antioxidants to effectively eliminate excessive free radicals at the site of generation. Here, we tested whether delivering catalase to the mitochondria, a site of free radical production in contracting muscle, could improve treadmill performance in C57Bl/6 mice. Recombinant adeno-associated virus serotype-9 (AV.RSV.MCAT was generated to express a mitochondria-targeted catalase gene. AV.RSV.MCAT was delivered to newborn C57Bl/6 mouse circulation at the dose of 10(12 vector genome particles per mouse. Three months later, we observed a approximately 2 to 10-fold increase of catalase protein and activity in skeletal muscle and the heart. Subcellular fractionation western blot and double immunofluorescence staining confirmed ectopic catalase expression in the mitochondria. Compared with untreated control mice, absolute running distance and body weight normalized running distance were significantly improved in AV.RSV.MCAT infected mice during exhaustive treadmill running. Interestingly, ex vivo contractility of the extensor digitorum longus muscle was not altered. Taken together, we have demonstrated that forced catalase expression in the mitochondria enhances exercise performance. Our result provides a framework for further elucidating the underlying mechanism. It also raises the hope of applying similar strategies to remove excessive, pathogenic free radicals in certain muscle diseases (such as Duchenne muscular dystrophy and ameliorate muscle disease.

  18. Immobilization and kinetics of catalase on calcium carbonate nanoparticles attached epoxy support.

    Science.gov (United States)

    Preety; Hooda, Vinita

    2014-01-01

    A novel hybrid epoxy/nano CaCO3 composite matrix for catalase immobilization was prepared by polymerizing epoxy resin in the presence of CaCO3 nanoparticles. The hybrid support was characterized using scanning electron microscopy and Fourier transform infrared spectroscopy. Catalase was successfully immobilized onto epoxy/nano CaCO3 support with a conjugation yield of 0.67 ± 0.01 mg/cm(2) and 92.63 ± 0.80 % retention of activity. Optimum pH and optimum temperature of free and immobilized catalases were found to be 7.0 and 35 °C. The value of Km for H2O2 was higher for immobilized enzyme (31.42 mM) than native enzyme (27.73 mM). A decrease in Vmax value from 1,500 to 421.10 μmol (min mg protein)(-1) was observed after immobilization. Thermal and storage stabilities of catalase improved immensely after immobilization. Immobilized enzyme retained three times than the activity of free enzyme when kept at 75 °C for 1 h and the half-life of enzyme increased five times when stored in phosphate buffer (0.01 M, pH 7.0) at 5 °C. The enzyme could be reused 30 times without any significant loss of its initial activity. Desorption of catalase from the hybrid support was minimum at pH 7.0.

  19. Novel Role of Endogenous Catalase in Macrophage Polarization in Adipose Tissue

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    Ye Seul Park

    2016-01-01

    Full Text Available Macrophages are important components of adipose tissue inflammation, which results in metabolic diseases such as insulin resistance. Notably, obesity induces a proinflammatory phenotypic switch in adipose tissue macrophages, and oxidative stress facilitates this switch. Thus, we examined the role of endogenous catalase, a key regulator of oxidative stress, in the activity of adipose tissue macrophages in obese mice. Catalase knockout (CKO exacerbated insulin resistance, amplified oxidative stress, and accelerated macrophage infiltration into epididymal white adipose tissue in mice on normal or high-fat diet. Interestingly, catalase deficiency also enhanced classical macrophage activation (M1 and inflammation but suppressed alternative activation (M2 regardless of diet. Similarly, pharmacological inhibition of catalase activity using 3-aminotriazole induced the same phenotypic switch and inflammatory response in RAW264.7 macrophages. Finally, the same phenotypic switch and inflammatory responses were observed in primary bone marrow-derived macrophages from CKO mice. Taken together, the data indicate that endogenous catalase regulates the polarization of adipose tissue macrophages and thereby inhibits inflammation and insulin resistance.

  20. Targeting catalase but not peroxiredoxins enhances arsenic trioxide-induced apoptosis in K562 cells.

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    Li-Li Song

    Full Text Available Despite considerable efficacy of arsenic trioxide (As2O3 in acute promyelocytic leukemia (APL treatment, other non-APL leukemias, such as chronic myeloid leukemia (CML, are less sensitive to As2O3 treatment. However, the underlying mechanism is not well understood. Here we show that relative As2O3-resistant K562 cells have significantly lower ROS levels than As2O3-sensitive NB4 cells. We compared the expression of several antioxidant enzymes in these two cell lines and found that peroxiredoxin 1/2/6 and catalase are expressed at high levels in K562 cells. We further investigated the possible role of peroxirdoxin 1/2/6 and catalase in determining the cellular sensitivity to As2O3. Interestingly, knockdown of peroxiredoxin 1/2/6 did not increase the susceptibility of K562 cells to As2O3. On the contrary, knockdown of catalase markedly enhanced As2O3-induced apoptosis. In addition, we provide evidence that overexpression of BCR/ABL cannot increase the expression of PRDX 1/2/6 and catalase. The current study reveals that the functional role of antioxidant enzymes is cellular context and treatment agents dependent; targeting catalase may represent a novel strategy to improve the efficacy of As2O3 in CML treatment.

  1. Functional and structural changes of human erythrocyte catalase induced by cimetidine: proposed model of binding.

    Science.gov (United States)

    Yazdi, Fatemeh; Minai-Tehrani, Dariush; Jahngirvand, Mahboubeh; Almasirad, Ali; Mousavi, Zahra; Masoud, Masoudeh; Mollasalehi, Hamidreza

    2015-06-01

    In erythrocyte, catalase plays an important role to protect cells from hydrogen peroxide toxicity. Hydrogen peroxide is a byproduct compound which is produced during metabolic pathway of cells. Cimetidine, a histamine H2 receptor antagonist, is used for gastrointestinal tract diseases and prevents the extra release of gastric acid. In this study, the effect of cimetidine on the activity of human erythrocyte catalase was investigated. Erythrocytes were broken by hypotonic solution. The supernatant was used for catalase assay and kinetics study. Lineweaver-Burk plot was performed to determine the type of inhibition. The kinetics data revealed that cimetidine inhibited the catalase activity by mixed inhibition. The IC50 (1.54 μM) and Ki (0.45 μM) values of cimetidine determined that the drug was bound to the enzyme with high affinity. Circular dichroism and fluorescence measurement showed that the binding of cimetidine to the enzyme affected the content of secondary structure of the enzyme as well as its conformational changes. Docking studies were carried out to detect the site in which the drug was bound to the enzyme. Molecular modeling and energy calculation of the binding showed that the cyanoguanidine group of the drug connected to Asp59 via two hydrogen bonds, while the imidazole group of the drug interacted with Phe64 in the enzyme by a hydrophobic interaction. In conclusion, cimetidine could bind to human erythrocyte catalase, and its interaction caused functional and conformational changes in the enzyme.

  2. Direct measurement of catalase activity in living cells and tissue biopsies.

    Science.gov (United States)

    Scaglione, Christine N; Xu, Qijin; Ramanujan, V Krishnan

    2016-01-29

    Spatiotemporal regulation of enzyme-substrate interactions governs the decision-making steps in biological systems. Enzymes, being functional units of every living cell, contribute to the macromolecular stability of cell survival, proliferation and hence are vital windows to unraveling the biological complexity. Experimental measurements capturing this dynamics of enzyme-substrate interactions in real time add value to this understanding. Furthermore these measurements, upon validation in realistic biological specimens such as clinical biopsies - can further improve our capability in disease diagnostics and treatment monitoring. Towards this direction, we describe here a novel, high-sensitive measurement system for measuring diffusion-limited enzyme-substrate kinetics in real time. Using catalase (enzyme) and hydrogen peroxide (substrate) as the example pair, we demonstrate that this system is capable of direct measurement of catalase activity in vitro and the measured kinetics follows the classical Michaelis-Menten reaction kinetics. We further demonstrate the system performance by measuring catalase activity in living cells and in very small amounts of liver biopsies (down to 1 μg total protein). Catalase-specific enzyme activity is demonstrated by genetic and pharmacological tools. Finally we show the clinically-relevant diagnostic capability of our system by comparing the catalase activities in liver biopsies from young and old mouse (liver and serum) samples. We discuss the potential applicability of this system in clinical diagnostics as well as in intraoperative surgical settings.

  3. ICAM-1 targeted catalase encapsulated PLGA-b-PEG nanoparticles against vascular oxidative stress.

    Science.gov (United States)

    Sari, Ece; Tunc-Sarisozen, Yeliz; Mutlu, Hulya; Shahbazi, Reza; Ucar, Gulberk; Ulubayram, Kezban

    2015-01-01

    Targeted delivery of therapeutics is the favourable idea, whereas it is possible to distribute the therapeutically active drug molecule only to the site of action. For this purpose, in this study, catalase encapsulated poly(D,L-lactide-co-glycolide)-block-poly(ethylene glycol) (PLGA-b-PEG) nanoparticles were developed and an endothelial target molecule (anti-ICAM-1) was conjugated to this carrier system in order to decrease the oxidative stress level in the target site. According to the enzymatic activity results, initial catalase activity of nanoparticles was increased from 27.39 U/mg to up to 45.66 U/mg by adding 5 mg/mL bovine serum albumin (BSA). After 4 h, initial catalase activity was preserved up to 46.98% while free catalase retained less than 4% of its activity in proteolytic environment. Furthermore, FITC labelled anti-ICAM-1 targeted catalase encapsulated nanoparticles (anti-ICAM-1/CatNPs) were rapidly taken up by cultured endothelial cells and concomitantly endothelial cells were resistant to H2O2 induced oxidative impairment.

  4. Development of a new catalase activity assay for biological samples using optical CUPRAC sensor

    Science.gov (United States)

    Bekdeşer, Burcu; Özyürek, Mustafa; Güçlü, Kubilay; Alkan, Fulya Üstün; Apak, Reşat

    2014-11-01

    A novel catalase activity assay was developed for biological samples (liver and kidney tissue homogenates) using a rapid and low-cost optical sensor-based ‘cupric reducing antioxidant capacity' (CUPRAC) method. The reagent, copper(II)-neocuproine (Cu(II)-Nc) complex, was immobilized onto a cation-exchanger film of Nafion, and the absorbance changes associated with the formation of the highly-colored Cu(I)-Nc chelate as a result of reaction with hydrogen peroxide (H2O2) was measured at 450 nm. When catalase was absent, H2O2 produced the CUPRAC chromophore, whereas catalase, being an effective H2O2 scavenger, completely annihilated the CUPRAC signal due to H2O2. Thus, the CUPRAC absorbance due to H2O2 oxidation concomitant with Cu(I)-Nc formation decreased proportionally with catalase. The developed sensor gave a linear response over a wide concentration range of H2O2 (0.68-78.6 μM). This optical sensor-based method applicable to tissue homogenates proved to be efficient for low hydrogen peroxide concentrations (physiological and nontoxic levels) to which the widely used UV method is not accurately responsive. Thus, conventional problems of the UV method arising from relatively low sensitivity and selectivity, and absorbance disturbance due to gaseous oxygen evolution were overcome. The catalase findings of the proposed method for tissue homogenates were statistically alike with those of HPLC.

  5. Fluorimetry as a Simple and Sensitive Method for Determination of Catalase

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    Mehdi Hedayati

    2014-02-01

    Full Text Available Background: Catalase enzyme plays an important role in the anti-oxidation defense of body so it is important to measure its activity. Nowadays catalase activity measurement is performed by expensive imported kits in various scientific fields. The purpose of this study was to design a sensitive fluorimetry method for measuring catalase activity with improved sensitivity, accuracy and speed. Materials and Methods: In this study, the reaction of hydrogen peroxide with peroxidase (as a reaction accelerator was used in fluorimetry for catalase activity measuring in serum samples in order to increase the sensitivity of the assay. The sensitivity and intra- and inter-assay accuracy, verification test, recovery and parallelism tests, comparison method and correlation and coherence investigation methods were also performed. In order to increase the accuracy and speed of reading, the assay was performed in microplates and reading was done in fluorimetry plates. Results: The percentage of intra- and inter-assay variation coefficients were measured 3.8- 6.6 % and 4.1-7.3%, respectively. Comparison of the results of mentioned method for 50 serum samples with common colorimetric method showed a good correlation (0.917. In assessing the accuracy, the recovery percent was obtained 91% to 107%. The test sensitivity was measured 0.02 IU/ml. Conclusion: The fluorimetry method by microplate reading has a sufficient precision, accuracy and efficiency for catalase activity measuring as well as speed of measurement. Thus it can be an alternative method to conventional imported colorimetric methods.

  6. Study on the role of catalase for uptake of metallic mercury Part 3 In vitro oxidation of metallic mercury by catalase and hydrogen peroxide generated by several oxidase system

    OpenAIRE

    劒持,堅志

    1984-01-01

    In vitro oxidation of metallic mercury by catalase and hydrogen peroxide generated by the glucose-glucose oxidase system, D-alanine-D-amino acid oxidase system and xanthine-xanthine oxidase-superoxide dismutase system was investigated. In vitro oxidation of metallic mercury by catalase and hydrogen peroxide generated by the reaction with glucose and glucose oxidase was observed in erythrocytes and crystalline beef liver catalase solution. The uptake depended on the concentration of glucose ox...

  7. Enzymatic exploration of catalase from a nanoparticle producing and biodecolorizing algae Shewanella xiamenensis BC01.

    Science.gov (United States)

    Ng, I-Son; Xu, Fangxin; Zhang, Xia; Ye, Chiming

    2015-05-01

    Shewanella xiamenensis (SXM) was found to produce nanoparticles (NPs) under aerobic condition. The oxidoreductase enzymatic activities including of catalase, manganese peroxidase, laccase, NADH dehydrogenase, flavin reductase, azoreductase and Fe reductase are first investigated. Catalase showed the greatest enzymatic activity among all oxidoreductases in SXM, which with strong activities in multiple substrates of ABTS, guaiacol and 2,6-DMP. The optimum temperature, pH, concentrations of H2O2 and 2,6-DMP for this enzyme were found to be 65 °C, pH 4.0, 128.7 mM and 10 mM, respectively. Finally, from the kinetic parameters and structure simulation of catalase, implied that SXM would potentially apply in bioremediation, microbe fuel cells (MFCs) and nano-biotechnology based on its distinguished enzymatic system.

  8. Layer-by-layer assembled multilayers using catalase-encapsulated gold nanoparticles

    Science.gov (United States)

    Kim, Sungwoo; Park, Jeongju; Cho, Jinhan

    2010-09-01

    We introduce a novel and versatile approach for the preparation of multilayers, based on catalase-encapsulated gold nanoparticles (CAT-AuNP), allowing electrostatic charge reversal and structural transformation through pH adjustment. CAT-AuNP, which are synthesized directly from CAT stabilizer, can be electrostatically assembled with anionic and cationic PEs as a result of the charge reversal of the catalase stabilizers through pH control. In particular, at pH 5.2, near the pI of catalase, dispersed CAT-AuNP are structurally transformed into colloidal or network CAT-AuNP nanocomposites. Furthermore, we demonstrate that the layer-by-layer assembled multilayers composed of PEs and CAT-AuNP induce an effective electron transfer between CAT and the electrode as well as a high loading of CAT and AuNP, and resultantly exhibit a highly catalytic activity toward H2O2.

  9. Insights into the selective binding and toxic mechanism of microcystin to catalase

    Science.gov (United States)

    Hu, Yuandong; Da, Liangjun

    2014-03-01

    Microcystin is a sort of cyclic nonribosomal peptides produced by cyanobacteria. It is cyanotoxin, which can be very toxic for plants and animals including humans. The present study evaluated the interaction of microcystin and catalase, under physiological conditions by means of fluorescence, three-dimensional (3D) fluorescence, circular dichroism (CD), Fourier Transform infrared (FT-IR) spectroscopy, and enzymatic reactionkinetic techniques. The fluorescence data showed that microcystin could bind to catalase to form a complex. The binding process was a spontaneous molecular interaction procedure, in which electrostatic interactions played a major role. Energy transfer and fluorescence studies proved the existence of a static binding process. Additionally, as shown by the three-dimensional fluorescence, CD and FT-IR results, microcystin could lead to conformational and microenvironmental changes of the protein, which may affect the physiological functions of catalase. The work provides important insights into the toxicity mechanism of microcystin in vivo.

  10. ENVIRONMENTAL EFFECTS ON SUPEROXIDE DISMUTASE AND CATALASE ACTIVITY AND EXPRESSION IN HONEY BEE.

    Science.gov (United States)

    Nikolić, Tatjana V; Purać, Jelena; Orčić, Snežana; Kojić, Danijela; Vujanović, Dragana; Stanimirović, Zoran; Gržetić, Ivan; Ilijević, Konstantin; Šikoparija, Branko; Blagojević, Duško P

    2015-12-01

    Understanding the cellular stress response in honey bees will significantly contribute to their conservation. The aim of this study was to analyze the response of the antioxidative enzymes superoxide dismutase and catalase in honey bees related to the presence of toxic metals in different habitats. Three locations were selected: (i) Tunovo on the mountain Golija, as control area, without industry and large human impact, (ii) Belgrade as urban area, and (iii) Zajača, as mining and industrial zone. Our results showed that the concentrations of lead (Pb) in whole body of bees vary according to habitat, but there was very significant increase of Pb in bees from investigated industrial area. Bees from urban and industrial area had increased expression of both Sod1 and Cat genes, suggesting adaptation to increased oxidative stress. However, in spite increased gene expression, the enzyme activity of catalase was lower in bees from industrial area suggesting inhibitory effect of Pb on catalase.

  11. Metabolic Damage and Premature Thymus Aging Caused by Stromal Catalase Deficiency.

    Science.gov (United States)

    Griffith, Ann V; Venables, Thomas; Shi, Jianjun; Farr, Andrew; van Remmen, Holly; Szweda, Luke; Fallahi, Mohammad; Rabinovitch, Peter; Petrie, Howard T

    2015-08-18

    T lymphocytes are essential mediators of immunity that are produced by the thymus in proportion to its size. The thymus atrophies rapidly with age, resulting in progressive diminution of new T cell production. This decreased output is compensated by duplication of existing T cells, but it results in gradual dominance by memory T cells and decreased ability to respond to new pathogens or vaccines. Here, we show that accelerated and irreversible thymic atrophy results from stromal deficiency in the reducing enzyme catalase, leading to increased damage by hydrogen peroxide generated by aerobic metabolism. Genetic complementation of catalase in stromal cells diminished atrophy, as did chemical antioxidants, thus providing a mechanistic link between antioxidants, metabolism, and normal immune function. We propose that irreversible thymic atrophy represents a conventional aging process that is accelerated by stromal catalase deficiency in the context of an intensely anabolic (lymphoid) environment.

  12. Synergistic effects between catalase inhibitors and modulators of nitric oxide metabolism on tumor cell apoptosis.

    Science.gov (United States)

    Scheit, Katrin; Bauer, Georg

    2014-10-01

    Inhibitors of catalase (such as ascorbate, methyldopa, salicylic acid and neutralizing antibodies) synergize with modulators of nitric oxide (NO) metabolism (such as arginine, arginase inhibitor, NO synthase-inducing interferons and NO dioxygenase inhibitors) in the singlet oxygen-mediated inactivation of tumor cell protective catalase. This is followed by reactive oxygen species (ROS)-dependent apoptosis induction. TGF-beta, NADPH oxidase-1, NO synthase, dual oxidase-1 and caspase-9 are characterized as essential catalysts in this process. The FAS receptor and caspase-8 are required for amplification of ROS signaling triggered by individual compounds, but are dispensable when the synergistic effect is established. Our findings explain the antitumor effects of catalase inhibitors and of compounds that target NO metabolism, as well as their synergy. These data may have an impact on epidemiological studies related to secondary plant compounds and open new perspectives for the establishment of novel antitumor drugs and for the improvement of established chemotherapeutics.

  13. Purification and Characterization of Catalase from Indigenous Fungi of Neurospora crassa InaCC F226

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    Pugoh Santoso

    2016-08-01

    Full Text Available Microbial catalase is an important industrial enzyme that catalyzes the decomposition of hydrogen peroxide into water and oxygen. This enzyme posseses great potential of application in food, textile and pharmaceutical industries. In this paper, Neurospora crassa InaCC F226 was used to produce catalase. The aim of this research was to purify the catalase producing by Neurospora crassa InaCC F226 using gel filtration column chromatography and characterization of temperature, pH, Vmax, Km and molecular weight. In this study, the Neurospora crassa was cultured on Vogel's medium for 5 days. The cells were harvested in the 50 mM buffer sodium phosphate (pH 7 containing 3% hydrogen peroxide. Supernatant containing crude enzymes of Catalase was separated from cells by centrifugation at 15 minutes on 13000 x g and 4oC. The purification result using gel filtration was specific activity 1464.9 U/mg, 10.7 fold and 21.7% yield. Characterization of the purified enzyme toward pH and temperature optimum was 7.0 (271.6 U/mL and 40oC (286.1 U/mL. The Km and Vmax found to be 8.8 mM and 5.7 s.mM-1.  The catalase activity increased by additiom of Fe+2, Ca+2 and inhibited by EDTA, Cu+2 and Mn+2. The molecular weight of  the catalase was 59.4 kDa

  14. Data supporting the spectrophotometric method for the estimation of catalase activity

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    Mahmoud Hussein Hadwan

    2016-03-01

    Full Text Available Here we provide raw and processed data and methods for the estimation of catalase activities. The method for presenting a simple and accurate colorimetric assay for catalase activities is described. This method is based on the reaction of undecomposed hydrogen peroxide with ammonium molybdate to produce a yellowish color, which has a maximum absorbance at 374 nm. The method is characterized by adding a correction factor to exclude the interference that arises from the presence of amino acids and proteins in serum. The assay acts to keep out the interferences that arose from measurement of absorbance at unsuitable wavelengths.

  15. Effect of helium-neon laser on activity and optical properties of catalase.

    Science.gov (United States)

    Artyukhov, V G; Basharina, O V; Pantak, A A; Sveklo, L S

    2000-06-01

    The effects of laser (632.8 nm) on functional and spectral properties of catalase at pH 6.0-7.4 were studied. Laser irradiation led to photoactivation of the enzyme at pH 7.1-7.4. Changes in the spectral properties of photomodified hemoprotein were found in the absorption spectrum of the protein component: apoenzyme displayed protective effects in relation to ferroporphyrin. Structural modifications of catalase induced by helium-neon laser irradiation correlated with its functional properties. These results can be used in clinical practice to design the individual management program.

  16. Superoxide anion production and superoxide dismutase and catalase activities in Coxiella burnetii.

    OpenAIRE

    Akporiaye, E T; Baca, O G

    1983-01-01

    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.

  17. Decrease in catalase activity of Folsomia candida fed a Bt rice diet

    DEFF Research Database (Denmark)

    Yuan, Yiyang; Ke, Xin; Chen, Fajun;

    2011-01-01

    Here we report the effects of three Bt-rice varieties and their non-Bt conventional isolines on biological traits including survival, reproduction, and the activities of three antioxidant enzymes superoxide dismutase, catalase and peroxidase, in the Collembolan, Folsomia candida. The reproduction...... was significantly lower when fed Kemingdao and Huahui1 than those feeding on their non-GM near-isogenic varieties Xiushui and Minghui63 respectively, this can be explained by the differences of plant compositions depended on variety of rice. The catalase activity of F. candida was significantly lower when fed...

  18. Computational study concerning the effect of some pesticides on the Proteus Mirabilis catalase activity

    Science.gov (United States)

    Isvoran, Adriana

    2016-03-01

    Assessment of the effects of the herbicides nicosulfuron and chlorsulfuron and the fungicides difenoconazole and drazoxlone upon catalase produced by soil microorganism Proteus mirabilis is performed using the molecular docking technique. The interactions of pesticides with the enzymes are predicted using SwissDock and PatchDock docking tools. There are correlations for predicted binding energy values for enzyme-pesticide complexes obtained using the two docking tools, all the considered pesticides revealing favorable binding to the enzyme, but only the herbicides bind to the catalytic site. These results suggest the inhibitory potential of chlorsulfuron and nicosulfuron on the catalase activity in soil.

  19. Catalase in testes and epididymidis of wistar rats fed zinc deficient diet

    OpenAIRE

    Bedwal S; Prasad S; Nair N; Saini M; Bedwal R

    2009-01-01

    Catalase activities have been evaluated in testes and caput and cauda epididymis of Wistar rats fed on zinc deficient diet for 2 and 4 weeks. The enzyme activity has been measured as chromic acetate formed by heating of dichromate (in acetic acid) in presence of H 2 O 2 with perchromic acid as an unstable intermediate. Observed non-significant increase in catalase activity in testes as well as in caput and cauda epididymis of 2 weeks experiments has been related to low levels of H 2 O 2 pro...

  20. KatB, a cyanobacterial Mn-catalase with unique active site configuration: Implications for enzyme function.

    Science.gov (United States)

    Bihani, Subhash C; Chakravarty, Dhiman; Ballal, Anand

    2016-04-01

    Manganese catalases (Mn-catalases), a class of H2O2 detoxifying proteins, are structurally and mechanistically distinct from the commonly occurring catalases, which contain heme. Active site of Mn-catalases can serve as template for the synthesis of catalase mimetics for therapeutic intervention in oxidative stress related disorders. However, unlike the heme catalases, structural aspects of Mn-catalases remain inadequately explored. The genome of the ancient cyanobacterium Anabaena PCC7120, shows the presence of two Mn-catalases, KatA and KatB. Here, we report the biochemical and structural characterization of KatB. The KatB protein (with a C-terminal his-tag) was over-expressed in Escherichia coli and purified by affinity chromatography. On the addition of Mn(2+) to the E. coli growth medium, a substantial increase in production of the soluble KatB protein was observed. The purified KatB protein was an efficient catalase, which was relatively insensitive to inhibition by azide. Crystal structure of KatB showed a hexameric assembly with four-helix bundle fold, characteristic of the Ferritin-like superfamily. With canonical Glu4His2 coordination geometry and two terminal water ligands, the KatB active site was distinctly different from that of other Mn-catalases. Interestingly, the KatB active site closely resembled the active sites of ruberythrin/bacterioferritin, bi-iron members of the Ferritin-like superfamily. The KatB crystal structure provided fundamental insights into the evolutionary relationship within the Ferritin-like superfamily and further showed that Mn-catalases can be sub-divided into two groups, each with a distinct active site configuration.

  1. Decrease in catalase activity of Folsomia candida fed a Bt rice diet

    Energy Technology Data Exchange (ETDEWEB)

    Yuan Yiyang, E-mail: yuanyy@ioz.ac.cn [State Key Laboratory of Integrated Management of Pest and Rodents, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101 (China); Graduate School, Chinese Academy of Sciences, Beijing 100039 (China); Ke Xin, E-mail: xinke@sibs.ac.cn [Shanghai Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai 200032 (China); Chen Fajun, E-mail: fajunchen@njau.edu.cn [College of Plant Protection, Department of Entomology, Nanjing Agricultural University, Nanjing 210095 (China); Krogh, Paul Henning, E-mail: phk@dmu.dk [Department of Bioscience, University of Aarhus, P.O. Box 314, Vejlsoevej 25, DK-8600 Silkeborg (Denmark); Ge Feng, E-mail: gef@ioz.ac.cn [State Key Laboratory of Integrated Management of Pest and Rodents, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101 (China)

    2011-12-15

    Here we report the effects of three Bt-rice varieties and their non-Bt conventional isolines on biological traits including survival, reproduction, and the activities of three antioxidant enzymes superoxide dismutase, catalase and peroxidase, in the Collembolan, Folsomia candida. The reproduction was significantly lower when fed Kemingdao and Huahui1 than those feeding on their non-GM near-isogenic varieties Xiushui and Minghui63 respectively, this can be explained by the differences of plant compositions depended on variety of rice. The catalase activity of F. candida was significantly lower when fed the Bt-rice variety Kemingdao compared to the near-isogenic non-Bt-rice variety Xiushui. This suggests that some Bt-rice varieties may impose environmental stress to collembolans. We emphasize that changes in activity of antioxidant enzymes of non-target organisms are important in understanding the ecological consequences for organisms inhabiting transgenic Bt-rice plantations. - Highlights: > We examine the effects of Bt-rice on Folsomia candida with laboratory test. > The reproduction of F. candida was decreased by two Bt-rice varieties. > Decreased reproduction caused by the differences of varieties or C/N ratio of rice. > The catalase activity was decreased by Bt-rice Kemingdao. > Some Bt-rice may impose environmental stress on NTOs. - The catalase of the collembolan (Folsomia candida) was decreased when fed Bt-rice, Kemingdao.

  2. Role of catalase in the virulence of Brucella melitensis in pregnant goats.

    Science.gov (United States)

    Gee, Jason M; Kovach, Michael E; Grippe, Vanessa K; Hagius, Sue; Walker, Joel V; Elzer, Philip H; Roop, R Martin

    2004-08-19

    An isogenic katE mutant derived from virulent Brucella melitensis 16M displays hypersensitivity to hydrogen peroxide in disk sensitivity assays but retains the capacity to colonize pregnant goats and induce abortion. These experimental findings indicate that although the sole periplasmic catalase of Brucella melitensis functions as an antioxidant, this enzyme does not play a critical role in virulence in the natural host.

  3. A study of the inhibition of catalase by dipotassium trioxohydroxytetrafluorotriborate K₂[B₃O₃F₄OH].

    Science.gov (United States)

    Islamovic, Safija; Galic, Borivoj; Milos, Mladen

    2014-10-01

    In the development of boronic acid-based enzyme inhibitors as potential pharmaceutical drugs, dipotassium trioxohydroxytetrafluorotriborate K2[B3O3F4OH] was listed as a promising new therapeutic for treatment of these diseases. The catalase-mediated conversion of hydrogen peroxide, in the presence and absence of K2[B3O3F4OH] was studied. The kinetics conformed to the Michaelis-Menten model. Lineweaver-Burk plots were linear and plotted the family of straight lines intersected on the abscissa indicating non-competitive inhibition of the catalase. It appears that in the absence of inhibitor, catalase operates the best at conditions around pH 7.1 and in the presence of K2[B3O3F4OH] the optimum is around pH 6.2. The uncatalyzed reaction of hydrogen peroxide decomposition generally has a value of activation energy of 75 kJ mole(-1), whereas catalase, in the absence of inhibitor, lowers the value to 11.2 kJ mole(-1), while in the presence 69 mmoles L(-1) of K2[B3O3F4OH] it was 37.8 kJ mole(-1).

  4. A manganese catalase from Thermomicrobium roseum with peroxidase and catecholase activity.

    Science.gov (United States)

    Baginski, Robin; Sommerhalter, Monika

    2017-01-01

    An enzyme with catechol oxidase activity was identified in Thermomicrobium roseum extracts via solution assays and activity-stained SDS-PAGE. Yet, the genome of T. roseum does not harbor a catecholase gene. The enzyme was purified with two anion exchange chromatography steps and ultimately identified to be a manganese catalase with additional peroxidase and catecholase activity. Catalase activity (6280 ± 430 IU/mg) clearly dominated over pyrogallol peroxidase (231 ± 53 IU/mg) and catecholase (3.07 ± 0.56 IU/mg) activity as determined at 70 °C. Most enzyme kinetic properties were comparable to previously characterized manganese catalase enzymes. Catalase activity was highest at alkaline pH values and showed inhibition by excess substrate and chloride. The apparent K m and k cat values were 20 mM and 2.02 × 10(4) s(-1) subunit(-1) at 25 °C and pH 7.0.

  5. Preparation and Characterization of Catalase-Loaded Solid Lipid Nanoparticles Protecting Enzyme against Proteolysis

    Directory of Open Access Journals (Sweden)

    Ce Qi

    2011-07-01

    Full Text Available Catalase-loaded solid lipid nanoparticles (SLNs were prepared by the double emulsion method (w/o/w and solvent evaporation techniques, using acetone/methylene chloride (1:1 as an organic solvent, lecithin and triglyceride as oil phase and Poloxmer 188 as a surfactant. The optimized SLN was prepared by lecithin: triglyceride ratio (5%, 20-second + 30-second sonication, and 2% Poloxmer 188. The mean particle size of SLN was 296.0 ± 7.0 nm, polydispersity index range and zeta potential were 0.322–0.354 and −36.4 ± 0.6, respectively, and the encapsulation efficiency reached its maximum of 77.9 ± 1.56. Catalase distributed between the solid lipid and inner aqueous phase and gradually released from Poloxmer coated SLNs up to 20% within 20 h. Catalase-loaded SLN remained at 30% of H2O2-degrading activity after being incubated with Proteinase K for 24 h, while free catalase lost activity within 1 h.

  6. Catalase and sodium fluoride mediated rehabilitation of enamel bleached with 37% hydrogen peroxide

    Directory of Open Access Journals (Sweden)

    Ruchi Thakur

    2015-01-01

    Full Text Available Background: Bleaching agents bring about a range of unwanted changes in the physical structure of enamel which needs to be restored qualitatively and timely. Catalase being an antioxidant ensures the effective removal of free radicals and improvement in fluoride mediated remineralization from the enamel microstructure which if retained may harm the integrity and affect the hardness of enamel. Materials and Methods: Thirty freshly extracted incisors were sectioned to 6 slabs which were divided into 5 groups: Group A, control; Group B, treatment with 37% hydrogen peroxide (HP; Group C, treatment with 37% HP and catalase, Group D, treatment with 37% HP and 5% sodium fluoride application, Group E, treatment with 37% HP followed by catalase and 5% sodium fluoride. Scanning electron microscope and microhardness analysis were done for all slabs. One-way ANOVA test was applied among different groups. Results: Vicker′s microhardness number (VHN of Group B and C was significantly lower. No significant difference between VHN of Group B and C. VHN of Group D was significantly higher than Group A, B, and C; but significantly lower than Group E. VHN of Group E was significantly higher than any other experimental group. One-way ANOVA revealed a highly significant P value (P = 0.0001 and so Tukey′s post-hoc Test for the group comparisons was employed. Conclusion: Subsequent treatment of bleached enamel with catalase and fluoride varnish separately results in repairing and significantly increasing the microhardness.

  7. Different thermal unfolding pathways of catalase in the presence of cationic surfactants.

    Science.gov (United States)

    Blanco, Elena; Ruso, Juan M; Prieto, Gerardo; Sarmiento, Félix

    2007-03-01

    In this paper we have corroborated the usefulness of spectroscopic techniques, such as UV-visible, in the study and thermodynamic characterization of the thermal unfolding of catalase as a function of the concentration and alkyl chain length of n-alkyltrimethylammonium bromides (CnTAB, n = 8, 10, and 12). For this reason, a thermodynamic model was used which included experimental data corresponding to the pre- and posttransition into the observable transition. It has been found that n-alkyltrimethylammonium bromides play two opposite roles in the folding and stability of catalase. They act as a structure stabilizer at a low molar concentration and as a destabilizer at a higher concentration. The maximum of the unfolding temperature has been found to decrease with the alkyl chain. The reason for this difference has been suggested to be the side chains involved. In the presence of C8TAB and C10TAB, Gibbs energies of unfolding (DeltaG(T)) decrease with concentration, whereas for C12TAB an increase has been observed. These findings can be explained by the fact that when differences in the hydrophobic nature of the surfactants exist, different pathways of unfolding may occur. Also, the presence of surfactants has been observed to affect the cold denaturation of catalase. Thermodynamic results suggest that the thermal denaturation of catalase in the presence of n-alkyltrimethylammonium bromides is a perfect transition between two states.

  8. Hydrogen peroxide homeostasis: activation of plant catalase by calcium/calmodulin

    Science.gov (United States)

    Yang, T.; Poovaiah, B. W.

    2002-01-01

    Environmental stimuli such as UV, pathogen attack, and gravity can induce rapid changes in hydrogen peroxide (H(2)O(2)) levels, leading to a variety of physiological responses in plants. Catalase, which is involved in the degradation of H(2)O(2) into water and oxygen, is the major H(2)O(2)-scavenging enzyme in all aerobic organisms. A close interaction exists between intracellular H(2)O(2) and cytosolic calcium in response to biotic and abiotic stresses. Studies indicate that an increase in cytosolic calcium boosts the generation of H(2)O(2). Here we report that calmodulin (CaM), a ubiquitous calcium-binding protein, binds to and activates some plant catalases in the presence of calcium, but calcium/CaM does not have any effect on bacterial, fungal, bovine, or human catalase. These results document that calcium/CaM can down-regulate H(2)O(2) levels in plants by stimulating the catalytic activity of plant catalase. Furthermore, these results provide evidence indicating that calcium has dual functions in regulating H(2)O(2) homeostasis, which in turn influences redox signaling in response to environmental signals in plants.

  9. Catalase C-262T polymorphism and risk of prostate cancer: evidence from meta-analysis.

    Science.gov (United States)

    Hu, Jieping; Feng, Fupeng; Zhu, Shimiao; Sun, Libin; Li, Gang; Jiang, Ning; Shang, Zhiqun; Niu, Yuanjie

    2015-03-10

    Catalase is an important endogenous antioxidant enzyme that detoxifies hydrogen peroxide to oxygen and water, thus limiting the deleterious effects of reactive oxygen species. Several studies investigated the role of the Catalase (CAT) C-262T gene polymorphism on the risk of prostate cancer (PCa), but get conflicting results. We performed a meta-analysis based on five studies, to determine whether Catalase C-262T polymorphism contributes to the risk of prostate cancer using odds ratios (OR) with 95% confidence intervals (CI). On the whole, our evidence indicates that CAT C-262T polymorphism significantly increases PCa risk in the allele comparison model (OR=1.094, 95% CI=1.015-1.178, P=0.018). In the stratified analysis by ethnicity, the same results are found among Caucasians (allele model, OR=1.090, 95% CI=1.009-1.177, P=0.028, dominant model, OR=1.108, 95% CI=1.023-1.201, P=0.012, recessive model, OR=1.379, 95% CI=1.158-1.641, P=0.000, homozygous model, OR=1.429, 95% CI=1.196-1.707, P=0.000, and heterozygote model, OR=1.224, 95% CI=1.020-1.469, P=0.030). In conclusion, this meta-analysis suggests a positive correlation between Catalase C-262T polymorphism and the development of PCa.

  10. Not so monofunctional-a case of thermostable Thermobifida fusca catalase with peroxidase activity

    NARCIS (Netherlands)

    Lončar, Nikola; Fraaije, Marco W

    2014-01-01

    Thermobifida fusca is a mesothermophilic organism known for its ability to degrade plant biomass and other organics, and it was demonstrated that it represents a rich resource of genes encoding for potent enzymes for biocatalysis. The thermostable catalase from T. fusca has been cloned and overexpre

  11. Impedance spectroscopy and conductometric biosensing for probing catalase reaction with cyanide as ligand and inhibitor.

    Science.gov (United States)

    Bouyahia, Naima; Hamlaoui, Mohamed Larbi; Hnaien, Mouna; Lagarde, Florence; Jaffrezic-Renault, Nicole

    2011-02-01

    In this work, a new biosensor was prepared through immobilization of bovine liver catalase in a photoreticulated poly (vinyl alcohol) membrane at the surface of a conductometric transducer. This biosensor was used to study the kinetics of catalase-H(2)0(2) reaction and its inhibition by cyanide. Immobilized catalase exhibited a Michaelis-Menten behaviour at low H(2)0(2) concentrations (biosensor response by increasing cyanide concentration was linear up to 50μM, with a cyanide detection limit of 6μM. In parallel, electrochemical characteristics of the catalase/PVA biomembrane and its interaction with cyanide were studied by cyclic voltammetry and impedance spectroscopy. Addition of the biomembrane onto the gold electrodes induced a significant increase of the interfacial polarization resistance R(P). On the contrary, cyanide binding resulted in a decrease of Rp proportional to KCN concentration in the 4 to 50μM range. Inhibition coefficient I(50) calculated by this powerful label-free and substrate-free technique (24.3μM) was in good agreement with that determined from the substrate-dependent conductometric biosensor (24.9μM).

  12. Do Superoxide Dismutase (SOD) and Catalase (CAT) protect Cells from DNA Damage Induced by Active Arsenicals?

    Science.gov (United States)

    Superoxide dismutase (SOD) catalyzes the conversion of superoxide to hydrogen peroxide, which can be converted to water and oxygen through the action of catalase. Heterozygous mice of strain B6: 129S7-SodltmlLeb/J were obtained from Jackson Laboratories and bred to produce offspr...

  13. HUMAN CATALASE IS IMPORTED AND ASSEMBLED IN PEROXISOMES OF SACCHAROMYCES-CEREVISIAE

    NARCIS (Netherlands)

    DEHOOP, MJ; HOLTMAN, WL; AB, G

    1993-01-01

    To study the conservation of peroxisomal targeting signals, we have determined the intracellular localization of human peroxisomal catalase when expressed in yeast. Using immunofluorescence, differential centrifugation and immuno-electron microscopy, we show that the protein is targeted to the perox

  14. Not so monofunctional-a case of thermostable Thermobifida fusca catalase with peroxidase activity

    NARCIS (Netherlands)

    Lončar, Nikola; Fraaije, Marco W

    2015-01-01

    Thermobifida fusca is a mesothermophilic organism known for its ability to degrade plant biomass and other organics, and it was demonstrated that it represents a rich resource of genes encoding for potent enzymes for biocatalysis. The thermostable catalase from T. fusca has been cloned and overexpre

  15. Overexpression of Catalase Enhances Benzo(a)pyrene Detoxification in Endothelial Microsomes

    Science.gov (United States)

    Yang, Fang; Yang, Hong; Ramesh, Aramandla; Goodwin, J. Shawn; Okoro, Emmanuel U.; Guo, ZhongMao

    2016-01-01

    We previously reported that overexpression of catalase upregulated xenobiotic- metabolizing enzyme (XME) expression and diminished benzo(a)pyrene (BaP) intermediate accumulation in mouse aortic endothelial cells (MAECs). Endoplasmic reticulum (ER) is the most active organelle involved in BaP metabolism. To examine the involvement of ER in catalase-induced BaP detoxification, we compared the level and distribution of XMEs, and the profile of BaP intermediates in the microsomes of wild-type and catalase transgenic endothelial cells. Our data showed that endothelial microsomes were enriched in cytochrome P450 (CYP) 1A1, CYP1B1 and epoxide hydrolase 1 (EH1), and contained considerable levels of NAD(P)H: quinone oxidoreductase-1 (NQO1) and glutathione S-transferase-pi (GSTP). Treatment of wild-type MAECs with 1μM BaP for 2 h increased the expression of microsomal CYP1A1, 1B1 and NQO1 by ~300, 64 and 116%, respectively. However, the same treatment did not significantly alter the expression of EH1 and GSTP. Overexpression of catalase did not significantly increase EH1, but upregulated BaP-induced expression of microsomal CYP1A1, 1B1, NQO1 and GSTP in the following order: 1A1>NQO1>GSTP>1B1. Overexpression of catalase did not alter the distribution of each of these enzymes in the microsomes. In contrast to our previous report showing lower level of BaP phenols versus BaP diols/diones in the whole-cell, this report demonstrated that the sum of microsomal BaP phenolic metabolites were ~60% greater than that of the BaP diols/diones after exposure of microsomes to BaP. Overexpression of catalase reduced the concentrations of microsomal BaP phenols and diols/diones by ~45 and 95%, respectively. This process enhanced the ratio of BaP phenol versus diol/dione metabolites in a potent manner. Taken together, upregulation of phase II XMEs and CYP1 proteins, but not EH1 in the ER might be the mechanism by which overexpression of catalase reduces the levels of all the BaP metabolites

  16. Purification, crystallization and preliminary crystallographic analysis of KatB, a manganese catalase from Anabaena PCC 7120.

    Science.gov (United States)

    Bihani, Subhash Chandra; Chakravarty, Dhiman; Ballal, Anand

    2013-11-01

    Catalases are enzymes that play an important role in the detoxification of hydrogen peroxide (H2O2) in aerobic organisms. Among catalases, haem-containing catalases are ubiquitously distributed and their enzymatic mechanism is very well understood. On the other hand, manganese catalases that contain a bimanganese core in the active site have been less well characterized and their mode of action is not fully understood. The genome of Anabaena PCC 7120 does not show the presence of a haem catalase-like gene; instead, two ORFs encoding manganese catalases (Mn-catalases) are present. Here, the crystallization and preliminary X-ray crystallographic analysis of KatB, one of the two Mn-catalases from Anabaena, are reported. KatB was crystallized using the hanging-drop vapour-diffusion method with PEG 400 as a precipitant and calcium acetate as an additive. Diffraction data were collected in-house on an Agilent SuperNova system using a microfocus sealed-tube X-ray source. The crystal diffracted to 2.2 Å resolution at 100 K. The tetragonal crystal belonged to space group P4(1)2(1)2 (or enantiomer), with unit-cell parameters a = b = 101.87, c = 138.86 Å. Preliminary X-ray diffraction analysis using the Matthews coefficient and self-rotation function suggests the presence of a trimer in the asymmetric unit.

  17. On the enzymatic activity of catalase : an iron L-edge X-ray absorption study of the active centre

    NARCIS (Netherlands)

    Bergmann, Nora; Bonhommeau, Sebastien; Lange, Kathrin M.; Greil, Stefanie M.; Eisebitt, Stefan; de Groot, Frank; Chergui, Majed; Aziz, Emad F.

    2010-01-01

    Catalase and methaemoglobin have very similar haem groups, which are both ferric, yet catalase decomposes hydrogen peroxide to water and oxygen very efficiently, while methaemoglobin does not. Structural studies have attributed this behaviour to their different distal environments. Here we present F

  18. Cytochemical localization of catalase and several hydrogen peroxide-producing oxidases in the nucleoids and matrix of rat liver peroxisomes

    NARCIS (Netherlands)

    Veenhuis, M.; Wendelaar Bonga, S.E.

    1979-01-01

    The distribution of catalase, amino acid oxidase, α-hydroxy acid oxidase, urate oxidase and alcohol oxidase was studied cytochemically in rat hepatocytes. The presence of catalase was demonstrated with the conventional diaminobenzidine technique. Oxidase activities were visualized with methods based

  19. Development of a catalase based biosensor for alcohol determination in beer samples.

    Science.gov (United States)

    Akyilmaz, Erol; Dinçkaya, Erhan

    2003-10-17

    An amperometric biosensor based on catalase enzyme for alcohol determination was developed. To construct the biosensor catalase was immobilized by using gelatin and glutaraldehyde on a Clark type dissolved oxygen (DO) probe covered with a teflon membrane which is sensitive for oxygen. The working principle of the biosensor depends on two reactions, which one is related to another, catalyzed by catalase enzyme. In the first reaction catalase catalyzes the degradation of hydrogen peroxide and oxygen is produced and also a steady-state DO concentration occurs in a few minutes. When ethanol added to the medium catalase catalyzes the degradation of both hydrogen peroxide and ethanol and this results in a new steady-state DO concentration. Difference for first and the last steady-state DO concentration occurred in the interval surface of DO probe membrane, which related to ethanol concentration, are detected by the biosensor. The biosensor response depends linearly on ethanol concentration between 0.05 and 1.0 mM with a detection limit of 0.05 mM and a response time of 3 min. In the optimization studies of the biosensor phosphate buffer (pH 7.0; 50 mM) and 35 degrees C were established as providing the optimum working conditions. In the characterization studies of the biosensor some parameters such as reproducibility, substrate specificity, operational and storage stability were carried out. Finally, by using the biosensor developed and enzimatic-spectrophotometric method alcohol concentration of some alcoholic drinks were determined and results were compared.

  20. The effect of superoxide dismutase mimetic and catalase on the quality of postthawed goat semen.

    Science.gov (United States)

    Shafiei, Mojtaba; Forouzanfar, Mohsen; Hosseini, Sayyed Morteza; Esfahani, Mohammad Hossein Nasr

    2015-05-01

    Manganese(III) meso-tetrakis(N-ethylpyridinium-2-yl)porphyrin chloride (MnTE) is a cell-permeable superoxide dismutase mimetic agent which can convert superoxide to hydrogen peroxide (H2O2). Supplementation of MnTE to a commercial semen extender can protect sperm from superoxide but not H2O2. Therefore, we proposed that addition of catalase (0.0, 200, or 400 IU/mL) in combination with MnTE (0.1 μM) may further improve the cryopreservation efficiency of goat semen in commercially optimized freezing media such as Andromed. Therefore, ejaculates were obtained from three adult bucks twice a week during the breeding season and diluted with Andromed supplemented with or without MnTE and catalase and were frozen in liquid nitrogen. Sperm parameters and reactive oxygen species contents were evaluated 2 hours after dilution (before freezing) and after freezing/thawing. The results revealed that all the treatments significantly (P ≤ 0.05) improved sperm motility, viability, and membrane integrity after freezing and reduced reactive oxygen species content compared with the control group, but maximum improvement was obtained in MnTE + 400 IU/mL catalase. In addition, supplementation with these antioxidants significantly (P ≤ 0.05) increases the cleavage rate after IVF. In conclusion, the results of present study suggest that addition of antioxidant MnTE or catalase to commercial optimized media, such as Andromed, improves total motility, membrane integrity, and viability of goat semen samples after thawing. But the degree of improvement for these parameters significantly (P ≤ 0.05) higher when MnTE and catalase were simultaneously added to the cryopreservation media.

  1. Novel nonsense mutation in the katA gene of a catalase-negative Staphylococcus aureus strain.

    Science.gov (United States)

    Lagos, Jaime; Alarcón, Pedro; Benadof, Dona; Ulloa, Soledad; Fasce, Rodrigo; Tognarelli, Javier; Aguayo, Carolina; Araya, Pamela; Parra, Bárbara; Olivares, Berta; Hormazábal, Juan Carlos; Fernández, Jorge

    2016-01-01

    We report the first description of a rare catalase-negative strain of Staphylococcus aureus in Chile. This new variant was isolated from blood and synovial tissue samples of a pediatric patient. Sequencing analysis revealed that this catalase-negative strain is related to ST10 strain, which has earlier been described in relation to S. aureus carriers. Interestingly, sequence analysis of the catalase gene katA revealed presence of a novel nonsense mutation that causes premature translational truncation of the C-terminus of the enzyme leading to a loss of 222 amino acids. Our study suggests that loss of catalase activity in this rare catalase-negative Chilean strain is due to this novel nonsense mutation in the katA gene, which truncates the enzyme to just 283 amino acids.

  2. Significance of membrane bioreactor design on the biocatalytic performance of glucose oxidase and catalase: Free vs. immobilized enzyme systems

    DEFF Research Database (Denmark)

    Morthensen, Sofie Thage; Meyer, Anne S.; Jørgensen, Henning;

    2017-01-01

    Membrane separation of xylose and glucose can be accomplished via oxidation of glucose to gluconic acid by enzymatic glucose oxidase catalysis. Oxygen for this reaction can be supplied via decomposition of hydrogen peroxide by enzymatic catalase catalysis. In order to maximize the biocatalytic...... productivity of glucose oxidase and catalase (gluconic acid yield per total amount of enzyme) the following system set-ups were compared: immobilization of glucose oxidase alone; co-immobilization of glucose oxidase and catalase; glucose oxidase and catalase free in the membrane bioreactor. Fouling......-induced enzyme immobilization in the porous support of an ultrafiltration membrane was used as strategy for entrapment of glucose oxidase and catalase. The biocatalytic productivity of the membrane reactor was found to be highly related to the oxygen availability, which in turn depended on the reactor...

  3. A Catalase-related Hemoprotein in Coral Is Specialized for Synthesis of Short-chain Aldehydes: DISCOVERY OF P450-TYPE HYDROPEROXIDE LYASE ACTIVITY IN A CATALASE.

    Science.gov (United States)

    Teder, Tarvi; Lõhelaid, Helike; Boeglin, William E; Calcutt, Wade M; Brash, Alan R; Samel, Nigulas

    2015-08-07

    In corals a catalase-lipoxygenase fusion protein transforms arachidonic acid to the allene oxide 8R,9-epoxy-5,9,11,14-eicosatetraenoic acid from which arise cyclopentenones such as the prostanoid-related clavulones. Recently we cloned two catalase-lipoxygenase fusion protein genes (a and b) from the coral Capnella imbricata, form a being an allene oxide synthase and form b giving uncharacterized polar products (Lõhelaid, H., Teder, T., Tõldsepp, K., Ekins, M., and Samel, N. (2014) PloS ONE 9, e89215). Here, using HPLC-UV, LC-MS, and NMR methods, we identify a novel activity of fusion protein b, establishing its role in cleaving the lipoxygenase product 8R-hydroperoxy-eicosatetraenoic acid into the short-chain aldehydes (5Z)-8-oxo-octenoic acid and (3Z,6Z)-dodecadienal; these primary products readily isomerize in an aqueous medium to the corresponding 6E- and 2E,6Z derivatives. This type of enzymatic cleavage, splitting the carbon chain within the conjugated diene of the hydroperoxide substrate, is known only in plant cytochrome P450 hydroperoxide lyases. In mechanistic studies using (18)O-labeled substrate and incubations in H2(18)O, we established synthesis of the C8-oxo acid and C12 aldehyde with the retention of the hydroperoxy oxygens, consistent with synthesis of a short-lived hemiacetal intermediate that breaks down spontaneously into the two aldehydes. Taken together with our initial studies indicating differing gene regulation of the allene oxide synthase and the newly identified catalase-related hydroperoxide lyase and given the role of aldehydes in plant defense, this work uncovers a potential pathway in coral stress signaling and a novel enzymatic activity in the animal kingdom.

  4. Molecular cloning, characterization and gene expression of an antioxidant enzyme catalase (MrCat) from Macrobrachium rosenbergii.

    Science.gov (United States)

    Arockiaraj, Jesu; Easwvaran, Sarasvathi; Vanaraja, Puganeshwaran; Singh, Arun; Othman, Rofina Yasmin; Bhassu, Subha

    2012-05-01

    In this study, we reported a full length of catalase gene (designated as MrCat), identified from the transcriptome database of freshwater prawn Macrobrachium rosenbergii. The complete gene sequence of the MrCat is 2504 base pairs in length, and encodes 516 amino acids. The MrCat protein contains three domains such as catalase 1 (catalase proximal heme-ligand signature) at 350-358, catalase 2 (catalase proximal active site signature) at 60-76 and catalase 3 (catalase family profile) at 20-499. The mRNA expressions of MrCat in healthy and the infectious hypodermal and hematopoietic necrosis virus (IHHNV) challenged M. rosenbergii were examined using quantitative real time polymerase chain reaction (qRT-PCR). The MrCat is highly expressed in digestive tract and all the other tissues (walking leg, gills, muscle, hemocyte, hepatopancreas, pleopods, brain and eye stalk) of M. rosenbergii taken for analysis. The expression is strongly up-regulated in digestive tract after IHHNV challenge. To understand its biological activity, the recombinant MrCat gene was constructed and expressed in Escherichia coli BL21 (DE3). The recombinant MrCat existed in high thermal stability and broad spectrum of pH, which showed over 95% enzyme activity between pH 5 and 10.5, and was stable from 40 °C to 70 °C, and exhibited 85-100% enzyme activity from 30 °C to 40 °C.

  5. Effect of a protease inhibitor on the stability of catalase in liver and blood from acatalasemic and normal mice.

    Directory of Open Access Journals (Sweden)

    Suzuki,Kazuhiko

    1991-10-01

    Full Text Available Effects of Gabexate mesilate (GM (([ethyl-4-(6-guanidino hexanoyloxy benzoate] methane sulfonate, a protease inhibitor, on the activities of catalase in liver, erythrocytes and reticulocytes from acatalasemic mice were examined. Preincubation without GM at 37 degrees C for 160 min lowered the catalase activities of liver, erythrocytes and reticulocytes from acatalasemic mice, to 24%, 40% and 10% of the initial levels, respectively. But, preincubation with GM at 37 degrees C for 160 min delayed the rapid decrease in activities of residual catalases in the liver, erythrocytes and reticulocytes of acatalasemic mice to 65%, 93% and 85% of the initial values, respectively. At 20 degrees C or below, no reduction in catalase activity of reticulocytes from acatalasemic mice occurred with or even without GM. At pH 5.0, the decrease in catalase activity of acatalasemic mice was small both in the presence and the absence of GM. In the alkaline range, the reduction in the enzyme activity of the mutant mice without GM was enhanced with increase in pH values up to 8.5. But the presence of GM during preincubation at pH 7.5, retained the catalase activity of acatalasemic mice, to 64% of the activity at pH 6.5. These data suggest that some factors affected by GM, might be responsible for the low stability and activity of catalase in the acatalasemic mice.

  6. Activation of Peroxisome Proliferator-Activated Receptor Alpha Improves Aged and UV-Irradiated Skin by Catalase Induction

    Science.gov (United States)

    Shin, Mi Hee; Lee, Se-Rah; Kim, Min-Kyoung; Shin, Chang-Yup

    2016-01-01

    Peroxisome proliferator-activated receptor alpha (PPARα) is a nuclear hormone receptor involved in the transcriptional regulation of lipid metabolism, fatty acid oxidation, and glucose homeostasis. Its activation stimulates antioxidant enzymes such as catalase, whose expression is decreased in aged human skin. Here we investigated the expression of PPARα in aged and ultraviolet (UV)-irradiated skin, and whether PPARα activation can modulate expressions of matrix metalloproteinase (MMP)-1 and procollagen through catalase regulation. We found that PPARα mRNA level was significantly decreased in intrinsically aged and photoaged human skin as well as in UV-irradiated skin. A PPARα activator, Wy14643, inhibited UV-induced increase of MMP-1 and decrease of procollagen expression and caused marked increase in catalase expression. Furthermore, production of reactive oxygen species (ROS) was suppressed by Wy14643 in UV-irradiated and aged dermal fibroblasts, suggesting that the PPARα activation-induced upregulation of catalase leads to scavenging of ROS produced due to UV irradiation or aging. PPARα knockdown decreased catalase expression and abolished the beneficial effects of Wy14643. Topical application of Wy14643 on hairless mice restored catalase activity and prevented MMP-13 and inflammatory responses in skin. Our findings indicate that PPARα activation triggers catalase expression and ROS scavenging, thereby protecting skin from UV-induced damage and intrinsic aging. PMID:27611371

  7. Understanding the role of the catalase/peroxide genes in H2O2 resistance of E. coli serotype O157:H7 biofilms

    Science.gov (United States)

    Introduction: Escherichia coli serotype O157:H7 defenses against H2O2 include the peroxiredoxin AhpC and three catalases: KatG (catalase-peroxidase), KatE (catalase), and the plasmid-encoded KatP (catalase/peroxidase). AhpC, KatG, and KatP are induced by OxyR in exponential phase, while KatE is indu...

  8. Overexpression of Ref-1 Inhibits Lead-induced Endothelial Cell Death via the Upregulation of Catalase.

    Science.gov (United States)

    Lee, Kwon Ho; Lee, Sang Ki; Kim, Hyo Shin; Cho, Eun Jung; Joo, Hee Kyoung; Lee, Eun Ji; Lee, Ji Young; Park, Myoung Soo; Chang, Seok Jong; Cho, Chung-Hyun; Park, Jin Bong; Jeon, Byeong Hwa

    2009-12-01

    The role of apurinic/apyrimidinic endonuclease1/redox factor-1 (Ref-1) on the lead (Pb)-induced cellular response was investigated in the cultured endothelial cells. Pb caused progressive cellular death in endothelial cells, which occurred in a concentration- and time-dependent manner. However, Ref-1 overexpression with AdRef-1 significantly inhibited Pb-induced cell death in the endothelial cells. Also the overexpression of Ref-1 significantly suppressed Pb-induced superoxide and hydrogen peroxide elevation in the endothelial cells. Pb exposure induced the downregulation of catalase, it was inhibited by the Ref-1 overexpression in the endothelial cells. Taken together, our data suggests that the overexpression of Ref-1 inhibited Pb-induced cell death via the upregulation of catalase in the cultured endothelial cells.

  9. Binding of chrysoidine to catalase: spectroscopy, isothermal titration calorimetry and molecular docking studies.

    Science.gov (United States)

    Yang, Bingjun; Hao, Fang; Li, Jiarong; Chen, Dongliang; Liu, Rutao

    2013-11-01

    Chrysoidine is an industrial azo dye and the presence of chrysoidine in water and food has become an environmental concern due to its negative effects on human beings. In this work, the interactions between chrysoidine and bovine liver catalase (BLC) were explored. Obvious loss in catalytic activity was observed after incubation of BLC with chrysoidine, and the inhibition effect of BLC was found to be of the non-competitive type. No profound conformational change of BLC occurs in the presence of chrysoidine as revealed by UV-vis absorption, circular dichroism and fluorescence spectroscopy studies. Isothermal titration calorimetry results indicate that catalase has two sets of binding sites for chrysoidine. Further, molecular docking simulations show that chrysoidine is located within the bottleneck in the main channel of the substrate to the active site of BLC, which explain the activity inhibition of BLC by chrysoidine.

  10. Catalase-Modified Carbon Electrodes: Persuading Oxygen To Accept Four Electrons Rather Than Two.

    Science.gov (United States)

    Sepunaru, Lior; Laborda, Eduardo; Compton, Richard G

    2016-04-18

    We successfully exploited the natural highly efficient activity of an enzyme (catalase) together with carbon electrodes to produce a hybrid electrode for oxygen reduction, very appropriate for energy transformation. Carbon electrodes, in principle, are cheap but poor oxygen reduction materials, because only two-electron reduction of oxygen occurs at low potentials, whereas four-electron reduction is key for energy-transformation technology. With the immobilization of catalase on the surface, the hydrogen peroxide produced electrochemically is decomposed back to oxygen by the enzyme; the enzyme natural activity on the surface regenerates oxygen, which is further reduced by the carbon electrode with no direct electron transfer between the enzyme and the electrode. Near full four-electron reduction of oxygen is realised on a carbon electrode, which is modified with ease by a commercially available enzyme. The value of such enzyme-modified electrode for energy-transformation devices is evident.

  11. High-catalase strains of Mycobacterium kansasii isolated from water in Texas.

    OpenAIRE

    Steadham, J E

    1980-01-01

    Isolation techniques with membrane-filtered potable water samples resulted in the isolation of potentially pathogenic high-catalase strains of Mycobacterium kansasii from 8 of 19 representative outlets in a small central Texas town. Mycobacterium gordonae was isolated from all samples, and Mycobacterium fortuitum was isolated from two samples. Data on chlorine levels are presented along with a possible explanation for the unusually high numbers of mycobacteria in these potable water samples. ...

  12. Immunolocalization of hypochlorite-induced, catalase-bound free radical formation in mouse hepatocytes

    OpenAIRE

    2006-01-01

    The establishment of oxidants as mediators of signal transduction has renewed the interest of investigators in oxidant production and metabolism. In particular, H2O2 has been demonstrated to play pivotal roles in mediating cell differentiation, proliferation and death. Intracellular concentrations of H2O2 are modulated by its rate of production and its rate of decomposition by catalase and peroxidases. In inflammation and infection some of the H2O2 is converted to hypochlorous acid, a key med...

  13. Changes in gene expression and catalase activity in Oryza sativa L. under abiotic stress.

    Science.gov (United States)

    Vighi, I L; Benitez, L C; do Amaral, M N; Auler, P A; Moraes, G P; Rodrigues, G S; da Maia, L C; Pinto, L S; Braga, E J B

    2016-11-03

    Different rice (Oryza sativa L.) genotypes were subjected to high salinity and low temperature (150 mM NaCl and 13°C, respectively) for 0, 6, 24, 48, or 72 h. We evaluated the simultaneous expression of the genes OsCATA, OsCATB, and OsCATC, correlated gene expression with enzyme activity, and verified the regulation of these genes through identification of cis-elements in the promoter region. The hydrogen peroxide content increased in a tolerant genotype and decreased in a sensitive genotype under both stress conditions. Lipid peroxidation increased in the tolerant genotype when exposed to cold, and in the sensitive genotype when exposed to high salinity. Catalase activity significantly increased in both genotypes when subjected to 13°C. In the tolerant genotype, OsCATA and OsCATB were the most responsive to high salinity and cold, while in the sensitive genotype, OsCATA and OsCATC responded positively to saline stress, as did OsCATA and OsCATB to low temperature. Cis-element analysis identified different regulatory sequences in the catalase promoter region of each genotype. The sensitive genotype maintained a better balance between hydrogen oxyacid levels, catalase activity, and lipid peroxidation under low temperature than the resistant genotype. OsCATA and OsCATB were the most responsive in the salt-tolerant genotype to cold, OsCATA and OsCATC were the most responsive to saline stress, and OsCATA and OsCATB were the most responsive to chilling stress in the sensitive genotype. There were positive correlations between catalase activity and OsCATB expression in the tolerant genotype under saline stress and in the sensitive genotype under cold stress.

  14. Molecular cloning, characterization and expression analysis of a catalase gene inPaphia textile

    Institute of Scientific and Technical Information of China (English)

    WU Xiangwei; LI Jiakai; TAN Jing; LIU Xiande

    2016-01-01

    Catalase is an important antioxidant protein that can protect organisms against various forms of oxidative damage by eliminating hydrogen peroxide. In this study, the catalase cDNA ofPaphia textile (PtCAT) was cloned using RT-PCR and rapid amplification of cDNA ends (RACE).PtCAT is 1 921 bp long and consists of a 5′-UTR of 50 bp, a 3′-UTR of 349 bp, and an ORF of 1 542 bp that encodes 513 amino acids with a molecular weight of 58.4 kD and an estimated isoelectric point of 8.2. Sequence alignment indicated that PtCAT contained a highly conserved catalytic signature motif (61FNRERIPERVVHAKGAG77), a proximal heme-ligand signature sequence (352RLFSYSDP359), and three catalytic amino acid residues (H72, N145, and Y356). PtCAT also contains two putative N-glycosylation sites (34NKT36 and437NFT439) and a peroxisome-targeting signal (511AQL513). Furthermore, PtCAT shares 53%–88% identity and 29%–89% similarity with other catalase amino acid sequences.PtCAT mRNA was present in all tested organs, including the heart, digestive gland, adductor muscle, gonad, gill, and mantle, but its expression was highest in the digestive gland. High-temperature-induced stress produced two expression patterns ofPtCAT mRNA: first, an initial up-regulation followed by a down-regulation in the heart, digestive gland, and gonad and, second, consistent down-regulation in all other organs. These results demonstrate that PtCAT is a typical member of the catalase family and might be involved in the responses to harmful environmental factors.

  15. The application of catalase for the elimination of hydrogen peroxide residues after bleaching of cotton fabrics

    Directory of Open Access Journals (Sweden)

    AMORIM ALEXANDRA M.

    2002-01-01

    Full Text Available Results of dyeing of cotton fabrics with a bifunctional reactive dye were significantly improved when the fabric after bleaching with hydrogen peroxide was treated with catalase for the elimination of hydrogen peroxide residues from the fabrics. Compared to processes with a varying number of washing steps, with and without commercial reducing agents, the consumption of water could be significantly reduced, without altering the final color shade.

  16. The monofunctional catalase KatE of Xanthomonas axonopodis pv. citri is required for full virulence in citrus plants.

    Directory of Open Access Journals (Sweden)

    María Laura Tondo

    Full Text Available BACKGROUND: Xanthomonas axonopodis pv. citri (Xac is an obligate aerobic phytopathogen constantly exposed to hydrogen peroxide produced by normal aerobic respiration and by the plant defense response during plant-pathogen interactions. Four putative catalase genes have been identified in silico in the Xac genome, designated as katE, catB, srpA (monofunctional catalases and katG (bifunctional catalase. METHODOLOGY/PRINCIPAL FINDINGS: Xac catalase activity was analyzed using native gel electrophoresis and semi-quantitative RT-PCR. We demonstrated that the catalase activity pattern was regulated in different growth stages displaying the highest levels during the stationary phase. KatE was the most active catalase in this phase of growth. At this stage cells were more resistant to hydrogen peroxide as was determined by the analysis of CFU after the exposition to different H(2O(2 concentrations. In addition, Xac exhibited an adaptive response to hydrogen peroxide, displaying higher levels of catalase activity and H(2O(2 resistance after treatment with sub-lethal concentrations of the oxidant. In the plant-like medium XVM2 the expression of KatE was strongly induced and in this medium Xac was more resistant to H(2O(2. A XackatE mutant strain was constructed by insertional mutagenesis. We observed that catalase induction in stationary phase was lost meanwhile the adaptive response to peroxide was maintained in this mutant. Finally, the XackatE strain was assayed in planta during host plant interaction rendering a less aggressive phenotype with a minor canker formation. CONCLUSIONS: Our results confirmed that in contrast to other Xanthomonas species, Xac catalase-specific activity is induced during the stationary phase of growth in parallel with the bacterial resistance to peroxide challenge. Moreover, Xac catalases expression pattern is modified in response to any stimuli associated with the plant or the microenvironment it provides. The catalase Kat

  17. Prevalence of Catalase (-21 A/T Gene Variant in South Indian (Tamil Population

    Directory of Open Access Journals (Sweden)

    A. Lourdhu Mary

    2014-01-01

    Full Text Available Catalase, an endogenous antioxidant enzyme, is responsible for regulating reactive species levels. Several epidemiologic studies have suggested that single nucleotide polymorphism in catalase gene may be associated with many diseases. The genotype of CAT (-21 A/T point mutation in promoter region of catalase gene was determined by polymerase chain based restriction fragment length polymorphism analysis in the DNA of 100 healthy volunteers. The frequency of CAT (-21 A/T gene polymorphism AA, AT, and TT genotypes was found to be 7, 23, and 70 percent, respectively. The mutant “T” allele frequency was found to be 0.82 among the south Indian (Tamil population. Chi square analysis showed that the study population lies within the Hardy-Weinberg equilibrium. The wild type genotype (AA was found to be very low (7% and the mutant genotype (AT/TT was found to be more prevalent (93% among the south Indian population. This suggests that the high prevalence of mutant genotype may increase the susceptibility to oxidative stress associated diseases.

  18. Role of phosphate on stability and catalase mimetic activity of cerium oxide nanoparticles.

    Science.gov (United States)

    Singh, Ragini; Singh, Sanjay

    2015-08-01

    Cerium oxide nanoparticles (CeNPs) have been recently shown to scavenge reactive oxygen and nitrogen species (ROS and RNS) in different experimental model systems. CeNPs (3+) and CeNPs (4+) have been shown to exhibit superoxide dismutase (SOD) and catalase mimetic activity, respectively. Due to their nanoscale dimension, CeNPs are expected to interact with the components of biologically relevant buffers and medium, which could alter their catalytic properties. We have demonstrated earlier that CeNPs (3+) interact with phosphate and lose the SOD activity. However, very little is known about the interaction of CeNPs (4+) with the phosphate and other anions, predominantly present in biological buffers and their effects on the catalase mimetic-activity of these nanoparticles. In this study, we report that catalase mimetic-activity of CeNPs (4+) is resistant to the phosphate anions, pH changes and composition of cell culture media. Given the abundance of phosphate anions in the biological system, it is likely that internalized CeNPs would be influenced by cytoplasmic and nucleoplasmic concentration of phosphate.

  19. Murine and human b locus pigmentation genes encode a glycoprotein (gp75) with catalase activity

    Energy Technology Data Exchange (ETDEWEB)

    Halaban, R.; Moellmann, G. (Yale Univ. School of Medicine, New Haven, CT (USA))

    1990-06-01

    Melanogenesis is regulated in large part by tyrosinase, and defective tyrosinase leads to albinism. The mechanisms for other pigmentation determinants (e.g., those operative in tyrosinase-positive albinism and in murine coat-color mutants) are not yet known. One murine pigmentation gene, the brown (b) locus, when mutated leads to a brown (b/b) or hypopigmentated (B{sup lt}/B{sup lt}) coat versus the wild-type black (B/B). The authors show that the b locus codes for a glycoprotein with the activity of a catalase (catalase B). Only the c locus protein is a tyrosinase. Because peroxides may be by-products of melanogenic activity and hydrogen peroxide in particular is known to destroy melanin precursors and melanin, they conclude that pigmentation is controlled not only by tyrosinase but also by a hydroperoxidase. The studies indicate that catalase B is identical with gp75, a known human melanosomal glycoprotein; that the b mutation is in a heme-associated domain; and that the B{sup lt} mutation renders the protein susceptible to rapid proteolytic degradation.

  20. Turning points in the evolution of peroxidase-catalase superfamily: molecular phylogeny of hybrid heme peroxidases.

    Science.gov (United States)

    Zámocký, Marcel; Gasselhuber, Bernhard; Furtmüller, Paul G; Obinger, Christian

    2014-12-01

    Heme peroxidases and catalases are key enzymes of hydrogen peroxide metabolism and signaling. Here, the reconstruction of the molecular evolution of the peroxidase-catalase superfamily (annotated in pfam as PF00141) based on experimentally verified as well as numerous newly available genomic sequences is presented. The robust phylogenetic tree of this large enzyme superfamily was obtained from 490 full-length protein sequences. Besides already well-known families of heme b peroxidases arranged in three main structural classes, completely new (hybrid type) peroxidase families are described being located at the border of these classes as well as forming (so far missing) links between them. Hybrid-type A peroxidases represent a minor eukaryotic subfamily from Excavates, Stramenopiles and Rhizaria sharing enzymatic and structural features of ascorbate and cytochrome c peroxidases. Hybrid-type B peroxidases are shown to be spread exclusively among various fungi and evolved in parallel with peroxidases in land plants. In some ascomycetous hybrid-type B peroxidases, the peroxidase domain is fused to a carbohydrate binding (WSC) domain. Both here described hybrid-type peroxidase families represent important turning points in the complex evolution of the whole peroxidase-catalase superfamily. We present and discuss their phylogeny, sequence signatures and putative biological function.

  1. Probing the two-domain structure of homodimeric prokaryotic and eukaryotic catalase-peroxidases.

    Science.gov (United States)

    Banerjee, Srijib; Zamocky, Marcel; Furtmüller, Paul G; Obinger, Christian

    2010-11-01

    Catalase-peroxidases (KatGs) are ancestral bifunctional heme peroxidases found in archaeons, bacteria and lower eukaryotes. In contrast to homologous cytochrome c peroxidase (CcP) and ascorbate peroxidase (APx) homodimeric KatGs have a two-domain monomeric structure with a catalytic N-terminal heme domain and a C-terminal domain of high sequence and structural similarity but without obvious function. Nevertheless, without its C-terminal counterpart the N-terminal domain exhibits neither catalase nor peroxidase activity. Except some hybrid-type proteins all other members of the peroxidase-catalase superfamily lack this C-terminal domain. In order to probe the role of the two-domain monomeric structure for conformational and thermal stability urea and temperature-dependent unfolding experiments were performed by using UV-Vis-, electronic circular dichroism- and fluorescence spectroscopy, as well as differential scanning calorimetry. Recombinant prokaryotic (cyanobacterial KatG from Synechocystis sp. PCC6803) and eukaryotic (fungal KatG from Magnaporthe grisea) were investigated. The obtained data demonstrate that the conformational and thermal stability of bifunctional KatGs is significantly lower compared to homologous monofunctional peroxidases. The N- and C-terminal domains do not unfold independently. Differences between the cyanobacterial and the fungal enzyme are relatively small. Data will be discussed with respect to known structure and function of KatG, CcP and APx.

  2. Terazosin-induced alterations in catalase expression and lipid peroxidation in the rat seminal vesicles.

    Science.gov (United States)

    Mitropoulos, D; Patris, E; Deliconstantinos, G; Kyroudi-Voulgari, A; Anastasiou, I; Perea, D

    2013-04-01

    Previous studies have shown that alpha1-adrenergic receptor antagonists may alter seminal vesicle contractility and impair fertility in male rats. This study was designed to investigate the effects of terazosin on the catalase expression in the seminal vesicles and the lipid peroxidation of the seminal fluid in normal adult rats. Wistar rats were treated with terazosin (1.2 mg kg(-1) body weight, given orally every second day) for 120 days. Catalase expression was assessed immunohistochemically in tissue sections of the seminal vesicles, and lipid peroxidation was estimated by measuring the malondialdehyde (MDA) levels in the seminal vesicles' fluid. The seminal vesicles in terazosin-treated rats were particularly distended in comparison with those of controls, and their secreting epithelium was suppressed. Cytoplasmic catalase expression in the secreting epithelial cells (% of cells) was increased in terazosin-treated specimens in comparison with controls (76.1 ± 17.1 versus 51.3 ± 25.1, P = 0.005). MDA levels (μm) were also higher in samples from treated subjects in comparison with controls (2.67 ± 1.19 versus 1.39 ± 0.19, P = 0.01). Although the direct effect of terazosin treatment on the seminal vesicles is that of impaired contractility, an indirect effect is that on fertility by increasing lipid peroxidation in the seminal fluid and/or through degrading of hydrogen peroxide that is essential for sperm capacitation.

  3. Human catalase gene polymorphism (CAT C-262T) and risk of male infertility.

    Science.gov (United States)

    Sabouhi, S; Salehi, Z; Bahadori, M H; Mahdavi, M

    2015-02-01

    Infertility is the failure of a couple to engender after endeavouring at least one full year of unprotected intercourse. It has been reported that reactive oxygen species contributed to pathogenesis of various disease. To inactivate ROS cells biosynthesise several antioxidant enzymes, one of them is catalase which contributes H2 O2 to H2 O and O2 . This study set out to delineate the association of catalase C-262T polymorphism with idiopathic male infertility. The study included 195 men with idiopathic infertility and 190 healthy volunteers. Genomic DNA was extracted from peripheral blood leucocytes. Genotype and allele frequencies were determined in patients and controls using allele-specific PCR (AS-PCR). The prevalence of genotype frequencies of the CAT CC/CT/TT was 31.79%, 65.12% and 3.07%, respectively, in infertile subjects, as against 24.73%, 55.26% and 20%, respectively, in healthy volunteers. Statistical analysis has emerged significant difference from the comparison of either genotype (P catalase C-262T polymorphism indicates that CAT-262T/T genotype confers less susceptibility to male infertility. Further studies with larger numbers of patients are required for further evaluation and confirmation of our finding.

  4. Spectroscopy, calorimetry and molecular simulation studies on the interaction of catalase with copper ion.

    Science.gov (United States)

    Hao, Fang; Jing, Mingyang; Zhao, Xingchen; Liu, Rutao

    2015-02-01

    In this research, the binding mechanism of Cu(2+) to bovine liver catalase (BLC) was studied by fluorescence spectroscopy, ultraviolet-visible (UV-vis) absorption spectroscopy, circular dichroism (CD) spectroscopy, isothermal titration calorimetry (ITC) and molecular docking methods. The cellar experiment was firstly carried out to investigate the inhibition effect of catalase. During the fluorescence quenching study, after correcting the inner filter effect (IFE), the fluorescence of BLC was found to be quenched by Cu(2+). The quenching mechanism was determined by fluorescence lifetime measurement, and was confirmed to be the dynamic mode. The secondary structure content of BLC was changed by the addition of Cu(2+), as revealed by UV-vis absorption and CD spectra, which further induces the decrease in BLC activity. Molecular simulation study indicates that Cu(2+) is located between two β-sheets and two random coils of BLC near to the heme group, and interacts with His 74 and Ser 113 residues near a hydrophilic area. The decrease of α-helix and the binding of His 74 are considered to be the major reason for the inhibition of BLC activity caused by Cu(2+). The ITC results indicate that the binding stoichiometry of Cu(2+) to catalase is 11.4. Moreover, the binding of Cu(2+) to BLC destroyed H-bonds, which was confirmed by the CD result.

  5. Structure Characteristic and Catalase Activity of Vitreoscilla Hemoglobin Bound with Membrane

    Institute of Scientific and Technical Information of China (English)

    ZHAO Hai-feng; WU Lei; LU Ming; HU Yu-lin; HAN Xiao; LI Zheng-qiang

    2011-01-01

    Lipid-bound [VHb(406)] and lipid-free [VHb(402)] wide type Vitreoscilla hemoglobin were separated from E. coli BL21(DE3). The Rz is 3.30 for [VHb(406)] and 3.15 for [VHb(402)], respectively. VHb(406) shows a characteristic absorption band at 626 nm, while VHb(402) shows one at 644 nm, and it has more a-helix than VHb(402). Data of Raman spectra experiment shows under the excitation wavelength 488 nm, v4 vibration of both VHb(402) and VHb(406) had a pure and strong signal at 1375 cm-l, which proves that iron porphyrin of both the samples is at their trivalence oxidation state. And no matter lipid binds VHb or not, the vinyl of porphyrin is at cis state. The catalase activity of Vitreoscilla hemoglobin was explored with L-dopa and H2O2 as substrates. The results indicate that VHb(406) has more catalase activity than VHb(402), VHb is a cell's oxygen modulator and its catalase ability is modulated by the membrane.

  6. Layer-by-layer assembled multilayers using catalase-encapsulated gold nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Sungwoo; Park, Jeongju [School of Advanced Materials Engineering, Kookmin University, Jeongneung-dong, Seongbuk-gu, Seoul 136-702 (Korea, Republic of); Cho, Jinhan, E-mail: jinhan71@korea.ac.kr [Department of Chemical and Biological Engineering, Korea University, Anam-dong, Seongbuk-gu, Seoul 136-701 (Korea, Republic of)

    2010-09-17

    We introduce a novel and versatile approach for the preparation of multilayers, based on catalase-encapsulated gold nanoparticles (CAT-Au{sub NP}), allowing electrostatic charge reversal and structural transformation through pH adjustment. CAT-Au{sub NP}, which are synthesized directly from CAT stabilizer, can be electrostatically assembled with anionic and cationic PEs as a result of the charge reversal of the catalase stabilizers through pH control. In particular, at pH 5.2, near the pI of catalase, dispersed CAT-Au{sub NP} are structurally transformed into colloidal or network CAT-Au{sub NP} nanocomposites. Furthermore, we demonstrate that the layer-by-layer assembled multilayers composed of PEs and CAT-Au{sub NP} induce an effective electron transfer between CAT and the electrode as well as a high loading of CAT and Au{sub NP}, and resultantly exhibit a highly catalytic activity toward H{sub 2}O{sub 2}.

  7. Specific function of the Met-Tyr-Trp adduct radical and residues Arg-418 and Asp-137 in the atypical catalase reaction of catalase-peroxidase KatG.

    Science.gov (United States)

    Zhao, Xiangbo; Khajo, Abdelahad; Jarrett, Sanchez; Suarez, Javier; Levitsky, Yan; Burger, Richard M; Jarzecki, Andrzej A; Magliozzo, Richard S

    2012-10-26

    Catalase activity of the dual-function heme enzyme catalase-peroxidase (KatG) depends on several structural elements, including a unique adduct formed from covalently linked side chains of three conserved amino acids (Met-255, Tyr-229, and Trp-107, Mycobacterium tuberculosis KatG numbering) (MYW). Mutagenesis, electron paramagnetic resonance, and optical stopped-flow experiments, along with calculations using density functional theory (DFT) methods revealed the basis of the requirement for a radical on the MYW-adduct, for oxyferrous heme, and for conserved residues Arg-418 and Asp-137 in the rapid catalase reaction. The participation of an oxyferrous heme intermediate (dioxyheme) throughout the pH range of catalase activity is suggested from our finding that carbon monoxide inhibits the activity at both acidic and alkaline pH. In the presence of H(2)O(2), the MYW-adduct radical is formed normally in KatG[D137S] but this mutant is defective in forming dioxyheme and lacks catalase activity. KatG[R418L] is also catalase deficient but exhibits normal formation of the adduct radical and dioxyheme. Both mutants exhibit a coincidence between MYW-adduct radical persistence and H(2)O(2) consumption as a function of time, and enhanced subunit oligomerization during turnover, suggesting that the two mutations disrupting catalase turnover allow increased migration of the MYW-adduct radical to protein surface residues. DFT calculations showed that an interaction between the side chain of residue Arg-418 and Tyr-229 in the MYW-adduct radical favors reaction of the radical with the adjacent dioxyheme intermediate present throughout turnover in WT KatG. Release of molecular oxygen and regeneration of resting enzyme are thereby catalyzed in the last step of a proposed catalase reaction.

  8. Unprecedented access of phenolic substrates to the heme active site of a catalase: substrate binding and peroxidase-like reactivity of Bacillus pumilus catalase monitored by X-ray crystallography and EPR spectroscopy.

    Science.gov (United States)

    Loewen, Peter C; Villanueva, Jacylyn; Switala, Jacek; Donald, Lynda J; Ivancich, Anabella

    2015-05-01

    Heme-containing catalases and catalase-peroxidases catalyze the dismutation of hydrogen peroxide as their predominant catalytic activity, but in addition, individual enzymes support low levels of peroxidase and oxidase activities, produce superoxide, and activate isoniazid as an antitubercular drug. The recent report of a heme enzyme with catalase, peroxidase and penicillin oxidase activities in Bacillus pumilus and its categorization as an unusual catalase-peroxidase led us to investigate the enzyme for comparison with other catalase-peroxidases, catalases, and peroxidases. Characterization revealed a typical homotetrameric catalase with one pentacoordinated heme b per subunit (Tyr340 being the axial ligand), albeit in two orientations, and a very fast catalatic turnover rate (kcat  = 339,000 s(-1) ). In addition, the enzyme supported a much slower (kcat  = 20 s(-1) ) peroxidatic activity utilizing substrates as diverse as ABTS and polyphenols, but no oxidase activity. Two binding sites, one in the main access channel and the other on the protein surface, accommodating pyrogallol, catechol, resorcinol, guaiacol, hydroquinone, and 2-chlorophenol were identified in crystal structures at 1.65-1.95 Å. A third site, in the heme distal side, accommodating only pyrogallol and catechol, interacting with the heme iron and the catalytic His and Arg residues, was also identified. This site was confirmed in solution by EPR spectroscopy characterization, which also showed that the phenolic oxygen was not directly coordinated to the heme iron (no low-spin conversion of the Fe(III) high-spin EPR signal upon substrate binding). This is the first demonstration of phenolic substrates directly accessing the heme distal side of a catalase.

  9. An SNP polymorphism (-844C/T) in the promoter of catalase gene leads to differential expression

    Institute of Scientific and Technical Information of China (English)

    LI Yanping; ZHANG Xin; WANG Zhimin; LU Daru; HANG Wei; JIN Li

    2004-01-01

    @@ Imbalance of redox state has been associated with human diseases, such as cardiovascular diseases, Huntington's disease and Alzheimer's disease[1]. Catalase, an important antioxidant enzyme, decomposes H2O2 into O2and H2O, therefore limiting the deleterious effect of the reactive oxygen species (ROS)[1 -4]. Previous study showed that a C-T polymorphism, located at -844 bp from the translational start site of catalase, is strongly associated with essential hypertension in an isolated Chinese population of Xiangchang country, Anhui Province[5]. To explore the impact of SNP-844C/T on the expression of catalase, human catalase promoters containing either SNP-844C or T variant were subcloned into the pGL3-Basic luciferase vector.

  10. Increasing the endogenous NO level causes catalase inactivation and reactivation of intercellular apoptosis signaling specifically in tumor cells.

    Science.gov (United States)

    Bauer, Georg

    2015-12-01

    Tumor cells generate extracellular superoxide anions and are protected against intercellular apoptosis-inducing HOCl- and NO/peroxynitrite signaling through the expression of membrane-associated catalase. This enzyme decomposes H2O2 and thus prevents HOCl synthesis. It efficiently interferes with NO/peroxynitrite signaling through oxidation of NO and decomposition of peroxynitrite. The regulatory potential of catalase at the crosspoint of ROS and RNS chemical biology, as well as its high local concentration on the outside of the cell membrane of tumor cells, establish tight control of intercellular signaling and thus prevent tumor cell apoptosis. Therefore, inhibition of catalase or its inactivation by singlet oxygen reactivate intercellular apoptosis-inducing signaling. Nitric oxide and peroxynitrite are connected with catalase in multiple and meaningful ways, as (i) NO can be oxidated by compound I of catalase, (ii) NO can reversibly inhibit catalase, (iii) peroxynitrite can be decomposed by catalase and (iv) the interaction between peroxynitrite and H2O2 leads to the generation of singlet oxygen that inactivates catalase. Therefore, modulation of the concentration of free NO through addition of arginine, inhibition of arginase, induction of NOS expression or inhibition of NO dioxygenase triggers an autoamplificatory biochemical cascade that is based on initial formation of singlet oxygen, amplification of superoxide anion/H2O2 and NO generation through singlet oxygen dependent stimulation of the FAS receptor and caspase-8. Finally, singlet oxygen is generated at sufficiently high concentration to inactivate protective catalase and to reactivate intercellular apoptosis-inducing ROS signaling. This regulatory network allows to establish several pathways for synergistic interactions, like the combination of modulators of NO metabolism with enhancers of superoxide anion generation, modulators of NO metabolism that act at different targets and between modulators of

  11. Effects of the seminal plasma zinc content and catalase activity on the semen quality of water buffalo (Bubalus bubalis) bulls.

    Science.gov (United States)

    Alavi-Shoushtari, S M; Rezai, S Asri; Ansari, M H Kh; Khaki, A

    2009-01-15

    In order to determine zinc and catalase content of seminal plasma in the buffalo and to study their associations with the semen characteristics, 54 semen samples were collected from 10 buffalo bulls; semen volume and sperm concentration, gross and progressive motility and viability were evaluated, seminal plasma was then harvested by centrifugation and its zinc content was estimated by atomic absorption spectrophotometer and its catalase activity determined by using a commercial kit. The zinc content of the seminal plasma (Mean +/- SEM) was recorded as 154.40 +/- 1.74 mg L(-1), while, the mean catalase value was 32.00 +/- 0.42 U mL(-1). The mean zinc values was highly correlated with sperm progressive motility and viability and with catalase values (p = 0.000 for all) and also was associated with gross motility (p = 0.020) and negatively with abnormal morphology (p = 0.049). The catalase values were highly associated with sperm progressive motility, viability and zinc content (p = 0.000 for all) and was associated with sperm gross motility (p = 0.024). For further clarification of these correlations, the samples were categorized in three groups of excellent (Ex, >90% motile, n = 33), good (Go, 80-89% motile, n = 15) and moderate (Mo, zinc and catalase values were recorded as 161.07 +/- 1.63 mg L(-1) and 33.41 +/- 0.34 U mL(-1) in Ex, 146.70 +/- 1.91 mg L(-1) and 31.01 +/- 0.67 in Go and 136.42 +/- 4.97 mg L(-1) and 26.51 +/- 0.87 U mL(-1) in Mo groups. The mean zinc value in Ex group was highly associated with sperm motility, viability and catalase values, in Go group was associated with catalase values and highly associated with sperm abnormal morphology and in Mo group it was highly associations with catalase values only. The mean catalase value in Ex group, was highly associated with sperm motility and viability, in Go group was associated with zinc content and in Mo groups was highly associated with the zinc content. These results show that seminal plasma zinc

  12. Purification, cloning, expression, and biochemical characterization of a monofunctional catalase, KatP, from Pigmentiphaga sp. DL-8.

    Science.gov (United States)

    Dong, Weiliang; Hou, Ying; Li, Shuhuan; Wang, Fei; Zhou, Jie; Li, Zhoukun; Wang, Yicheng; Huang, Fei; Fu, Lei; Huang, Yan; Cui, Zhongli

    2015-04-01

    Catalases are essential components of the cellular equipment used to cope with oxidative stress. The monofunctional catalase KatP was purified from Pigmentiphaga sp. using ammonium sulfate precipitation (ASP), diethylaminoethyl ion exchange chromatography (IEC), and hydrophobic interaction chromatography (HIC). The purified catalase formed polymer with an estimated monomer molecular mass of 54kDa, which were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and zymogram analysis. KatP exhibited a specific catalytic activity of 73,000U/mg, which was higher than that of catalase-1 of Comamonas terrigena N3H (55,900U/mg). Seven short tryptic fragments of this catalase were obtained by electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF MS/MS), and the gene, katP, was cloned by PCR amplification and overexpressed in Escherichia coli BL21 (DE3). Based on the complete amino acid sequence, KatP was identified as a clade 3 monofunctional catalase. The specific activities of recombinant KatP for hydrogen peroxide (690,000U/mg) increased 9-fold over that of the parent strain. The Km and Vmax of recombinant KatP were 9.48mM and 81.2mol/minmg, respectively. The optimal pH and temperature for KatP were 7.0 and 37°C, respectively, and the enzyme displayed abroad pH-stable range of 4.0-11.0. The enzyme was inhibited by Zn(2+), Cu(2+), Cr(2+), and Mn(2+), whereas Fe(3+) and Mg(2+) stimulated KatP enzymatic activity. Interestingly, the catalase activity of recombinant KatP displayed high stability under different temperature and pH conditions, suggesting that KatP is a potential candidate for the production of catalase.

  13. Direct electrochemistry and electrocatalytic activity of catalase immobilized onto electrodeposited nano-scale islands of nickel oxide.

    Science.gov (United States)

    Salimi, Abdollah; Sharifi, Ensiyeh; Noorbakhsh, Abdollah; Soltanian, Saied

    2007-02-01

    Cyclic voltammetry was used for simultaneous formation and immobilization of nickel oxide nano-scale islands and catalase on glassy carbon electrode. Electrodeposited nickel oxide may be a promising material for enzyme immobilization owing to its high biocompatibility and large surface. The catalase films assembled on nickel oxide exhibited a pair of well defined, stable and nearly reversible CV peaks at about -0.05 V vs. SCE at pH 7, characteristic of the heme Fe (III)/Fe (II) redox couple. The formal potential of catalase in nickel oxide film were linearly varied in the range 1-12 with slope of 58.426 mV/pH, indicating that the electron transfer is accompanied by single proton transportation. The electron transfer between catalase and electrode surface, (k(s)) of 3.7(+/-0.1) s(-1) was greatly facilitated in the microenvironment of nickel oxide film. The electrocatalytic reduction of hydrogen peroxide at glassy carbon electrode modified with nickel oxide nano-scale islands and catalase enzyme has been studied. The embedded catalase in NiO nanoparticles showed excellent electrocatalytic activity toward hydrogen peroxide reduction. Also the modified rotating disk electrode shows good analytical performance for amperometric determination of hydrogen peroxide. The resultant catalase/nickel oxide modified glassy carbon electrodes exhibited fast amperometric response (within 2 s) to hydrogen peroxide reduction (with a linear range from 1 microM to 1 mM), excellent stability, long term life and good reproducibility. The apparent Michaelis-Menten constant is calculated to be 0.96(+/-0.05)mM, which shows a large catalytic activity of catalase in the nickel oxide film toward hydrogen peroxide. The excellent electrochemical reversibility of redox couple, high stability, technical simplicity, lake of need for mediators and short preparations times are advantages of this electrode. Finally the activity of biosensor for nitrite reduction was also investigated.

  14. Catalase -262C>T polymorphisms in Hungarian vitiligo patients and in controls: further acatalasemia mutations in Hungary.

    Science.gov (United States)

    Kósa, Zsuzsanna; Fejes, Zsolt; Nagy, Teréz; Csordás, Melinda; Simics, Enikő; Remenyik, Eva; Góth, László

    2012-04-01

    Catalase is the main regulator of hydrogen peroxide metabolism. In vitiligo patients there are conflicting data on its activity and no data on the effect of -262C>T polymorphism in the catalase gene. Blood catalase activity, -262C>T polymorphism and acatalasemia mutations were examined in 75 vitiligo patients and in 162 controls, in Hungary. We measured blood catalase activity and conducted analyses with PCR-SSCP, polyacrylamide gel electrophoresis and silver staining in combination with RFLP and nucleotide sequencing. Comparison of the wild (CC) genotype and the mutant (TT) genotype in the vitiligo patients revealed a non significant (P > 0.19) increase in blood catalase. Male controls with the CT genotype had significantly (P mutation (37C>T in exon 9) and the 113G>A (exon 9) mutation in Hungary are further proofs of genetic heterogeneity origin of acatalasemia mutations. In conclusion, the -262 C>T polymorphism has a reverse effect on blood catalase in vitiligo patients and in controls. In controls the mutant genotypes and alleles are more frequent in Hungary than in several other populations. The new acatalasemia mutations are further examples of heterogeneity of acatalasemia.

  15. A comprehensive evaluation of catalase-like activity of different classes of redox-active therapeutics.

    Science.gov (United States)

    Tovmasyan, Artak; Maia, Clarissa G C; Weitner, Tin; Carballal, Sebastián; Sampaio, Romulo S; Lieb, Dominik; Ghazaryan, Robert; Ivanovic-Burmazovic, Ivana; Ferrer-Sueta, Gerardo; Radi, Rafael; Reboucas, Julio S; Spasojevic, Ivan; Benov, Ludmil; Batinic-Haberle, Ines

    2015-09-01

    Because of the increased insight into the biological role of hydrogen peroxide (H2O2) under physiological and pathological conditions and the role it presumably plays in the action of natural and synthetic redox-active drugs, there is a need to accurately define the type and magnitude of reactions that may occur with this intriguing and key species of redoxome. Historically, and frequently incorrectly, the impact of catalase-like activity has been assigned to play a major role in the action of many redox-active drugs, mostly SOD mimics and peroxynitrite scavengers, and in particular MnTBAP(3-) and Mn salen derivatives. The advantage of one redox-active compound over another has often been assigned to the differences in catalase-like activity. Our studies provide substantial evidence that Mn(III) N-alkylpyridylporphyrins couple with H2O2 in actions other than catalase-related. Herein we have assessed the catalase-like activities of different classes of compounds: Mn porphyrins (MnPs), Fe porphyrins (FePs), Mn(III) salen (EUK-8), and Mn(II) cyclic polyamines (SOD-active M40403 and SOD-inactive M40404). Nitroxide (tempol), nitrone (NXY-059), ebselen, and MnCl2, which have not been reported as catalase mimics, were used as negative controls, while catalase enzyme was a positive control. The dismutation of H2O2 to O2 and H2O was followed via measuring oxygen evolved with a Clark oxygen electrode at 25°C. The catalase enzyme was found to have kcat(H2O2)=1.5×10(6)M(-1) s(-1). The yield of dismutation, i.e., the maximal amount of O2 evolved, was assessed also. The magnitude of the yield reflects an interplay between the kcat(H2O2) and the stability of compounds toward H2O2-driven oxidative degradation, and is thus an accurate measure of the efficacy of a catalyst. The kcat(H2O2) values for 12 cationic Mn(III) N-substituted (alkyl and alkoxyalkyl) pyridylporphyrin-based SOD mimics and Mn(III) N,N'-dialkylimidazolium porphyrin, MnTDE-2-ImP(5+), ranged from 23 to 88M(-1) s

  16. Role of oxyR from Sinorhizobium meliloti in Regulating the Expression of Catalases

    Institute of Scientific and Technical Information of China (English)

    Li LUO; Ming-Sheng QI; Shi-Yi YAO; Hai-Ping CHENG; Jia-Bi ZHU; Guan-Qiao YU

    2005-01-01

    The process of symbiotic nitrogen fixation results in the generation of reactive oxygen species such as the superoxide anion (O2-) and hydrogen peroxide (H2O2). The response of rhizobia to these toxic oxygen species is an important factor in nodulation and nitrogen fixation. In Sinorhizobium meliloti, one oxyR homologue and three catalase genes, katA, katB, and katC were detected by sequence analysis. This oxyR gene is located next to and divergently from katA on the chromosome. To investigate the possible roles of oxyR in regulating the expression of catalases at the transcriptional level in S. meliloti, an insertion mutant of this gene was constructed. The mutant was more sensitive and less adaptive to H2O2 than the wild type strain, and total catalase/peroxidase activity was reduced approximately fourfold with the OxyR mutation relative to controls. The activities of KatA and KatB and the expression of katA::lacZ and katB::lacZ promoter fusions were increased in the mutant strain compared with the parental strain grown in the absence of H2O2,indicating that katA and katB are repressed by OxyR. However, when exposed to H2O2, katA expression was also increased in both S. meliloti and Escherichia coli. When exposed to H2O2, OxyR is converted from a reduced to an oxidized form in E. coli. We concluded that the reduced form of OxyR functions as a repressor of katA and katB expression. Thus, in the presence of H2O2, reduced OxyR is converted to the oxidized form of OxyR that then results in increased katA expression. We further showed that oxyR expression is autoregulated via negative feedback.

  17. Effect of catalase on the liquid storage of mithun (Bos frontalis) semen

    Institute of Scientific and Technical Information of China (English)

    P Peruma; JK Chamuah; C Rajkhowa

    2013-01-01

    Objective: To assess the effect of catalase (CAT) on sperm motility, viability, total sperm abnormality, acrosomal and plasma membrane integrity, enzymatic profiles such as aspartate amino transaminase (AST), alanine amino transaminase (ALT), biochemical profiles such as cholesterol efflux and malonaldehyde (MDA) production and antioxidant profiles such as reduced glutathione (GSH), superoxide dismutase (SOD) and total antioxidant capacity (TAC). Methods: Total numbers of 50 ejaculates were collected twice a week from eight mithun bulls and semen was split into four equal aliquots, diluted with the TEYC extender. Group 1: semen without additives (control), group 2 to group 4: semen was diluted with 50 U/mL, 100 U/mL and 150 U/mL of catalase, respectively. These seminal, enzymatic, biochemical and antioxidant profiles were assessed at 5 ℃ for 0, 6, 12, 24 and 30 h of incubation. Results: Inclusion of catalase into diluent resulted in significant (P < 0.05) decrease in percentages of dead spermatozoa, abnormal spermatozoa and acrosomal abnormalities at different hours of storage periods as compared with control group. Additionally, CAT at 50 and 150 U/mL were inferior to CAT 100 U/mL treatments as regards to these characteristics and CAT at 100 U/mL has significant improvement in quality of mithun semen in in- vitro stored for up to 30 h.Conclusions:It was concluded that the possible protective effects of CAT on sperm parameters are it prevent efflux of cholesterol from cell membrane, MDA production and protect the function of antioxidants during preservation.

  18. Post-transcriptional regulator Hfq binds catalase HPII: crystal structure of the complex.

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    Koji Yonekura

    Full Text Available We report a crystal structure of Hfq and catalase HPII from Escherichia coli. The post-transcriptional regulator Hfq plays a key role in the survival of bacteria under stress. A small non-coding RNA (sRNA DsrA is required for translation of the stationary phase sigma factor RpoS, which is the central regulator of the general stress response. Hfq facilitates efficient translation of rpoS mRNA, which encodes RpoS. Hfq helps in the function of other specific proteins involved in RNA processing, indicating its versatility in the cell. However, structural information regarding its interactions with partners is missing. Here we obtained crystals of Hfq and HPII complexes from cell lysates following attempts to overexpress a foreign membrane protein. HPII is one of two catalases in E. coli and its mRNA is transcribed by an RNA polymerase holoenzyme containing RpoS, which in turn is under positive control of small non-coding RNAs and of the RNA chaperone Hfq. This sigma factor is known to have a pronounced effect on the expression of HPII. The crystal structure reveals that a Hfq hexamer binds each subunit of a HPII tetramer. Each subunit of the Hfq hexamer exhibits a unique binding mode with HPII. The hexamer of Hfq interacts via its distal surface. The proximal and distal surfaces are known to specifically bind different sRNAs, and binding of HPII could affect Hfq function. Hfq-HPII complexation has no effect on catalase HPII activity.

  19. A Catalase-related Hemoprotein in Coral Is Specialized for Synthesis of Short-chain Aldehydes

    Science.gov (United States)

    Teder, Tarvi; Lõhelaid, Helike; Boeglin, William E.; Calcutt, Wade M.; Brash, Alan R.; Samel, Nigulas

    2015-01-01

    In corals a catalase-lipoxygenase fusion protein transforms arachidonic acid to the allene oxide 8R,9-epoxy-5,9,11,14-eicosatetraenoic acid from which arise cyclopentenones such as the prostanoid-related clavulones. Recently we cloned two catalase-lipoxygenase fusion protein genes (a and b) from the coral Capnella imbricata, form a being an allene oxide synthase and form b giving uncharacterized polar products (Lõhelaid, H., Teder, T., Tõldsepp, K., Ekins, M., and Samel, N. (2014) PloS ONE 9, e89215). Here, using HPLC-UV, LC-MS, and NMR methods, we identify a novel activity of fusion protein b, establishing its role in cleaving the lipoxygenase product 8R-hydroperoxy-eicosatetraenoic acid into the short-chain aldehydes (5Z)-8-oxo-octenoic acid and (3Z,6Z)-dodecadienal; these primary products readily isomerize in an aqueous medium to the corresponding 6E- and 2E,6Z derivatives. This type of enzymatic cleavage, splitting the carbon chain within the conjugated diene of the hydroperoxide substrate, is known only in plant cytochrome P450 hydroperoxide lyases. In mechanistic studies using 18O-labeled substrate and incubations in H218O, we established synthesis of the C8-oxo acid and C12 aldehyde with the retention of the hydroperoxy oxygens, consistent with synthesis of a short-lived hemiacetal intermediate that breaks down spontaneously into the two aldehydes. Taken together with our initial studies indicating differing gene regulation of the allene oxide synthase and the newly identified catalase-related hydroperoxide lyase and given the role of aldehydes in plant defense, this work uncovers a potential pathway in coral stress signaling and a novel enzymatic activity in the animal kingdom. PMID:26100625

  20. Highly Active and Stable Large Catalase Isolated from a Hydrocarbon Degrading Aspergillus terreus MTCC 6324

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    Preety Vatsyayan

    2016-01-01

    Full Text Available A hydrocarbon degrading Aspergillus terreus MTCC 6324 produces a high level of extremely active and stable cellular large catalase (CAT during growth on n-hexadecane to combat the oxidative stress caused by the hydrocarbon degrading metabolic machinery inside the cell. A 160-fold purification with specific activity of around 66 × 105 U mg−1 protein was achieved. The native protein molecular mass was 368 ± 5 kDa with subunit molecular mass of nearly 90 kDa, which indicates that the native CAT protein is a homotetramer. The isoelectric pH (pI of the purified CAT was 4.2. BLAST aligned peptide mass fragments of CAT protein showed its highest similarity with the catalase B protein from other fungal sources. CAT was active in a broad range of pH 4 to 12 and temperature 25°C to 90°C. The catalytic efficiency (Kcat/Km of 4.7 × 108 M−1 s−1 within the studied substrate range and alkaline pH stability (half-life, t1/2 at pH 12~15 months of CAT are considerably higher than most of the extensively studied catalases from different sources. The storage stability (t1/2 of CAT at physiological pH 7.5 and 4°C was nearly 30 months. The haem was identified as haem b by electrospray ionization tandem mass spectroscopy (ESI-MS/MS.

  1. A bio-mimetic zinc/tau protein as an artificial catalase.

    Science.gov (United States)

    Asadollahi, Kazem; Jasemi, Neda Sadat Kazemein; Riazi, Gholam Hossein; Katuli, Fatemeh Hedayati; Yazdani, Fahimeh; Sartipnia, Nasrin; Moosavi, Mohammad Amin; Rahimi, Arash; Falahati, Mojtaba

    2016-11-01

    In this study, the catalase-like activity of monomeric tau protein was reported in the presence of of zinc (Zn(II)) ions at low pH value. Monomeric tau protein contains two SH groups that are a target of disulfide bond formation. However these SH groups are able to interact with Zn(II) ion at pH 7.2 which creates a thiol bond as a mimetic model of chloroperoxidase active site which performs catalase like activity at low pH. Zn(II)/tau protein complex decomposed H2O2 with a high rate (Vm) as well as an efficient turn oven number (kcat) at pH 3. This remarkable catalase like activity is may be attributed to the conformational reorientation of protein at low pH. Circular dichroism (CD) studies did not demonstrate any secondary structural changes of tau protein after addition of Zn(II) ions at pH 7.2. In addition, tau protein shows identical CD bands at pH 7.2 and 3. Moreover, fluorescence quenching of tau by Zn(II) at pH 7.2 was initiated by complex formation rather than by dynamic collision. A significant red shift (6nm) was observed in the emission maximum of the fluorescence spectra when the protein was dissolved at pH 3 compared to pH 7.2. This conformational change can provide information regarding the rearrangements of the protein structure and exposure of Cys-Zn(II) group to the solvent which induces easy access of active site to H2O2 molecules and corresponding enhanced catalytic activity of Zn(II)/tau protein complex. This study introduces tau protein as a bio-inspired high performing scaffold for transition metal encapsulation and introducing an engineered apoprotein-induced biomimetic enzyme.

  2. A Single Nucleotide Polymorphism in Catalase Is Strongly Associated with Ovarian Cancer Survival.

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    Jimmy Belotte

    Full Text Available Ovarian cancer is the deadliest of all gynecologic cancers. Recent evidence demonstrates an association between enzymatic activity altering single nucleotide polymorphisms (SNP with human cancer susceptibility. We sought to evaluate the association of SNPs in key oxidant and antioxidant enzymes with increased risk and survival in epithelial ovarian cancer. Individuals (n = 143 recruited were divided into controls, (n = 94: healthy volunteers, (n = 18, high-risk BRCA1/2 negative (n = 53, high-risk BRCA1/2 positive (n = 23 and ovarian cancer cases (n = 49. DNA was subjected to TaqMan SNP genotype analysis for selected oxidant and antioxidant enzymes. Of the seven selected SNP studied, no association with ovarian cancer risk (Pearson Chi-square was found. However, a catalase SNP was identified as a predictor of ovarian cancer survival by the Cox regression model. The presence of this SNP was associated with a higher likelihood of death (hazard ratio (HR of 3.68 (95% confidence interval (CI: 1.149-11.836 for ovarian cancer patients. Kaplan-Meier survival analysis demonstrated a significant median overall survival difference (108 versus 60 months, p<0.05 for those without the catalase SNP as compared to those with the SNP. Additionally, age at diagnosis greater than the median was found to be a significant predictor of death (HR of 2.78 (95% CI: 1.022-7.578. This study indicates a strong association with the catalase SNP and survival of ovarian cancer patients, and thus may serve as a prognosticator.

  3. MicroRNA-30b-mediated regulation of catalase expression in human ARPE-19 cells.

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    Rashidul Haque

    Full Text Available BACKGROUND: Oxidative injury to retinal pigment epithelium (RPE and retinal photoreceptors has been linked to a number of retinal diseases, including age-related macular degeneration (AMD. Reactive oxygen species (ROS-mediated gene expression has been extensively studied at transcriptional levels. Also, the post-transcriptional control of gene expression at the level of translational regulation has been recently reported. However, the microRNA (miRNA/miR-mediated post-transcriptional regulation in human RPE cells has not been thoroughly looked at. Increasing evidence points to a potential role of miRNAs in diverse physiological processes. METHODOLOGY/PRINCIPAL FINDINGS: We demonstrated for the first time in a human retinal pigment epithelial cell line (ARPE-19 that the post-transcriptional control of gene expression via miRNA modulation regulates human catalase, an important and potent component of cell's antioxidant defensive network, which detoxifies hydrogen peroxide (H(2O(2 radicals. Exposure to several stress-inducing agents including H(2O(2 has been reported to alter miRNA expression profile. Here, we demonstrated that a sublethal dose of H(2O(2 (200 µM up-regulated the expression of miR-30b, a member of the miR-30 family, which inhibited the expression of endogenous catalase both at the transcript and protein levels. However, antisense (antagomirs of miR-30b was not only found to suppress the miR-30b mimics-mediated inhibitions, but also to dramatically increase the expression of catalase even under an oxidant environment. CONCLUSIONS/SIGNIFICANCE: We propose that a microRNA antisense approach could enhance cytoprotective mechanisms against oxidative stress by increasing the antioxidant defense system.

  4. A catalase from the freshwater mussel Cristaria plicata with cloning, identification and protein characterization.

    Science.gov (United States)

    Yang, Xilan; Li, Gang; Wen, Chungen; Hu, Baoqing; Deng, Lirong; Pei, Pengzu; Xie, Yanhai

    2011-09-01

    Catalase is an important antioxidant protein which can protect organisms against various oxidative stresses by eliminating hydrogen peroxide. The catalase cDNA of Cristaria plicata (cpCAT) was cloned from the haemocytes using degenerate primers by the method of 3' and 5' rapid amplification of cDNA ends PCR. The gene is 4863 bp long and has a total of two introns and three exons. The precise size and location of the introns and exons have been determined. In addition the full-length cDNA of cpCAT contained 2618 bp, The cDNA contained a 5' untranslated region (UTR) of 136 nucleotides, the 3' UTR of 979 bp with a canonical polyadenylation signal sequence AATAAA and a polyA tail, and an open reading frame (ORF) of 1503 bp, encoding 501 amino acid residues with 56.86 kDa predicted molecular weight. The theoretical isoelectric point was 6.77. BLAST analysis showed that the deduced amino acid sequence of cpCAT had significant homology to catalases from animals, plants and bacteria. The deduced amino acid sequence of cpCAT had characteristic features of catalase family such as catalytic site motif (61FNRERIPERVVHAKGAG77), heme-ligand signature motif (351RLYSYSDTH359), two glycosylation sites (N145, N436), NADPH binding site and the three catalytic amino acid residues (His72, Asn145 and Tyr355). It had no signal peptide. The phylogenetic tree indicated that cpCAT gene was very close to the gene of scallops, Chlamys farreri. The enzymatic activity of purified recombinant cpCAT was 11194.4 ± 40.4 U/mg, it might resist against H(2)O(2). The recombinant enzyme held higher thermal stability, the optimum temperature was 25 °C, it retained more than 82% activity between 25 and 60 °C. The stability of the recombinant enzyme were higher between pH 5 and 10, and the optimal pH value was 7.0. When cpCAT was treated with 2-4 moL/L urea and 1%-3% SDS, the activity was also stable, it kept more than 80% activity.

  5. X-ray diffraction study of Penicillium Vitale catalase in the complex with aminotriazole

    Energy Technology Data Exchange (ETDEWEB)

    Borovik, A. A.; Grebenko, A. I.; Melik-Adamyan, V. R., E-mail: mawr@ns.crys.ras.ru [Russian Academy of Sciences, Shubnikov Institute of Crystallography (Russian Federation)

    2011-07-15

    The three-dimensional structure of the enzyme catalase from Penicillium vitale in a complex with the inhibitor aminotriazole was solved and refined by protein X-ray crystallography methods. An analysis of the three-dimensional structure of the complex showed that the inhibition of the enzyme occurs as a result of the covalent binding of aminotriazole to the amino-acid residue His64 in the active site of the enzyme. An investigation of the three-dimensional structure of the complex resulted in the amino-acid residues being more precisely identified. The binding sites of saccharide residues and calcium ions in the protein molecule were found.

  6. Synthesis and activity of Helicobacter pylori urease and catalase at low pH.

    OpenAIRE

    Bauerfeind, P; Garner, R; Dunn, B E; Mobley, H L

    1997-01-01

    BACKGROUND: Helicobacter pylori produces large amounts of urease presumably to be prepared for the rare event of a sudden acid exposure. The hypothesis that H pylori is acid sensitive and protein production is inhibited by low pH was examined. METHODS: H pylori or its soluble enzymes were incubated buffered or unbuffered at a pH ranging from 2-7 in the presence of 5 mM urea for 30 minutes. After exposure, urease and catalase activities of whole cells, supernatants, and soluble enzyme preparat...

  7. Catalase-negative Staphylococcus aureus isolated from a diabetic foot ulcer

    Directory of Open Access Journals (Sweden)

    MR Zali

    2010-12-01

    Full Text Available We report a catalase-negative Staphylococcus aureus isolated from a 56-year-old male diabetic patient with foot ulcer who attended our surgery ward. Species identification was confirmed by Gram staining, standard biochemical tests and PCR amplification of the nuc and fem genes. Antibiotic susceptibility showed that the strain was sensitive to imepenem, chloramphenicol, amoxicillin, vancomycin and resistant to oxacillin, penicillin, ceftriaxone, erythromycin, clindamycin, and amikacin. Clinicians and microbiologists must be encouraged to identify and report these atypical strains and the infections associated with them in order to establish their role in pathogenesis.

  8. X-ray diffraction study of Penicillium Vitale catalase in the complex with aminotriazole

    Science.gov (United States)

    Borovik, A. A.; Grebenko, A. I.; Melik-Adamyan, V. R.

    2011-07-01

    The three-dimensional structure of the enzyme catalase from Penicillium vitale in a complex with the inhibitor aminotriazole was solved and refined by protein X-ray crystallography methods. An analysis of the three-dimensional structure of the complex showed that the inhibition of the enzyme occurs as a result of the covalent binding of aminotriazole to the amino-acid residue His64 in the active site of the enzyme. An investigation of the three-dimensional structure of the complex resulted in the amino-acid residues being more precisely identified. The binding sites of saccharide residues and calcium ions in the protein molecule were found.

  9. Distribution of a Nocardia brasiliensis Catalase Gene Fragment in Members of the Genera Nocardia, Gordona, and Rhodococcus

    Science.gov (United States)

    Vera-Cabrera, Lucio; Johnson, Wendy M.; Welsh, Oliverio; Resendiz-Uresti, Francisco L.; Salinas-Carmona, Mario C.

    1999-01-01

    An immunodominant protein from Nocardia brasiliensis, P61, was subjected to amino-terminal and internal sequence analysis. Three sequences of 22, 17, and 38 residues, respectively, were obtained and compared with the protein database from GenBank by using the BLAST system. The sequences showed homology to some eukaryotic catalases and to a bromoperoxidase-catalase from Streptomyces violaceus. Its identity as a catalase was confirmed by analysis of its enzymatic activity on H2O2 and by a double-staining method on a nondenaturing polyacrylamide gel with 3,3′-diaminobenzidine and ferricyanide; the result showed only catalase activity, but no peroxidase. By using one of the internal amino acid sequences and a consensus catalase motif (VGNNTP), we were able to design a PCR assay that generated a 500-bp PCR product. The amplicon was analyzed, and the nucleotide sequence was compared to the GenBank database with the observation of high homology to other bacterial and eukaryotic catalases. A PCR assay based on this target sequence was performed with primers NB10 and NB11 to confirm the presence of the NB10-NB11 gene fragment in several N. brasiliensis strains isolated from mycetoma. The same assay was used to determine whether there were homologous sequences in several type strains from the genera Nocardia, Rhodococcus, Gordona, and Streptomyces. All of the N. brasiliensis strains presented a positive result but only some of the actinomycetes species tested were positive in the PCR assay. In order to confirm these findings, genomic DNA was subjected to Southern blot analysis. A 1.7-kbp band was observed in the N. brasiliensis strains, and bands of different molecular weight were observed in cross-reacting actinomycetes. Sequence analysis of the amplicons of selected actinomycetes showed high homology in this catalase fragment, thus demonstrating that this protein is highly conserved in this group of bacteria. PMID:10325357

  10. Cardiac-specific catalase overexpression rescues anthrax lethal toxin-induced cardiac contractile dysfunction: role of oxidative stress and autophagy

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    Kandadi Machender R

    2012-11-01

    Full Text Available Abstract Background Lethal and edema toxins secreted by Bacillus anthracis during anthrax infection were found to incite serious cardiovascular complications. However, the underlying mechanisms in anthrax lethal toxin-induced cardiac anomalies remain unknown. This study was designed to evaluate the impact of antioxidant enzyme catalase in anthrax lethal toxin-induced cardiomyocyte contractile dysfunction. Methods Wild type (WT and cardiac-specific catalase overexpression mice were challenged with lethal toxin (2 μg/g, intraperotineally (i.p.. Cardiomyocyte contractile and intracellular Ca2+ properties were assessed 18 h later using an IonOptix edge-detection system. Proteasome function was assessed using chymotrypsin-like and caspase-like activities. GFP-LC3 puncta and Western blot analysis were used to evaluate autophagy and protein ubiquitination. Results Lethal toxin exposure suppressed cardiomyocyte contractile function (suppressed peak shortening, maximal velocity of shortening/re-lengthening, prolonged duration of shortening/re-lengthening, and impaired intracellular Ca2+ handling, the effects of which were alleviated by catalase. In addition, lethal toxin triggered autophagy, mitochondrial and ubiquitin-proteasome defects, the effects of which were mitigated by catalase. Pretreatment of cardiomyocytes from catalase mice with the autophagy inducer rapamycin significantly attenuated or ablated catalase-offered protection against lethal toxin-induced cardiomyocyte dysfunction. On the other hand, the autophagy inhibitor 3-MA ablated or significantly attenuated lethal toxin-induced cardiomyocyte contractile anomalies. Conclusions Our results suggest that catalase is protective against anthrax lethal toxin-induced cardiomyocyte contractile and intracellular Ca2+ anomalies, possibly through regulation of autophagy and mitochondrial function.

  11. N-glycosylation-negative catalase: a useful tool for exploring the role of hydrogen peroxide in the endoplasmic reticulum.

    Science.gov (United States)

    Lortz, S; Lenzen, S; Mehmeti, I

    2015-03-01

    Disulfide bond formation during protein folding of nascent proteins is associated with the generation of H2O2 in the endoplasmic reticulum (ER). Approaches to quantifying H2O2 directly within the ER failed because of the oxidative environment in the ER lumen, and ER-specific catalase expression to detoxify high H2O2 concentrations resulted in an inactive protein owing to N-glycosylation. Therefore, the N-glycosylation motifs at asparagine-244 and -439 of the human catalase protein were deleted by site-directed mutagenesis. The ER-targeted expression of these variants revealed that the deletion of the N-glycosylation motif only at asparagine-244 (N244) was associated with the maintenance of full enzymatic activity in the ER. Expression of catalase N244 in the ER (ER-Catalase N244) was ER-specific and protected the cells significantly against exogenously added H2O2. With the expression of ER-Catalase N244, a highly effective H2O2 inactivation within the ER was achieved for the first time. Catalase has a high H2O2-inactivation capacity without the need of reducing cofactors, which might interfere with the ER redox homeostasis, and is not involved in protein folding. With these characteristics ER-Catalase N244 is an ideal tool to explore the impact of ER-generated H2O2 on the generation of disulfide bonds or to study the induction of ER-stress pathways through protein folding overload and accumulation of H2O2.

  12. Antioxidant catalase rescues against high fat diet-induced cardiac dysfunction via an IKKβ-AMPK-dependent regulation of autophagy.

    Science.gov (United States)

    Liang, Lei; Shou, Xi-Ling; Zhao, Hai-Kang; Ren, Gu-Qun; Wang, Jian-Bang; Wang, Xi-Hui; Ai, Wen-Ting; Maris, Jackie R; Hueckstaedt, Lindsay K; Ma, Ai-Qun; Zhang, Yingmei

    2015-02-01

    Autophagy, a conservative degradation process for long-lived and damaged proteins, participates in a variety of biological processes including obesity. However, the precise mechanism of action behind obesity-induced changes in autophagy still remains elusive. This study was designed to examine the role of the antioxidant catalase in high fat diet-induced changes in cardiac geometry and function as well as the underlying mechanism of action involved with a focus on autophagy. Wild-type (WT) and transgenic mice with cardiac overexpression of catalase were fed low or high fat diet for 20 weeks prior to assessment of myocardial geometry and function. High fat diet intake triggered obesity, hyperinsulinemia, and hypertriglyceridemia, the effects of which were unaffected by catalase transgene. Myocardial geometry and function were compromised with fat diet intake as manifested by cardiac hypertrophy, enlarged left ventricular end systolic and diastolic diameters, fractional shortening, cardiomyocyte contractile capacity and intracellular Ca²⁺ mishandling, the effects of which were ameliorated by catalase. High fat diet intake promoted reactive oxygen species production and suppressed autophagy in the heart, the effects of which were attenuated by catalase. High fat diet intake dampened phosphorylation of inhibitor kappa B kinase β(IKKβ), AMP-activated protein kinase (AMPK) and tuberous sclerosis 2 (TSC2) while promoting phosphorylation of mTOR, the effects of which were ablated by catalase. In vitro study revealed that palmitic acid compromised cardiomyocyte autophagy and contractile function in a manner reminiscent of fat diet intake, the effect of which was significantly alleviated by inhibition of IKKβ, activation of AMPK and induction of autophagy. Taken together, our data revealed that the antioxidant catalase counteracts against high fat diet-induced cardiac geometric and functional anomalies possibly via an IKKβ-AMPK-dependent restoration of myocardial

  13. Atividade relativa da catalase de losna-branca (Parthenium hysterophorus comparada à de outras espécies daninhas Catalase relative activity of ragweed (Parthenium hysterophorus compared to that of other weed species

    Directory of Open Access Journals (Sweden)

    S.J.P. Carvalho

    2012-06-01

    Full Text Available Este trabalho foi desenvolvido com o objetivo de avaliar a atividade relativa da catalase em extrato aquoso de losna-branca (Parthenium hysterophorus, bem como comparála à atividade da catalase de outras espécies daninhas. O trabalho constou de três fases, que envolveram a padronização do método, comparação da atividade relativa da catalase de plantas da família Asteraceae e comparação com outras 11 espécies daninhas, sendo estas: Euphorbia heterophylla, Alternanthera tenella, Cenchrus echinatus, Panicum maximum, Amaranthus viridis, Ipomoea hederifolia, Galinsoga parviflora, Bidens pilosa, Sonchus oleraceus, Cyperus rotundus e Commelina benghalensis. Observou-se resposta linear crescente da reação entre extrato aquoso de losna-branca e peróxido de hidrogênio, em razão da concentração do extrato vegetal. Em todas as fases, a atividade relativa da catalase de extrato de losna-branca foi superior à atividade da catalase das demais espécies daninhas. Com os dados obtidos nas três fases, conclui-se que a maior atividade relativa observada para a catalase da losnabranca contribui significativamente para a tolerância dessa espécie ao herbicida paraquat. Essa maior atividade pode ser consequência da maior concentração enzimática nas células ou devido à maior atividade intrínseca da enzima (afinidade enzima-substrato, havendo necessidade de estudos mais precisos para essa conclusão.This work was carried out to evaluate catalase relative activity of ragweed (Parthenium hysterophorus aqueous extract, as well as to compare it with catalase activity of other weed species. It consisted of three phases, involving method standardization, comparison of the catalase relative activity in Asteraceae family plants and that of ragweed catalase activity with the following 11 weed species: Euphorbia heterophylla, Alternanthera tenella, Cenchrus echinatus, Panicum maximum, Amaranthus viridis, Ipomoea hederifolia, Galinsoga parviflora

  14. 精神分裂症患者与过氧化氢酶(Catalase)基因多态性%The Association between Schizophrenics and Catalase Gene Polymorphism

    Institute of Scientific and Technical Information of China (English)

    赵昌烈; 李光哲; 崔成虎; 许妍姬

    2009-01-01

    目的:探讨过氧化氢酶(Catalase)基因多态性与精神分裂症的相关性.方法:采用Amp-RFLP(amplication-restriction fregment lengh polymorphis)方法对精神分裂症患者和对照组的过氧化氢酶基因相关性进行了研究.结果:167例精神分裂症患者与155例对照组之间过氧化氢酶基因型频率和等位基因频率无统计学意义,76例女性精神分裂症患者与60例对照组之间过氧化氢酶基因型频率(χ2=11.096,df=2,P=0.004)有显著性差异.结论:过氧化氢酶多态性与精神分裂症的遗传学并不存在关联性,但是研究表明过氧化氢酶基因型频率与女性精神分裂症患者有显著性差异.

  15. Mn(II) complexes with bipyridine, phenanthroline and benzoic acid: Biological and catalase-like activity

    Indian Academy of Sciences (India)

    Ibrahim Kani; Özlem Atlier; Kiymet Güven

    2016-04-01

    Five mononuclear Mn(II) complexes, [Mn(phen)2(ClO4)2] (1), [Mn(phen)3](ClO4)2(H2CO3)2(2), [Mn(bipy)2(ClO4)2] (3), [Mn(bipy)3](ClO4)2) (4), and Mn(phen)2(ba)(H2O)](ClO4)(CH3OH) (5), where bipy = 2,2’-bipyridine, phen = 1,10-phenanthroline, and ba = benzoic acid were prepared and characterized by Xray, IR and UV-Vis spectroscopies, and their catalase-like and biological activities were studied. The presence of two different types and the number of chelating NN-donor neutral ligands allowed for analysis of their effects on the catalase and biological activities. It was observed that the presence and number of phen ligands improved the activity more than the bipy ligand. Complexes 1 and 2, which contain more basic phen ligands, disproportionate H2O2 faster than complexes 3 and 4, which contain less basic bipy ligands. The in vitro antimicrobial activities of all the complexes were also tested against seven bacterial strains by microdilution tests. All the bacterial isolates demonstrated sensitivity to the complexes and the antifungal (anticandidal) activities of the Mn(II) complexes were remarkably higher than the reference drug ketoconazole.

  16. Motor-based microprobe powered by bio-assembled catalase for motion detection of DNA.

    Science.gov (United States)

    Xie, Yuzhe; Fu, Shizhe; Wu, Jie; Lei, Jianping; Ju, Huangxian

    2017-01-15

    A motor-based microprobe is proposed using a tubular microengine powered by bio-assembled enzyme as catalyst and exploited for washing-free detection of DNA through motion readout. The microprobe is fabricated by assembling a catalase layer on the inner surface of poly(3,4-ethylenedioxythiophene)/Au (PEDOT/Au) microtube through DNA conjugate, which is responsible for the biocatalytic bubble propulsion. The sensing concept of the microprobe relies on the target-induced release of catalase through the DNA strand-replacement hybridization, which decreases the amount of enzyme assembled on microtube to slow down the movement of the microprobe. Therefore, the motion speed is negatively correlated with the target concentration. At the optimal conditions, the microprobe can conveniently distinguish the concentration of specific DNA in a range of 0.5-10µM without any washing and separation step. This microprobe can be prepared in batch with good reproducibility and stability, and its motion speed can be conveniently visualized by optical microscope. The proposed motor-based microprobe and its dynamic sensing method provide a novel platform for the development of intelligent microprobe and clinical diagnostic strategy.

  17. Identification and characteristic analysis of the catalase gene from Locusta migratoria.

    Science.gov (United States)

    Zhang, Xueyao; Li, Yahong; Wang, Junxiu; Zhang, Tingting; Li, Tao; Dong, Wei; Ma, Enbo; Zhang, Jianzhen

    2016-09-01

    Catalase (CAT) is a ubiquitous antioxidant enzyme in almost all living organisms exposed to atmosphere, which involved in decomposing harmful hydrogen peroxide, into oxygen and water. In this study, a full-length cDNA (1524bp) encoding the catalase gene (LmCAT) from Locusta migratoria was cloned (accession number KT716445). The open reading frame of the LmCAT gene encoded 507 amino acids and shared 57.8%-97.8% amino acid identities with other insect CATs. The coding region was interrupted by 9 introns, while its promoter region contained 15 putative binding sites for 5 kinds of transcriptional regulation factors. For the stage-specific expression profile, LmCAT was highly expressed in the fourth-instar nymphs. For the tissue-specific expression profile, the LmCAT transcripts were highest in the fat bodies, and relatively abundant in the gastric caecum, Malpighian tubules, ovary and integument. Moreover, the result showed that quercetin could significantly induce the expression level of LmCAT. The expression of LmCAT could be silenced by RNAi, but the moralities were not significantly different between control and RNAi groups. Our results would provide valuable information for further study on the ROS regulation mechanism in insect.

  18. Molecular Cloning and Expression Analysis of a Catalase Gene (NnCAT) from Nelumbo nucifera.

    Science.gov (United States)

    Dong, Chen; Zheng, Xingfei; Diao, Ying; Wang, Youwei; Zhou, Mingquan; Hu, Zhongli

    2015-11-01

    Rapid amplification cDNA end (RACE) assay was established to achieve the complete cDNA sequence of a catalase gene (NnCAT) from Nelumbo nucifera. The obtained full-length cDNA was 1666 bp in size and contained a 1476-bp open reading frame. The 3D structural model of NnCAT was constructed by homology modeling. The putative NnCAT possessed all the main characteristic amino acid residues and motifs of catalase (CAT) protein family, and the phylogenetic analysis revealed that NnCAT grouped together with high plants. Moreover, recombinant NnCAT showed the CAT activity (758 U/mg) at room temperature, holding high activity during temperature range of 20-50 °C, then the optimal pH of recombinant protein was assessed from pH 4 to pH 11. Additionally, real-time PCR assay demonstrated that NnCAT mRNA was expressed in various tissues of N. nucifera, with the highest expression in young leaf and lowest level in the root, and mRNA level of NnCAT was significantly augmented in response to short-time mechanical wounding. Different expression pattern of NnCAT gene suggested that NnCAT probably played a defensive role in the initial stages of oxidative stress, regulating the level of reactive oxygen species (ROS) by extracellular stimuli such as short-time mechanical wounding.

  19. Comprehensive Functional Analysis of the Catalase Gene Family in Arabidopsis thaliana

    Institute of Scientific and Technical Information of China (English)

    Yan-Yan Du; Peng-Cheng Wang; Jia Chen; Chun-Peng Song

    2008-01-01

    In Arabidopsis, catalase (CAT) genes encode a small family of proteins including CAT1, CAT2 and CAT3, which catalyze the decomposition of hydrogen peroxide (H2O2) and play an important role in controlling homeostasis of reactive oxygen species (ROS). Here, we analyze the expression profiles and activities of three catalases under different treatments including drought, cold, oxidative stresses, abscisic acid and salicylic acid in Arabidopsis. Our results reveal that CAT1 is an important player in the removal of H2O2 generated under various environmental stresses. CAT2 and CAT3 are major H2O2 scavengers that contribute to ROS homeostasis in light or darkness, respectively. In addition, CAT2 is activated by cold and drought stresses and CAT3 is mainly enhanced by abscisic acid and oxidative treatments as well as at the senescence stage. These results, together with previous data, suggest that the network of transcriptional control explains how CATs and other scavenger enzymes such as peroxidase and superoxide dismutase may be coordinately regulated during development, but differentially expressed in response to different stresses for controlling ROS homeostasis.

  20. Ability of recombinant human catalase to suppress inflammation of the murine lung induced by influenza A.

    Science.gov (United States)

    Shi, Xunlong; Shi, Zhihui; Huang, Hai; Zhu, Hongguang; Zhou, Pei; Zhu, Haiyan; Ju, Dianwen

    2014-06-01

    Influenza A virus pandemics and emerging antiviral resistance highlight the urgent need for novel generic pharmacological strategies that reduce both viral replication and inflammation of the lung. We have previously investigated the therapeutic efficacy of recombinant human catalase (rhCAT) against viral pneumonia in mice, but the protection mechanisms involved were not explored. In the present study, we have performed a more in-depth analysis covering survival, lung inflammation, immune cell responses, production of cytokines, and inflammation signaling pathways in mice. Male imprinting control region mice were infected intranasally with high pathogenicity (H1N1) influenza A virus followed by treatment with recombinant human catalase. The administration of rhCAT resulted in a significant reduction in inflammatory cell infiltration (e.g., macrophages and neutrophils), inflammatory cytokine levels (e.g., IL-2, IL-6, TNF-α, IFN-γ), the level of the intercellular adhesion molecule 1 chemokine and the mRNA levels of toll-like receptors TLR-4, TLR-7, and NF-κB, as well as partially maintaining the activity of the antioxidant enzymes system. These findings indicated that rhCAT might play a key protective role in viral pneumonia of mice via suppression of inflammatory immune responses.

  1. Copper suppresses abscisic acid catabolism and catalase activity, and inhibits seed germination of rice.

    Science.gov (United States)

    Ye, Nenghui; Li, Haoxuan; Zhu, Guohui; Liu, Yinggao; Liu, Rui; Xu, Weifeng; Jing, Yu; Peng, Xinxiang; Zhang, Jianhua

    2014-11-01

    Although copper (Cu) is an essential micronutrient for plants, a slight excess of Cu in soil can be harmful to plants. Unfortunately, Cu contamination is a growing problem all over the world due to human activities, and poses a soil stress to plant development. As one of the most important biological processes, seed germination is sensitive to Cu stress. However, little is known about the mechanism of Cu-induced inhibition of seed germination. In the present study, we investigated the relationship between Cu and ABA which is the predominant regulator of seed germination. Cu at a concentration of 30 µM effectively inhibited germination of rice caryopsis. ABA content in germinating seeds under copper stress was also higher than that under control conditions. Quantitative real-time PCR (qRT-PCR) revealed that Cu treatment reduced the expression of OsABA8ox2, a key gene of ABA catabolism in rice seeds. In addition, both malondialdehyde (MDA) and H2O2 contents were increased by Cu stress in the germinating seeds. Antioxidant enzyme assays revealed that only catalase activity was reduced by excess Cu, which was consistent with the mRNA profile of OsCATa during seed germination under Cu stress. Together, our results demonstrate that suppression of ABA catabolism and catalase (CAT) activity by excess Cu leads to the inhibition of seed germination of rice.

  2. Catalase, carbonic anhydrase and xanthine oxidase activities in patients with mycosis fungoides.

    Science.gov (United States)

    Cengiz, Fatma Pelin; Beyaztas, Serap; Gokce, Basak; Arslan, Oktay; Guler, Ozen Ozensoy

    2015-04-01

    Mycosis fungoides (MF) is the most common form of cutaneous T-cell lymphoma. In several studies the relationship between catalase (CAT), human cytosolic carbonic anhydrases (CA; hCA-I and hCA-II) and xanthine oxidase (XO) enzyme activities have been investigated in various types of cancers but carbonic anhydrase, catalase and xanthine oxidase activities in patients with MF have not been previously reported. Therefore, in this preliminary study we aim to investigate CAT, CA and XO activities in patients with MF. This study enrolled 32 patients with MF and 26 healthy controls. According to the results, CA and CAT activities were significantly lower in patients with mycosis fungoides than controls (p < 0.001) (p < 0.001). There was no significant difference in XO activity between patient and control group (p = 0.601). Within these findings, we believe these enzyme activity levels might be a potentially important finding as an additional diagnostic biochemical tool for MF.

  3. Effects of some environmental parameters on catalase activity measured in the mussel (Mytilus galloprovincialis) exposed to lindane

    Energy Technology Data Exchange (ETDEWEB)

    Khessiba, Asma [Laboratoire de Bio-surveillance de l' Environnement, Unite d' Ecologie Cotiere, Faculte des Sciences de Bizerte, 7021, Zarzouna (Tunisia); Romeo, Michele [UMR INRA-UNSA 1112, ROSE - Reponse des Organismes aux Stress Environnementaux, Faculte des Sciences, BP 71, 06108, Nice Cedex 2 (France)]. E-mail: romeo@unice.fr; Aissa, Patricia [Laboratoire de Bio-surveillance de l' Environnement, Unite d' Ecologie Cotiere, Faculte des Sciences de Bizerte, 7021, Zarzouna (Tunisia)

    2005-01-01

    Mussels (Mytilus galloprovincialis), collected from the Bizerta lagoon, were acclimated for four days to various conditions of temperature, salinity, photoperiod and food supply and then exposed to lindane at a concentration of 40 {mu}g l{sup -1}. Catalase activity, which is a biomarker of exposure to an oxidative stress, was measured in the whole soft tissues of control and assay groups. In control mussels, high temperature, high salinity and light duration significantly increased catalase activity whereas this activity decreased when food, composed of freeze-dried, algae was available. When mussels were treated with lindane, catalase activities were higher than in controls. This increase was significant with temperature, salinity and light duration. The food supply did not change catalase activity, which was always higher compared to controls. Oxidative stress was shown in mussels exposed to lindane. The results highlight the need of considering abiotic parameters in biomonitoring studies, and especially when using catalase as a biomarker. - Oxidative stress in mussels exposed to lindane was also influenced by a number of abiotic parameters.

  4. Uncaria tomentosa extracts protect human erythrocyte catalase against damage induced by 2,4-D-Na and its metabolites.

    Science.gov (United States)

    Bukowska, Bożena; Bors, Milena; Gulewicz, Krzysztof; Koter-Michalak, Maria

    2012-06-01

    The effect of ethanolic and aqueous extracts from leaves and bark of Uncaria tomentosa was studied, with particular attention to catalase activity (CAT - EC. 1.11.1.6). We observed that all tested extracts, at a concentration of 250 μg/mL were not toxic to erythrocyte catalase because they did not decreased its activity. Additionally, we investigated the protective effect of extracts on changes in CAT activity in the erythrocytes incubated with sodium salt of 2,4-dichlorophenoxyacetic acid (2,4-D-Na) and its metabolites i.e., 2,4-dichlorophenol (2,4-DCP) and catechol. Previous investigations showed that these chemicals decreased activity of erythrocyte catalase (Bukowska et al., 2000; Bukowska and Kowalska, 2004). The erythrocytes were divided into two portions. The first portion was incubated for 1 and 5h at 37°C with 2,4-D-Na, 2,4-DCP and catechol, and second portion was preincubated with extracts for 10 min and then incubated with xenobiotics for 1 and 5h. CAT activity was measured in the first and second portion of the erythrocytes. We found a protective effect of the extracts from U. tomentosa on the activity of catalase incubated with xenobiotics studied. Probably, phenolic compounds contained in U. tomentosa scavenged free radicals, and therefore protected active center (containing -SH groups) of catalase.

  5. Lower Serum Catalase Level is Associated with Preterm Labor among Pregnant Women at Sanglah Hospital Denpasar, Bali-Indonesia

    Directory of Open Access Journals (Sweden)

    I Ketut Surya Negara

    2016-09-01

    Full Text Available Background: Preterm labor is still become a serious health problem in Obstetric and Perinatology with no sensitive biomarker currently approved. Several studies show that decrease antioxidant activity may play significant role in preterm labor. However, only few studies had been conducted to evaluate blood catalase level in preterm labor and assess its role in preterm labor. Objective: The aim this study was to identify the differences of maternal serum catalase level in preterm labor compared with preterm pregnancy. Methods: An observational analytic cross sectional study was conducted from February to December 2014 using pregnant women with 28-36 weeks’ gestational age. Blood catalase level was evaluated by colorimetric method and the data was analyzed by SPPS for Windows 17.0 program. Results: 12 subjects were enrolled and divided into preterm and control group. No significant differences between mean age, gestational age, and parity between preterm and control group. However, blood catalase level was significantly lower in preterm group compared with control group (81.82 ± 20.38 vs 159.38 ± 35.79; p=0.001. Conclusion: Serum maternal catalase level were significantly lower in preterm labor compared with preterm normal pregnancy.

  6. PRODUCING OF ENZYME PREPARATION AND ANALYSIS OF ENZYME PREPARATION OF PEROXIDASE AND CATALASE OF SOME SPECIES OF BASIDIOMYCETES

    Directory of Open Access Journals (Sweden)

    Fedotov O.V.

    2013-04-01

    Full Text Available A method for obtaining of enzyme preparations of enzyme preparations (EP of peroxidases and catalases fungal extracellular and inracellular origin from cultures of Basidiomycetes was developed. The strains Flammulina velutipes F-vv, Agrocybe cylindracea167; Fistulina hepatica Fh-08 and Pleurotus ostreatus P-208 and P-01 were used as producers of oxidoreductases. Strains were grown on modified glucose-peptone media. Fractionation was carried out by salting out the enzymes with ammonium sulfate at 40-70% saturation of peroxidases and 80% of saturation - for catalase. These solutions protein fractions was further purified by dialysis and gel filtration on Molselekt granules G-50 and G-75. The enzyme solution was subjected to freeze-drying. The individual characteristics of the enzyme preparations were found. The individual characteristics of the enzyme preparations are the activity of enzymes, the protein content and amino-acid composition of enzyme preparations. It was established that strain F. velutipes F-vv was an active producer of intracellular and strain of A. cylindracea 167 was an active producer of extracellular peroxidase. The strains of P. ostreatus P-01 and P-208 were the active producers of extracellular catalase, and the strainsof F. hepatica Fh-08 were active producers of intracellular catalase. The developed methods for producing of enzymes catalase and peroxidase preparations of extra-and intracellular origin provided new antioxidant enzymes, which have their own properties and application prospects in various sectors of industry and science research.

  7. A factor converting viable but nonculturable Vibrio cholerae to a culturable state in eukaryotic cells is a human catalase.

    Science.gov (United States)

    Senoh, Mitsutoshi; Hamabata, Takashi; Takeda, Yoshifumi

    2015-08-01

    In our previous work, we demonstrated that viable but nonculturable (VBNC) Vibrio cholerae O1 and O139 were converted to culturable by coculture with eukaryotic cells. Furthermore, we isolated a factor converting VBNC V. cholerae to culturable (FCVC) from a eukaryotic cell line, HT-29. In this study, we purified FCVC by successive column chromatographies comprising UNO Q-6 anion exchange, Bio-Scale CHT2-1 hydroxyapatite, and Superdex 200 10/300 GL. Homogeneity of the purified FCVC was demonstrated by SDS-PAGE. Nano-LC MS/MS analysis showed that the purified FCVC was a human catalase. An experiment of RNAi knockdown of catalase mRNA from HT-29 cells and treatment of the purified FCVC with a catalase inhibitor, 3-amino-1,2,4-triazole confirmed that the FCVC was a catalase. A possible role of the catalase in converting a VBNC V. cholerae to a culturable state in the human intestine is discussed.

  8. Important role of catalase in the cellular response of the budding yeast Saccharomyces cerevisiae exposed to ionizing radiation.

    Science.gov (United States)

    Nishimoto, Takuto; Furuta, Masakazu; Kataoka, Michihiko; Kishida, Masao

    2015-03-01

    Ionizing radiation indirectly causes oxidative stress in cells via reactive oxygen species (ROS), such as hydroxyl radicals (OH(-)) generated by the radiolysis of water. We investigated how the catalase function was affected by ionizing radiation and analyzed the phenotype of mutants with a disrupted catalase gene in Saccharomyces cerevisiae exposed to radiation. The wild-type yeast strain and isogenic mutants with disrupted catalase genes were exposed to various doses of (60)Co gamma-rays. There was no difference between the wild-type strain and the cta1 disruption mutant following exposure to gamma-ray irradiation. In contrast, there was a significant decrease in the ctt1 disruption mutant, suggesting that this strain exhibited decreased survival on gamma-ray exposure compared with other strains. In all three strains, stationary phase cells were more tolerant to the exposure of gamma-rays than exponential phase cells, whereas the catalase activity in the wild-type strain and cta1 disruption mutant was higher in the stationary phase than in the exponential phase. These data suggest a correlation between catalase activity and survival following gamma-ray exposure. However, this correlation was not clear in the ctt1 disruption mutant, suggesting that other factors are involved in the tolerance to ROS induced by irradiation.

  9. Immunological detection of alkaline-diaminobenzidine-negative peroxisomes of the nematode Caenorhabditis elegans: Purification and unique pH optima of peroxisomal catalase

    OpenAIRE

    Togo, Summanuna H.; Maebuchi, Motohiro; Yokota, Sadaki; Bun-ya, Masanori; Kawahara, Akira; Kamiryo, Tatsuyuki

    2000-01-01

    We purified catalase-2 of the nematode Caenorhabditis elegans and identified peroxisomes in this organism. The peroxisomes of C. elegans were not detectable by cytochemical staining using 3,3'-diaminobenzidine, a commonly used method depending on the peroxidase activity of peroxisomal catalase at pH 9 in which genuine peroxidases are inactive. The cDNA sequences of C. elegans predict two catalases very similar to each other throughout the molecule, except for the short C-terminal sequence; ca...

  10. siRNA-based Analysis of the Abrogation of the Protective Function of Membrane-associated Catalase of Tumor Cells.

    Science.gov (United States)

    Bauer, Georg

    2017-02-01

    Tumor cells, in contrast to non-malignant cells, show sustained expression of membrane-associated NADPH oxidase-1 and therefore generate extracellular superoxide anions and their dismutation product H2O2 In order to prevent intercellular reactive oxygen species/reactive nitrogen species (ROS/RNS)-dependent apoptosis-inducing signaling, tumor cells need to express membrane-associated catalase that interferes with HOCl and nitric oxide/peroxynitrite signaling. Catalase is attached to tumor cells through the activity of transglutaminase-2 and is prevented from superoxide anion-dependent inhibition through coexpression of membrane-associated superoxide dismutase. Therefore, specific inhibition of membrane-associated catalase should reactivate intercellular ROS/RNS-dependent apoptosis-inducing signaling. These processes are analyzed here through small interfering RNA-mediated knockdown of essential signaling compounds. This allows to establish a rather comprehensive picture of intercellular ROS/RNS signaling that may be instrumental for future therapeutic approaches.

  11. Cloning of catalase and expression patterns of catalase and selenium-dependent glutathione peroxidase fromExopalaemon carinicauda in response to low salinity stress

    Institute of Scientific and Technical Information of China (English)

    REN Hai; LI Jian; LI Jitao; YING Yu; GE Hongxing; LI Dongli; YU Tianji

    2015-01-01

    Catalase (CAT) and selenium-dependent glutathione peroxidase (Se-GPx) play a vital role in protecting organisms against various oxidative stresses by eliminating H2O2. The objective of this paper is to evaluate the roles of these antioxidant molecules in the ridgetail white prawnExopalaemon carinicauda in response to low salinity stress. A complementary DNA (cDNA) containing the complete coding sequence of CAT was cloned from the hepatopancreas using reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends. The full-length cDNA of CAT (2 649 bp) contains a 5'-untranslated region (UTR) of 78 bp, a 3'- UTR of 1 017 bp, with a poly (A) tail, and an open reading frame of 1 554 bp encoding a 517-amino-acid polypeptide with predicted molecular mass of 58.46 kDa and estimated isoelectric point of 6.64. This CAT sequence contained the proximal active site signature (60FDRERIPERVVHAKGAG76), proximal heme-ligand signature sequence (350RLFSYPDTH358) and three catalytic amino acid residues (His71, Asn144 and Tyr354). Sequence comparison showed that the CAT deduced amino acid sequence ofE.carinicauda shared 68%-92% of identities with those of other species. Quantitative real-time PCR analysis revealed that CAT mRNA was widely expressed in the hepatopancreas (highest), hemocyte, eyestalk, heart, gill, muscle, ovary and stomach. Under low salinity stress, CAT and GPx mRNA expression levels both in the gill and hepatopancreas increased significantly at the first 48 h and 6 h respectively, indicating a tissue- and time-dependent antioxidant response inE.carinicauda. All these results indicate thatE.carinicauda CAT is a member of the CAT family and might be involved in the acute response against low salinity stress.

  12. The catalase C-262T gene polymorphism and cancer risk: a systematic review and meta-analysis.

    Science.gov (United States)

    Shen, Yongchun; Li, Diandian; Tian, Panwen; Shen, Konglong; Zhu, Jing; Feng, Mei; Wan, Chun; Yang, Ting; Chen, Lei; Wen, Fuqiang

    2015-04-01

    Many studies suggest that catalase C-262T gene polymorphism is associated with cancer risk, but with inconsistent results. This study aimed to summarize the overall association between catalase C-262T polymorphism and cancer risk. Literature search was performed in PubMed, Embase, and other databases, studies regarding the association between catalase C-262T polymorphism and cancer risk were identified, and data were retrieved and analyzed by using Review Manager 5.0.24 and STATA 12.0. A total of 18 publications with 22 case-control studies, including 9777 cancer patients and 12,223 controls, met the inclusion criteria. Meta-analysis results showed significant association between catalase C-262 T polymorphism and cancer risk (TT vs CT + CC: odds ratio [OR] = 1.17, 95% confidence interval [CI] = 1.03-1.31, P = 0.01). Subgroup analyses stratified by cancer types suggested the catalase C-262T polymorphism was significantly associated with an increased prostate cancer risk (TT vs CT + CC: OR = 1.61, 95% CI = 1.17-2.22, P = 0.004); for subgroup analyses stratified by ethnicity, no associations between this polymorphism and Asians or whites were identified (CT + TT vs CC: OR = 1.11, 95% CI = 0.98-1.26, P = 0.09 for whites; OR = 1.19, 95% CI = 0.78-1.80, P = 0.42 for Asians). In summary, the catalase C-262T polymorphism may be a risk factor for cancer with cancer type-specific effects. Further studies should be performed to confirm these findings.

  13. Genetic polymorphisms in catalase and CYP1B1 determine DNA adduct formation by benzo(a)pyrene ex vivo.

    Science.gov (United States)

    Schults, Marten A; Chiu, Roland K; Nagle, Peter W; Wilms, Lonneke C; Kleinjans, Jos C; van Schooten, Frederik J; Godschalk, Roger W

    2013-03-01

    Genetic polymorphisms can partially explain the large inter-individual variation in DNA adduct levels following exposure to polycyclic aromatic hydrocarbons. Effects of genetic polymorphisms on DNA adduct formation are difficult to assess in human studies because exposure misclassification attenuates underlying relationships. Conversely, ex vivo studies offer the advantage of controlled exposure settings, allowing the possibility to better elucidate genotype-phenotype relationships and gene-gene interactions. Therefore, we exposed lymphocytes of 168 non-smoking volunteers ex vivo to the environmental pollutant benzo(a)pyrene (BaP) and BaP-related DNA adducts were quantified. Thirty-four genetic polymorphisms were assessed in genes involved in carcinogen metabolism, oxidative stress and DNA repair. Polymorphisms in catalase (CAT, rs1001179) and cytochrome P450 1B1 (CYP1B1, rs1800440) were significantly associated with DNA adduct levels, especially when combined. Moreover, reverse transcription-polymerase chain reaction (RT-PCR) analysis in a subset of 30 subjects revealed that expression of catalase correlated strongly with expression of CYP1B1 (R = 0.92, P CYP1B1 and how they simultaneously affect BaP-related DNA adduct levels, catalase expression was transiently knocked down in the human lung epithelial cell line A549. Although catalase knockdown did not immediately change CYP1B1 gene expression, recovery of catalase expression 8 h after the knockdown coincided with a 2.2-fold increased expression of CYP1B1 (P polymorphism in the promoter region of CAT may determine the amount and activity of catalase, which may subsequently regulate the expression of CYP1B1. As a result, both genetic polymorphisms modulate DNA adduct levels in lymphocytes by BaP ex vivo.

  14. Evaluation of Malondialdehyde, Superoxide Dismutase and Catalase Activity in Fetal Cord Blood of Depressed Mothers

    Science.gov (United States)

    Camkurt, Mehmet Akif; Fındıklı, Ebru; Bakacak, Murat; Tolun, Fatma İnanç; Karaaslan, Mehmet Fatih

    2017-01-01

    Objective The umbilical cord consists of two arteries and one vein and it functions in the transport between the maternal and fetal circulation. Biochemical analysis of fetal cord blood (FCB) during delivery could be beneficial in terms of understanding the fetal environment. In this study, we aimed to investigate oxidative parameters like malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT) levels in FCB during delivery. Methods We collected FCB samples during caesarean section. Our study included 33 depressed mothers and 37 healthy controls. We investigated MDA, SOD, and CAT levels in FCB samples. Results We found no significant difference between groups in terms of MDA (p=0.625), SOD (p=0.940), and CAT (p=0.413) levels. Conclusion Our study reveals probable protective effects of the placenta from oxidative stress. Future studies should include larger samples. PMID:28138108

  15. Catalase-positive microperoxisomes in rat soleus and extensor digitorum longus muscle fiber types

    Science.gov (United States)

    Riley, Danny A.; Bain, James L. W.; Ellis, Stanley

    1988-01-01

    The size, distribution, and content of catalase-reactive microperoxisomes were investigated cytochemically in three types of muscle fibers from the soleus and the extensor digitorum longus (EDL) of male rats. Muscle fibers were classified on the basis of the mitochondrial content and distribution, the Z-band widths, and the size and shape of myofibrils as the slow-twitch oxidative (SO), the fast-twitch oxidative glycolytic (FOG), and the fast-twitch glycolytic (FG) fibers. It was found that both the EDL and soleus SO fibers possessed the largest microperoxisomes. A comparison of microperoxisome number per muscle fiber area or the microperoxisome area per fiber area revealed following ranking, starting from the largest number and the area-ratio values: soleus SO, EDL SO, EDL FOG, and EDL FG.

  16. Francisella tularensis Catalase Restricts Immune Function by Impairing TRPM2 Channel Activity.

    Science.gov (United States)

    Shakerley, Nicole L; Chandrasekaran, Akshaya; Trebak, Mohamed; Miller, Barbara A; Melendez, J Andrés

    2016-02-19

    As an innate defense mechanism, macrophages produce reactive oxygen species that weaken pathogens and serve as secondary messengers involved in immune function. The Gram-negative bacterium Francisella tularensis utilizes its antioxidant armature to limit the host immune response, but the mechanism behind this suppression is not defined. Here we establish that F. tularensis limits Ca(2+) entry in macrophages, thereby limiting actin reorganization and IL-6 production in a redox-dependent fashion. Wild type (live vaccine strain) or catalase-deficient F. tularensis (ΔkatG) show distinct profiles in their H2O2 scavenging rates, 1 and 0.015 pm/s, respectively. Murine alveolar macrophages infected with ΔkatG display abnormally high basal intracellular Ca(2+) concentration that did not increase further in response to H2O2. Additionally, ΔkatG-infected macrophages displayed limited Ca(2+) influx in response to ionomycin, as a result of ionophore H2O2 sensitivity. Exogenously added H2O2 or H2O2 generated by ΔkatG likely oxidizes ionomycin and alters its ability to transport Ca(2+). Basal increases in cytosolic Ca(2+) and insensitivity to H2O2-mediated Ca(2+) entry in ΔkatG-infected cells are reversed by the Ca(2+) channel inhibitors 2-aminoethyl diphenylborinate and SKF-96365. 2-Aminoethyl diphenylborinate but not SKF-96365 abrogated ΔkatG-dependent increases in macrophage actin remodeling and IL-6 secretion, suggesting a role for H2O2-mediated Ca(2+) entry through the transient receptor potential melastatin 2 (TRPM2) channel in macrophages. Indeed, increases in basal Ca(2+), actin polymerization, and IL-6 production are reversed in TRPM2-null macrophages infected with ΔkatG. Together, our findings provide compelling evidence that F. tularensis catalase restricts reactive oxygen species to temper macrophage TRPM2-mediated Ca(2+) signaling and limit host immune function.

  17. Cu(II)-disulfide complexes display simultaneous superoxide dismutase- and catalase-like activities.

    Science.gov (United States)

    Aliaga, Margarita E; Andrade-Acuña, Daniela; López-Alarcón, Camilo; Sandoval-Acuña, Cristián; Speisky, Hernán

    2013-12-01

    Superoxide is a potentially toxic by-product of cellular metabolism. We have addressed here the in vitro ability of complexes formed between copper(II) ions and various biologically-occurring disulfides (RSSR: oxidized glutathione, cystine, homocystine and α-lipoic acid) to react with superoxide. The studied complexes were found to react with superoxide (generated by a xanthine/xanthine oxidase system) at rate constants (kCu(II)-RSSR) close to 10(6)M(-1)s(-1), which are three orders of magnitude lower than that reported for superoxide dismutase (SOD) but comparable to that of several other copper-containing complexes reported as SOD mimetics. The interaction between the tested Cu(II)-RSSR and superoxide, led to the generation and recovery of concentrations of hydrogen peroxide and oxygen that were, respectively, below and above those theoretically-expected from a sole SOD mimetic action. Interestingly, oxygen was generated when the Cu(II)-RSSR complexes were directly incubated with hydrogen peroxide. Taken together, these results reveal that the Cu(II)-RSSR complexes not only have the capacity to dismutate superoxide but also to simultaneously act like catalase mimetic molecules. When added to superoxide-overproducing mitochondria (condition attained by its exposure to diclofenac), three of the tested complexes were able (2-4μM), not only to totally restore, but also to lower below the basal level the mitochondrial production of superoxide. The present study is first in reporting on the potential of Cu(II)-disulfide complexes to act as SOD and catalase like molecules, suggesting a potential for these types of molecules to act as such under physiological and/or oxidative-stress conditions.

  18. Differential effects of superoxide dismutase and superoxide dismutase/catalase mimetics on human breast cancer cells.

    Science.gov (United States)

    Shah, Manisha H; Liu, Guei-Sheung; Thompson, Erik W; Dusting, Gregory J; Peshavariya, Hitesh M

    2015-04-01

    Reactive oxygen species (ROS) such as superoxide and hydrogen peroxide (H2O2) have been implicated in development and progression of breast cancer. In the present study, we have evaluated the effects of the superoxide dismutase (SOD) mimetic MnTmPyP and the SOD/catalase mimetic EUK 134 on superoxide and H2O2 formation as well as proliferation, adhesion, and migration of MCF-7 and MDA-MB-231 cells. Superoxide and H2O2 production was examined using dihydroethidium and Amplex red assays, respectively. Cell viability and adhesion were measured using a tetrazolium-based MTT assay. Cell proliferation was determined using trypan blue assay. Cell cycle progression was analyzed using flow cytometry. Clonal expansion of a single cell was performed using a colony formation assay. Cell migration was measured using transwell migration assay. Dual luciferase assay was used to determine NF-κB reporter activity. EUK 134 effectively reduced both superoxide and H2O2, whereas MnTmPyP removed superoxide but enhanced H2O2 formation. EUK 134 effectively attenuated viability, proliferation, clonal expansion, adhesion, and migration of MCF-7 and MDA-MB-231 cells. In contrast, MnTmPyP only reduced clonal expansion of MCF-7 and MDA-MB-231 cells but had no effect on adhesion and cell cycle progression. Tumor necrosis factor-alpha-induced NF-κB activity was reduced by EUK 134, whereas MnTmPyP enhanced this activity. These data indicate that the SOD mimetic MnTmPyP and the SOD/catalase mimetic EUK 134 exert differential effects on breast cancer cell growth. Inhibition of H2O2 signaling using EUK 134-like compound might be a promising approach to breast cancer therapy.

  19. Investigation on the toxic interaction of superparamagnetic iron oxide nanoparticles with catalase

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Zehua [Shandong Key Laboratory of Water Pollution Control and Resource Reuse, School of Environmental Science and Engineering, China–America CRC for Environment and Health, Shandong Province, Shandong University, 27# Shanda South Road, Jinan 250100 (China); Liu, Hongwei [Shandong Key Laboratory of Water Pollution Control and Resource Reuse, School of Environmental Science and Engineering, China–America CRC for Environment and Health, Shandong Province, Shandong University, 27# Shanda South Road, Jinan 250100 (China); Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085 (China); Hu, Xinxin; Song, Wei [Shandong Key Laboratory of Water Pollution Control and Resource Reuse, School of Environmental Science and Engineering, China–America CRC for Environment and Health, Shandong Province, Shandong University, 27# Shanda South Road, Jinan 250100 (China); Liu, Rutao, E-mail: rutaoliu@sdu.edu.cn [Shandong Key Laboratory of Water Pollution Control and Resource Reuse, School of Environmental Science and Engineering, China–America CRC for Environment and Health, Shandong Province, Shandong University, 27# Shanda South Road, Jinan 250100 (China)

    2015-03-15

    Superparamagnetic iron oxide nanoparticles (SPIONs) have been investigated for various applications in targeted drug delivery and magnetic resonance imaging. Given their clinical relevance, there is a need to understand these particles' potential cytotoxic effects and possible mechanisms of cytotoxicity. Using a variety of spectroscopic techniques, we investigated the interaction of SPIONs with catalase (CAT) in an aqueous environment. Catalase is an important enzyme that protects cells and tissues from oxidative damage by reactive oxygen species (ROS). Therefore, in this work, CAT served as a model protein for examining the physiological effects of SPIONs due to is function in eliminating H{sub 2}O{sub 2}. Synchronous fluorescence spectroscopy results showed that SPIONs have little effect on tryptophan residues in CAT. Data from circular dichroism (CD) and UV–vis spectroscopies showed that CAT α-helical content decreased from 32.4% to 29.1% in the presence of SPIONs. Moreover, a ca. 10% decrease in CAT activity was observed in the presence of SPIONs at a 20:1 particle:protein ratio. These results show that SPIONs can interact with proteins to alter both their structure and function. Further studies with CAT or other toxicologically relevant enzymes may be used for elucidating the mechanisms of SPION cytotoxicity. - Highlights: • This work established the binding mode of SPIONs with CAT on molecular level. • The interaction mechanism was explored by multiple spectroscopic techniques. • SPIONs can loosen the skeleton of protein and increase the exposure of amide moieties in the hydrophobic pocket. • SPIONs can inhibit CAT activity and trigger conformational changes in CAT.

  20. Toward "stable-on-the-table" enzymes: improving key properties of catalase by covalent conjugation with poly(acrylic acid).

    Science.gov (United States)

    Riccardi, Caterina M; Cole, Kyle S; Benson, Kyle R; Ward, Jessamyn R; Bassett, Kayla M; Zhang, Yiren; Zore, Omkar V; Stromer, Bobbi; Kasi, Rajeswari M; Kumar, Challa V

    2014-08-20

    Several key properties of catalase such as thermal stability, resistance to protease degradation, and resistance to ascorbate inhibition were improved, while retaining its structure and activity, by conjugation to poly(acrylic acid) (PAA, Mw 8000) via carbodiimide chemistry where the amine groups on the protein are appended to the carboxyl groups of the polymer. Catalase conjugation was examined at three different pH values (pH 5.0, 6.0, and 7.0) and at three distinct mole ratios (1:100, 1:500, and 1:1000) of catalase to PAA at each reaction pH. The corresponding products are labeled as Cat-PAA(x)-y, where x is the protein to polymer mole ratio and y is the pH used for the synthesis. The coupling reaction consumed about 60-70% of the primary amines on the catalase; all samples were completely water-soluble and formed nanogels, as evidenced by gel electrophoresis and electron microscopy. The UV circular dichroism (CD) spectra indicated substantial retention of protein secondary structure for all samples, which increased to 100% with increasing pH of the synthesis and polymer mole fraction. Soret CD bands of all samples indicated loss of ∼50% of band intensities, independent of the reaction pH. Catalytic activities of the conjugates increased with increasing synthesis pH, where 55-80% and 90-100% activity was retained for all samples synthesized at pH 5.0 and pH 7.0, respectively, and the Km or Vmax values of Cat-PAA(100)-7 did not differ significantly from those of the free enzyme. All conjugates synthesized at pH 7.0 were thermally stable even when heated to ∼85-90 °C, while native catalase denatured between 55 and 65 °C. All conjugates retained 40-90% of their original activities even after storing for 10 weeks at 8 °C, while unmodified catalase lost all of its activity within 2 weeks, under similar storage conditions. Interestingly, PAA surrounding catalase limited access to the enzyme from large molecules like proteases and significantly increased

  1. Expression, Tag Cleavage and Identification of Catalase/GST Fusion Protein of Helicobacterpylori%幽门杆菌Catalase/GST融合蛋白的表达、标签切除及鉴定

    Institute of Scientific and Technical Information of China (English)

    姜茵; 奚月; 李妍

    2012-01-01

    旨在利用GST融合基因表达系统表达幽门螺杆菌Catalase融合蛋白,并利用凝血酶切除GST标签.将重组质粒Catalase/pGEX-4T-1转化大肠杆菌BL21( DE3)感受态中,用IPTG进行诱导表达,菌体经反复冻融、溶菌酶裂解及超声破菌后,Catalase/GST融合蛋白以部分可溶性的形式表达在上清中.采用谷胱甘肽琼脂糖树脂Glutathione Sepharose 4B对其进行纯化,得到Catalase/GST融合蛋白,再用凝血酶进行GST标签的切除,所得产物进行Western blotting鉴定.高效表达出Catalase/GST融合蛋白的相对分子质量约85 kD,凝血酶成功地切除了GST标签,Western blotting证实Catalase蛋白能被鼠抗Catalase单克隆抗体识别.

  2. Thermo-alkali-stable catalases from newly isolated Bacillus sp. for the treatment and recycling of textile bleaching effluents.

    Science.gov (United States)

    Paar, A; Costa, S; Tzanov, T; Gudelj, M; Robra, K H; Cavaco-Paulo, A; Gübitz, G M

    2001-08-23

    Three thermoalkaliphilic bacteria, which were grown at pH 9.3-10 and 60-65 degrees C were isolated out of a textile wastewater drain. The unknown micro-organisms were identified as thermoalkaliphilic Bacillus sp. Growth conditions were studied and catalase activities and stabilities compared. Catalases from Bacillus SF showed high stabilities at 60 degrees C and pH 9 (t1/2=38 h) and thus this strain was chosen for further investigations, such as electron microscopy, immobilization of catalase and hydrogen peroxide degradation studies. Degradation of hydrogen peroxide with an immobilized catalase from Bacillus SF enabled the reuse of the water for the dyeing process. In contrast, application of the free enzyme for treatment of bleaching effluents, caused interaction between the denaturated protein and the dye, resulting in reduced dye uptake, and a higher color difference of 1.3DeltaE* of dyed fabrics compared to 0.9DeltaE* when using the immobilized enzyme.

  3. Novel immobilization process of a thermophilic catalase: efficient purification by heat treatment and subsequent immobilization at high temperature.

    Science.gov (United States)

    Xu, Juan; Luo, Hui; López, Claudia; Xiao, Jing; Chang, Yanhong

    2015-10-01

    The main goal of the present work is to investigate a novel process of purification and immobilization of a thermophilic catalase at high temperatures. The catalase, originated from Bacillus sp., was overexpressed in a recombinant Escherichia coli BL21(DE3)/pET28-CATHis and efficiently purified by heat treatment, achieving a threefold purification. The purified catalase was then immobilized onto an epoxy support at different temperatures (25, 40, and 55 °C). The immobilizate obtained at higher temperatures reached its maximum activity in a shorter time than that obtained at lower temperatures. Furthermore, immobilization at higher temperatures required a lower ionic strength than immobilization at lower temperatures. The characteristics of immobilized enzymes prepared at different temperatures were investigated. The high-temperature immobilizate (55 °C) showed the highest thermal stability, followed by the 40 °C immobilizate. And the high-temperature immobilizate (55 °C) had slightly higher operational stability than the 25 °C immobilizate. All of the immobilized catalase preparations showed higher stability than the free enzyme at alkaline pH 10.0, while the alkali resistance of the 25 °C immobilizate was slightly better than that of the 40 and 55 °C immobilizates.

  4. Erythrocyte Catalase Activity in More Frequent Microcytic Hypochromic Anemia: Beta-Thalassemia Trait and Iron Deficiency Anemia

    Directory of Open Access Journals (Sweden)

    Sandra Stella Lazarte

    2015-01-01

    Full Text Available Most common microcytic hypochromic anemias are iron deficiency anemia (IDA and β-thalassemia trait (BTT, in which oxidative stress (OxS has an essential role. Catalase causes detoxification of H2O2 in cells, and it is an indispensable antioxidant enzyme. The study was designed to measure erythrocyte catalase activity (ECAT in patients with IDA (10 or BTT (21, to relate it with thalassemia mutation type (β0 or β+ and to compare it with normal subjects (67. Ninety-eight individuals were analyzed since September 2013 to June 2014 in Tucumán, Argentina. Total blood count, hemoglobin electrophoresis at alkaline pH, HbA2, catalase, and iron status were performed. β-thalassemic mutations were determined by real-time PCR. Normal range for ECAT was 70,0–130,0 MU/L. ECAT was increased in 14% (3/21 of BTT subjects and decreased in 40% (4/10 of those with IDA. No significant difference (p=0,245 was shown between normal and BTT groups, while between IDA and normal groups the difference was proved to be significant (p=0,000. In β0 and β+ groups, no significant difference (p=0,359 was observed. An altered ECAT was detected in IDA and BTT. These results will help to clarify how the catalase activity works in these anemia types.

  5. Erythrocyte Catalase Activity in More Frequent Microcytic Hypochromic Anemia: Beta-Thalassemia Trait and Iron Deficiency Anemia

    Science.gov (United States)

    Lazarte, Sandra Stella; Mónaco, María Eugenia; Jimenez, Cecilia Laura; Ledesma Achem, Miryam Emilse; Terán, Magdalena María; Issé, Blanca Alicia

    2015-01-01

    Most common microcytic hypochromic anemias are iron deficiency anemia (IDA) and β-thalassemia trait (BTT), in which oxidative stress (OxS) has an essential role. Catalase causes detoxification of H2O2 in cells, and it is an indispensable antioxidant enzyme. The study was designed to measure erythrocyte catalase activity (ECAT) in patients with IDA (10) or BTT (21), to relate it with thalassemia mutation type (β0 or β+) and to compare it with normal subjects (67). Ninety-eight individuals were analyzed since September 2013 to June 2014 in Tucumán, Argentina. Total blood count, hemoglobin electrophoresis at alkaline pH, HbA2, catalase, and iron status were performed. β-thalassemic mutations were determined by real-time PCR. Normal range for ECAT was 70,0–130,0 MU/L. ECAT was increased in 14% (3/21) of BTT subjects and decreased in 40% (4/10) of those with IDA. No significant difference (p = 0,245) was shown between normal and BTT groups, while between IDA and normal groups the difference was proved to be significant (p = 0,000). In β0 and β+ groups, no significant difference (p = 0,359) was observed. An altered ECAT was detected in IDA and BTT. These results will help to clarify how the catalase activity works in these anemia types. PMID:26527217

  6. Multiple abiotic stress tolerance of the transformants yeast cells and the transgenic Arabidopsis plants expressing a novel durum wheat catalase.

    Science.gov (United States)

    Feki, Kaouthar; Kamoun, Yosra; Ben Mahmoud, Rihem; Farhat-Khemakhem, Ameny; Gargouri, Ali; Brini, Faiçal

    2015-12-01

    Catalases are reactive oxygen species scavenging enzymes involved in response to abiotic and biotic stresses. In this study, we described the isolation and functional characterization of a novel catalase from durum wheat, designed TdCAT1. Molecular Phylogeny analyses showed that wheat TdCAT1 exhibited high amino acids sequence identity to other plant catalases. Sequence homology analysis showed that TdCAT1 protein contained the putative calmodulin binding domain and a putative conserved internal peroxisomal targeting signal PTS1 motif around its C-terminus. Predicted three-dimensional structural model revealed the presence of four putative distinct structural regions which are the N-terminal arm, the β-barrel, the wrapping and the α-helical domains. TdCAT1 protein had the heme pocket that was composed by five essential residues. TdCAT1 gene expression analysis showed that this gene was induced by various abiotic stresses in durum wheat. The expression of TdCAT1 in yeast cells and Arabidopsis plants conferred tolerance to several abiotic stresses. Compared with the non-transformed plants, the transgenic lines maintained their growth and accumulated more proline under stress treatments. Furthermore, the amount of H2O2 was lower in transgenic lines, which was due to the high CAT and POD activities. Taken together, these data provide the evidence for the involvement of durum wheat catalase TdCAT1 in tolerance to multiple abiotic stresses in crop plants.

  7. The Antitumor Effect of Single-domain Antibodies Directed Towards Membrane-associated Catalase and Superoxide Dismutase.

    Science.gov (United States)

    Bauer, Georg; Motz, Manfred

    2016-11-01

    Neutralizing single-domain antibodies directed towards catalase or superoxide dismutase (SOD) caused efficient reactivation of intercellular reactive oxygen species/reactive nitrogen species (ROS/RNS)-dependent apoptosis-inducing signaling specifically in human tumor cells. Single-domain antibodies targeted tumor cell-specific membrane-associated SOD and catalase, but not the corresponding intracellular enzymes. They were shown to be about 200-fold more effective than corresponding classical recombinant antigen-binding fragments and more than four log steps more efficient than monoclonal antibodies. Combined addition of single-domain antibodies against catalase and SOD caused a remarkable synergistic effect. Proof-of-concept experiments in immunocompromised mice using human tumor xenografts and single-domain antibodies directed towards SOD showed an inhibition of tumor growth. Neutralizing single-domain antibodies directed to catalase and SOD also caused a very strong synergistic effect with the established chemotherapeutic agent taxol, indicating an overlap of signaling pathways. This effect might also be useful in order to avoid unwanted side-effects and to drastically lower the costs for taxol-based therapy.

  8. Horizontal gene transfer confers adaptive advantages to phytopathogenic fungi: a case study of catalase-peroxidase in Fusarium verticillioides

    Science.gov (United States)

    Horizontal gene transfer (HGT), the exchange and stable integration of genetic material between different evolutionary lineages, is widely observed in fungi. We hypothesize that successful stabilization of HGT elements provides adaptive advantages (e.g., virulence). Catalase/peroxidases (KatGs) are ...

  9. Differential mRNA expression of Sporothrix schenckii catalase gene in two growth phases and effect factors

    Institute of Scientific and Technical Information of China (English)

    WANG Xiao-hui; LI Ruo-yu; CAO Cun-wei; WANG Yu-hong; LIU Wei; QIAO Jian-jun; LI Yu-zhen

    2008-01-01

    @@ Sporothrix schenckii (S. schenckii), a dimorphic fungus, is the etiological agent of sporotrichosis. After entrance of microconidia or mycelial fragments into a mammalian host, the fungus differentiates into the parasitic yeast form. Meanwhile, several defensive signals would be triggered when the innate immune system was stimulated by 5. schenckii invasion and microbe-specific phagocytosis. The success of 5. schenckii infection partly depends on its ability to avoid oxidative damage from reactive oxygen species (ROS) released by polymorphonuclear leukocytes and activated macrophages. However, the antioxidant defense mechanisms of S. schenckii remains unknown. Catalases, one of the central enzymes involved in scavenging ROS via converting hydrogen peroxide (H2O2) to water and molecular oxygen, play an essential role in protecting intracellular pathogenic fungi against ROS1,2 and regulating growth and development in some fungi.3'4 No catalase in 5. schenckii has been identified and characterized previously. Recently we have cloned a catalase homologous gene from the yeast form of 5. schenckii and designated it Sscat. In this report, we used quantitative real-time polymerase chain reaction (PCR) to measure and compare mRNA expression of Sscat in the mycelium-to-yeast transition and H2O2-challenge induced microenvironments in vitro. The results presented may help in understanding the role of 5. schenckii catalase in the fungus-host interaction with respect to ROS.

  10. Cloning and expression of the catalase-peroxidase gene from the hyperthermophilic archaeon Archaeoglobus fulgidus and characterization of the enzyme

    NARCIS (Netherlands)

    Kengen, S.W.M.; Bikker, F.; Vos, de W.M.; Oost, van der J.

    2001-01-01

    A putative perA gene from Archaeoglobus fulgidus was cloned and expressed in Escherichia coli BL21(DE3), and the recombinant catalase-peroxidase was purified to homogeneity. The enzyme is a homodimer with a subunit molecular mass of 85 kDa. UV-visible spectroscopic analysis indicated the presence of

  11. Isolation of a novel peroxisomal catalase gene from sugarcane, which is responsive to biotic and abiotic stresses.

    Directory of Open Access Journals (Sweden)

    Yachun Su

    Full Text Available Catalase is an iron porphyrin enzyme, which serves as an efficient scavenger of reactive oxygen species (ROS to avoid oxidative damage. In sugarcane, the enzymatic activity of catalase in a variety (Yacheng05-179 resistant to the smut pathogen Sporisorium scitamineum was always higher than that of the susceptible variety (Liucheng03-182, suggesting that catalase activity may have a positive correlation with smut resistance in sugarcane. To understand the function of catalase at the molecular level, a cDNA sequence of ScCAT1 (GenBank Accession No. KF664183, was isolated from sugarcane infected by S. scitamineum. ScCAT1 was predicted to encode 492 amino acid residues, and its deduced amino acid sequence shared a high degree of homology with other plant catalases. Enhanced growth of ScCAT1 in recombinant Escherichia coli Rosetta cells under the stresses of CuCl2, CdCl2 and NaCl indicated its high tolerance. Q-PCR results showed that ScCAT1 was expressed at relatively high levels in the bud, whereas expression was moderate in stem epidermis and stem pith. Different kinds of stresses, including S. scitamineum challenge, plant hormones (SA, MeJA and ABA treatments, oxidative (H2O2 stress, heavy metal (CuCl2 and hyper-osmotic (PEG and NaCl stresses, triggered a significant induction of ScCAT1. The ScCAT1 protein appeared to localize in plasma membrane and cytoplasm. Furthermore, histochemical assays using DAB and trypan blue staining, as well as conductivity measurement, indicated that ScCAT1 may confer the sugarcane immunity. In conclusion, the positive response of ScCAT1 to biotic and abiotic stresses suggests that ScCAT1 is involved in protection of sugarcane against reactive oxidant-related environmental stimuli.

  12. Apparent Catalase Synthesis in Sunflower Cotyledons during the Change in Microbody Function: A Mathematical Approach for the Quantitative Evaluation of Density-labeling Data.

    Science.gov (United States)

    Betsche, T; Gerhardt, B

    1978-10-01

    Density-labeling with 10 mm K(15)NO(3)/70% (2)H(2)O has been used to investigate catalase synthesis in different developmental stages of sunflower (Helianthus annuus L.) cotyledons. A mathematical approach is introduced for the quantitative evaluation of the density-labeling data. The method allows, in the presence of preexisting enzyme activity, calculation of this synthesized activity (apparent enzyme synthesis) which results from the balance between actual enzyme synthesis and the degradation of newly synthesized enzyme at a given time. During greening of the cotyledons, when the catalase activity declines and the population of leaf peroxisomes is formed, the apparent catalase synthesis is lower than, or at best equal to, that occurring during a developmental stage when the leaf peroxisome population is established and catalase synthesis and degradation of total catalase are in equilibrium. This result suggests a formation, in fatty cotyledons, of the leaf peroxisomes by transformation of the glyoxysomes rather than by de novo synthesis.

  13. Effect of covalent attachment of neomycin on conformational and aggregation properties of catalase.

    Science.gov (United States)

    Hashemnia, S; Mokhtari, Z; Tashkhourian, J; Moosavi-Movahedi, A A

    2015-04-01

    The carboxylic groups of glutamic acid and aspartic acid residues of catalase (CAT) were chemically modified using the treatment of the enzyme with 1-ethyl-3-(3'-dimethylamino) carbodiimide hydrochloride (EDC) and neomycin. The effect of covalent attachment of neomycin on the enzymatic activity, conformational and aggregation properties of CAT was investigated. The modification of CAT with different concentrations of neomycin showed two different types of behavior, depending up on the concentration range of neomycin. In the concentration range from 0.0 to 5.2 mM, neomycin-modified CAT, compared to the native enzyme exhibited higher a-helix content, reduced surface hydrophobicity, little enhancement in CAT activity and a better protection against thermal aggregation, whereas at concentrations greater than 5.2 mM, the modified enzyme exhibited a significant decrease in CAT activity and an increase in random coil content which may result in disorder in the protein structure and increase in thermal aggregation. This modification is a rapid and simple approach to investigate the role of aspartate and glutamate residues in the structure, function and folding of CAT.

  14. Activity of catalase adsorbed to carbon nanotubes: effects of carbon nanotube surface properties.

    Science.gov (United States)

    Zhang, Chengdong; Luo, Shuiming; Chen, Wei

    2013-09-15

    Nanomaterials have been studied widely as the supporting materials for enzyme immobilization. However, the interactions between enzymes and carbon nanotubes (CNT) with different morphologies and surface functionalities may vary, hence influencing activities of the immobilized enzyme. To date how the adsorption mechanisms affect the activities of immobilized enzyme is not well understood. In this study the adsorption of catalase (CAT) on pristine single-walled carbon nanotubes (SWNT), oxidized single-walled carbon nanotubes (O-SWNT), and multi-walled carbon nanotubes (MWNT) was investigated. The adsorbed enzyme activities decreased in the order of O-SWNT>SWNT>MWNT. Fourier transforms infrared spectroscopy (FTIR) and circular dichrois (CD) analyses reveal more significant loss of α-helix and β-sheet of MWNT-adsorbed than SWNT-adsorbed CAT. The difference in enzyme activities between MWNT-adsorbed and SWNT-adsorbed CAT indicates that the curvature of surface plays an important role in the activity of immobilized enzyme. Interestingly, an increase of β-sheet content was observed for CAT adsorbed to O-SWNT. This is likely because as opposed to SWNT and MWNT, O-SWNT binds CAT largely via hydrogen bonding and such interaction allows the CAT molecule to maintain the rigidity of enzyme structure and thus the biological function.

  15. Imipramine enhances neuroprotective effect of PEP-1-Catalase against ischemic neuronal damage.

    Science.gov (United States)

    Kim, Dae Won; Kim, Duk-Soo; Kim, Mi Jin; Kwon, Soon Won; Ahn, Eun Hee; Jeong, Hoon Jae; Sohn, Eun Jeong; Dutta, Suman; Lim, Soon Sung; Cho, Sung-Woo; Lee, Kil Soo; Park, Jinseu; Eum, Won Sik; Hwang, Hyun Sook; Choi, Soo Young

    2011-10-01

    The protein transduction domains have been reported to have potential to deliver the exogenous molecules, including proteins, to living cells. However, poor transduction of proteins limits therapeutic application. In this study, we examined whether imipramine could stimulate the transduction efficiency of PEP-1 fused proteins into astrocytes. PEP-1-catalase (PEP-1- CAT) was transduced into astrocytes in a time- and dose-dependent manner, reducing cellular toxicity induced by H(2)O(2). Additionally, the group of PEP-1-CAT (+) imipramine showed enhancement of transduction efficiency and therefore increased cellular viability than that of PEP-1-CAT alone. In the gerbil ischemia models, PEP-1-CAT displayed significant neuroprotection in the CA1 region of the hippocampus. Interestingly, PEP-1-CAT (+) imipramine prevented neuronal cell death and lipid peroxidation more markedly than PEP-1-CAT alone. Therefore, our results suggest that imipramine can be used as a drug to enhance the transduction of PEP-1 fusion proteins to cells or animals and their efficacies against various disorders.

  16. Nanospherical Brush as Catalase Container for Enhancing the Detection Sensitivity of Competitive Plasmonic ELISA.

    Science.gov (United States)

    Huang, Xiaolin; Chen, Rui; Xu, Hengyi; Lai, Weihua; Xiong, Yonghua

    2016-02-01

    Plasmonic enzyme-linked immunosorbent assay (pELISA) based on catalase (CAT)-mediated gold nanoparticle growth shows great potential for the determination of disease-related biomarkers at ultralow concentrations by using sandwich formats. However, the relatively low sensitivity of this strategy using competitive formats limits its adoption for hapten detection. Herein, we present an improved competitive pELISA for ultrasensitive detection of ochratoxin A (OTA), where silica nanoparticles carrying poly(acrylic acid) brushes (SiO2@PAA) were used to decrease the affinity of competing antigens to anti-OTA monoclonal antibodies and amplify the signal as a "CAT container" (SiO2@PAA@CAT). The developed competitive pELISA exhibits extremely high sensitivity for OTA with detection limits of 10(-18) and 5 × 10(-20) g/mL by the naked eye and microplate reader, respectively. These values are at least 7 orders of magnitude lower than that of competitive CAT-based pELISA (10(-11) g/mL by the naked eye) and 8 orders of magnitude lower than that of horseradish peroxidase-based conventional ELISA (10(-11) g/mL by the microplate reader), respectively. Reliability and robustness of the proposed method were evaluated using actual agricultural products and human serum samples. This study demonstrated the potential of this modified method in practical applications involving the ultrasensitive detection of mycotoxins or other haptens.

  17. Catalase-Based Modified Graphite Electrode for Hydrogen Peroxide Detection in Different Beverages

    Directory of Open Access Journals (Sweden)

    Giovanni Fusco

    2016-01-01

    Full Text Available A catalase-based (NAF/MWCNTs nanocomposite film modified glassy carbon electrode for hydrogen peroxide (H2O2 detection was developed. The developed biosensor was characterized in terms of its bioelectrochemical properties. Cyclic voltammetry (CV technique was employed to study the redox features of the enzyme in the absence and in the presence of nanomaterials dispersed in Nafion® polymeric solution. The electron transfer coefficient, α, and the electron transfer rate constant, ks, were found to be 0.42 and 1.71 s−1, at pH 7.0, respectively. Subsequently, the same modification steps were applied to mesoporous graphite screen-printed electrodes. Also, these electrodes were characterized in terms of their main electrochemical and kinetic parameters. The biosensor performances improved considerably after modification with nanomaterials. Moreover, the association of Nafion with carbon nanotubes retained the biological activity of the redox protein. The enzyme electrode response was linear in the range 2.5–1150 μmol L−1, with LOD of 0.83 μmol L−1. From the experimental data, we can assess the possibility of using the modified biosensor as a useful tool for H2O2 determination in packaged beverages.

  18. Study on the interaction of catalase with pesticides by flow injection chemiluminescence and molecular docking.

    Science.gov (United States)

    Tan, Xijuan; Wang, Zhuming; Chen, Donghua; Luo, Kai; Xiong, Xunyu; Song, Zhenghua

    2014-08-01

    The interaction mechanisms of catalase (CAT) with pesticides (including organophosphates: disulfoton, isofenphos-methyl, malathion, isocarbophos, dimethoate, dipterex, methamidophos and acephate; carbamates: carbaryl and methomyl; pyrethroids: fenvalerate and deltamethrin) were first investigated by flow injection (FI) chemiluminescence (CL) analysis and molecular docking. By homemade FI-CL model of lg[(I0-I)/I]=lgK+nlg[D], it was found that the binding processes of pesticides to CAT were spontaneous with the apparent binding constants K of 10(3)-10(5) L mol(-1) and the numbers of binding sites about 1.0. The binding abilities of pesticides to CAT followed the order: fenvalerate>deltamethrin>disulfoton>isofenphos-methyl>carbaryl>malathion>isocarbophos>dimethoate>dipterex>acephate>methomyl>methamidophos, which was generally similar to the order of determination sensitivity of pesticides. The thermodynamic parameters revealed that CAT bound with hydrophobic pesticides by hydrophobic interaction force, and with hydrophilic pesticides by hydrogen bond and van der Waals force. The pesticides to CAT molecular docking study showed that pesticides could enter into the cavity locating among the four subdomains of CAT, giving the specific amino acid residues and hydrogen bonds involved in CAT-pesticides interaction. It was also found that the lgK values of pesticides to CAT increased regularly with increasing lgP, Mr, MR and MV, suggesting that the hydrophobicity and steric property of pesticide played essential roles in its binding to CAT.

  19. Analysis of oxidative stress status, catalase and catechol-O-methyltransferase polymorphisms in Egyptian vitiligo patients.

    Directory of Open Access Journals (Sweden)

    Dina A Mehaney

    Full Text Available Vitiligo is the most common depigmentation disorder of the skin. Oxidative stress is implicated as one of the probable events involved in vitiligo pathogenesis possibly contributing to melanocyte destruction. Evidence indicates that certain genes including those involved in oxidative stress and melanin synthesis are crucial for development of vitiligo. This study evaluates the oxidative stress status, the role of catalase (CAT and catechol-O-Methyltransferase (COMT gene polymorphisms in the etiology of generalized vitiligo in Egyptians. Total antioxidant capacity (TAC and malondialdehyde (MDA levels as well as CAT exon 9 T/C and COMT 158 G/A polymorphisms were determined in 89 patients and 90 age and sex-matched controls. Our results showed significantly lower TAC along with higher MDA levels in vitiligo patients compared with controls. Meanwhile, genotype and allele distributions of CAT and COMT polymorphisms in cases were not significantly different from those of controls. Moreover, we found no association between both polymorphisms and vitiligo susceptibility. In conclusion, the enhanced oxidative stress with the lack of association between CAT and COMT polymorphisms and susceptibility to vitiligo in our patients suggest that mutations in other genes related to the oxidative pathway might contribute to the etiology of generalized vitiligo in Egyptian population.

  20. Catalase-Based Modified Graphite Electrode for Hydrogen Peroxide Detection in Different Beverages.

    Science.gov (United States)

    Fusco, Giovanni; Bollella, Paolo; Mazzei, Franco; Favero, Gabriele; Antiochia, Riccarda; Tortolini, Cristina

    2016-01-01

    A catalase-based (NAF/MWCNTs) nanocomposite film modified glassy carbon electrode for hydrogen peroxide (H2O2) detection was developed. The developed biosensor was characterized in terms of its bioelectrochemical properties. Cyclic voltammetry (CV) technique was employed to study the redox features of the enzyme in the absence and in the presence of nanomaterials dispersed in Nafion® polymeric solution. The electron transfer coefficient, α, and the electron transfer rate constant, ks , were found to be 0.42 and 1.71 s(-1), at pH 7.0, respectively. Subsequently, the same modification steps were applied to mesoporous graphite screen-printed electrodes. Also, these electrodes were characterized in terms of their main electrochemical and kinetic parameters. The biosensor performances improved considerably after modification with nanomaterials. Moreover, the association of Nafion with carbon nanotubes retained the biological activity of the redox protein. The enzyme electrode response was linear in the range 2.5-1150 μmol L(-1), with LOD of 0.83 μmol L(-1). From the experimental data, we can assess the possibility of using the modified biosensor as a useful tool for H2O2 determination in packaged beverages.

  1. Catalase and ascorbate peroxidase-representative H2O2-detoxifying heme enzymes in plants.

    Science.gov (United States)

    Anjum, Naser A; Sharma, Pallavi; Gill, Sarvajeet S; Hasanuzzaman, Mirza; Khan, Ekhlaque A; Kachhap, Kiran; Mohamed, Amal A; Thangavel, Palaniswamy; Devi, Gurumayum Devmanjuri; Vasudhevan, Palanisamy; Sofo, Adriano; Khan, Nafees A; Misra, Amarendra Narayan; Lukatkin, Alexander S; Singh, Harminder Pal; Pereira, Eduarda; Tuteja, Narendra

    2016-10-01

    Plants have to counteract unavoidable stress-caused anomalies such as oxidative stress to sustain their lives and serve heterotrophic organisms including humans. Among major enzymatic antioxidants, catalase (CAT; EC 1.11.1.6) and ascorbate peroxidase (APX; EC 1.11.1.11) are representative heme enzymes meant for metabolizing stress-provoked reactive oxygen species (ROS; such as H2O2) and controlling their potential impacts on cellular metabolism and functions. CAT mainly occurs in peroxisomes and catalyzes the dismutation reaction without requiring any reductant; whereas, APX has a higher affinity for H2O2 and utilizes ascorbate (AsA) as specific electron donor for the reduction of H2O2 into H2O in organelles including chloroplasts, cytosol, mitochondria, and peroxisomes. Literature is extensive on the glutathione-associated H2O2-metabolizing systems in plants. However, discussion is meager or scattered in the literature available on the biochemical and genomic characterization as well as techniques for the assays of CAT and APX and their modulation in plants under abiotic stresses. This paper aims (a) to introduce oxidative stress-causative factors and highlights their relationship with abiotic stresses in plants; (b) to overview structure, occurrence, and significance of CAT and APX in plants;

  2. A Single Nucleotide Polymorphism in Catalase Is Strongly Associated with Ovarian Cancer Survival.

    Science.gov (United States)

    Belotte, Jimmy; Fletcher, Nicole M; Saed, Mohammed G; Abusamaan, Mohammed S; Dyson, Gregory; Diamond, Michael P; Saed, Ghassan M

    2015-01-01

    Ovarian cancer is the deadliest of all gynecologic cancers. Recent evidence demonstrates an association between enzymatic activity altering single nucleotide polymorphisms (SNP) with human cancer susceptibility. We sought to evaluate the association of SNPs in key oxidant and antioxidant enzymes with increased risk and survival in epithelial ovarian cancer. Individuals (n = 143) recruited were divided into controls, (n = 94): healthy volunteers, (n = 18), high-risk BRCA1/2 negative (n = 53), high-risk BRCA1/2 positive (n = 23) and ovarian cancer cases (n = 49). DNA was subjected to TaqMan SNP genotype analysis for selected oxidant and antioxidant enzymes. Of the seven selected SNP studied, no association with ovarian cancer risk (Pearson Chi-square) was found. However, a catalase SNP was identified as a predictor of ovarian cancer survival by the Cox regression model. The presence of this SNP was associated with a higher likelihood of death (hazard ratio (HR) of 3.68 (95% confidence interval (CI): 1.149-11.836)) for ovarian cancer patients. Kaplan-Meier survival analysis demonstrated a significant median overall survival difference (108 versus 60 months, pcancer patients, and thus may serve as a prognosticator.

  3. A superoxide dismutase/catalase mimetic nanomedicine for targeted therapy of inflammatory bowel disease.

    Science.gov (United States)

    Zhang, Qixiong; Tao, Hui; Lin, Yongyao; Hu, Ying; An, Huijie; Zhang, Dinglin; Feng, Shibin; Hu, Houyuan; Wang, Ruibing; Li, Xiaohui; Zhang, Jianxiang

    2016-10-01

    Oxidative stress, resulting from excessive generation of reactive oxygen species (ROS), plays a pivotal role in the initiation and progression of inflammatory bowel disease (IBD). To develop an efficacious and safe nanotherapy against IBD, we designed and developed a superoxide dismutase/catalase mimetic nanomedicine comprising a hydrogen peroxide-eliminating nanomatrix and a free radical scavenger Tempol (Tpl). To this end, an oxidation-responsive β-cyclodextrin material (OxbCD) was synthesized, and a Tpl-loaded OxbCD nanoparticle (Tpl/OxbCD NP) was produced. Hydrolysis of OxbCD NP could be triggered by hydrogen peroxide, leading to on-demand release of loaded Tpl molecules from Tpl/OxbCD NP. OxbCD NP was able to efficiently accumulate in the inflamed colon in mice, thereby dramatically reducing nonspecific distribution after oral delivery. In three mouse colitis models, oral administration of Tpl/OxbCD NP notably mitigated manifestations relevant to colitis, and significantly suppressed expression of proinflammatory mediators, with the efficacy superior over free Tpl or a control nanomedicine based on poly(lactide-co-glycolide) (PLGA). Accordingly, by scavenging multiple components of ROS, Tpl/OxbCD NP may effectively reduce ulcerative colitis in mice, and it can be intensively developed as a translational nanomedicine for the management of IBD and other inflammatory diseases.

  4. Catalase eliminates reactive oxygen species and influences the intestinal microbiota of shrimp.

    Science.gov (United States)

    Yang, Hui-Ting; Yang, Ming-Chong; Sun, Jie-Jie; Guo, Fang; Lan, Jiang-Feng; Wang, Xian-Wei; Zhao, Xiao-Fan; Wang, Jin-Xing

    2015-11-01

    Intestinal innate immune response is an important defense mechanism of animals and humans against external pathogens. The mechanism of microbiota homeostasis in host intestines has been well studied in mammals and Drosophila. The reactive oxygen species (ROS) and antimicrobial peptides have been reported to play important roles in homeostasis. However, how to maintain the microbiota homeostasis in crustacean intestine needs to be elucidated. In this study, we identified a novel catalase (MjCAT) involved in ROS elimination in kuruma shrimp, Marsupenaeus japonicus. MjCAT mRNA was widely distributed in hemocytes, heart, hepatopancreas, gills, stomach, and intestine. After the shrimp were challenged with pathogenic bacteria via oral infection, the expression level of MjCAT was upregulated, and the enzyme activity was increased in the intestine. ROS level was also increased in the intestine at early time after oral infection and recovered rapidly. When MjCAT was knocked down by RNA interference (RNAi), high ROS level maintained longer time, and the number of bacteria number was declined in the shrimp intestinal lumen than those in the control group, but the survival rate of the MjCAT-RNAi shrimp was declined. Further study demonstrated that the intestinal villi protruded from epithelial lining of the intestinal wall were damaged by the high ROS level in MjCAT-knockdown shrimp. These results suggested that MjCAT participated in the intestinal host-microbe homeostasis by regulating ROS level.

  5. Combined effects of water temperature and copper ion concentration on catalase activity in Crassostrea ariakensis

    Science.gov (United States)

    Wang, Hui; Yang, Hongshuai; Liu, Jiahui; Li, Yanhong; Liu, Zhigang

    2015-07-01

    A central composite experimental design and response surface method were used to investigate the combined effects of water temperature (18-34°C) and copper ion concentration (0.1-1.5 mg/L) on the catalase (CAT) activity in the digestive gland of Crassostrea ariakensis. The results showed that the linear effects of temperature were significant ( P0.05), and the quadratic effects of copper ion concentration were significant ( P0.05), and the effect of temperature was greater than that of copper ion concentration. A model equation of CAT enzyme activity in the digestive gland of C. ariakensis toward the two factors of interest was established, with R 2, Adj. R 2 and Pred. R 2 values as high as 0.943 7, 0.887 3 and 0.838 5, respectively. These findings suggested that the goodness of fit to experimental data and predictive capability of the model were satisfactory, and could be practically applied for prediction under the conditions of the study. Overall, the results suggest that the simultaneous variation of temperature and copper ion concentration alters the activity of the antioxidant enzyme CAT by modulating active oxygen species metabolism, which may be utilized as a biomarker to detect the effects of copper pollution.

  6. Bisphenol S Interacts with Catalase and Induces Oxidative Stress in Mouse Liver and Renal Cells.

    Science.gov (United States)

    Zhang, Rui; Liu, Rutao; Zong, Wansong

    2016-08-31

    Bisphenol S (BPS) is present in multitudinous consumer products and detected in both food and water. It also has been a main substitute for bisphenol A (BPA) in the food-packaging industry. Yet, the toxicity of BPS is not fully understood. The present study of the toxicity of BPS was divided into two parts. First, oxidative stress, cell viability, apoptosis level, and catalase (CAT) activity in mouse hepatocytes and renal cells were investigated after BPS exposure. After 12 h of incubation with BPS, all of these parameters of hepatocytes and renal cells changed by >15% as the concentration of BPS ranged from 0.1 to 1 mM. Second, the direct interaction between BPS and CAT on the molecule level was investigated by multiple spectral methods and molecular docking investigations. BPS changed the structure and the activity of CAT through binding to the Gly 117 residue on the substrate channel of the enzyme. The main binding forces were hydrogen bond and hydrophobic force.

  7. Association of Catalase and Glutathione Peroxidase 1 Polymorphisms with Chronic Hepatitis C Outcome.

    Science.gov (United States)

    Sousa, Vanessa C S D; Carmo, Rodrigo F; Vasconcelos, Luydson R S; Aroucha, Dayse C B L; Pereira, Leila M M B; Moura, Patrícia; Cavalcanti, Maria S M

    2016-05-01

    The hepatic damage caused by hepatitis C virus (HCV) infection is associated with the host immune response and viral regulatory factors. Catalase (CAT) and glutathione peroxidase 1 (GPX1) are antioxidant enzymes located in the peroxisomes and mitochondria, respectively, and are responsible for the control of intracellular hydrogen peroxide levels. Polymorphisms in CAT (C-262T) and GPX1 (Pro198Leu) are correlated with serum levels and enzyme activity. This study aimed to investigate the association of genetic polymorphisms of CAT C-262T (rs1001179) and GPX1 Pro198Leu (rs1050450) with different stages of liver fibrosis and development of hepatocellular carcinoma (HCC). This study included 445 patients with chronic hepatitis C, of whom 139 patients had mild fibrosis (F0-F1), 200 had moderate/severe fibrosis (F2-F4), and 106 had HCC. Genotyping of SNPs was performed by real-time PCR using TaqMan probes. The Pro/Pro genotype of GPX1 was significantly associated with fibrosis severity, HCC, Child Pugh score, and BCLC staging. Additionally, patients carrying both CT+TT genotypes in the CAT gene and the Pro/Pro genotype in the GPX1 gene had higher risk for developing moderate/severe fibrosis or HCC (p = 0.009, OR 2.40 and p = 0.002, OR 3.56, respectively). CAT and GPX1 polymorphisms may be implicated in the severity of liver fibrosis and HCC caused by HCV.

  8. A rapid and sensitive alcohol oxidase/catalase conductometric biosensor for alcohol determination.

    Science.gov (United States)

    Hnaien, M; Lagarde, F; Jaffrezic-Renault, N

    2010-04-15

    A new conductometric biosensor has been developed for the determination of short chain primary aliphatic alcohols. The biosensor assembly was prepared through immobilization of alcohol oxidase from Hansenula sp. and bovine liver catalase in a photoreticulated poly(vinyl alcohol) membrane at the surface of interdigitated microelectrodes. The local conductivity increased rapidly after alcohol addition, reaching steady-state within 10 min. The sensitivity was maximal for methanol (0.394+/-0.004 microS microM(-1), n=5) and decreased by increasing the alcohol chain length. The response was linear up to 75 microM for methanol, 70 microM for ethanol and 65 microM for 1-propanol and limits of detection were 0.5 microM, 1 microM and 3 microM, respectively (S/N=3). No significant loss of the enzyme activities was observed after 3 months of storage at 4 degrees C in a 20mM phosphate buffer solution pH 7.2 (two or three measurements per week). After 4 months, 95% of the initial signal still remained. The biosensor response to ethanol was not significantly affected by acetic, lactic, ascorbic, malic, oxalic, citric, tartaric acids or glucose. The bi-enzymatic sensor was successfully applied to the determination of ethanol in different alcoholic beverages.

  9. Catalase overexpression in aortic smooth muscle prevents pathological mechanical changes underlying abdominal aortic aneurysm formation.

    Science.gov (United States)

    Maiellaro-Rafferty, Kathryn; Weiss, Daiana; Joseph, Giji; Wan, William; Gleason, Rudolph L; Taylor, W Robert

    2011-08-01

    The causality of the associations between cellular and mechanical mechanisms of abdominal aortic aneurysm (AAA) formation has not been completely defined. Because reactive oxygen species are established mediators of AAA growth and remodeling, our objective was to investigate oxidative stress-induced alterations in aortic biomechanics and microstructure during subclinical AAA development. We investigated the mechanisms of AAA in an angiotensin II (ANG II) infusion model of AAA in apolipoprotein E-deficient (apoE(-/-)) mice that overexpress catalase in vascular smooth muscle cells (apoE(-/-)xTg(SMC-Cat)). At baseline, aortas from apoE(-/-)xTg(SMC-Cat) exhibited increased stiffness and the microstructure was characterized by 50% more collagen content and less elastin fragmentation. ANG II treatment for 7 days in apoE(-/-) mice altered the transmural distribution of suprarenal aortic circumferential strain (quantified by opening angle, which increased from 130 ± 1° at baseline to 198 ± 8° after 7 days of ANG II treatment) without obvious changes in the aortic microstructure. No differences in aortic mechanical behavior or suprarenal opening angle were observed in apoE(-/-)xTg(SMC-Cat) after 7 days of ANG II treatment. These data suggest that at the earliest stages of AAA development H(2)O(2) is functionally important and is involved in the control of local variations in remodeling across the vessel wall. They further suggest that reduced elastin integrity at baseline may predispose the abdominal aorta to aneurysmal mechanical remodeling.

  10. Catalase prevents maternal diabetes-induced perinatal programming via the Nrf2-HO-1 defense system.

    Science.gov (United States)

    Chang, Shiao-Ying; Chen, Yun-Wen; Zhao, Xin-Ping; Chenier, Isabelle; Tran, Stella; Sauvé, Alexandre; Ingelfinger, Julie R; Zhang, Shao-Ling

    2012-10-01

    We investigated whether overexpression of catalase (CAT) in renal proximal tubular cells (RPTCs) could prevent the programming of hypertension and kidney disease in the offspring of dams with maternal diabetes. Male offspring of nondiabetic and diabetic dams from two transgenic (Tg) lines (Hoxb7-green fluorescent protein [GFP]-Tg [controls] and Hoxb7/CAT-GFP-Tg, which overexpress CAT in RPTCs) were studied from the prenatal period into adulthood. Nephrogenesis, systolic blood pressure, renal hyperfiltration, kidney injury, and reactive oxygen species (ROS) generation were assessed. Gene expression of transforming growth factor-β1 (TGF-β1), nuclear factor erythroid 2p45-related factor-2 (Nrf2), and heme oxygenase-1 (HO-1) was tested in both in vitro and in vivo studies. Renal dysmorphogenesis was observed in offspring of Hoxb7-GFP-Tg dams with severe maternal diabetes; the affected male offspring displayed higher renal ROS generation and developed hypertension and renal hyperfiltration as well as renal injury with heightened TGF-β1 expression in adulthood. These changes were ameliorated in male offspring of diabetic Hoxb7/CAT-GFP-Tg dams via the Nrf2-HO-1 defense system. CAT promoted Nrf2 nuclear translocation and HO-1 gene expression, seen in both in vitro and in vivo studies. In conclusion, CAT overexpression in the RPTCs ameliorated maternal diabetes-induced perinatal programming, mediated, at least in part, by triggering the Nrf2-HO-1 defense system.

  11. Self-cloning significantly enhances the production of catalase in Bacillus subtilis WSHDZ-01.

    Science.gov (United States)

    Xu, Sha; Guo, Yaqiong; Du, Guocheng; Zhou, Jingwen; Chen, Jian

    2014-08-01

    The katA gene that encodes catalase (CAT) in Bacillus subtilis WSHDZ-01 was overexpressed in B. subtilis WB600 and B. subtilis WSHDZ-01. The CAT yield in both transformed strains was significantly improved compared to that in the wild-type WSHDZ-01 in shake flask culture. When cultured in a 3-L stirred tank reactor (STR), the recombinant CAT activity in B. subtilis WSHDZ-01 could be improved by 419 %, reaching up to 39,117 U/mL and was 8,149.4 U/mg dry cell weight, which is the highest activity reported in Bacillus sp. However, the recombinant CAT in B. subtilis WB600 cultured in a 3-L STR was not significantly improved by any of the common means for process optimization, and the highest CAT activity was 3,673.5 U/mg dry cell weight. The results suggest that self-cloning of the complete expression cassette in the original strain is a reasonable strategy to improve the yield of wild-type enzymes.

  12. Improving of catalase stability properties by encapsulation in alginate/Fe3O4 magnetic composite beads for enzymatic removal of H2O2.

    Science.gov (United States)

    Doğaç, Yasemin Ispirli; Çinar, Mürvet; Teke, Mustafa

    2015-01-01

    The aim of this study was enhancing of stability properties of catalase enzyme by encapsulation in alginate/nanomagnetic beads. Amounts of carrier (10-100 mg) and enzyme concentrations (0.25-1.5 mg/mL) were analyzed to optimize immobilization conditions. Also, the optimum temperature (25-50°C), optimum pH (3.0-8.0), kinetic parameters, thermal stability (20-70°C), pH stability (4.0-9.0) operational stability (0-390 min), and reusability were investigated for characterization of the immobilized catalase system. The optimum pH levels of both free and immobilized catalase were 7.0. At the thermal stability studies, the magnetic catalase beads protected 90% activity, while free catalase maintained only 10% activity at 70°C. The thermal profile of magnetic catalase beads was spread over a large area. Similarly, this system indicated the improving of the pH stability. The reusability, which is especially important for industrial applications, was also determined. Thus, the activity analysis was done 50 times in succession. Catalase encapsulated magnetic alginate beads protected 83% activity after 50 cycles.

  13. Characterizing dose-responses of catalase to nitrofurazone exposure in model ciliated protozoan Euplotes vannus for ecotoxicity assessment: enzyme activity and mRNA expression.

    Science.gov (United States)

    Li, Jiqiu; Zhou, Liang; Lin, Xiaofeng; Yi, Zhenzhen; Al-Rasheid, Khaled A S

    2014-02-01

    In environmental studies, some biological responses, known as biomarkers, have been used as a powerful bioassay tool for more than four decades. Disparity between enzyme activity and mRNA abundance leads to correlation equivocality, which makes the application of biomarkers for environmental risk assessment more complicated. This study investigates this disparity in the case of catalase when used as a biomarker for detecting ecotoxicity induced by antibiotics in aquatic ecosystems. In particular, dose-responses for catalase activity and mRNA expression abundance were investigated in Euplotes vannus which were exposed to graded doses of nitrofurazone for several discrete durations, and dose-response models were developed to characterize the dose-response dynamics. Significant differences were found in both catalase activity and mRNA expression abundance among the E. vannus treated with nitrofurazone. Catalase activity showed a hormetic-like effect in terms of dose-response, characterized by a biphasic relationship which was more clearly evident after a longer exposure period, while mRNA expression abundance increased linearly with the exposure duration. Additionally, the correlation between catalase activity and mRNA expression abundance reversed along with the duration of exposure to nitrofurazone. Taken together, our results demonstrate that catalase mRNA expression offers a more straightforward dose-response model than enzyme activity. Our findings suggest that both catalase enzyme activity and mRNA expression abundance can be used jointly as bioassay tools for detecting ecotoxicity induced by nitrofurazone in aquatic ecosystems.

  14. Immunodetection of a brown planthopper (Nilaparvata lugens Stål) salivary catalase-like protein into tissues of rice, Oryza sativa.

    Science.gov (United States)

    Petrova, A; Smith, C M

    2014-02-01

    Saliva plays an important role in host plant-phloem-feeding insect molecular interactions. To better elucidate the role of insect saliva, a series of experiments were conducted to establish if catalase from the salivary glands of the brown planthopper (BPH; Nilaparvata lugens Stål) was secreted into rice host plant tissue during feeding. Catalase is the main enzyme that decomposes hydrogen peroxide (H2O2) at high concentrations. H2O2 is a part of the free radicals system that mediates important physiological roles including signalling and defence. Previous studies have suggested that H2O2 is involved in the rice endogenous response to BPH feeding. If, the BPH secretes catalase into host plant tissue this will counter the effects of H2O2, from detoxification to interfering with plant signalling and defence mechanisms. When BPHs were fed on a hopper-resistant rice variety for 24 h, catalase activity in the salivary glands increased 3.5-fold compared with hoppers fed on a susceptible rice variety. Further supporting evidence of the effects of BPH catalase was demonstrated by immunodetection analyses where results from two independent sources: BPH-infested rice tissue and BPH-probed artificial diets, suggest that the BPH secretes catalase-like protein during feeding. The possible physiological roles of BPH-secreted catalase are discussed.

  15. Dynamics of erythrocyte count, hemoglobin, and catalase activity in rat blood in hypokinesia, muscular activity and restoration

    Science.gov (United States)

    Taneyeva, G. V.; Potapovich, G. M.; Voloshko, N. A.; Uteshev, A. B.

    1980-01-01

    Tests were conducted to prove that muscular exertion (in this instance swimming) of different duration and intensity, as well as hypodynamia, result in an increase of hemoglobin and number of red blood cells in peripheral blood rats. Catalase activity increased with an increase in the duration of swimming, but only up to 6 hr; with 7-9 hr of swimming as well as in hypodynamia, catalase activity decreased. It was also observed that under hypodynamia as well as in 3, 5 and 6 hr exertion (swimming) the color index of blood decreased. Pressure chamber treatment (for 8 min each day for one week), alternating a 2 min negative pressure up to 35 mm Hg with 1 min positive pressure, increased the erythrocyte count and hemoglobin content.

  16. The use of glucose oxidase and catalase for the enzymatic reduction of the potential ethanol content in wine.

    Science.gov (United States)

    Röcker, Jessica; Schmitt, Matthias; Pasch, Ludwig; Ebert, Kristin; Grossmann, Manfred

    2016-11-01

    Due to the increase of sugar levels in wine grapes as one of the impacts of climate change, alcohol reduction in wines becomes a major focus of interest. This study combines the use of glucose oxidase and catalase activities with the aim of rapid conversion of glucose into non-fermentable gluconic acid. The H2O2 hydrolysing activity of purified catalase is necessary in order to stabilize glucose oxidase activity. After establishing the adequate enzyme ratio, the procedure was applied in large-scale trials (16L- and 220L-scale) of which one was conducted in a winery under industrial wine making conditions. Both enzyme activity and wine flavour were clearly influenced by the obligatory aeration in the different trials. With the enzyme treatment an alcohol reduction of 2%vol. was achieved after 30h of aeration. However the enzyme treated wines were significantly more acidic and less typical.

  17. Catalase expression in MCF-7 breast cancer cells is mainly controlled by PI3K/Akt/mTor signaling pathway.

    Science.gov (United States)

    Glorieux, Christophe; Auquier, Julien; Dejeans, Nicolas; Sid, Brice; Demoulin, Jean-Baptiste; Bertrand, Luc; Verrax, Julien; Calderon, Pedro Buc

    2014-05-15

    Catalase is an antioxidant enzyme that catalyzes mainly the transformation of hydrogen peroxide into water and oxygen. Although catalase is frequently down-regulated in tumors the underlying mechanism remains unclear. Few transcription factors have been reported to directly bind the human catalase promoter. Among them FoxO3a has been proposed as a positive regulator of catalase expression. Therefore, we decided to study the role of the transcription factor FoxO3a and the phosphatidylinositol-3 kinase (PI3K) signaling pathway, which regulates FoxO3a, in the expression of catalase. To this end, we developed an experimental model of mammary breast MCF-7 cancer cells that acquire resistance to oxidative stress, the so-called Resox cells, in which catalase is overexpressed as compared with MCF-7 parental cell line. In Resox cells, Akt expression is decreased but its phosphorylation is enhanced when compared with MCF-7 cells. A similar profile is observed for FoxO3a, with less total protein but more phosphorylated FoxO3a in Resox cells, correlating with its higher Akt activity. The modulation of FoxO3a expression by knockdown and overexpression strategies did not affect catalase expression, neither in MCF-7 nor in Resox cells. Inhibition of PI3K and mTOR by LY295002 and rapamycin, respectively, decreases the phosphorylation of downstream targets (i.e. GSK3β and p70S6K) and leads to an increase of catalase expression only in MCF-7 but not in Resox cells. In conclusion, FoxO3a does not appear to play a critical role in the regulation of catalase expression in both cancer cells. Only MCF-7 cells are sensitive and dependent on PI3K/Akt/mTOR signaling.

  18. Salicylic acid is a modulator of catalase isozymes in chickpea plants infected with Fusarium oxysporum f. sp. ciceri.

    Science.gov (United States)

    Gayatridevi, S; Jayalakshmi, S K; Sreeramulu, K

    2012-03-01

    The relationship between salicylic acid level catalases isoforms chickpea cv. ICCV-10 infected with Fusarium oxysporum f. sp. ciceri was investigated. Pathogen-treated chickpea plants showed high levels of SA compared with the control. Two isoforms of catalases in shoot extract (CAT-IS and CAT-IIS) and single isoform in root extract (CAT-R) were detected in chickpea. CAT-IS and CAT-R activities were inhibited in respective extracts treated with pathogen whereas, CAT-IIS activity was not inhibited. These isoforms were purified and their kinetic properties studied in the presence or absence of SA. The molecular mass determined by SDS-PAGE of CAT-IS, CAT-IIS and CAT-R was found to be 97, 40 and 66 kDa respectively. Kinetic studies indicated that Km and V(max) of CAT-IS were 0.2 mM and 300 U/mg, 0.53 mM and 180 U/mg for CAT-IIS and 0.25 mM and 280 U/mg for CAT-R, respectively. CAT-IS and CAT-R were found to be more sensitive to SA and 50% of their activities were inhibited at 6 and 4 μM respectively, whereas CAT-IIS was insensitive to SA up to 100 μM. Quenching of the intrinsic tryptophan fluorescence of purified catalases were used to quantitate SA binding; the estimated K(d) value for CAT-IS, CAT-IIS and CAT-R found to be 2.3 μM, 3.1 mM and 2.8 μM respectively. SA is a modulator of catalase isozymes activity, supports its role in establishment of SAR in chickpea plants infected with the pathogen.

  19. Effects of Soy-Germ Protein on Catalase Activity of Plasma and Erythocyte of Metabolic Syndrome Women

    Directory of Open Access Journals (Sweden)

    Hery Winarsi

    2015-01-01

    Full Text Available Oxidative stress always accompany patients with metabolic syndrome (MS. Several researchers reported that soy-protein is able to decrease oxidative stress level. However, there is no report so far about soy-germ protein in relation to its potential to the decrease oxidative stress level of MS patients. The aim of this study was to explore the potential of soy-germ protein on activity of catalase enzyme in blood’s plasma as well as erythrocytes of MS patients. Double-blind randomized clinical trial was used as an experimental study. Thirty respondents were included in this study with MS, normal level blood sugar, low-HDL cholesterol but high in triglyceride, 40-65 years old, Body Mass Index > 25 kg/m2, live in Purwokerto and agreed to sign the informed consent. They were randomly grouped into 3 different groups, 10 each: Group I, was given special milk that contains soy-germ protein and Zn; Group II, soy-germ protein, while Group III was placebo; for two consecutive months. Data were taken from blood samples in 3 different periods i.e. 0, 1, and 2 months after treatment. Two months after treatment, there was an increase from 5.36 to 20.17 IU/mg (P = 0.028 in activity of catalase enzyme in blood’s plasma respondents who consumed milk containing soy-germ protein with or without Zn. A similar trend of catalase activity, but at a lower level, was also noticed in erythrocyte; which increased from 88.31 to 201.11 IU/mg (P = 0.013. The increase in activity of catalase enzyme in blood’s plasma was 2.2 times higher than that in erythrocytes.

  20. PRODUCING OF ENZYME PREPARATION AND ANALYSIS OF ENZYME PREPARATION OF PEROXIDASE AND CATALASE OF SOME SPECIES OF BASIDIOMYCETES

    OpenAIRE

    Fedotov O. V.; Voloshko T.E.

    2013-01-01

    A method for obtaining of enzyme preparations of enzyme preparations (EP) of peroxidases and catalases fungal extracellular and inracellular origin from cultures of Basidiomycetes was developed. The strains Flammulina velutipes F-vv, Agrocybe cylindracea167; Fistulina hepatica Fh-08 and Pleurotus ostreatus P-208 and P-01 were used as producers of oxidoreductases. Strains were grown on modified glucose-peptone media. Fractionation was carried out by salting out the enzymes with ammonium sulfat...

  1. Two-dimensional HYSCORE spectroscopy of superoxidized manganese catalase: a model for the oxygen-evolving complex of photosystem II.

    Science.gov (United States)

    Coates, Christopher S; Milikisiyants, Sergey; Chatterjee, Ruchira; Whittaker, Mei M; Whittaker, James W; Lakshmi, K V

    2015-04-16

    The solar water-splitting protein complex, photosystem II (PSII), catalyzes one of the most energetically demanding reactions in Nature by using light energy to drive a catalyst capable of oxidizing water. The water oxidation reaction takes place at the tetra-nuclear manganese calcium-oxo (Mn4Ca-oxo) cluster at the heart of the oxygen-evolving complex (OEC) of PSII. Previous studies have determined the magnetic interactions between the paramagnetic Mn4Ca-oxo cluster and its environment in the S2 state of the OEC. The assignments for the electron-nuclear magnetic interactions that were observed in these studies were facilitated by the use of synthetic dimanganese di-μ-oxo complexes. However, there is an immense need to understand the effects of the protein environment on the coordination geometry of the Mn4Ca-oxo cluster in the OEC of PSII. In the present study, we use a proteinaceous model system to examine the protein ligands that are coordinated to the dimanganese catalytic center of manganese catalase from Lactobacillus plantarum. We utilize two-dimensional hyperfine sublevel correlation (2D HYSCORE) spectroscopy to detect the weak magnetic interactions of the paramagnetic dinuclear manganese catalytic center of superoxidized manganese catalase with the nitrogen and proton atoms of the surrounding protein environment. We obtain a complete set of hyperfine interaction parameters for the protons of a water molecule that is directly coordinated to the dinuclear manganese center. We also obtain a complete set of hyperfine and quadrupolar interaction parameters for two histidine ligands as well as a coordinated azide ligand, in azide-treated superoxidized manganese catalase. On the basis of the values of the hyperfine interaction parameters of the dimanganese model, manganese catalase, and those of the S2 state of the OEC of PSII, for the first time, we discuss the impact of a proteinaceous environment on the coordination geometry of multinuclear manganese clusters.

  2. Catalase Expression Is Modulated by Vancomycin and Ciprofloxacin and Influences the Formation of Free Radicals in Staphylococcus aureus Cultures.

    Science.gov (United States)

    Wang, Ying; Hougaard, Anni B; Paulander, Wilhelm; Skibsted, Leif H; Ingmer, Hanne; Andersen, Mogens L

    2015-09-01

    Detection of free radicals in biological systems is challenging due to their short half-lives. We have applied electron spin resonance (ESR) spectroscopy combined with spin traps using the probes PBN (N-tert-butyl-α-phenylnitrone) and DMPO (5,5-dimethyl-1-pyrroline N-oxide) to assess free radical formation in the human pathogen Staphylococcus aureus treated with a bactericidal antibiotic, vancomycin or ciprofloxacin. While we were unable to detect ESR signals in bacterial cells, hydroxyl radicals were observed in the supernatant of bacterial cell cultures. Surprisingly, the strongest signal was detected in broth medium without bacterial cells present and it was mitigated by iron chelation or by addition of catalase, which catalyzes the decomposition of hydrogen peroxide to water and oxygen. This suggests that the signal originates from hydroxyl radicals formed by the Fenton reaction, in which iron is oxidized by hydrogen peroxide. Previously, hydroxyl radicals have been proposed to be generated within bacterial cells in response to bactericidal antibiotics. We found that when S. aureus was exposed to vancomycin or ciprofloxacin, hydroxyl radical formation in the broth was indeed increased compared to the level seen with untreated bacterial cells. However, S. aureus cells express catalase, and the antibiotic-mediated increase in hydroxyl radical formation was correlated with reduced katA expression and catalase activity in the presence of either antibiotic. Therefore, our results show that in S. aureus, bactericidal antibiotics modulate catalase expression, which in turn influences the formation of free radicals in the surrounding broth medium. If similar regulation is found in other bacterial species, it might explain why bactericidal antibiotics are perceived as inducing formation of free radicals.

  3. Imparting functionality to biocatalysts via embedding enzymes into nanoporous materials by a de novo approach: size-selective sheltering of catalase in metal-organic framework microcrystals.

    Science.gov (United States)

    Shieh, Fa-Kuen; Wang, Shao-Chun; Yen, Chia-I; Wu, Chang-Cheng; Dutta, Saikat; Chou, Lien-Yang; Morabito, Joseph V; Hu, Pan; Hsu, Ming-Hua; Wu, Kevin C-W; Tsung, Chia-Kuang

    2015-04-01

    We develop a new concept to impart new functions to biocatalysts by combining enzymes and metal-organic frameworks (MOFs). The proof-of-concept design is demonstrated by embedding catalase molecules into uniformly sized ZIF-90 crystals via a de novo approach. We have carried out electron microscopy, X-ray diffraction, nitrogen sorption, electrophoresis, thermogravimetric analysis, and confocal microscopy to confirm that the ~10 nm catalase molecules are embedded in 2 μm single-crystalline ZIF-90 crystals with ~5 wt % loading. Because catalase is immobilized and sheltered by the ZIF-90 crystals, the composites show activity in hydrogen peroxide degradation even in the presence of protease proteinase K.

  4. A novel pyrogallol red-based assay to assess catalase activity: Optimization by response surface methodology.

    Science.gov (United States)

    Abderrahim, Mohamed; Arribas, Silvia M; Condezo-Hoyos, Luis

    2017-05-01

    Pyrogallol red (PGR) was identified as a novel optical probe for the detection of hydrogen peroxide (H2O2) based on horseradish peroxidase (HRP)-catalyzed oxidation. Response surface methodology (RSM) was applied as a tool to optimize the concentrations of PGR (100µmolL(-1)), HRP (1UmL(-1)) and H2O2 (250µmolL(-1)) and used to develop a sensitive PGR-based catalase (CAT) activity assay (PGR-CAT assay). N-ethylmaleimide -NEM- (102mmolL(-1)) was used to avoid interference produced by thiol groups while protecting CAT activity. Incubation time (30min) for samples or CAT used as standard and H2O2 as well as signal stability (stable between 5 and 60min) were also evaluated. PGR-CAT assay was linear within the range of 0-4UmL(-1) (R(2)=0.993) and very sensitive with limits of detection (LOD) of 0.005UmL(-1) and quantitation (LOQ) of 0.01UmL(-1). PGR-CAT assay showed an adequate intra-day RSD=0.6-9.5% and inter-day RSD=2.4-8.9%. Bland-Altman analysis and Passing-Bablok and Pearson correlation analysis showed good agreement between CAT activity as measured by the PRG-CAT assay and the Amplex Red assay. The PGR-CAT assay is more sensitive than all the other colorimetric assays reported, particularly the Amplex Red assay, and the cost of PGR is a small fraction (about 1/1000) of that of an Amplex Red probe, so it can be expected to find wide use among scientists studying CAT activity in biological samples.

  5. Response of Superoxide Dismutase, Catalase, and ATPase Activity in Bacteria Exposed to Acetamiprid

    Institute of Scientific and Technical Information of China (English)

    XIAO-HUA YAO; HANG MIN; ZHEN-MEI LV

    2006-01-01

    To investigate how acetamiprid, a new insecticide, affects the activity of superoxide dismutase (SOD),catalase (CAT), and ATPase and the SOD isozyme patterns in two G- bacteria, E. coli K12 and Pse. FH2, and one G+ bacterum,B. subtilis. Methods The SOD, CAT, and ATPase specific activities of cell lysates were determined spectrophotometrically at 550 nm, 240 nm, and 660 nm, respectively, with kits A001, A016, and A007. SOD isozyme patterns were detected by native PAGE analysis. Results SOD and CAT activities in the tested bacteria increased significantly in a concentration-dependent manner after different concentrations of acetamiprid were applied. The activity of SOD in B. subtilis and Pse. FH2 was stimulated and reached the highest level after treatment with 100 mg/L acetamiprid for 0.5 h. For Pse. FH2, there was another stimulation of SOD activity after acetamiprid application for about 8.0 h and the second stimulation was stronger than the first.The stimulation by acetamiprid showed a relative lag for E. coli K12. Acetamiprid seemed to exhibit a similar effect on CAT activity of the two G- bacteria and had an evident influence on ATPase activity in the three bacteria within a relatively short period. Only one SOD isozyme was detectable in Pse. FH2 and B. subtilis, while different isozyme compositions in E. coli could be detected by native PAGE analysis. Conclusion Acetamiprid causes a certain oxidative stress on the three bacteria which may not only elevate SOD and CAT activities but also generate new SOD isozymes to antagonize oxidative stress.However, this oxidative stress lasts for a relatively short time and does not cause a long-term damage.

  6. Pregnancy rates in cattle with cryopreserved sexed spermatozoa: effects of laser intensity, staining conditions and catalase.

    Science.gov (United States)

    Schenk, J L; Seidel, G E

    2007-01-01

    The overall aim of this research was to improve fertility of cattle inseminated with sexed spermatozoa by improving sperm sorting procedures. Six field trials were conducted in which 4,264 heifers were inseminated into the uterine body with cryopreserved sexed or unsexed control spermatozoa. Pregnancy or calving rates with doses of 2 x 10(6) sexed spermatozoa ranged from 32 to 51%; these averaged 69% of the pregnancy rates with 20 x 10(6) unsexed, control spermatozoa (range 53 to 79% of controls). Fertility of sexed spermatozoa was especially low on farms where control fertility was low. Accuracy of sexing ranged from 86 to 91%. Laser power of 150 mW for interrogating spermatozoa did not result in lower pregnancy rates (43%) than when power was decreased as much as possible for a particular sorting batch (50 to 130 mW) to still achieve sexing accuracy (38% pregnant). Addition of catalase to fluids containing spermatozoa was beneficial when thawed spermatozoa were incubated in vitro for 2 h but had no effect on pregnancy rates. There also was no effect on pregnancy rates between two concentrations of Hoechst 33342 for staining spermatozoa. Freezing 2 x 10(6) sexed spermatozoa at 20 x 10(6)/ml resulted in a slightly higher rate of pregnancy (P < 0.05) than at 10 x 10(6)/ml. The information obtained in these trials, along with other improvements, notably lowering pressure in the sorting system from 50 to 40 psi, has been used to improve procedures for sexing spermatozoa commercially.

  7. Effect of soil contamination with azadirachtin on dehydrogenase and catalase activity of soil

    Directory of Open Access Journals (Sweden)

    Rıdvan Kızılkaya

    2012-07-01

    Full Text Available nsecticides are used in modern agriculture in large quantities to control pests and increase crop yield. Their use, however, has resulted in the disruption of ecosystems because of the effects on non-target soil microorganisms, some environmental problems, and decreasing soil fertility. These negative effects of synthetic pesticides on the environment have led to the search for alternative means of pest control. One such alternative is use of natural plant products such as azadirachtin that have pesticidal activity. The aim of this experiment was to study the effect of soil contamination by azadirachtin (C35H44O16 on dehydrogenase (DHA and catalase activity (CA of soil under field conditions in Perm, Russia. The tests were conducted on loamy soil (pHH2O 6.7, ECH2O 0.213 dSm-1, organic carbon 0.99%, to which the following quantities of azadirachtin were added: 0, 15, 30 and 60 mL da-1 of soil. Experimental design was randomized plot design with three replications. The DHA and CA analyses were performed 7, 14 and 21 days after the field experiment was established. The results of field experiment showed that azadirachtin had a positive influence on the DHA and CA at different soil sampling times. The increased doses of azadirachtin applied resulted in the higher level of DHA and CA in soil. The soil DHA and CA showed the highest activity on the 21th day after 60 mL azadirachtin da-1 application doses.

  8. Mutation of Arabidopsis CATALASE2 results in hyponastic leaves by changes of auxin levels.

    Science.gov (United States)

    Gao, Xiang; Yuan, Hong-Mei; Hu, Ye-Qin; Li, Jing; Lu, Ying-Tang

    2014-01-01

    Auxin and H2 O2 play vital roles in plant development and environmental responses; however, it is unclear whether and how H2 O2 modulates auxin levels. Here, we investigate this question using cat2-1 mutant, which exhibits reduced catalase activity and accumulates high levels of H2 O2 under photorespiratory conditions. At a light intensity of 150 μmol m(-2) s(-1) , the mutant exhibited up-curled leaves that have increased H2 O2 contents and decreased auxin levels. At low light intensities (30 μmol m(-2) s(-1)), the leaves of the mutant were normal, but exhibited reduced H2 O2 contents and elevated auxin levels. These findings suggest that H2 O2 modulates auxin levels. When auxin was directly applied to cat2-1 leaves, the up-curled leaves curled downwards. In addition, transformation of cat2-1 plants with pCAT2:iaaM, which increases auxin levels, rescued the hyponastic leaf phenotype. Using qRT-PCR, we demonstrated that the transcription of auxin synthesis-related genes and of genes that regulate leaf curvature is suppressed in cat2-1. Furthermore, application of glutathione rescued the up-curled leaves of cat2-1 and increased auxin levels, but did not change H2 O2 levels. Thus, the hyponastic leaves of cat2-1 reveal crosstalk between H2 O2 and auxin signalling that is mediated by changes in glutathione redox status.

  9. Transcriptional inhibition of the Catalase gene in phosphine-induced oxidative stress in Drosophila melanogaster.

    Science.gov (United States)

    Liu, Tao; Li, Li; Zhang, Fanhua; Wang, Yuejin

    2015-10-01

    Phosphine (PH3) is a toxic substance to pest insects and is therefore commonly used in pest control. The oxidative damage induced by PH3 is considered to be one of the primary mechanisms of its toxicity in pest insects; however, the precise mode of PH3 action in this process is still unclear. In this study, we evaluated the responses of several oxidative biomarkers and two of the main antioxidant enzymes, catalase (CAT) and superoxide dismutase (SOD), after fumigation treatment with PH3 in Drosophila melanogaster as a model system. The results showed that larvae exposed to sub-lethal levels of PH3 (0.028 mg/L) exhibited lower aerobic respiration rates and higher levels of hydrogen peroxide (H2O2) and lipid peroxidation (LPO). Furthermore, unlike SOD, the activity and expression of CAT and its encoding gene were downregulated by PH3 in a time- and dose-dependent manner. Finally, the responses of six potential transcription factors of PH3 were determined by real-time polymerase chain reaction to explore the regulation mechanism of DmCAT by PH3. There were no significant effects of PH3 on three nuclear factor-kappa B homologs (DORSAL, DIF, and RELISH) or two activator protein-1 genes (JUN and FOS), while dramatic inhibition of DNA replication-related element factor (DREF) expression was observed after fumigation with PH3, suggesting that PH3 could inhibit the expression of DmCAT via the DRE/DREF system. These results confirmed that PH3 induces oxidative stress and targets CAT by downregulating its encoding gene in Drosophila. Our results provide new insight into the signal transduction mechanism between PH3 and its target genes.

  10. Hydrogen peroxide-induced astaxanthin biosynthesis and catalase activity in Xanthophyllomyces dendrorhous.

    Science.gov (United States)

    Liu, Yuan Shuai; Wu, Jian Yong

    2006-12-01

    Xanthophyllomyces dendrorhous (formerly Phaffia rhodozyma) in shake-flask cultures was exposed to 10-20 mmol/L H(2)O(2) at various culture stages, and the astaxanthin production was significantly increased by H(2)O(2) fed at 0 or 24 h (exponential phase), but only slightly at 48 h (near stationary phase). The astaxanthin production was enhanced most significantly with double feeding of 10 mmol/L H(2)O(2) at 0 and 24 h, reaching a cellular content of 1.30 mg/g cell and a volumetric yield of 10.4 mg/L, which were 83 and 65% higher, respectively, than those of the control (0.71 mg/g cell and 6.3 mg/L). The intracellular catalase (CAT) activity was also increased after H(2)O(2) treatment. The increases in CAT and astaxanthin of cells could be detected within 4 h of H(2)O(2) treatment. The increase in the astaxanthin content of cells was concomitant with a notable decrease in the beta-carotene content. The older yeast cells at late culture stage (120 h), due perhaps in part to their higher astaxanthin contents, were more tolerant to H(2)O(2) toxicity than the younger cells (24 h). No enhancement of the astaxanthin biosynthesis was attained when H(2)O(2) was added to the yeast culture together with a sufficient amount of exogenous CAT. The results suggest that astaxanthin biosynthesis in X. dendrorhous can be stimulated by H(2)O(2) as an antioxidative response.

  11. The Catalase –262C/T Promoter Polymorphism and Diabetic Complications in Caucasians with Type 2 Diabetes

    Directory of Open Access Journals (Sweden)

    Kátia Gonçalves dos Santos

    2006-01-01

    Full Text Available Catalase is a central antioxidant enzyme constituting the primary defense against oxidative stress. In this study, we investigated whether the functional –262C/T polymorphism in the promoter of catalase gene is associated with the presence of diabetic retinopathy (DR, diabetic nephropathy (DN and ischemic heart disease (IHD in 520 Caucasian-Brazilians with type 2 diabetes. The –262C/T polymorphism was also examined in 100 Caucasian blood donors. Patients underwent a clinical and laboratory evaluation consisting of a questionnaire, physical examination, assessment of diabetic complications and laboratory tests. Genotype analysis was performed using the polymerase chain reaction followed by digestion with restriction enzyme. The genotype and allele frequencies of the –262C/T polymorphism in patients with type 2 diabetes were very similar to those of blood donors (T allele frequency = 0.20 and 0.18, respectively. Likewise, there were no differences in either genotype or allele frequencies between type 2 diabetic patients with or without DR, DN or IHD. Thus, our results do not support the hypothesis that the –262C/T polymorphism is related to the development of DR, DN or IHD in patients with type 2 diabetes. Further studies are necessary to elucidate the role of catalase gene polymorphisms in the pathogenesis of diabetic complications.

  12. The effect of catalase on recovery of heat-injured DNA-repair mutants of Escherichia coli.

    Science.gov (United States)

    Mackey, B M; Seymour, D A

    1987-06-01

    The apparent sensitivity of Escherichia coli K12 to mild heat was increased by recA (def), recB and polA, but not by uvrA, uvrB or recF mutations. However, addition of catalase to the rich plating medium used to assess viability restored counts of heat-injured recA, recB and polA strains to wild-type levels. E. coli p3478 polA was sensitized by heat to a concentration of hydrogen peroxide similar to that measured in autoclaved recovery medium. The apparent heat sensitivity of DNA-repair mutants is thus due to heat-induced sensitivity to the low levels of peroxide present in rich recovery media. It is proposed that DNA damage in heated cells could occur indirectly by an oxidative mechanism. The increased peroxide sensitivity of heat-injured cells was not due to a decrease in total catalase activity but may be related specifically to inactivation of the inducible catalase/peroxidase (HPI).

  13. [Heterologous extracellular expression and initial characterization of the peroxisomal catalase from the methylotrophic yeast Hansenula polymorpha in Pichia pastoris].

    Science.gov (United States)

    Tian, Y -S; Xu, H; Peng, R -H; Yao, Q -H

    2013-01-01

    Catalase is well known to eliminate H2O2 in cells and reduces the toxicity of peroxide compounds. A catalase gene HpCat1 of methylotrophic yeast Hansenula polymorpha without the part coding the native signal peptide was cloned into expression vector pYM3165 and then integrated into genome of Pichia pastoris GS115 by electroporation. The result of the enzyme activity assay and SDS-PAGE demonstrated that the recombinant protein (HpCAT1) of H. polymorpha was extracellularly expressed in P. pastoris. The expressed catalase was recovered from the culture supernatant of P. pastoris GS 115 and purified by (NH4) 2SO4 fractionation and Ni-NTA affinity chromatography. The main biochemical properties of the recombinant protein HpCAT1, such as thermodependence and thermostability, pH optimum and pH stability, as well as the effect of metal ions and chemicals, were characterized. With H2O2 as the substrate, HpCAT1 displayed pH and tem- perature optima of approximately 2.6 and 45°C,respectively. The recombinant HpCAT1 activity was inhibited by 1 mM Hg2+ and Cu2+, but was highly enhanced by 1.0 mM Fe2+.

  14. Impact of Microscale and Pilot-Scale Freeze-Drying on Protein Secondary Structures: Sucrose Formulations of Lysozyme and Catalase.

    Science.gov (United States)

    Peters, Björn-Hendrik; Leskinen, Jari T T; Molnár, Ferdinand; Ketolainen, Jarkko

    2015-11-01

    Microscale (MS) freeze-drying offers rapid process cycles for early-stage formulation development. The effects of the MS approach on the secondary structures of two model proteins, lysozyme and catalase, were compared with pilot-scale (PS) vial freeze-drying. The secondary structures were assessed by attenuated total reflection Fourier transformed infrared spectroscopy. Formulations were made with increasing sucrose-protein ratios. Freeze-drying protocols involved regular cooling without thermal treatment and annealing with MS and PS equipment, and cooling rate variations with the MS. Principal component analysis of smoothed second-derivative amide I spectra revealed sucrose-protein ratio-dependent shifts toward α-helical structures. Transferability of sucrose-protein formulations from MS to PS vial freeze-drying was evidenced at regular cooling rates. Local differences in protein secondary structures between the bottom and top of sucrose-catalase samples could be detected at the sucrose-catalase ratios of 1 and 2, this being related to the initial filling height and ice crystal morphology. Annealing revealed temperature, protein, formulation, and sample location-dependent effects influencing surface morphology at the top, or causing protein secondary structure perturbation at the bottom. With the MS approach, protein secondary structure differences at different cooling rates could be detected for sucrose-lysozyme samples at the sucrose-lysozyme ratio of 1.

  15. A volumetric meter chip for point-of-care quantitative detection of bovine catalase for food safety control.

    Science.gov (United States)

    Cui, Xingye; Hu, Jie; Choi, Jane Ru; Huang, Yalin; Wang, Xuemin; Lu, Tian Jian; Xu, Feng

    2016-09-07

    A volumetric meter chip was developed for quantitative point-of-care (POC) analysis of bovine catalase, a bioindicator of bovine mastitis, in milk samples. The meter chip displays multiplexed quantitative results by presenting the distance of ink bar advancement that is detectable by the naked eye. The meter chip comprises a poly(methyl methacrylate) (PMMA) layer, a double-sided adhesive (DSA) layer and a glass slide layer fabricated by the laser-etching method, which is typically simple, rapid (∼3 min per chip), and cost effective (∼$0.2 per chip). Specially designed "U shape" reaction cells are covered by an adhesive tape that serves as an on-off switch, enabling the simple operation of the assay. As a proof of concept, we employed the developed meter chip for the quantification of bovine catalase in raw milk samples to detect catalase concentrations as low as 20 μg/mL. The meter chip has great potential to detect various target analytes for a wide range of POC applications.

  16. Identification of Festuca arundinacea Schreb Cat1 Catalase Gene and Analysis of its Expression Under Abiotic Stresses

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Abiotic stresses, such as drought, high salinity, and cold/freezing, lead plants to produce excess reactive oxygen species. Catalase, a unique hydrogen peroxide-scavenging enzyme, plays a very important role in plants. To characterize the catalase involved in plant response to abiotic stresses, we constructed a cDNA library from 4 ℃-treated Festuca arundinacea Schreb seedlings and isolated a catalase gene from this library.The cDNA (FaCat1, 1 735 bp) contained an open reading frame of 1 479 bp. BLAST analysis indicated that the deduced amino acid sequence showed 96% identity with that from wheat TaCat1 and 87% identity with that from maize ZmCat2. Northern blotting analysis showed an obvious increase of FaCat1 transcripts in leaves in contrast with roots. Time-course analysis of the expression of FaCat1 in F. arundinacea leaves showed that FaCat1 expression was upregulated in cold- and salt-stressed leaves, with the FaCat1 transcripts accumulating mostly at 4 or 2 h after cold or salt stress, respectively. No significant changes in FaCat1 transcription were observed in dried leaves and inhibition of FaCat1 transcription was found in abscisic acid (ABA)-treated leaves,indicating that the FaCat1 gene is differentially expressed during cold, high salt, drought, and ABA treatment in F. arundinacea leaves.

  17. Manganese L-edge X-ray absorption spectroscopy of manganese catalase from Lactobacillus plantarum and mixed valence manganese complexes

    Energy Technology Data Exchange (ETDEWEB)

    Grush, M.M.; Chen, J.; George, S.J. [Univ. of California, Davis, CA (United States)] [and others

    1996-01-10

    The first Mn L-edge absorption spectra of a Mn metalloprotein are presented in this paper. Both reduced and superoxidized Mn catalase have been examined by fluorescence-detected soft X-ray absorption spectroscopy, and their Mn L-edge spectra are dramatically different. The spectrum of reduced Mn(II)Mn(II) catalase has been interpreted by ligand field atomic multiplet calculations and by comparison to model compound spectra. The analysis finds a 10 Dq value of nearly 1.1 eV, consistent with coordination by predominately nitrogen and oxygen donor ligands. For interpretation of mixed valence Mn spectra, an empirical simulation procedure based on the addition of homovalent model compound spectra has been developed and was tested on a variety of Mn complexes and superoxidized Mn catalase. This routine was also used to determine the oxidation state composition of the Mn in [Ba{sub 8}Na{sub 2}ClMn{sub 16}(OH){sub 8}(CO{sub 3}){sub 4}L{sub 8}] .53 H{sub 2}O (L=1,3-diamino-2-hydroxypropane-N,N,N`N`-tetraacetic acid). 27 refs., 6 figs.

  18. Catalase (KatA plays a role in protection against anaerobic nitric oxide in Pseudomonas aeruginosa.

    Directory of Open Access Journals (Sweden)

    Shengchang Su

    Full Text Available Pseudomonas aeruginosa (PA is a common bacterial pathogen, responsible for a high incidence of nosocomial and respiratory infections. KatA is the major catalase of PA that detoxifies hydrogen peroxide (H2O2, a reactive oxygen intermediate generated during aerobic respiration. Paradoxically, PA displays elevated KatA activity under anaerobic growth conditions where the substrate of KatA, H2O2, is not produced. The aim of the present study is to elucidate the mechanism underlying this phenomenon and define the role of KatA in PA during anaerobiosis using genetic, biochemical and biophysical approaches. We demonstrated that anaerobic wild-type PAO1 cells yielded higher levels of katA transcription and expression than aerobic cells, whereas a nitrite reductase mutant ΔnirS produced ∼50% the KatA activity of PAO1, suggesting that a basal NO level was required for the increased KatA activity. We also found that transcription of the katA gene was controlled, in part, by the master anaerobic regulator, ANR. A ΔkatA mutant and a mucoid mucA22 ΔkatA bacteria demonstrated increased sensitivity to acidified nitrite (an NO generator in anaerobic planktonic and biofilm cultures. EPR spectra of anaerobic bacteria showed that levels of dinitrosyl iron complexes (DNIC, indicators of NO stress, were increased significantly in the ΔkatA mutant, and dramatically in a ΔnorCB mutant compared to basal levels of DNIC in PAO1 and ΔnirS mutant. Expression of KatA dramatically reduced the DNIC levels in ΔnorCB mutant. We further revealed direct NO-KatA interactions in vitro using EPR, optical spectroscopy and X-ray crystallography. KatA has a 5-coordinate high spin ferric heme that binds NO without prior reduction of the heme iron (Kd ∼6 μM. Collectively, we conclude that KatA is expressed to protect PA against NO generated during anaerobic respiration. We proposed that such protective effects of KatA may involve buffering of free NO when potentially toxic

  19. Modification of catalase and MAPK in Vicia faba cultivated in soil with high natural radioactivity and treated with a static magnetic field.

    Science.gov (United States)

    Haghighat, Nazanin; Abdolmaleki, Parviz; Ghanati, Faezeh; Behmanesh, Mehrdad; Payez, Atefeh

    2014-03-01

    The effects of a static magnetic field (SMF) and high natural radioactivity (HR) on catalase and MAPK genes in Vicia faba were investigated. Soil samples with high natural radioactivity were collected from Ramsar in north Iran where the annual radiation absorbed dose from background radiation is higher than 20mSv/year. The specific activity of the radionuclides of (232)Th, (236)Ra, and (40)K was measured using gamma spectrometry. The seeds were planted either in the soil with high natural radioactivity or in the control soils and were then exposed to a SMF of 30mT for 8 days; 8h/day. Levels of expression of catalase and MAPK genes, catalase activity and H2O2 content were evaluated. The results demonstrated significant differences in the expression of catalase and MAPK genes in SMF- and HR-treated plants compared to the controls. An increase in catalase activity was accompanied by increased expression of its gene and accumulation of H2O2. Relative expression of the MAPK gene in treated plants, however, was lower than those of the controls. The results suggest that the response of V. faba plants to SMF and HR may be mediated by modification of catalase and MAPK.

  20. The effect of catalase on migration and invasion of lung cancer cells by regulating the activities of cathepsin S, L, and K.

    Science.gov (United States)

    Tsai, Ju-Ying; Lee, Mon-Juan; Dah-Tsyr Chang, Margaret; Huang, Haimei

    2014-04-15

    Abundant clinical evidences indicate that up-regulation of several cathepsins in many human cancers is correlated with malignant progression and poor patient prognosis. In addition, a decrease in catalase activity or accumulation of hydrogen peroxide correlates with cancer metastasis. Recent studies indicate that cathepsin activation and expression can be modulated via H2O2 treatment. However, the actual relationship between catalase and cathepsins is not yet fully understood. In the present study, we found that catalase expression (or activity) was higher, while intracellular and extracellular Cat S, Cat L, and Cat K activities were lower in the non-invasive CL1-0 cells compared to the highly invasive CL1-5 cells. After CL1-0 cells were transfected with catalase-shRNA, the corresponding ROS (H2O2) level and Cat S, Cat L, or Cat K expression (or activity) was up-regulated, accompanied by an increase in cell migration and invasion. On the other hand, ROS (H2O2) level, cathepsin S, L, and K activities, cell migration and invasion were decreased in catalase-overexpressed CL1-5 cells. It is suggested that catalase may regulate cathepsin activity by controlling the production of ROS (H2O2), leading to variation in migration and invasion ability of lung cancer cells.

  1. Prognostic significance of catalase expression and its regulatory effects on hepatitis B virus X protein (HBx) in HBV-related advanced hepatocellular carcinomas.

    Science.gov (United States)

    Cho, Mi-Young; Cheong, Jae Youn; Lim, Wonchung; Jo, Sujin; Lee, Youngsoo; Wang, Hee-Jung; Han, Kyou-Hoon; Cho, Hyeseong

    2014-12-15

    Hepatitis B virus X protein (HBx) plays a role in liver cancer development. We previously showed that ROS increased HBx levels and here, we investigated the role of antioxidants in the regulation of HBx expression and their clinical relevance. We found that overexpression of catalase induced a significant loss in HBx levels. The cysteine null mutant of HBx (Cys-) showed a dramatic reduction in its protein stability. In clonogenic proliferation assays, Huh7-X cells produced a significant number of colonies whereas Huh7-Cys- cells failed to generate them. The Cys at position 69 of HBx was crucial to maintain its protein stability and transactivation function in response to ROS. Among 50 HBV-related hepatocellular carcinoma (HCC) specimens, 72% of HCCs showed lower catalase levels than those of surrounding non-tumor tissues. In advanced stage IV, catalase levels in non-tumor tissues were increased whereas those in tumors were further reduced. Accordingly, patients with a high T/N ratio for catalase showed significantly longer survival than those with a low T/N ratio. Together, catalase expression in HCC patients can be clinically useful for prediction of patient survival, and restoration of catalase expression in HCCs could be an important strategy for intervention in HBV-induced liver diseases.

  2. Cardiac-specific overexpression of catalase prevents diabetes-induced pathological changes by inhibiting NF-κB signaling activation in the heart.

    Science.gov (United States)

    Cong, Weitao; Ruan, Dandan; Xuan, Yuanhu; Niu, Chao; Tao, Youli; Wang, Yang; Zhan, Kungao; Cai, Lu; Jin, Litai; Tan, Yi

    2015-12-01

    Catalase is an antioxidant enzyme that specifically catabolizes hydrogen peroxide (H2O2). Overexpression of catalase via a heart-specific promoter (CAT-TG) was reported to reduce diabetes-induced accumulation of reactive oxygen species (ROS) and further prevent diabetes-induced pathological abnormalities, including cardiac structural derangement and left ventricular abnormity in mice. However, the mechanism by which catalase overexpression protects heart function remains unclear. This study found that activation of a ROS-dependent NF-κB signaling pathway was downregulated in hearts of diabetic mice overexpressing catalase. In addition, catalase overexpression inhibited the significant increase in nitration levels of key enzymes involved in energy metabolism, including α-oxoglutarate dehydrogenase E1 component (α-KGD) and ATP synthase α and β subunits (ATP-α and ATP-β). To assess the effects of the NF-κB pathway activation on heart function, Bay11-7082, an inhibitor of the NF-κB signaling pathway, was injected into diabetic mice, protecting mice against the development of cardiac damage and increased nitrative modifications of key enzymes involved in energy metabolism. In conclusion, these findings demonstrated that catalase protects mouse hearts against diabetic cardiomyopathy, partially by suppressing NF-κB-dependent inflammatory responses and associated protein nitration.

  3. The effect of alcohol and hydrogen peroxide on liver hepcidin gene expression in mice lacking antioxidant enzymes, glutathione peroxidase-1 or catalase.

    Science.gov (United States)

    Harrison-Findik, Duygu Dee; Lu, Sizhao

    2015-05-06

    This study investigates the regulation of hepcidin, the key iron-regulatory molecule, by alcohol and hydrogen peroxide (H2O2) in glutathione peroxidase-1 (gpx-1(-/-)) and catalase (catalase(-/-)) knockout mice. For alcohol studies, 10% ethanol was administered in the drinking water for 7 days. Gpx-1(-/-) displayed significantly higher hepatic H2O2 levels than catalase(-/-) compared to wild-type mice, as measured by 2'-7'-dichlorodihydrofluorescein diacetate (DCFH-DA). The basal level of liver hepcidin expression was attenuated in gpx-1(-/-) mice. Alcohol increased H2O2 production in catalase(-/-) and wild-type, but not gpx-1(-/-), mice. Hepcidin expression was inhibited in alcohol-fed catalase(-/-) and wild-type mice. In contrast, alcohol elevated hepcidin expression in gpx-1(-/-) mice. Gpx-1(-/-) mice also displayed higher level of basal liver CHOP protein expression than catalase(-/-) mice. Alcohol induced CHOP and to a lesser extent GRP78/BiP expression, but not XBP1 splicing or binding of CREBH to hepcidin gene promoter, in gpx-1(-/-) mice. The up-regulation of hepatic ATF4 mRNA levels, which was observed in gpx-1(-/-) mice, was attenuated by alcohol. In conclusion, our findings strongly suggest that H2O2 inhibits hepcidin expression in vivo. Synergistic induction of CHOP by alcohol and H2O2, in the absence of gpx-1, stimulates liver hepcidin gene expression by ER stress independent of CREBH.

  4. Efeito tóxico dos praguicidas maneb e paraquat sobre a atividade da enzima antioxidante catalase em ratos

    Directory of Open Access Journals (Sweden)

    M. D. Arbo

    2009-01-01

    Full Text Available

    Os radicais livres estão envolvidos em um grande número de enfermidades do ser humano. O cérebro tem níveis baixos de enzimas antioxidantes e um conteúdo lípidico elevado, tornando-se muito susceptível ao ataque de espécies reativas de oxigênio. Neste trabalho avaliou- se a lipoperoxidação em hipocampo e a atividade da enzima catalase em estriado e hipocampo de ratos tratados com o fungicida maneb (30 mg/kg e o herbicida paraquat (10 mg/kg. Não houve alteração na lipoperoxidação nem na atividade enzimática no hipocampo dos animais tratados com ambos os praguicidas, porém foi observada uma inibição da catalase no estriado dos ratos tratados com maneb e com paraquat. Com estes resultados pode-se sugerir, de forma preliminar, uma ação tóxica maior sobre centros dopaminérgicos. Estudos sobre a toxicidade destes compostos são essenciais na compreensão do papel destes praguicidas e dos radicais livres na etiologia das doenças. Palavras-chave: catalase; paraquat; maneb; estriado; hipocampo; radicais livres.

  5. Characterization of OxyR as a negative transcriptional regulator that represses catalase production in Corynebacterium diphtheriae.

    Directory of Open Access Journals (Sweden)

    Ju-Sim Kim

    Full Text Available Corynebacterium diphtheriae and Corynebacterium glutamicum each have one gene (cat encoding catalase. In-frame Δcat mutants of C. diphtheriae and C. glutamicum were hyper-sensitive to growth inhibition and killing by H(2O(2. In C. diphtheriae C7(β, both catalase activity and cat transcription decreased ~2-fold during transition from exponential growth to early stationary phase. Prototypic OxyR in Escherichia coli senses oxidative stress and it activates katG transcription and catalase production in response to H(2O(2. In contrast, exposure of C. diphtheriae C7(β to H(2O(2 did not stimulate transcription of cat. OxyR from C. diphtheriae and C. glutamicum have 52% similarity with E. coli OxyR and contain homologs of the two cysteine residues involved in H(2O(2 sensing by E. coli OxyR. In-frame ΔoxyR deletion mutants of C. diphtheriae C7(β, C. diphtheriae NCTC13129, and C. glutamicum were much more resistant than their parental wild type strains to growth inhibition by H(2O(2. In the C. diphtheriae C7(β ΔoxyR mutant, cat transcripts were about 8-fold more abundant and catalase activity was about 20-fold greater than in the C7(β wild type strain. The oxyR gene from C. diphtheriae or C. glutamicum, but not from E. coli, complemented the defect in ΔoxyR mutants of C. diphtheriae and C. glutamicum and decreased their H(2O(2 resistance to the level of their parental strains. Gel-mobility shift, DNaseI footprint, and primer extension assays showed that purified OxyR from C. diphtheriae C7(β bound, in the presence or absence of DTT, to a sequence in the cat promoter region that extends from nucleotide position -55 to -10 with respect to the +1 nucleotide in the cat ORF. These results demonstrate that OxyR from C. diphtheriae or C. glutamicum functions as a transcriptional repressor of the cat gene by a mechanism that is independent of oxidative stress induced by H(2O(2.

  6. Characterization of OxyR as a negative transcriptional regulator that represses catalase production in Corynebacterium diphtheriae.

    Science.gov (United States)

    Kim, Ju-Sim; Holmes, Randall K

    2012-01-01

    Corynebacterium diphtheriae and Corynebacterium glutamicum each have one gene (cat) encoding catalase. In-frame Δcat mutants of C. diphtheriae and C. glutamicum were hyper-sensitive to growth inhibition and killing by H(2)O(2). In C. diphtheriae C7(β), both catalase activity and cat transcription decreased ~2-fold during transition from exponential growth to early stationary phase. Prototypic OxyR in Escherichia coli senses oxidative stress and it activates katG transcription and catalase production in response to H(2)O(2). In contrast, exposure of C. diphtheriae C7(β) to H(2)O(2) did not stimulate transcription of cat. OxyR from C. diphtheriae and C. glutamicum have 52% similarity with E. coli OxyR and contain homologs of the two cysteine residues involved in H(2)O(2) sensing by E. coli OxyR. In-frame ΔoxyR deletion mutants of C. diphtheriae C7(β), C. diphtheriae NCTC13129, and C. glutamicum were much more resistant than their parental wild type strains to growth inhibition by H(2)O(2). In the C. diphtheriae C7(β) ΔoxyR mutant, cat transcripts were about 8-fold more abundant and catalase activity was about 20-fold greater than in the C7(β) wild type strain. The oxyR gene from C. diphtheriae or C. glutamicum, but not from E. coli, complemented the defect in ΔoxyR mutants of C. diphtheriae and C. glutamicum and decreased their H(2)O(2) resistance to the level of their parental strains. Gel-mobility shift, DNaseI footprint, and primer extension assays showed that purified OxyR from C. diphtheriae C7(β) bound, in the presence or absence of DTT, to a sequence in the cat promoter region that extends from nucleotide position -55 to -10 with respect to the +1 nucleotide in the cat ORF. These results demonstrate that OxyR from C. diphtheriae or C. glutamicum functions as a transcriptional repressor of the cat gene by a mechanism that is independent of oxidative stress induced by H(2)O(2).

  7. Oxidative stress, hemoglobin content, superoxide dismutase and catalase activity influenced by sulphur baths and mud packs in patients with osteoarthritis

    Directory of Open Access Journals (Sweden)

    Jokić Aleksandar

    2010-01-01

    Full Text Available Background/Aim. It is weel-known that sulphur baths and mud paks demonstrate beneficial effects on patients suffering from degenerative knee and hip osteoarthritis (OA through the increased activity of protective antioxidant enzymes. The aim of this study was to assess lipid peroxidation level, i.e. malondialdehyde concetration, in individuals with knee and/or hip osteoarthritis (OA, as well as to determine the influence of sulphur baths and mud packs application on the activity of superoxide dismutase (SOD and catalase (CAT in order to minimize or eliminate excessive free radical species production (oxidative stress. Methods. Thirty one patiens with knee and/or hip OA of both sexes were included in the study. All OA patients received mud pack and sulphur bath for 20 minutes a day, for 6 consecutive days a week, over 3 weeks. Blood lipid peroxidation, ie malondialdehyde concentration, superoxide dismutase and catalase activity were measured spectrophotometrically, before, on day 5 during the treatment and at the end of spa cure. Healthy volunteers (n = 31 were the controls. Results. The sulphur baths and mud packs treatment of OA patients caused a significant decrease in plasma malondialdehyde concentration compared to the controls ( p < 0.001. The mean SOD activity before the terapy was 1 836.24 U/gHb, on day 5 it rose to 1 942.15 U/gHb and after the spa cure dropped to 1 745.98 U/gHb. Catalase activity before the therapy was 20.56 kU/gHb and at the end of the terapy decreased to 16.16 kU/gHb. The difference in catalase activity before and after the therapy was significant (p < 0.001, and also significant as compared to control (p < 0.001. At the end of the treatment significant increase of hemoglobin level and significant decrease of pain intensity were noticed. Conclusion. A combined 3-week treatment by sulphur bath and mud packs led to a significant decrease of lipid peroxidation in plasma, as well as pain intensity in the patients with OA

  8. Changes in catalase activity in leaves of woody and bushy plants in the conditions of air pollution by compounds of fluorine, sulfur and nitrogen

    Directory of Open Access Journals (Sweden)

    Y. Prysedskyj

    2016-08-01

    Full Text Available Recently environmental pollution by industrial waste products has become a significant environmental factor that essentially limits the vital functions of plants and reduces their species diversity. The antioxidant system is of special importance for tolerance reactions of plants to stressful environmental conditions, in particular, contamination by industrial pollutants. One of the constituents of this system is oxidoreductase, including catalase. Consequently, we have conducted experiments to determine how the nature of the complex compounds of fluorine, nitrogen and sulfur influences catalase activity in leaves of selected species of trees and shrubs. The investigation was made according to the complete factorial experiment that allowed us to study the effect of these pollutants both individually and in combination. We used the iodometric method to determine the level of catalase activity. Statistical analysis of the obtained results was performed by means of dispersion analysis with the comparison according to the Duncan method. The results of the research showed the possible impact of pollutants on the activity of catalase, which depends on the resilience of the plants, structure and duration of potency of the pollutants. With less resilient plant species (Sorbus aucuparia L., Fraxinus lanceolata Borkh. air pollution with a combination of fluorine, sulfur and nitrogen in most cases caused a reduction of catalase activity. Thus, in S. aucuparia a 5-hour exposure to low concentrations of pollutants (HF – 0.2 ml/m3, NH3 – 1.2 ml/m3, SO2 and H2SO4 – 0.9–1.0 ml/m3 caused an inhibition of catalase activity by 40.5%, and a ten-hour exposure caused a 61.4% inhibition compared with the control plants. With increased concentrations of pollutants  catalase function was inhibited by 35.8–73.6%, depending on the duration of their fumigation. For F. lanceolata, the pollutants’ effect on catalase activity caused a decrease in function of this

  9. Catalase activity is stimulated by H2O2 in rich culture medium and is required for H2O2 resistance and adaptation in yeast

    Directory of Open Access Journals (Sweden)

    Dorival Martins

    2014-01-01

    Full Text Available Catalases are efficient scavengers of H2O2 and protect cells against H2O2 stress. Examination of the H2O2 stimulon in Saccharomyces cerevisiae revealed that the cytosolic catalase T (Ctt1 protein level increases 15-fold on H2O2 challenge in synthetic complete media although previous work revealed that deletion of the CCT1 or CTA1 genes (encoding peroxisomal/mitochondrial catalase A does not increase the H2O2 sensitivity of yeast challenged in phosphate buffer (pH 7.4. This we attributed to our observation that catalase activity is depressed when yeast are challenged with H2O2 in nutrient-poor media. Hence, we performed a systematic comparison of catalase activity and cell viability of wild-type yeast and of the single catalase knockouts, ctt1∆ and cta1∆, following H2O2 challenge in nutrient-rich medium (YPD and in phosphate buffer (pH 7.4. Ctt1 but not Cta1 activity is strongly induced by H2O2 when cells are challenged in YPD but suppressed when cells are challenged in buffer. Consistent with the activity results, exponentially growing ctt1∆ cells in YPD are more sensitive to H2O2 than wild-type or cta1∆ cells, whereas in buffer all three strains exhibit comparable H2O2 hypersensitivity. Furthermore, catalase activity is increased during adaptation to sublethal H2O2 concentrations in YPD but not in buffer. We conclude that induction of cytosolic Ctt1 activity is vital in protecting yeast against exogenous H2O2 but this activity is inhibited by H2O2 when cells are challenged in nutrient-free media.

  10. Catalase activity is stimulated by H(2)O(2) in rich culture medium and is required for H(2)O(2) resistance and adaptation in yeast.

    Science.gov (United States)

    Martins, Dorival; English, Ann M

    2014-01-01

    Catalases are efficient scavengers of H2O2 and protect cells against H2O2 stress. Examination of the H2O2 stimulon in Saccharomyces cerevisiae revealed that the cytosolic catalase T (Ctt1) protein level increases 15-fold on H2O2 challenge in synthetic complete media although previous work revealed that deletion of the CCT1 or CTA1 genes (encoding peroxisomal/mitochondrial catalase A) does not increase the H2O2 sensitivity of yeast challenged in phosphate buffer (pH 7.4). This we attributed to our observation that catalase activity is depressed when yeast are challenged with H2O2 in nutrient-poor media. Hence, we performed a systematic comparison of catalase activity and cell viability of wild-type yeast and of the single catalase knockouts, ctt1∆ and cta1∆, following H2O2 challenge in nutrient-rich medium (YPD) and in phosphate buffer (pH 7.4). Ctt1 but not Cta1 activity is strongly induced by H2O2 when cells are challenged in YPD but suppressed when cells are challenged in buffer. Consistent with the activity results, exponentially growing ctt1∆ cells in YPD are more sensitive to H2O2 than wild-type or cta1∆ cells, whereas in buffer all three strains exhibit comparable H2O2 hypersensitivity. Furthermore, catalase activity is increased during adaptation to sublethal H2O2 concentrations in YPD but not in buffer. We conclude that induction of cytosolic Ctt1 activity is vital in protecting yeast against exogenous H2O2 but this activity is inhibited by H2O2 when cells are challenged in nutrient-free media.

  11. Layer by layer assembly of catalase and amine-terminated ionic liquid onto titanium nitride nanoparticles modified glassy carbon electrode: Study of direct voltammetry and bioelectrocatalytic activity

    Energy Technology Data Exchange (ETDEWEB)

    Saadati, Shagayegh [Department of Chemistry, University of Kurdistan, P.O. Box 416, Sanandaj (Iran, Islamic Republic of); Salimi, Abdollah, E-mail: absalimi@uok.ac.ir [Department of Chemistry, University of Kurdistan, P.O. Box 416, Sanandaj (Iran, Islamic Republic of); Research Center for Nanotechnology, University of Kurdistan, P.O. Box 416, Sanandaj (Iran, Islamic Republic of); Hallaj, Rahman; Rostami, Amin [Department of Chemistry, University of Kurdistan, P.O. Box 416, Sanandaj (Iran, Islamic Republic of)

    2012-11-13

    Highlights: Black-Right-Pointing-Pointer Catalase and amine-terminated ionic liquid were immobilized to GC/TiNnp with LBL assembly method. Black-Right-Pointing-Pointer First a thin layer of NH{sub 2}-IL is covalently attached to GC/TiNnp electrode using electro-oxidation. Black-Right-Pointing-Pointer With alternative assemble of IL and catalase with positive and negative charged, multilayer was formed. Black-Right-Pointing-Pointer Immobilized catalase shows excellent electrocatalytic activity toward H{sub 2}O{sub 2} reduction. Black-Right-Pointing-Pointer Biosensor response is directly correlated to the number of bilayers. - Abstract: A novel, simple and facile layer by layer (LBL) approach is used for modification of glassy carbon (GC) electrode with multilayer of catalase and nanocomposite containing 1-(3-Aminopropyl)-3-methylimidazolium bromide (amine terminated ionic liquid (NH{sub 2}-IL)) and titanium nitride nanoparticles (TiNnp). First a thin layer of NH{sub 2}-IL is covalently attached to GC/TiNnp electrode using electro-oxidation method. Then, with alternative self assemble positively charged NH{sub 2}-IL and negatively charged catalase a sensitive H{sub 2}O{sub 2} biosensor is constructed, whose response is directly correlated to the number of bilayers. The surface coverage of active catalase per bilayer, heterogeneous electron transfer rate constant (k{sub s}) and Michaelis-Menten constant (K{sub M}) of immobilized catalase were 3.32 Multiplication-Sign 10{sup -12} mol cm{sup -2}, 5.28 s{sup -1} and 1.1 mM, respectively. The biosensor shows good stability, high reproducibility, long life-time, and fast amperometric response with the high sensitivity of 380 {mu}A mM{sup -1} cm{sup -2} and low detection limit of 100 nM at concentration range up to 2.1 mM.

  12. Mechanism of pH-switchable peroxidase and catalase-like activities of gold, silver, platinum and palladium.

    Science.gov (United States)

    Li, Junnan; Liu, Wenqi; Wu, Xiaochun; Gao, Xingfa

    2015-04-01

    Despite being increasingly used as artificial enzymes, little has been known for the origin of the pH-switchable peroxidase-like and catalase-like activities of metals. Using calculations and experiments, we report the mechanisms for both activities and their pH-switchability for metals Au, Ag, Pd and Pt. The calculations suggest that both activities are intrinsic properties of metals, regardless of the surfaces and intersections of facets exposed to environments. The pre-adsorbed OH groups on the surfaces, which are only favorably formed in basic conditions, trigger the switch between both activities and render the pH-switchability. The adsorption energies between H2O2 and metals can be used as convenient descriptors to predict the relative enzyme-like activities of the metals with similar surface morphologies. The results agree with the enzyme-mimic activities that have been experimentally reported for Au, Ag, Pt and predict that Pd should have the similar properties. The prediction, as well as the predicted activity order for the four metals, has been verified by the experimental tests. The results thus provide an in-depth insight into the peroxidase-like and catalase-like activities of the metals and will guide the de novo design, synthesis and application of artificial enzymes based on inorganic materials.

  13. Dental Fluorosis and Catalase Immunoreactivity of the Brain Tissues in Rats Exposed to High Fluoride Pre- and Postnatally.

    Science.gov (United States)

    Güner, Şirin; Uyar-Bozkurt, Süheyla; Haznedaroğlu, Eda; Menteş, Ali

    2016-11-01

    This study evaluated dental fluorosis of the incisors and immunoreactivity in the brain tissues of rats given chronic fluoride doses pre- and postnatally. Female rats were given drinking water with 0, 30 or 100 ppm fluoride ad libitum throughout gestation and the nursing period. In addition, 63 male offspring were treated with the same water regimens as the mothers after weaning and were followed for 1, 3 or 5 months. The upper and lower incisors were collected, and all teeth were examined under a stereomicroscope and scored by two blinded examiners using a modified rodent enamel fluorosis index. Cortical, hippocampal and cerebellar brain samples were evaluated morphologically and immunohistochemically. All fluoride-treated pups were born with low body weight (p = 0.001). All animals from the fluoride groups had enamel fluorosis with defects of various degrees. The increase in the dental fluorosis scores in the fluoride treatment groups was significant (p catalase immunoreactivity in the 30- and 100-ppm fluoride groups was significantly higher than that in the controls after 1, 3 and 5 months (p catalase immunoreactivity in the brain tissues, which may reflect the neurobehavioral toxicity of fluoride.

  14. Superoxide dismutase/catalase mimetic EUK-134 prevents diaphragm muscle weakness in monocrotalin-induced pulmonary hypertension

    Science.gov (United States)

    Tatebayashi, Daisuke; Lee, Jaesik; Westerblad, Håkan; Lanner, Johanna T.

    2017-01-01

    Patients with pulmonary hypertension (PH) suffer from inspiratory insufficiency, which has been associated with intrinsic contractile dysfunction in diaphragm muscle. Here, we examined the role of redox stress in PH-induced diaphragm weakness by using the novel antioxidant, EUK-134. Male Wistar rats were randomly divided into control (CNT), CNT + EUK-134 (CNT + EUK), monocrotaline-induced PH (PH), and PH + EUK groups. PH was induced by a single intraperitoneal injection of monocrotaline (60 mg/kg body weight). EUK-134 (3 mg/kg body weight/day), a cell permeable mimetic of superoxide dismutase (SOD) and catalase, was daily intraperitoneally administered starting one day after induction of PH. After four weeks, diaphragm muscles were excised for mechanical and biochemical analyses. There was a decrease in specific tetanic force in diaphragm bundles from the PH group, which was accompanied by increases in: protein expression of NADPH oxidase 2/gp91phox, SOD2, and catalase; 3-nitrotyrosine content and aggregation of actin; glutathione oxidation. Treatment with EUK-134 prevented the force decrease and the actin modifications in PH diaphragm bundles. These data show that redox stress plays a pivotal role in PH-induced diaphragm weakness. Thus, antioxidant treatment can be a promising strategy for PH patients with inspiratory failure. PMID:28152009

  15. [Influence of low dozes of ionizing radiation on accumulation of melanin pigments and catalase and superoxidedismutase activities in Cladosporium cladosporioides].

    Science.gov (United States)

    Tuhaĭ, T I

    2007-01-01

    Influence of low dozes of ionizing radiation on melanin pigments synthesis and activity of antioxidant enzymes catalase and superoxidedismutase of two strains of Cladosporium cladosporioides 4, (isolated from radioactive soil) and 396 (control) were investigated. It was shown, that in C. cladosporioides 4 under the exposure of ionizing radiation an increase of melanin synthesis in a stationary growth phase and increase of superoxidedismutase activity in a logarithmic phase were observed; in the control strain C. cladosporioides 396 activation of melanin synthesis and superoxide dismutase activity in both growth phases was revealed. It was established that in C. cladosporioides 4 the endocellular catalase activity in a logarithmic phase is 3.2 times higher, than in control strain. Under the action of ionizing radiation a 2-fold increase of this enzyme activity unlike the control strain in which the activity inhibition was revealed. The obtained results testify to the complex response of antioxidant systems and melanin to the action of low dozes of radiation which depends on the growth phase and presence of radioadaptation properties in the investigated fungi.

  16. Neonatal Persistent Exposure to 6-Propyl-2-thiouracil, a Thyroid-Disrupting Chemical, Differentially Modulates Expression of Hepatic Catalase and C/EBP-β in Adult Rats.

    Science.gov (United States)

    Bunker, Suresh Kumar; Dandapat, Jagneshwar; Sahoo, Sunil Kumar; Roy, Anita; Chainy, Gagan B N

    2016-02-01

    Persistent exposure of rats to 6-propyl-2-thiouracil (PTU) from birth resulted in decreases in plasma thyroid hormone (TH) levels and hepatic expression of catalase and CCAAT enhancer binding protein β (C/EBP-β). Catalase promoter region (-185 to +52) that contains binding sites for C/EBP-β showed an augmentation in the methylation level along with a change in methylation pattern of CpG islands in response to PTU treatment. PTU withdrawal on 30 days of birth restored TH levels and C/EBP-β to control rats in adulthood. Although catalase expression was restored to some extent in adult rats in response to PTU withdrawal, a permanent change in its promoter CpG methylation pattern was recorded. The results suggest that downregulation of adult hepatic catalase gene in response to persistent neonatal PTU exposure may not solely be attributed to thyroid-disrupting properties of PTU. It is possible that besides thyroid-disrupting behavior, PTU may impair expression of hepatic catalase by altering methylation pattern of its promoter.

  17. Contact-dependent depletion of hydrogen peroxide by catalase is a novel mechanism of myeloid-derived suppressor cell induction operating in human hepatic stellate cells.

    Science.gov (United States)

    Resheq, Yazid J; Li, Ka-Kit; Ward, Stephen T; Wilhelm, Annika; Garg, Abhilok; Curbishley, Stuart M; Blahova, Miroslava; Zimmermann, Henning W; Jitschin, Regina; Mougiakakos, Dimitrios; Mackensen, Andreas; Weston, Chris J; Adams, David H

    2015-03-15

    Myeloid-derived suppressor cells (MDSC) represent a unique cell population with distinct immunosuppressive properties that have been demonstrated to shape the outcome of malignant diseases. Recently, human hepatic stellate cells (HSC) have been reported to induce monocytic-MDSC from mature CD14(+) monocytes in a contact-dependent manner. We now report a novel and unexpected mechanism by which CD14(+)HLADR(low/-) suppressive cells are induced by catalase-mediated depletion of hydrogen peroxide (H2O2). Incubation of CD14(+) monocytes with catalase led to a significant induction of functional MDSC compared with media alone, and H2O2 levels inversely correlated with MDSC frequency (r = -0.6555, p Catalase was detected in primary HSC and a stromal cell line, and addition of the competitive catalase inhibitor hydroxylamine resulted in a dose-dependent impairment of MDSC induction and concomitant increase of H2O2 levels. The NADPH-oxidase subunit gp91 was significantly increased in catalase-induced MDSC as determined by quantitative PCR outlining the importance of oxidative burst for the induction of MDSC. These findings represent a so far unrecognized link between immunosuppression by MDSC and metabolism. Moreover, this mechanism potentially explains how stromal cells can induce a favorable immunological microenvironment in the context of tissue oxidative stress such as occurs during cancer therapy.

  18. Extraction of superoxide dismutase, catalase, and carbonic anhydrase from stroma-free red blood cell hemolysate for the preparation of the nanobiotechnological complex of polyhemoglobin-superoxide dismutase-catalase-carbonic anhydrase.

    Science.gov (United States)

    Guo, C; Gynn, M; Chang, T M S

    2015-06-01

    We report a novel method to simultaneously extract superoxide dismutase (SOD), catalase (CAT), and carbonic anhydrase (CA) from the same sample of red blood cells (RBCs). This avoids the need to use expensive commercial enzymes, thus enabling a cost-effective process for large-scale production of a nanobiotechnological polyHb-SOD-CAT-CA complex, with enhancement of all three red blood cell functions. An optimal concentration of phosphate buffer for ethanol-chloroform treatment results in good recovery of CAT, SOD, and CA after extraction. Different concentrations of the enzymes can be used to enhance the activity of polyHb-SOD-CAT-CA to 2, 4, or 6 times that of RBC.

  19. Three-phase partitioning as a rapid and easy method for the purification and recovery of catalase from sweet potato tubers (Solanum tuberosum).

    Science.gov (United States)

    Duman, Yonca Avcı; Kaya, Erdem

    2013-07-01

    Three-phase partitioning (TPP) was used to purify and recover catalase from potato crude extract. The method consists of ammonium sulfate saturation, t-butanol addition, and adjustment of pH, respectively. The best catalase recovery (262 %) and 14.1-fold purification were seen in the interfacial phase in the presence of 40 % (w/v) ammonium sulfate saturation with 1.0:1.0 crude extract/t-butanol ratio (v/v) at pH 7 in a single step. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the enzyme showed comparatively purification and protein molecular weight was nearly found to be 56 kDa. This study shows that TPP is a simple, economical, and quick method for the recovering of catalase and can be used for the purification process.

  20. [Total Peroxidase and Catalase Activity of Luminous Basidiomycetes Armillaria borealis and Neonothopanus nambi in Comparison with the Level of Light Emission].

    Science.gov (United States)

    Mogil'naya, O A; Ronzhin, N O; Medvedeva, S E; Bondar, V S

    2015-01-01

    The peroxidase and catalase activities in the mycelium of luminous basidiomycetes Armillaria borealis and Neonothopanus nambi in normal conditions and under stress were compared. An increase in the luminescence level was observed under stress, as well as an increase in peroxidase and catalase activities. Moreover, the peroxidase activity in extracts of A. borealis mycelium was found to be almost one and a half orders of magnitude higher, and the catalase activity more than two orders of magnitude higher in comparison with the N. nambi mycelium. It can be suggested that the difference between the brightly luminescent and dimly luminescent mycelium of N. nambi is due to the content of H2O2 or other peroxide compounds.

  1. Piper betle shows antioxidant activities, inhibits MCF-7 cell proliferation and increases activities of catalase and superoxide dismutase

    Directory of Open Access Journals (Sweden)

    Abrahim Noor

    2012-11-01

    Full Text Available Abstract Background Breast cancer is the most common form of cancer and the focus on finding chemotherapeutic agents have recently shifted to natural products. Piper betle is a medicinal plant with various biological activities. However, not much data is available on the anti-cancer effects of P. betle on breast cancer. Due to the current interest in the potential effects of antioxidants from natural products in breast cancer treatment, we investigated the antioxidant activities of the leaves of P. betle and its inhibitory effect on the proliferation of the breast cancer cell line, MCF-7. Methods The leaves of P. betle were extracted with solvents of varying polarities (water, methanol, ethyl acetate and hexane and their phenolic and flavonoid content were determined using colorimetric assays. Phenolic composition was characterized using HPLC. Antioxidant activities were measured using FRAP, DPPH, superoxide anion, nitric oxide and hyroxyl radical scavenging assays. Biological activities of the extracts were analysed using MTT assay and antioxidant enzyme (catalase, superoxide dismutase, glutathione peroxidase assays in MCF-7 cells. Results Overall, the ethyl acetate extract showed the highest ferric reducing activity and radical scavenging activities against DPPH, superoxide anion and nitric oxide radicals. This extract also contained the highest phenolic content implying the potential contribution of phenolics towards the antioxidant activities. HPLC analyses revealed the presence of catechin, morin and quercetin in the leaves. The ethyl acetate extract also showed the highest inhibitory effect against the proliferation of MCF-7 cells (IC50=65 μg/ml. Treatment of MCF-7 cells with the plant extract increased activities of catalase and superoxide dismutase. Conclusions Ethyl acetate is the optimal solvent for the extraction of compounds with antioxidant and anti-proliferative activities. The increased activities of catalase and superoxide

  2. In vitro and in vivo inhibitory effects of some fungicides on catalase produced and purified from white-rot fungus Phanerochaete chrysosporium.

    Science.gov (United States)

    Kavakçıoğlu, Berna; Tarhan, Leman

    2014-10-01

    In this study, in vitro and in vivo effects of some commonly used fungicides, antibiotics, and various chemicals on isolated and purified catalase from Phanerochaete chrysosporium were investigated. The catalase was purified 129.10-fold by using 60% ammonium sulfate and 60% ethanol precipitations, DEAE-cellulose anion exchange and Sephacryl-S-200 gel filtration chromatographies from P. chrysosporium growth in carbon- and nitrogen-limited medium for 12 days. The molecular weight of native purified catalase from P. chrysosporium was found to be 290 ± 10 kDa, and sodium dodecyl sulfate (SDS)-PAGE results indicated that enzyme consisted of four apparently identical subunits, with a molecular weight of 72.5 ± 2.5 kDa. Kinetic characterization studies showed that optimum pH and temperature, Km and Vmax values of the purified catalase which were stable in basic region and at comparatively high temperatures were 7.5, 30°C, 289.86 mM, and 250,000 U/mg, respectively. The activity of purified catalase from P. chrysosporium was significantly inhibited by dithiothreitol (DTT), 2-mercaptoethanol, iodoacetamide, EDTA, and sodium dodecyl sulfate (SDS). It was found that while antibiotics had no inhibitory effects, 45 ppm benomyl, 144 ppm captan, and 47.5 ppm chlorothalonil caused 14.52, 10.82, and 38.86% inhibition of purified catalase, respectively. The inhibition types of these three fungicides were found to be non-competitive inhibition with the Ki values of 1.158, 0.638, and 0.145 mM and IC50 values of 0.573, 0.158, 0.010 mM, respectively. The results of in vivo experiments also showed that benomyl, captan and chlorothalonil caused 15.25, 1.96, and 36.70% activity decreases after 24-h treatments compared to that of the control.

  3. Purification, biochemical characterization, and implications of an alkali-tolerant catalase from the spacecraft-associated and oxidation-resistant Acinetobacter gyllenbergii 2P01AA.

    Science.gov (United States)

    Muster, N; Derecho, I; Dallal, F; Alvarez, R; McCoy, K B; Mogul, R

    2015-04-01

    Herein, we report on the purification, characterization, and sequencing of catalase from Acinetobacter gyllenbergii 2P01AA, an extremely oxidation-resistant bacterium that was isolated from the Mars Phoenix spacecraft assembly facility. The Acinetobacter are dominant members of the microbial communities that inhabit spacecraft assembly facilities and consequently may serve as forward contaminants that could impact the integrity of future life-detection missions. Catalase was purified by using a 3-step chromatographic procedure, where mass spectrometry provided respective subunit and intact masses of 57.8 and 234.6 kDa, which were consistent with a small-subunit tetrameric catalase. Kinetics revealed an extreme pH stability with no loss in activity between pH 5 and 11.5 and provided respective kcat/Km and kcat values of ∼10(7) s(-1) M(-1) and 10(6) s(-1), which are among the highest reported for bacterial catalases. The amino acid sequence was deduced by in-depth peptide mapping, and structural homology suggested that the catalases from differing strains of A. gyllenbergii differ only at residues near the subunit interfaces, which may impact catalytic stability. Together, the kinetic, alkali-tolerant, and halotolerant properties of the catalase from A. gyllenbergii 2P01AA are significant, as they are consistent with molecular adaptations toward the alkaline, low-humidity, and potentially oxidizing conditions of spacecraft assembly facilities. Therefore, these results support the hypothesis that the selective pressures of the assembly facilities impact the microbial communities at the molecular level, which may have broad implications for future life-detection missions.

  4. Effect of various levels of catalase antioxidant in semen extenders on lipid peroxidation and semen quality after the freeze-thawing bull semen

    Directory of Open Access Journals (Sweden)

    Reza Asadpour

    2011-11-01

    Full Text Available The objective of this study was to evaluate effect of different concentrations of catalase in two extenders on motility, viability and lipid peroxidation bull spermatozoa during semen freezing process. Thirty ejaculates collected from ten Holstein bulls were pooled and evaluated at 37 °C. Pool ejaculated was split into two main experimental groups, 1 and 2. In experiment 1, specimen was diluted to a final concentration of 30 × 106 spermatozoa with citrate-egg yolk and in experiment 2; specimen was diluted with tris-egg yolk extender to the same concentration. In both experiments diluted semen was divided into three aliquots, including a control and two test groups. Each aliquot was rediluted with an equal volume of extender either without (control or with one of the antioxidants contained one of the following antioxidants: catalase (CAT; 100 IU mL-1 catalase (CAT; 200 IU mL-1 and control group. No significant differences were observed in sperm viability and motility following addition of catalase enzyme at concentration of 100 IU mL-1 and 200 IU mL-1 to citrate-egg yolk extender. But the highest sperm viability was achieved by addition of 100 IU mL-1 and 200 IU mL-1 catalase to tris-egg yolk semen extender compared with the control group (P < 0.05. Malondialdehyde levels did not change with addition of catalase in both extenders compared with the control group. The obtained results provide a new approach to the cryopreservation of bull semen, and could positively contribute to intensive cattle production.

  5. Direct and indirect inactivation of tumor cell protective catalase by salicylic acid and anthocyanidins reactivates intercellular ROS signaling and allows for synergistic effects.

    Science.gov (United States)

    Scheit, Katrin; Bauer, Georg

    2015-03-01

    Salicylic acid and anthocyanidins are known as plant-derived antioxidants, but also can provoke paradoxically seeming prooxidant effects in vitro. These prooxidant effects are connected to the potential of salicylic acid and anthocyanidins to induce apoptosis selectively in tumor cells in vitro and to inhibit tumor growth in animal models. Several epidemiological studies have shown that salicylic acid and its prodrug acetylsalicylic acid are tumor-preventive for humans. The mechanism of salicylic acid- and anthocyanidin-dependent antitumor effects has remained enigmatic so far. Extracellular apoptosis-inducing reactive oxygen species signaling through the NO/peroxynitrite and the HOCl signaling pathway specifically induces apoptosis in transformed cells. Tumor cells have acquired resistance against intercellular reactive oxygen species signaling through expression of membrane-associated catalase. Here, we show that salicylic acid and anthocyanidins inactivate tumor cell protective catalase and thus reactive apoptosis-inducing intercellular reactive oxygen species signaling of tumor cells and the mitochondrial pathway of apoptosis Salicylic acid inhibits catalase directly through its potential to transform compound I of catalase into the inactive compound II. In contrast, anthocyanidins provoke a complex mechanism for catalase inactivation that is initiated by anthocyanidin-mediated inhibition of NO dioxygenase. This allows the formation of extracellular singlet oxygen through the reaction between H(2)O(2) and peroxynitrite, amplification through a caspase8-dependent step and subsequent singlet oxygen-mediated inactivation of catalase. The combination of salicylic acid and anthocyanidins allows for a remarkable synergistic effect in apoptosis induction. This effect may be potentially useful to elaborate novel therapeutic approaches and crucial for the interpretation of epidemiological results related to the antitumor effects of secondary plant compounds.

  6. Electrospun regenerated cellulose nanofibrous membranes surface-grafted with polymer chains/brushes via the atom transfer radical polymerization method for catalase immobilization.

    Science.gov (United States)

    Feng, Quan; Hou, Dayin; Zhao, Yong; Xu, Tao; Menkhaus, Todd J; Fong, Hao

    2014-12-10

    In this study, an electrospun regenerated cellulose (RC) nanofibrous membrane with fiber diameters of ∼200-400 nm was prepared first; subsequently, 2-hydroxyethyl methacrylate (HEMA), 2-dimethylaminoethyl methacrylate (DMAEMA), and acrylic acid (AA) were selected as the monomers for surface grafting of polymer chains/brushes via the atom transfer radical polymerization (ATRP) method. Thereafter, four nanofibrous membranes (i.e., RC, RC-poly(HEMA), RC-poly(DMAEMA), and RC-poly(AA)) were explored as innovative supports for immobilization of an enzyme of bovine liver catalase (CAT). The amount/capacity, activity, stability, and reusability of immobilized catalase were evaluated, and the kinetic parameters (Vmax and Km) for immobilized and free catalase were determined. The results indicated that the respective amounts/capacities of immobilized catalase on RC-poly(HEMA) and RC-poly(DMAEMA) nanofibrous membranes reached 78 ± 3.5 and 67 ± 2.7 mg g(-1), which were considerably higher than the previously reported values. Meanwhile, compared to that of free CAT (i.e., 18 days), the half-life periods of RC-CAT, RC-poly(HEMA)-CAT, RC-poly(DMAEMA)-CAT, and RC-poly(AA)-CAT were 49, 58, 56, and 60 days, respectively, indicating that the storage stability of immobilized catalase was also significantly improved. Furthermore, the immobilized catalase exhibited substantially higher resistance to temperature variation (tested from 5 to 70 °C) and lower degree of sensitivity to pH value (tested from 4.0 and 10.0) than the free catalase. In particular, according to the kinetic parameters of Vmax and Km, the nanofibrous membranes of RC-poly(HEMA) (i.e., 5102 μmol mg(-1) min(-1) and 44.89 mM) and RC-poly(DMAEMA) (i.e., 4651 μmol mg(-1) min(-1) and 46.98 mM) had the most satisfactory biocompatibility with immobilized catalase. It was therefore concluded that the electrospun RC nanofibrous membranes surface-grafted with 3-dimensional nanolayers of polymer chains/brushes would be

  7. Direct Electrochemistry of Catalase Incorporated in Collagen Films%Cat-Collagen薄膜电极的直接电化学

    Institute of Scientific and Technical Information of China (English)

    李敏; 陈志敏; 辛先荣

    2006-01-01

    研究了过氧化氢酶(Catalase,简称Cat)在胶原蛋白(Collagen)薄膜电极中的电化学行为.在pH 7.0的缓冲溶液中,Cat-Collagen薄膜电极在-0.47 V(vs.SCE)附近有一对可逆的氧化还原峰,为Cat血红素辅基Fe(Ⅲ)/Fe(Ⅱ)电对的特征峰.体系的pH对循环伏安峰的位置有明显的影响,且Cat-Collagen薄膜电极中Cat血红素辅基发生在一个电子转移的同时还可能伴随着一个质子转移.

  8. Infantile Refsum disease: deficiency of catalase-containing particles (peroxisomes), alkyldihydroxyacetone phosphate synthase and peroxisomal beta-oxidation enzyme proteins.

    Science.gov (United States)

    Wanders, R J; Schutgens, R B; Schrakamp, G; van den Bosch, H; Tager, J M; Schram, A W; Hashimoto, T; Poll-Thé, B T; Saudubrau, J M

    1986-08-01

    In recent years a number of biochemical abnormalities have been described in patients with the infantile form of Refsum disease, including the accumulation of very long chain fatty acids, trihydroxycoprostanoic acid and pipecolic acid. In this paper we show that catalase-containing particles (peroxisomes), alkyl dihydroxyacetone phosphate synthase and acyl-CoA oxidase protein are deficient in patients with infantile Refsum disease. These findings suggest that in the infantile form of Refsum disease, as in the cerebro-hepato-renal (Zellweger) syndrome the multiplicity of biochemical abnormalities is due to a deficiency of peroxisomes and hence to a generalized loss of peroxisomal functions. As a consequence the infantile form of Refsum disease can be diagnosed biochemically by methods already available for the prenatal and postnatal diagnosis of the cerebro-hepato-renal (Zellweger) syndrome.

  9. [Protective action figurations for superoxide dismutase - chondroitin sulfate - catalase bienzyme conjugate after its medicative administration in endotoxin shock].

    Science.gov (United States)

    Maksimenko, A V; Vavaeva, A V; Zvyagintseva, M A; Abramov, A A; Timoshin, A A; Vavaev, A V; Lakomkin, V L

    2016-03-01

    Previously it found that the bienzymatic conjugate superoxide dismutase-chondroitin sulfate, catalase (SOD-CHS-CAT) increased the survival rate of rats with endotoxic shock caused by the administration of lipopolysaccharide (LPS). This effect was observed both in preventive (before LPS) and therapeutic conjugate administration (after the administration of LPS). This study shows that the development of endotoxic shock is accompanied by increased levels of NO in the liver, lungs, kidneys, heart; administration of the SOD-CHS-CAT conjugate insignificantly influenced this parameter. At the same time, the changes in blood urea and creatinine suggest the protective effect of the conjugate on renal function, while diverse changes in biochemical parameters studied complicate the formation of the agreed conclusions on the state of other organs.

  10. SHORT-ROOT Deficiency Alleviates the Cell Death Phenotype of the Arabidopsis catalase2 Mutant under Photorespiration-Promoting Conditions.

    Science.gov (United States)

    Waszczak, Cezary; Kerchev, Pavel I; Mühlenbock, Per; Hoeberichts, Frank A; Van Der Kelen, Katrien; Mhamdi, Amna; Willems, Patrick; Denecker, Jordi; Kumpf, Robert P; Noctor, Graham; Messens, Joris; Van Breusegem, Frank

    2016-08-01

    Hydrogen peroxide (H2O2) can act as a signaling molecule that influences various aspects of plant growth and development, including stress signaling and cell death. To analyze molecular mechanisms that regulate the response to increased H2O2 levels in plant cells, we focused on the photorespiration-dependent peroxisomal H2O2 production in Arabidopsis thaliana mutants lacking CATALASE2 (CAT2) activity (cat2-2). By screening for second-site mutations that attenuate the PSII maximum efficiency (Fv'/Fm') decrease and lesion formation linked to the cat2-2 phenotype, we discovered that a mutation in SHORT-ROOT (SHR) rescued the cell death phenotype of cat2-2 plants under photorespiration-promoting conditions. SHR deficiency attenuated H2O2-dependent gene expression, oxidation of the glutathione pool, and ascorbate depletion in a cat2-2 genetic background upon exposure to photorespiratory stress. Decreased glycolate oxidase and catalase activities together with accumulation of glycolate further implied that SHR deficiency impacts the cellular redox homeostasis by limiting peroxisomal H2O2 production. The photorespiratory phenotype of cat2-2 mutants did not depend on the SHR functional interactor SCARECROW and the sugar signaling component ABSCISIC ACID INSENSITIVE4, despite the requirement for exogenous sucrose for cell death attenuation in cat2-2 shr-6 double mutants. Our findings reveal a link between SHR and photorespiratory H2O2 production that has implications for the integration of developmental and stress responses.

  11. Copper and resveratrol attenuates serum catalase, glutathione peroxidase, and element values in rats with DMBA-induced mammary carcinogenesis.

    Science.gov (United States)

    Skrajnowska, Dorota; Bobrowska-Korczak, Barbara; Tokarz, Andrzej; Bialek, Slawomir; Jezierska, Ewelina; Makowska, Justyna

    2013-12-01

    In this paper, a hypothesis was assessed whether or not the intoxication with copper and supplementation with copper plus resveratrol would result in changes in the activities of catalase and glutathione peroxidase and moreover if the characteristic changes would appear in concentrations of copper, iron, calcium, magnesium, and zinc in the serum of rats with chemically induced carcinogenesis. Female Sprague-Dawley rats were divided into study groups which, apart from the standard diet, were treated with copper (42.6 mg Cu/kg food as CuSO4·5H2O) or copper plus resveratrol (0.2 mg/kg body) via gavage for a period from 40 days until 20 weeks of age. In cancer groups, the rats were treated with a dose of 80 mg/body weight of 7,12-dimethyl-1,2-benz[a]anthracene (DMBA) given in rapeseed oil at 50 and 80 days of age to induce mammary carcinogenesis. The control groups included the rats kept in the same conditions and fed with the same diet as the animals from the study groups, but not DMBA-treated. The activity of catalase significantly decreased in groups of rats with mammary carcinogenesis that were supplemented with copper (p copper plus resveratrol (p cancer groups of nonsupplemented rats, the increase of glutathione peroxidase activity was observed. The process of carcinogenesis and the applied supplementation significantly altered the concentrations of trace elements in serum, in particular as concerns iron and copper. The mean serum iron levels in rats with breast cancer were significantly lower than those in the control groups (p copper levels significantly decreased in the groups of rats with mammary carcinogenesis that were supplemented with copper or copper plus resveratrol in comparison with the control groups that received the same diets (p copper and zinc/iron ratios in blood may be used as one of the prognostic factors in breast cancer research.

  12. Fluconazole and amphotericin-B resistance are associated with increased catalase and superoxide dismutase activity in Candida albicans and Candida dubliniensis

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    Carlos Eduardo Blanco Linares

    2013-12-01

    Full Text Available Introduction Candida dubliniensis, a new species of Candida that has been recovered from several sites in healthy people, has been associated with recurrent episodes of oral candidiasis in AIDS and HIV-positive patients. This species is closely related to C. albicans. The enzymatic activity of C. dubliniensis in response to oxidative stress is of interest for the development of drugs to combat C. dubliniensis. Methods Fluconazole- and amphotericin B-resistant strains were generated as described by Fekete-Forgács et al. (2000. Superoxide dismutase (SOD and catalase assays were performed as described by McCord and Fridovich (1969 and Aebi (1984, respectively. Results We demonstrated that superoxide dismutase (SOD and catalase activities were significantly higher (p<0.05 in the fluconazole- and amphotericin B-resistant strains of C. dubliniensis and C. albicans than in the sensitive strains. The catalase and SOD activities were also significantly (p<0.01 higher in the sensitive and resistant C. albicans strains than in the respective C. dubliniensis strains. Conclusions These data suggest that C. albicans is better protected from oxidative stress than C. dubliniensis and that fluconazole, like amphotericin B, can induce oxidative stress in Candida; oxidative stress induces an adaptive response that results in a coordinated increase in catalase and SOD activities.

  13. Blockade by metal complexing agents and by catalase of the effects of arachidonic acid on platelets: relevance to the study of anti-inflammatory mechanisms.

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    Vargaftig, B B; Tranier, Y; Chignard, M

    1975-08-01

    Metal-chelating agents inhibited platelet aggregation and the accompanying generation of rabbit aorta contracting and PG-like activities, when platelets were challenged with arachidonic acid. Inhibition required the presence of the chelating agents in the medium, and was insured by reagents avid for free or protein-bound copper. Catalase also prevented aggregation and generation of pharmacologically active substances; its activity was reversed by aminothiol agents and by Cu2+ and Zn2+, shown previously to potentiate the platelet effects of arachidonic acid. Inhibition by indomethacin was not prevented by amino-thiol drugs nor by Cu2+ or Zn2+. The catalase-induced inhibition was not affected by scavenging of thiol groups; this rules out, as a mechanism of action of catalase, the increased destruction of popoperoxides by glutathione peroxidase, which requires reduced glutathione as hydrogen donor. The results are compatible with the hypothesis that the agent that mediates platelet aggregation by arachidonic acid is a popoperoxide, requiring the presence either of H2O2 or of a similarly catalase-sensitive substance to be generated.

  14. The respiratory burst activity and expression of catalase in white shrimp, Litopenaeus vannamei, during long-term exposure to pH stress.

    Science.gov (United States)

    Wang, Wei-Na; Li, Bao-Sheng; Liu, Jin-Jian; Shi, Lei; Alam, M J; Su, Shi-Juan; Wu, Juan; Wang, Lei; Wang, An-Li

    2012-08-01

    In this study, changes of reactive oxygen species (ROS) and the mRNA expression of catalase of the Pacific white shrimp, Litopenaeus vannamei, exposed to pH (5.4, 6.7, 8.0, and 9.3) stress was investigated at different stress time (24, 48, 72, 96, and 120 h). Level of malondialdehyde (MDA) in shrimp also were assessed. The results revealed that acidic (pH 5.4 and 6.7) or alkaline exposure (pH 9.3) induced production of ROS hemocytes and increase of MDA level in shrimp. Moreover, the catalase mRNA expression in hepatopancreas of L. vannamei was up-regulated in 24 h at pH 5.4, in 72 h at pH 6.7 and in 48 h at pH 9.3, whereas was down-regulated significantly after 72 h acidic (pH 5.4 and 6.7) or alkaline (pH 9.4) exposure. In the present study, there was the relationship between ROS and catalase mRNA expression under normal acidic and alkaline conditions. At pH 8, the increase of catalase transcripts due to up-regulation by ROS, whereas MDA level did not significantly change, suggesting activation of corresponding protective mechanisms of detoxifying ROS is essential for the proper functioning of cells and the survival of shrimps.

  15. VCP Is an integral component of a novel feedback mechanism that controls intracellular localization of catalase and H2O2 Levels.

    Directory of Open Access Journals (Sweden)

    Katsuhiro Murakami

    Full Text Available Catalase is a key antioxidant enzyme that catalyzes the decomposition of hydrogen peroxide (H2O2 to water and oxygen, and it appears to shuttle between the cytoplasm and peroxisome via unknown mechanisms. Valosin-containing protein (VCP belongs to the AAA class of ATPases and is involved in diverse cellular functions, e.g. cell cycle and protein degradation, etc. Here we show that VCP and PEX19, a protein essential for peroxisome biogenesis, interact with each other. Knockdown of either VCP or PEX19 resulted in a predominantly cytoplasmic redistribution of catalase, and loss of VCP ATPase activity also increased its cytoplasmic redistribution. Moreover, VCP knockdown decreased intracellular ROS levels in normal and H2O2-treated cells, and an oxidation-resistant VCP impaired the ROS-induced cytoplasmic redistribution of catalase. These observations reveal a novel feedback mechanism, in which VCP can sense H2O2 levels, and regulates them by controlling the localization of catalase.

  16. Singlet oxygen treatment of tumor cells triggers extracellular singlet oxygen generation, catalase inactivation and reactivation of intercellular apoptosis-inducing signaling.

    Science.gov (United States)

    Riethmüller, Michaela; Burger, Nils; Bauer, Georg

    2015-12-01

    Intracellular singlet oxygen generation in photofrin-loaded cells caused cell death without discrimination between nonmalignant and malignant cells. In contrast, extracellular singlet oxygen generation caused apoptosis induction selectively in tumor cells through singlet oxygen-mediated inactivation of tumor cell protective catalase and subsequent reactivation of intercellular ROS-mediated apoptosis signaling through the HOCl and the NO/peroxynitrite signaling pathway. Singlet oxygen generation by extracellular photofrin alone was, however, not sufficient for optimal direct inactivation of catalase, but needed to trigger the generation of cell-derived extracellular singlet oxygen through the interaction between H2O2 and peroxynitrite. Thereby, formation of peroxynitrous acid, generation of hydroxyl radicals and formation of perhydroxyl radicals (HO2(.)) through hydroxyl radical/H2O2 interaction seemed to be required as intermediate steps. This amplificatory mechanism led to the formation of singlet oxygen at a sufficiently high concentration for optimal inactivation of membrane-associated catalase. At low initial concentrations of singlet oxygen, an additional amplification step needed to be activated. It depended on singlet oxygen-dependent activation of the FAS receptor and caspase-8, followed by caspase-8-mediated enhancement of NOX activity. The biochemical mechanisms described here might be considered as promising principle for the development of novel approaches in tumor therapy that specifically direct membrane-associated catalase of tumor cells and thus utilize tumor cell-specific apoptosis-inducing ROS signaling.

  17. A natural xanthone increases catalase activity but decreases NF-kappa B and lipid peroxidation in U-937 and HepG2 cell lines.

    Science.gov (United States)

    Sahoo, Binay K; Zaidi, Adeel H; Gupta, Pankaj; Mokhamatam, Raveendra B; Raviprakash, Nune; Mahali, Sidhartha K; Manna, Sunil K

    2015-10-05

    Mangiferin, a C-glycosyl xanthone, has shown anti-inflammatory, antioxidant, and anti-tumorigenic activities. In the present study, we investigated the molecular mechanism for the antioxidant property of mangiferin. Considering the role of nuclear transcription factor kappa B (NF-κB) in inflammation and tumorigenesis, we hypothesized that modulating its activity will be a viable therapeutic target in regulating the redox-sensitive ailments. Our results show that mangiferin blocks several inducers, such as tumor necrosis factor (TNF), lypopolysaccharide (LPS), phorbol-12-myristate-13-acetate (PMA) or hydrogen peroxide (H2O2) mediated NF-κB activation via inhibition of reactive oxygen species generation. In silico docking studies predicted strong binding energy of mangiferin to the active site of catalase (-9.13 kcal/mol), but not with other oxidases such as myeloperoxidase, glutathione peroxidase, or inducible nitric oxide synthase. Mangiferin increased activity of catalase by 44%, but had no effect on myeloperoxidase activity in vitro. Fluorescence spectroscopy further revealed the binding of mangiferin to catalase at the single site with binding constant and binding affinity of 3.1×10(-7) M(-1) and 1.046 respectively. Mangiferin also inhibits TNF-induced lipid peroxidation and thereby protects apoptosis. Hence, mangiferin with its ability to inhibit NF-κB and increase the catalase activity may prove to be a potent therapeutic.

  18. Coexpressed Catalase Protects Chimeric Antigen Receptor-Redirected T Cells as well as Bystander Cells from Oxidative Stress-Induced Loss of Antitumor Activity.

    Science.gov (United States)

    Ligtenberg, Maarten A; Mougiakakos, Dimitrios; Mukhopadhyay, Madhura; Witt, Kristina; Lladser, Alvaro; Chmielewski, Markus; Riet, Tobias; Abken, Hinrich; Kiessling, Rolf

    2016-01-15

    Treatment of cancer patients by adoptive T cell therapy has yielded promising results. In solid tumors, however, T cells encounter a hostile environment, in particular with increased inflammatory activity as a hallmark of the tumor milieu that goes along with abundant reactive oxygen species (ROS) that substantially impair antitumor activity. We present a strategy to render antitumor T cells more resilient toward ROS by coexpressing catalase along with a tumor specific chimeric Ag receptor (CAR) to increase their antioxidative capacity by metabolizing H2O2. In fact, T cells engineered with a bicistronic vector that concurrently expresses catalase, along with the CAR coexpressing catalase (CAR-CAT), performed superior over CAR T cells as they showed increased levels of intracellular catalase and had a reduced oxidative state with less ROS accumulation in both the basal state and upon activation while maintaining their antitumor activity despite high H2O2 levels. Moreover, CAR-CAT T cells exerted a substantial bystander protection of nontransfected immune effector cells as measured by CD3ζ chain expression in bystander T cells even in the presence of high H2O2 concentrations. Bystander NK cells, otherwise ROS sensitive, efficiently eliminate their K562 target cells under H2O2-induced oxidative stress when admixed with CAR-CAT T cells. This approach represents a novel means for protecting tumor-infiltrating cells from tumor-associated oxidative stress-mediated repression.

  19. 1,25-Dihydroxyvitamin D3 and its analogues increase catalase at the mRNA, protein and activity level in a canine transitional carcinoma cell line.

    Science.gov (United States)

    Middleton, R P; Nelson, R; Li, Q; Blanton, A; Labuda, J A; Vitt, J; Inpanbutr, N

    2015-12-01

    Antioxidant enzymes, such as catalase, superoxide dismutases (SOD), MnSOD and Cu/ZnSOD, protect cells by scavenging reactive oxygen species (ROS). Numerous studies have reported the anti-cancer effects of 1,25-dihydroxyvitamin D3 (calcitriol) and its related analogues, seocalcitol and analogue V. In this study, canine bladder transitional cell carcinoma (cbTCC) cells were used to determine effects of calcitriol and its related analogues on antioxidant enzyme gene expression, protein expression and activity. Catalase mRNA was increased in response to calcitriol (10(-7) M), and seocalcitol (10(-7) and 10(-9) M). MnSOD mRNA was decreased in response to calcitriol at 10(-7) M. Catalase was significantly increased in response to calcitriol (10(-7) and 10(-9) M), and seocalcitol (10(-9) M). Catalase enzymatic activity increased in response to calcitriol, seocalcitol and analogue V (10(-9) M). In addition, global gene expression analysis identified the involvement of mitogen-activated protein kinase (MAPK) signalling in cbTCC's response to calcitriol and seocalcitol treatment.

  20. The Effect of Atrazine on Soil Catalase Activity%阿特拉津对土壤过氧化氢酶活性的影响

    Institute of Scientific and Technical Information of China (English)

    那日苏; 林泮; 张爽; 陈鹏; 王慧芳; 张凤杰

    2014-01-01

    采用室内培养方法,研究了阿特拉津胁迫对土壤中过氧化氢酶的活性的影响。结果发现:在培养期间,阿特拉津对土壤过氧化氢酶表现为明显的抑制作用。3种模型拟合阿特拉津浓度与酶活性关系均达到显著相关关系。土壤阿特拉津对过氧化氢酶的生态阈值的平均值为103 mg/kg。%This article adopts indoor culture method to analyze the effects of atrazine stress on catalase activity in soil . The results show that during the culture ,atrazine has a remarkable inhibiting effect on soil catalase .The three model-fittings show the obvious significant correlation between soil catalase activity and atrazine concentration .Soil atrazine's mean ecological threshold to catalase is103 mg/kg .

  1. Modulatory effect of pineapple peel extract on lipid peroxidation, catalase activity and hepatic biomarker levels in blood plasma of alcohol-induced oxidative stressed rats

    Institute of Scientific and Technical Information of China (English)

    Okafor OY; Erukainure OL; Ajiboye JA; Adejobi RO; Owolabi FO; Kosoko SB

    2011-01-01

    Objective: To investigate the ability of the methanolic extract of pineapple peel to modulate alcohol-induced lipid peroxidation, changes in catalase activities and hepatic biochemical marker levels in blood plasma. Methods: Oxidative stress was induced by oral administration of ethanol (20% w/v) at a dosage of 5 mL/kg bw in rats. After 28 days of treatment, the rats were fasted overnight and sacrificed by cervical dislocation. Blood was collected with a 2 mL syringe by cardiac puncture and was centrifuged at 3000 rpm for 10 min. The plasma was analyzed to evaluate malondialdehyde (MDA), catalase activity, aspartate aminotransferase (AST), alkaline phosphatase (ALP) and alanine aminotransferase (ALT) concentrations. Results: Administration of alcohol caused a drastic increase (87.74%) in MDA level compared with the control. Pineapple peel extract significantly reduced the MDA level by 60.16% at 2.5 mL/kg bw. Rats fed alcohol only had the highest catalase activity, treatment with pineapple peel extract at 2.5 mL/kg bw however, reduced the activity. Increased AST, ALP and ALT activities were observed in rats fed alcohol only respectively, treatment with pineapple peel extract drastically reduced their activities. Conclusions: The positive modulation of lipid peroxidation, catalase activities as well as hepatic biomarker levels of blood plasma by the methanolic extract of pineapple peels under alcohol-induced oxidative stress is an indication of its protective ability in the management of alcohol-induced toxicity.

  2. Identification of a member of the catalase multigene family on wheat chromosome 7A associated with flour b* colour and biological significance of allelic variation.

    Science.gov (United States)

    Li, Dora A; Walker, Esther; Francki, Michael G

    2015-12-01

    Carotenoids (especially lutein) are known to be the pigment source for flour b* colour in bread wheat. Flour b* colour variation is controlled by a quantitative trait locus (QTL) on wheat chromosome 7AL and one gene from the carotenoid pathway, phytoene synthase, was functionally associated with the QTL on 7AL in some, but not all, wheat genotypes. A SNP marker within a sequence similar to catalase (Cat3-A1snp) derived from full-length (FL) cDNA (AK332460), however, was consistently associated with the QTL on 7AL and implicated in regulating hydrogen peroxide (H2O2) to control carotenoid accumulation affecting flour b* colour. The number of catalase genes on chromosome 7AL was investigated in this study to identify which gene may be implicated in flour b* variation and two were identified through interrogation of the draft wheat genome survey sequence consisting of five exons and a further two members having eight exons identified through comparative analysis with the single catalase gene on rice chromosome 6, PCR amplification and sequencing. It was evident that the catalase genes on chromosome 7A had duplicated and diverged during evolution relative to its counterpart on rice chromosome 6. The detection of transcripts in seeds, the co-location with Cat3-A1snp marker and maximised alignment of FL-cDNA (AK332460) with cognate genomic sequence indicated that TaCat3-A1 was the member of the catalase gene family associated with flour b* colour variation. Re-sequencing identified three alleles from three wheat varieties, TaCat3-A1a, TaCat3-A1b and TaCat3-A1c, and their predicted protein identified differences in peroxisomal targeting signal tri-peptide domain in the carboxyl terminal end providing new insights into their potential role in regulating cellular H2O2 that contribute to flour b* colour variation.

  3. Revealing mechanisms of selective, concentration-dependent potentials of 4-hydroxy-2-nonenal to induce apoptosis in cancer cells through inactivation of membrane-associated catalase.

    Science.gov (United States)

    Bauer, Georg; Zarkovic, Neven

    2015-04-01

    Tumor cells generate extracellular superoxide anions and are protected against superoxide anion-mediated intercellular apoptosis-inducing signaling by the expression of membrane-associated catalase. 4-Hydroxy-2-nonenal (4-HNE), a versatile second messenger generated during lipid peroxidation, has been shown to induce apoptosis selectively in malignant cells. The findings described in this paper reveal the strong, concentration-dependent potential of 4-HNE to specifically inactivate extracellular catalase of tumor cells both indirectly and directly and to consequently trigger apoptosis in malignant cells through superoxide anion-mediated intercellular apoptosis-inducing signaling. Namely, 4-HNE caused apoptosis selectively in NOX1-expressing tumor cells through inactivation of their membrane-associated catalase, thus reactivating subsequent intercellular signaling through the NO/peroxynitrite and HOCl pathways, followed by the mitochondrial pathway of apoptosis. Concentrations of 4-HNE of 1.2 µM and higher directly inactivated membrane-associated catalase of tumor cells, whereas at lower concentrations, 4-HNE triggered a complex amplificatory pathway based on initial singlet oxygen formation through H2O2 and peroxynitrite interaction. Singlet-oxygen-dependent activation of the FAS receptor and caspase-8 increased superoxide anion generation by NOX1 and amplification of singlet oxygen generation, which allowed singlet-oxygen-dependent inactivation of catalase. 4-HNE and singlet oxygen cooperate in complex autoamplificatory loops during this process. The finding of these novel anticancer pathways may be useful for understanding the role of 4-HNE in the control of malignant cells and for the optimization of ROS-dependent therapeutic approaches including antioxidant treatments.

  4. 琼脂固定化过氧化氢酶的催化活性%Catalytic activity of agar-immobilized catalase

    Institute of Scientific and Technical Information of China (English)

    张俊; 李云平; 应坤

    2012-01-01

    采用琼脂包埋法对过氧化氢酶进行固定化,考察了固定化过氧化氢酶催化过氧化氢降解的活性,确定了最佳催化反应条件.结果表明,经琼脂包埋法固定化后,过氧化氢酶仍保留较高的催化活性,其催化过氧化氢分解反应的最佳条件为温度35℃、pH 9.0.与此同时,固定化过氧化氢酶具有更强的温度适应能力和更宽的pH作用范围,并具有一定的重复使用性能.%Catalase was immobilized by encapsulation with agar. The catalytic activity of as-obtained immobilized catalase for the degradation of hydrogen peroxide was evaluated, and the optimal condition for the catalytic reaction was established. Results indicate that agar-immobi-lized catalase retains good catalytic activity. The optimal temperature and pH value for the agar-immobilized catalase to catalyze the decomposition and degradation of H2O2 are suggested as 35 °C and 9.0. In the meantime, agar-immobilized catalase possesses improved ability to adapt to temperature and pH, and it can be reused for several times.

  5. Cytoplasmic expression of recombinant interleukin-2 and interleukin-4 proteins results in hydrogen peroxide accumulation and reduction in catalase activity in Escherichia coli

    Directory of Open Access Journals (Sweden)

    M.S Hejazi

    2009-08-01

    Full Text Available Background and the purpose of the study: The Reactive oxygen species (ROS is induced in the cells following various stresses but the effect of recombinant protein expression on ROS generation has not been studied yet. In this study, H2O2 concentration and catalase activity variations and their correlation with cell growth following cytoplasmic expression of human interleukin-2 (hIL-2 and mouse interleukin-4 (mIL-4 in Escherichia coli were investigated. Additionally, the effect of recombinant protein expression under different conditions was compared to the effect of foreign DNA introduction on these factors. Methods: Plasmids pEThIL-2 and pETmIL-4 were used for expression of human interleukin-2 (hIL-2 and mouse interleukin-4 (mIL-4 inside the cytoplasm of the cells. Having confirmed protein expression using SDS-PAGE analysis and ELISA assay, H2O2 concentration and catalase activity were measured at various ODs. Results and major conclusion: Empty vector introduction increased significantly H2O2 concentration of the cells. However, H2O2 concentration in hIL-2 and mIL-4 expressing cells was significantly higher than its concentration in empty vector transformed cells. Catalase activity was reduced in foreign DNA introduced cells. It was more lowered following expression of recombinant proteins. Results of this study revealed the relationship between foreign DNA introduction and protein expression with H2O2 elevation and catalase activity reduction. There was also correlation between H2O2 elevation and reduction in catalase activity with the cell growth depression.

  6. Inhibitory effects of deferasirox on the structure and function of bovine liver catalase: a spectroscopic and theoretical study.

    Science.gov (United States)

    Moradi, M; Divsalar, A; Saboury, A A; Ghalandari, B; Harifi, A R

    2015-01-01

    Deferasirox (DFX), as an oral chelator, is used for treatment of transfusional iron overload. In this study, we have investigated the effects of DFX as an iron chelator, on the function and structure of bovine liver catalase (BLC) by different spectroscopic methods of UV-visible, fluorescence, and circular dichroism (CD) at two temperatures of 25 and 37 °C. In vitro kinetic studies showed that DFX can inhibit the enzymatic activity in a competitive manner. KI value was calculated 39 nM according to the Lineweaver-Burk plot indicating a high rate of inhibition of the enzyme. Intrinsic fluorescence data showed that increasing the drug concentrations leads to a significant decrease in the intrinsic emission of the enzyme indicating a significant change in the three-dimensional environment around the chromophores of the enzyme structure. By analyzing the fluorescence quenching data, it was found that the BLC has two binding sites for DFX and the values of binding constant at 25 and 37 °C were calculated 1.7 × 10(7) and 3 × 10(7) M(-1), respectively. The static type of quenching mechanism is involved in the quenching of intrinsic emission of enzyme. The thermodynamic data suggest that hydrophobic interactions play a major role in the binding reaction. UV-vis spectroscopy results represented the changes in tryptophan (Trp) absorption and Soret band spectra, which indicated changes in Trp and heme group position caused by the drug binding. Also, CD data represented that high concentrations of DFX lead to a significant decreasing in the content of β-sheet and random coil accompanied an increasing in α-helical content of the protein. The molecular docking results indicate that docking may be an appropriate method for prediction and confirmation of experimental results and also useful for determining the binding mechanism of proteins and drugs. According to above results, it can be concluded that the DFX can chelate the Fe(III) on the enzyme active site leading

  7. Increased activities of both superoxide dismutase and catalase were indicators of acute depressive episodes in patients with major depressive disorder.

    Science.gov (United States)

    Tsai, Meng-Chang; Huang, Tiao-Lai

    2016-01-30

    Oxidative stress may play an important role in the pathophysiology of major depressive disorder (MDD). The aim of this study was to investigate the serum levels of oxidative stress biomarkers and S100B in patients with MDD in an acute phase, and evaluate the changes in superoxide dismutase (SOD), protein carbonyl content (PCC), glutathione peroxidase (GPX), 8-hydroxy 2'-deoxyguanosine after treatment (8-OHdG), catalase (CAT), thiobarbituric acid reactive substances (TBARS) and S100B. We consecutively enrolled 21 MDD inpatients in an acute phase and 40 healthy subjects. Serum oxidative stress markers were measured with assay kits. Serum SOD and CAT activities in MDD patients in an acute phase were significantly higher than those of healthy subjects, and serum PCC levels were significantly lower. The HAM-D scores had a significantly positive association with S100B levels. Eighteen depressed patients were followed up, and there was no significant difference among all of the markers after treatment. In conclusion, our results suggest that increased activities of both SOD and CAT might be indicators of acute depressive episodes in MDD patients.

  8. The effect of seedling chilling on glutathione content, catalase and peroxidase activity in Brassica oleracea L. var. italica

    Directory of Open Access Journals (Sweden)

    Renata Wojciechowska

    2013-09-01

    Full Text Available The study was designed to determine the possible relationship between Brassica oleracea var. italica seedlings stored at 2°C in the dark for seven and fourteen days, respectively, and the level of certain antioxidant parameters in particular organs. A parallel objective of the experiment was to determine if the reaction of seedlings to low temperature might be persistent in fully developed plants until harvest time. After 14 days of chilling a significant increase in the glutathione content was observed in the seedling leaves in comparison to the non-chilled plants. During vegetation in field conditions this effect was maintained in leaves up to the stage of formation of flower buds. At harvest the highest content of glutathione was demonstrated in broccoli heads, obtained from plants, which were previously chilled in the seedling phase for two weeks. Peroxidase activity in broccoli seedlings increased each year of the three-year study due to the duration of the cooling time, whereas in the case of catalase the changes were not so distinct. At harvest time the activity of both enzymes in the leaves and flower buds fluctuated according to the particular year of study.

  9. Oxidative status, in vitro iron-induced lipid oxidation and superoxide dismutase, catalase and glutathione peroxidase activities in rhea meat.

    Science.gov (United States)

    Terevinto, A; Ramos, A; Castroman, G; Cabrera, M C; Saadoun, A

    2010-04-01

    Rhea (Rhea americana) muscles Obturatorius medialis (OM) Iliotibialis lateralis (IL) and Iliofibularis (I), obtained from farmed animals, were evaluated regarding their oxidative/antioxidant status. The mean level of thiobarbituric acid reactive substances (TBARS) expressed as malonaldehyde (MDA) content was of 0.84 mg MDA/kg wet tissue for the three muscles. TBARS level was significantly higher in IL than OM and I, with the two latter showing similar levels. The mean level of carbonyl proteins expressed as dinitrophenylhydrazine (DNPH) was 1.59 nmol DNPH mg(-1). Carbonyl protein levels were significantly different (Pmuscles (IL>OM>I). Iron-induced TBARS generation was not significantly different between the three muscles at any time, nor for each muscle during the 5 h of the experiment. Superoxide dismutase activity in IL muscle was significantly higher (Pmuscle. However, the difference between IL and OM muscles was not significant. The differences between the three muscles became not significant when the results were expressed by mg of protein contained in the extract, instead by g of wet tissue. No differences were found for catalase (micromol of discomposed H(2)O(2) min(-1) g(-1) wet tissue or by mg of protein contained in the extract) and glutathione peroxidase (micromol ol of oxidized NADPH min(-1) g(-1) of wet tissue or by mg of protein contained in the extract) activities between the three muscles.

  10. Increases of Catalase and Glutathione Peroxidase Expressions by Lacosamide Pretreatment Contributes to Neuroprotection Against Experimentally Induced Transient Cerebral Ischemia.

    Science.gov (United States)

    Choi, Hyun Young; Park, Joon Ha; Chen, Bai Hui; Shin, Bich Na; Lee, Yun Lyul; Kim, In Hye; Cho, Jeong-Hwi; Lee, Tae-Kyeong; Lee, Jae-Chul; Won, Moo-Ho; Ahn, Ji Hyeon; Tae, Hyun-Jin; Yan, Bing Chun; Hwang, In Koo; Cho, Jun Hwi; Kim, Young-Myeong; Kim, Sung Koo

    2016-09-01

    Lacosamide is a new antiepileptic drug which is widely used to treat partial-onset seizures. In this study, we examined the neuroprotective effect of lacosamide against transient ischemic damage and expressions of antioxidant enzymes such as Zn-superoxide dismutase (SOD1), Mn-superoxide dismutase (SOD2), catalase (CAT) and glutathione peroxidase (GPX) in the hippocampal cornu ammonis 1 (CA1) region following 5 min of transient global cerebral ischemia in gerbils. We found that pre-treatment with 25 mg/kg lacosamide protected CA1 pyramidal neurons from transient global cerebral ischemic insult using hematoxylin-eosin staining and neuronal nuclear antigen immunohistochemistry. Transient ischemia dramatically changed expressions of SOD1, SOD2 and GPX, not CAT, in the CA1 pyramidal neurons. Lacosamide pre-treatment increased expressions of CAT and GPX, not SOD1 and 2, in the CA1 pyramidal neurons compared with controls, and their expressions induced by lacosamide pre-treatment were maintained after transient cerebral ischemia. In brief, pre-treatment with lacosamide protected hippocampal CA1 pyramidal neurons from ischemic damage induced by transient global cerebral ischemia, and the lacosamide-mediated neuroprotection may be closely related to increases of CAT and GPX expressions by lacosamide pre-treatment.

  11. In vitro effect of sodium fluoride on malondialdehyde concentration and on superoxide dismutase, catalase, and glutathione peroxidase in human erythrocytes.

    Science.gov (United States)

    Gutiérrez-Salinas, José; García-Ortíz, Liliana; Morales González, José A; Hernández-Rodríguez, Sergio; Ramírez-García, Sotero; Núñez-Ramos, Norma R; Madrigal-Santillán, Eduardo

    2013-01-01

    The aim of this paper was to describe the in vitro effect of sodium fluoride (NaF) on the specific activity of the major erythrocyte antioxidant enzymes, as well as on the membrane malondialdehyde concentration, as indicators of oxidative stress. For this purpose, human erythrocytes were incubated with NaF (0, 7, 28, 56, and 100 μg/mL) or NaF (100 μg/mL) + vitamin E (1, 2.5, 5 and 10 μg/mL). The malondialdehyde (MDA) concentration on the surface of the erythrocytes was determined, as were the enzymatic activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GlPx). Our results demonstrated that erythrocytes incubated with increasing NaF concentrations had an increased MDA concentration, along with decreased activity of antioxidant enzymes. The presence of vitamin E partially reversed the toxic effects of NaF on erythrocytes. These findings suggest that NaF induces oxidative stress in erythrocytes in vitro, and this stress is partially reversed by the presence of vitamin E.

  12. Investigation on the interaction between isorhamnetin and bovine liver catalase by spectroscopic techniques under different pH conditions.

    Science.gov (United States)

    Yang, Yumin; Li, Daojin

    2016-08-01

    The binding of isorhamnetin to bovine liver catalase (BLC) was first investigated at 302, 310 and 318 K at pH 7.4 using spectroscopic methods including fluorescence spectra, circular dichroism (CD) and UV-vis absorption. Spectrophotometric observations are rationalized mainly in terms of a static quenching process. The binding constants and binding sites were evaluated by fluorescence quenching methods. Enzymatic activity of BLC in the absence and presence of isorhamnetin was measured using a UV/vis spectrophotometer. The result revealed that the binding of isorhamnetin to BLC led to a reduction in the activity of BLC. The positive entropy change and enthalpy change indicated that the interaction of isorhamnetin with BLC was mainly driven by hydrophobic forces. The distance r between the donor (BLC) and acceptor (isorhamnetin) was estimated to be 2.99 nm according to fluorescence resonance energy transfer. Fluorescence, synchronous fluorescence, and CD spectra showed no obvious change in the conformation of BLC upon the binding of isorhamnetin. In addition, the influence of pH on the binding of isorhamnetin to BLC was investigated and the binding ability of the drug to BLC deceased under other pH conditions (pH 9.0, 6.5, 5.0, 3.5, or 2.0) as compared with that at pH 7.4. Copyright © 2016 John Wiley & Sons, Ltd.

  13. Isolation and polyphasic characterization of a novel hyper catalase producing thermophilic bacterium for the degradation of hydrogen peroxide.

    Science.gov (United States)

    Sooch, Balwinder Singh; Kauldhar, Baljinder Singh; Puri, Munish

    2016-11-01

    A newly isolated microbial strain of thermophilic genus Geobacillus has been described with emphasis on polyphasic characterization and its application for degradation of hydrogen peroxide. The validation of this thermophilic strain of genus Geobacillus designated as BSS-7 has been demonstrated by polyphasic taxonomy approaches through its morphological, biochemical, fatty acid methyl ester profile and 16S rDNA sequencing. This thermophilic species of Geobacillus exhibited growth at broad pH and temperature ranges coupled with production of extraordinarily high quantities of intracellular catalase, the latter of which as yet not been reported in any member of this genus. The isolated thermophilic bacterial culture BSS-7 exhibited resistance against a variety of organic solvents. The immobilized whole cells of the bacterium successfully demonstrated the degradation of hydrogen peroxide (H2O2) in a packed bed reactor. This strain has potential application in various analytical and diagnostic methods in the form of biosensors and biomarkers in addition to applications in the textile, paper, food and pharmaceutical industries.

  14. Coupled expression of Cu/Zn-superoxide dismutase and catalase in cassava improves tolerance against cold and drought stresses.

    Science.gov (United States)

    Xu, Jia; Duan, Xiaoguang; Yang, Jun; Beeching, John R; Zhang, Peng

    2013-06-01

    Recently we reported that the joint expression of cassava Cu/Zn superoxide dismutase (MeCu/ZnSOD) and catalase (MeCAT1) prolonged the shelf life of cassava storage-roots by the stabilization of reactive oxygen species (ROS) homeostasis after harvest. Since oxidative damage is a major feature of plants exposed to environmental stresses, transgenic cassava showing increased expression of the cytosolic MeCu/ZnSOD and the peroxisomal MeCAT1 should have improved resistance against other abiotic stresses. After cold treatment, the transgenic cassava maintained higher SOD and CAT activities and lower malendialdehyde content than those of wild type plants (WT). Detached leaves of transgenic cassava also showed slower transpirational water loss than those of WT. When plants were not watered for 30 d, transgenic lines exhibited a significant increase in water retention ability, accumulated 13% more proline and 12% less malendialdehyde than WT's, and showed enhanced activity of SOD and CAT. These results imply that manipulation of the antioxidative mechanism allows the development of staple crops with improved tolerance to abiotic stresses.

  15. Ascorbate Peroxidase and Catalase Activities and Their Genetic Regulation in Plants Subjected to Drought and Salinity Stresses.

    Science.gov (United States)

    Sofo, Adriano; Scopa, Antonio; Nuzzaci, Maria; Vitti, Antonella

    2015-06-12

    Hydrogen peroxide (H2O2), an important relatively stable non-radical reactive oxygen species (ROS) is produced by normal aerobic metabolism in plants. At low concentrations, H2O2 acts as a signal molecule involved in the regulation of specific biological/physiological processes (photosynthetic functions, cell cycle, growth and development, plant responses to biotic and abiotic stresses). Oxidative stress and eventual cell death in plants can be caused by excess H2O2 accumulation. Since stress factors provoke enhanced production of H2O2 in plants, severe damage to biomolecules can be possible due to elevated and non-metabolized cellular H2O2. Plants are endowed with H2O2-metabolizing enzymes such as catalases (CAT), ascorbate peroxidases (APX), some peroxiredoxins, glutathione/thioredoxin peroxidases, and glutathione sulfo-transferases. However, the most notably distinguished enzymes are CAT and APX since the former mainly occurs in peroxisomes and does not require a reductant for catalyzing a dismutation reaction. In particular, APX has a higher affinity for H2O2 and reduces it to H2O in chloroplasts, cytosol, mitochondria and peroxisomes, as well as in the apoplastic space, utilizing ascorbate as specific electron donor. Based on recent reports, this review highlights the role of H2O2 in plants experiencing water deficit and salinity and synthesizes major outcomes of studies on CAT and APX activity and genetic regulation in drought- and salt-stressed plants.

  16. Physiological analyses indicate superoxide dismutase, catalase, and phytochelatins play important roles in Pb tolerance in Eremochloa ophiuroides.

    Science.gov (United States)

    Li, Xi; Cen, Huameng; Chen, Youxiang; Xu, Siying; Peng, Lingli; Zhu, Hanmingyue; Li, Yiqiao

    2016-01-01

    Phytoremediation is considered to be a promising approach to restore or stabilize soil contaminated by lead (Pb). Turfgrasses, due to their high biomass yields, are considered to be suitable for use in phytoextraction of soil contaminated with heavy metal. It has been demonstrated that centipedegrass (Eremochloa ophiuroides (Munro) Hack., Poaceae) is a good turfgrass for restore of soil contaminated by Pb. However, the enhanced tolerant mechanisms in metallicolous (M) centipedegrass accessions remain unknown. In this study, we made a comparative study of growth performance, Pb accumulation, antioxidant levels, and phytochelatin concentrations in roots and shoots from M and nonmetallicolous (NM) centipedegrass accessions. Results showed that turf quality and growth rate were less repressed in M accessions than in NM accession. Pb stress caused generation of reactive oxygen species in centipedegrass with relatively lower levels in M accessions. Antioxidant activity analysis indicated that superoxide dismutase and catalase played important roles in Pb tolerance in M accessions. M accessions accumulated more Pb in roots and shoots. Greatly increased phytochelatins and less repressed sulfur contents in roots and shoots of M accessions indicated that they correlated with Pb accumulation and tolerance in centipedegrass.

  17. Spectroscopic investigations on the effect of N-Acetyl-L-cysteine-Capped CdTe Quantum Dots on catalase

    Science.gov (United States)

    Sun, Haoyu; Yang, Bingjun; Cui, Erqian; Liu, Rutao

    2014-11-01

    Quantum dots (QDs) are recognized as some of the most promising semiconductor nanocrystals in biomedical applications. However, the potential toxicity of QDs has aroused wide public concern. Catalase (CAT) is a common enzyme in animal and plant tissues. For the potential application of QDs in vivo, it is important to investigate the interaction of QDs with CAT. In this work, the effect of N-Acetyl-L-cysteine-Capped CdTe Quantum Dots with fluorescence emission peak at 612 nm (QDs-612) on CAT was investigated by fluorescence, synchronous fluorescence, fluorescence lifetime, ultraviolet-visible (UV-vis) absorption and circular dichroism (CD) techniques. Binding of QDs-612 to CAT caused static quenching of the fluorescence, the change of the secondary structure of CAT and the alteration of the microenvironment of tryptophan residues. The association constants K were determined to be K288K = 7.98 × 105 L mol-1 and K298K = 7.21 × 105 L mol-1. The interaction between QDs-612 and CAT was spontaneous with 1:1 stoichiometry approximately. The CAT activity was also inhibited for the bound QDs-612. This work provides direct evidence about enzyme toxicity of QDs-612 to CAT in vitro and establishes a new strategy to investigate the interaction between enzyme and QDs at a molecular level, which is helpful for clarifying the bioactivities of QDs in vivo.

  18. The impact of heterologous catalase expression and superoxide dismutase overexpression on enhancing the oxidative resistance in Lactobacillus casei.

    Science.gov (United States)

    Lin, Jinzhong; Zou, Yexia; Cao, Kunlin; Ma, Chengjie; Chen, Zhengjun

    2016-05-01

    Two heme-dependent catalase genes were amplified from genomic DNA of Lactobacillus plantarum WCFS1 (KatE1) and Lactobacillus brevis ATCC 367 (KatE2), respectively, and a manganese-containing superoxide dismutase from Lactobacillus casei MCJΔ1 (MnSOD) were cloned into plasmid pELX1, yielding pELX1-KatE1, pELX1-KatE2 and pELX1-MnSOD, then the recombinant plasmids were transferred into L. casei MCJΔ1. The strains of L. casei MCJΔ1/pELX1-KatE1 and L. casei MCJΔ1/pELX1-KatE2 were tolerant at 2 mM H2O2. The survival rates of L. casei MCJΔ1/pELX1-KatE1 and L. casei MCJΔ1/pELX1-KatE2 were 270-fold and 300-fold higher than that of the control strain on a short-term H2O2 exposure, and in aerated condition, the survival cells counts were 146- and 190-fold higher than that of the control strain after 96 h of incubation. Furthermore, L. casei MCJΔ1/pELX1-MnSOD was the best in three recombinants which was superior in the living cell viability during storage when co-storage with Lactobacillus delbrueckii subsp. lactis LBCH-1.

  19. Engineering of a novel tri-functional enzyme with MnSOD, catalase and cell-permeable activities.

    Science.gov (United States)

    Luangwattananun, Piriya; Yainoy, Sakda; Eiamphungporn, Warawan; Songtawee, Napat; Bülow, Leif; Ayudhya, Chartchalerm Isarankura Na; Prachayasittikul, Virapong

    2016-04-01

    Cooperative function of superoxide dismutase (SOD) and catalase (CAT), in protection against oxidative stress, is known to be more effective than the action of either single enzyme. Chemical conjugation of the two enzymes resulted in molecules with higher antioxidant activity and therapeutic efficacy. However, chemical methods holds several drawbacks; e.g., loss of enzymatic activity, low homogeneity, time-consuming, and the need of chemical residues removal. Yet, the conjugated enzymes have never been proven to internalize into target cells. In this study, by employing genetic and protein engineering technologies, we reported designing and production of a bi-functional protein with SOD and CAT activities for the first time. To enable cellular internalization, cell penetrating peptide from HIV-1 Tat (TAT) was incorporated. Co-expression of CAT-MnSOD and MnSOD-TAT fusion genes allowed simultaneous self-assembly of the protein sequences into a large protein complex, which is expected to contained one tetrameric structure of CAT, four tetrameric structures of MnSOD and twelve units of TAT. The protein showed cellular internalization and superior protection against paraquat-induced cell death as compared to either complex bi-functional protein without TAT or to native enzymes fused with TAT. This study not only provided an alternative strategy to produce multifunctional protein complex, but also gained an insight into the development of therapeutic agent against oxidative stress-related conditions.

  20. Retinol increases catalase activity and protein content by a reactive species-dependent mechanism in Sertoli cells.

    Science.gov (United States)

    Gelain, Daniel Pens; de Bittencourt Pasquali, Matheus Augusto; Zanotto-Filho, Alfeu; de Souza, Luiz Fernando; de Oliveira, Ramatis Birnfeld; Klamt, Fábio; Moreira, José Cláudio Fonseca

    2008-07-10

    Vitamin A (retinol) is widely used as an antioxidant in therapeutic interventions and dietary supplementations. However, the redox properties of retinoids have been the subject of intense debate in the last few years, as recent works observed deleterious effects caused by retinol supplementation in clinical trials. In the present work, we show that retinol treatment (7 microM, 24 h) led to catalase (EC 1.11.1.6; CAT) activation in cultured Sertoli cells by increasing its protein content in a reactive species-dependent manner. Retinol treatment also increased cell lipoperoxidation, assessed by determination of thiobarbituric acid-reactive substances (TBARS), and intracellular reactive species production, determined by the real-time dihydrochlorofluorescein (DCFH-DA) assay. However, no alterations on CAT mRNA expression (assessed by RT-PCR) were observed, indicating an effect independent of CAT gene-transcription regulation. Importantly, all the effects induced by retinol were inhibited by the antioxidant Trolox, a hydrophilic analogue of alpha-tocopherol. These results show for the first time that retinol increases CAT activity by a redox-dependent modulation of its protein content in a cell culture model. CAT activity or expression are widely used as indexes of oxidative stress in biological systems; since no changes in CAT mRNA expression were detected in these conditions, the use of CAT gene-transcription activation when assessing oxidative stress should be re-evaluated.

  1. A synthetic superoxide dismutase/catalase mimetic EUK-207 mitigates radiation dermatitis and promotes wound healing in irradiated rat skin.

    Science.gov (United States)

    Doctrow, Susan R; Lopez, Argelia; Schock, Ashley M; Duncan, Nathan E; Jourdan, Megan M; Olasz, Edit B; Moulder, John E; Fish, Brian L; Mäder, Marylou; Lazar, Jozef; Lazarova, Zelmira

    2013-04-01

    In the event of a radionuclear attack or nuclear accident, the skin would be the first barrier exposed to radiation, though skin injury can progress over days to years following exposure. Chronic oxidative stress has been implicated as being a potential contributor to the progression of delayed radiation-induced injury to skin and other organs. To examine the causative role of oxidative stress in delayed radiation-induced skin injury, including impaired wound healing, we tested a synthetic superoxide dismutase (SOD)/catalase mimetic, EUK-207, in a rat model of combined skin irradiation and wound injury. Administered systemically, beginning 48 hours after irradiation, EUK-207 mitigated radiation dermatitis, suppressed indicators of tissue oxidative stress, and enhanced wound healing. Evaluation of gene expression in irradiated skin at 30 days after exposure revealed a significant upregulation of several key genes involved in detoxication of reactive oxygen and nitrogen species. This gene expression pattern was primarily reversed by EUK-207 therapy. These results demonstrate that oxidative stress has a critical role in the progression of radiation-induced skin injury, and that the injury can be mitigated by appropriate antioxidant compounds administered 48 hours after exposure.

  2. Response of peroxidase and catalase to acid rain stress during seed germination of rice, wheat, and rape

    Institute of Scientific and Technical Information of China (English)

    Lihong WANG; Xiaohua HUANG; Qing ZHOU

    2008-01-01

    Seed germination of plants with various acid-resistance display different responses to acid rain. To understand the reason why such differences occur, the effects of simulated acid rain (pH 2.5-5.0) on the activities of peroxidase (ROD) and catalase (CAT) during seed ger-mination of rice (O. sativa),-wheat (T. aestivum), and rape (B. chinensis var. oleifera) were investigated. Results indi-cated that the maximum change in activities of CAT and POD by acid rain treatment with different acidity and time in relation to the referent treatment without acid rain, was in the order: rice (28.8%, 31.7%)wheat (4.0)>rape (5.0). Moreover, the change in activity of POD was higher than that of CAT, which showed that POD was more sensitive to acid rain stress than CAT. The difference in the ability of POD and CAT in removing free radicals was one reason why the germina-tion indexes of these three species behaved differently.

  3. Evaluation of Both Malondialdehyde and Catalase Enzymes In Semen, Tissue And Blood In Adult Men With Grade 3 Varicocele

    Directory of Open Access Journals (Sweden)

    Ercan Malkoc

    2013-04-01

    Full Text Available This work aimed to assess malondialdehyde (MDA and catalase (CAT levels in semen, blood and tissue and to investigate their relationship with spermiogram parameters in the presence of grade 3 varicocele patients. Following the partial removal of the varicose vein during varicocelectomy, both MDA and CAT levels were assessed in the tissue and semen fluid. Additionally MDA levels were measured in blood samples collected from varicose veins during varicocelectomy. A total of 88 patients, mean age 21.8, were enrolled in the study. While progressive motility (A + B was <32% in 11 (12.5%, and #8805; 32 in 77 (87.5% patients; the total motility (A + B + C was <40 in 11 (12.5%, and and #8805; 40% in 77 (87.5% patients. Sperm count was <5 million/ml in 13 patients (14.8%, <15million/ml in 29 (32.9%, and and #8805; 15million/ml in 40 (45.5% patients. When patients were subgrouped according to the spermia, sperm count and sperm motility ratios, MDA and CAT levels did not differ significantly except for semen MDA activity which was significantly higher in patients with a sperm count lower than 15 million per ml (p=0.044. This finding suggests that MDA levels may affect the sperm counts. [Dis Mol Med 2013; 1(2.000: 26-30

  4. Growth rate, catalase and superoxide dismutase activities in rock carp (Procypris rabaudi Tchang) exposed to supersaturated total dissolved gas

    Institute of Scientific and Technical Information of China (English)

    Xiao-qing LIU; Ke-feng LI; Jun DU; Jia LI; Ran LI

    2011-01-01

    Total dissolved gas supersaturation (TDGS) appears when the pressures of gases in a solution exceed the barometric pressures.TDGS is often caused by flood discharge at dams.It may lead to gas bubble disease (GBD) for fish and biochemical responses of selected fish and other aquatic organisms.The purpose of this study was to determine the impact of long-term TDGS levels on the growth and biochemical responses of rock carp (Procypris rabaudi Tchang) dwelling in the upper reaches of the Yangtze River.Three-year-old rock carp were exposed to TDGS levels at 100%,104%,108%,112%,and 116% for 42 d.Samples were taken every 7 d after the start of the trial in order to determine catalase (CAT) and superoxide dismutase (SOD) activities in gill and muscle tissues.Samples were taken at Days 0 and 42 of exposure to determine growth rate.Little effect was found on growth rate in all treatment groups.SOD and CAT activities varied in different tissues,according to time of exposure and TDGS levels.The biochemical response of fish exposed to TDGS was more obvious in gill tissue than in muscle tissue.Surveys of SOD and CAT activities in different tissues offer important information about the effect of TDGS on the rare fish in the Yangtze River,and may help evaluate the risk to the aquatic eco-environment and aquatic ecosystem in the downstream of the Yangtze River.

  5. Direct electrochemistry of catalase at amine-functionalized graphene/gold nanoparticles composite film for hydrogen peroxide sensor

    Energy Technology Data Exchange (ETDEWEB)

    Huang Kejing, E-mail: kejinghuang@163.co [College of Chemistry and Chemical Engineering, Xinyang Normal University, 237 Chang' an Road, Xinyang, He' nan 464000 (China); Niu Dejun; Liu Xue; Wu Zhiwei; Fan Yang; Chang Yafang; Wu Yingying [College of Chemistry and Chemical Engineering, Xinyang Normal University, 237 Chang' an Road, Xinyang, He' nan 464000 (China)

    2011-02-28

    Direct electrochemistry and electrocatalysis of catalase (Cat) was studied based on a nano-composite film consisting of amine functionalized graphene and gold nanoparticles (AuNPs) modified glassy carbon electrode. Graphene was synthesized chemically by Hummers and Offeman method and then was functionalized with amino groups via chemical modification of carboxyl groups introduced on the graphene surface. The nano-composite film showed an obvious promotion of the direct electron transfer between Cat and the underlying electrode, which attributed to the synergistic effect of graphene-NH{sub 2} and AuNPs. The resultant bioelectrode retained its biocatalytic activity and offered fast and sensitive H{sub 2}O{sub 2} quantification. Under the optimized experimental conditions, hydrogen peroxide was detected in the concentration range from 0.3 to 600 {mu}M with a detection limit of 50 nM at S/N = 3. The biosensor exhibited some advantages, such as short time respond (2 s), high sensitivity (13.4 {mu}A/mM) and good reproducibility (RSD = 5.8%).

  6. Ascorbate Peroxidase and Catalase Activities and Their Genetic Regulation in Plants Subjected to Drought and Salinity Stresses

    Directory of Open Access Journals (Sweden)

    Adriano Sofo

    2015-06-01

    Full Text Available Hydrogen peroxide (H2O2, an important relatively stable non-radical reactive oxygen species (ROS is produced by normal aerobic metabolism in plants. At low concentrations, H2O2 acts as a signal molecule involved in the regulation of specific biological/physiological processes (photosynthetic functions, cell cycle, growth and development, plant responses to biotic and abiotic stresses. Oxidative stress and eventual cell death in plants can be caused by excess H2O2 accumulation. Since stress factors provoke enhanced production of H2O2 in plants, severe damage to biomolecules can be possible due to elevated and non-metabolized cellular H2O2. Plants are endowed with H2O2-metabolizing enzymes such as catalases (CAT, ascorbate peroxidases (APX, some peroxiredoxins, glutathione/thioredoxin peroxidases, and glutathione sulfo-transferases. However, the most notably distinguished enzymes are CAT and APX since the former mainly occurs in peroxisomes and does not require a reductant for catalyzing a dismutation reaction. In particular, APX has a higher affinity for H2O2 and reduces it to H2O in chloroplasts, cytosol, mitochondria and peroxisomes, as well as in the apoplastic space, utilizing ascorbate as specific electron donor. Based on recent reports, this review highlights the role of H2O2 in plants experiencing water deficit and salinity and synthesizes major outcomes of studies on CAT and APX activity and genetic regulation in drought- and salt-stressed plants.

  7. Perfil de susceptibilidade a antimicrobianos em amostras de cocos Gram-positivos, catalase negativos, isoladas de mastite subclínica bubalina Profile of antimicrobial susceptibility in strains of Gram positive cocos, negative catalase, isolated from buffalo subclinical mastitis

    Directory of Open Access Journals (Sweden)

    Maria C.E. Vianni

    2003-06-01

    Full Text Available Estudou-se o perfil de susceptibilidade a antimicrobianos em cocos Gram-positivos catalase negativos (21 amostras de Lactococcus garvieae e 6 de Enterococcus gallinarum, isoladas do leite de fêmeas com mastite subclínica e pertencentes a uma população composta por seis rebanhos bubalinos localizados no Estado do Rio de Janeiro. O teste utilizado foi o da difusão de discos em agar Müller Hinton, segundo recomendações do National Committee for Clinical Laboratory Standards - NCCLS, tendo sido testados discos com ampicilina (10mg, cefalotina (30mg, cefotaxima (30mg, cefoxitina (30mg, cloranfenicol (30mg, eritromicina (15mg, gentamicina (10mg, nitrofurantoína (300mg, norfloxacina (10mg, penicilina (10 UI, tetraciclina (30mg e vancomicina (30mg. Os resultados evidenciaram que em se tratando de Lactococcus garvieae, o antimicrobiano mais eficiente foi o nitrofurantoína com 85,71% de sensibilidade, seguido da cefotaxima (61,90%, vancomicina (52,38%, norfloxacina (47,62% e cefalotina (47,62%. A maior resistência foi desenvolvida frente a penicilina e ampicilina, com 95,24% de resistênciapara os dois antimicrobianos testados. O perfil de susceptibilidade desenvolvido pelas amostras de Enterococcus gallinarum, mostrou baixa sensibilidade frente aos antimicrobianos testados, onde os maiores índices foram observados frente eritromicina e gentamicina, com 33,34% de sensibilidade para ambos; quanto à resistência desenvolvida, foi possível observar 100% de resistência com relação a vancomicina e tetraciclina, seguindo-se cloranfenicol, penicilina, ampicilina, cefoxitina, cefalotina, cefotaxima, norfloxacina e nitrofurantoína, todas evidenciando uma resistência de 83,33% das amostras testadas.The susceptibility of antimicrobials was studied in Gram positive and catalase negative cocci (21 samples of Lactococcus garvieae and 6 Enterococcus gallinarum, isolated from the milk of cows with subclinical mastitis, belonging to six buffalo herds in

  8. Hepatoprotective effects of Poly-[hemoglobin-superoxide dismutase-catalase-carbonic anhydrase] on alcohol-damaged primary rat hepatocyte culture in vitro.

    Science.gov (United States)

    Jiang, Wenhua; Bian, Yuzhu; Wang, Zhenghui; Chang, Thomas Ming Swi

    2017-02-01

    We have prepared a novel nanobiotherapeutic, Poly-[hemoglobin-superoxide dismutase-catalase-carbonic anhydrase], which not only transports both oxygen and carbon dioxide but also a therapeutic antioxidant. Our previous study in a severe sustained 90 min hemorrhagic shock rat model shows that it has a hepatoprotective effect. We investigate its hepatoprotective effect further in this present report using an alcohol-damaged primary hepatocyte culture model. Results show that it significantly reduced ethanol-induced AST release, lipid peroxidation, and ROS production in rat primary hepatocytes culture. It also significantly enhanced the viability of ethanol-treated hepatocytes. Thus, the result shows that Poly-[hemoglobin-superoxide dismutase-catalase-carbonic anhydrase] also has some hepatoprotective effects against alcohol-induced injury in in vitro rat primary hepatocytes cell culture. This collaborate our previous observation of its hepatoprotective effect in a severe sustained 90-min hemorrhagic shock rat model.

  9. Change in the Content of Salicylic Acid and in the Activities of Phenylalanine Ammonia-Lyase and Catalase in Wheat Seedling Roots Under the Effect of Azospirillum Lectins

    Directory of Open Access Journals (Sweden)

    Alen'kina S.A.

    2012-05-01

    Full Text Available We investigated the time course of changes in the endogenous content of salicylic acid, the ratio between the acid's free and bound forms, and changes in the activities of phenylalanine ammonia-lyase and catalase in wheat seedling roots under the effect of the lectins of two strains of the associative nitrogen-fixing bacterium Azospirillum: A. brasilense Sp7 and its mutant defective in lectin activity, A. brasilense Sp7.2.3. Differences in plant response to the action of the lectins from these two strains were established. On the basis of the obtained data, a model was proposed for lectin-assisted induction of resistance, according to which the lectin effect on the roots of seedlings results in accumulation of free salicylic acid, which inhibits catalase activity, ultimately leading to accumulation of hydrogen peroxide and to formation of induced resistance.

  10. Electrospun Poly(acrylonitrile-co-acrylic acid) Nanofibrous Membranes for Catalase Immobilization:Effect of Porphyrin Filling on the Enzyme Activity

    Institute of Scientific and Technical Information of China (English)

    KE Bei-bei; WAN Ling-shu; HUANG Xiao-jun; XU Zhi-kang

    2011-01-01

    Porphyrin-filled nanofibrous membranes were facilely prepared by electrospinning of the mixtures of poly(acryionitrile-co-acrylic acid)(PANCAA) and porphyrins. 5,10,15,20-Tetraphenyiporphyrin(TPP) and its metalloderivatives(ZnTPP and CuTPP) were studied as filling mediators for the immobilization of redox enzyme. Results indicate that the introduction of TPP, ZnTPP and CuTPP improves the retention activity of the immobilized catalase.Among these three porphyrins, the ZnTPP-filled PANCAA nanofibrous membrane exhibits an activity retention of 93%, which is an exciting improvement. This improvement is attributed to both the strong catalase-porphyrin affinity and the possible facilitated electron transfer induced by the porphyrin as evidenced by quartz crystal microbalance (QCM) and fluorescence spectroscopy studies.

  11. Correlation between acid, TBA, peroxide and iodine values, catalase and glutathione peroxidase activities of chicken, cattle and camel meat during refrigerated storage

    Directory of Open Access Journals (Sweden)

    Hamid Reza Gheisari

    2011-08-01

    Full Text Available The aim of this study was correlation determination between fat putrefaction indices and antioxidative enzymes in chicken, cattle and camel meat during refrigerated storage. Longissimus dorsi muscle of three Iranian dromedary one humped camel and three Holstein cattle and breast muscle of three broiler breeder chicken were obtained from the carcasses 3 days postmortem. The samples were ground and stored at 4 °C for 0, 2, or 4 days. Peroxide, TBA, acid and iodine values, catalase and glutathione peroxidase (GSH-Px activities of the muscles were performed in each storage time. Catalase and GSH-Px activities were much higher in camel than in chicken and cattle and higher in cattle than in chicken. TBA value was lower in chicken than in camel. Camel had higher acid value than cattle. Chicken showed the highest and camel had the lowest iodine values. Catalase and GSH-Px activities and iodine values were quite stable during refrigerated storage. Acid values increased significantly over storage days in cattle. During the 4-day storage period, TBA and peroxide values increased. GSH-Px activity showed negative correlation with acid and TBA values in chicken and cattle. Acid value (for chicken and cattle and peroxide value (for 3 animal species showed positive correlation with TBA content. Iodine value had positive correlation with catalase activity in cattle and negative correlation with peroxide and TBA values in camel. In conclusion, our results indicate that peroxide and TBA values can be used as lipid quality indices in chicken, cattle and camel meat during 4 day storage in refrigerator. [Vet. World 2011; 4(4.000: 153-157

  12. Isolation and characterization of a catalase gene "HuCAT3" from pitaya (Hylocereus undatus) and its expression under abiotic stress.

    Science.gov (United States)

    Nie, Qiong; Gao, Guo-Li; Fan, Qing-jie; Qiao, Guang; Wen, Xiao-Peng; Liu, Tao; Peng, Zhi-Jun; Cai, Yong-Qiang

    2015-05-25

    Abiotic stresses usually cause H2O2 accumulation, with harmful effects, in plants. Catalase may play a key protective role in plant cells by detoxifying this excess H2O2. Pitaya (Hylocereus undatus) shows broad ecological adaptation due to its high tolerance to abiotic stresses, e.g. drought, heat and poor soil. However, involvement of the pitaya catalase gene (HuCAT) in tolerance to abiotic stresses is unknown. In the present study, a full-length HuCAT3 cDNA (1870 bp) was isolated from pitaya based on our previous microarray data and RACE method. The cDNA sequence and deduced amino acid sequence shared 73-77% and 75-80% identity with other plant catalases, respectively. HuCAT3 contains conserved catalase family domain and catalytic sites. Pairwise comparison and phylogenetic analysis indicated that HuCAT3 is most similar to Eriobotrya japonica CAT, followed by Dimocarpus longan CAT and Nicotiana tabacum CAT1. Expression profile analysis demonstrated that HuCAT3 is mainly expressed in green cotyledons and mature stems, and was regulated by H2O2, drought, cold and salt stress, whereas, its expression patterns and maximum expression levels varied with stress types. HuCAT activity increased as exposure to the tested stresses, and the fluctuation of HuCAT activity was consistent with HuCAT3 mRNA abundance (except for 0.5 days upon drought stress). HuCAT3 mRNA elevations and HuCAT activities changes under cold stress were also in conformity with the cold tolerances among the four genotypes. The obtained results confirmed a major role of HuCAT3 in abiotic stress response of pitaya. This may prove useful in understanding pitaya's high tolerance to abiotic stresses at molecular level.

  13. A teleostan homolog of catalase from black rockfish (Sebastes schlegelii): insights into functional roles in host antioxidant defense and expressional responses to septic conditions.

    Science.gov (United States)

    Elvitigala, Don Anushka Sandaruwan; Priyathilaka, Thanthrige Thiunuwan; Whang, Ilson; Nam, Bo-Hye; Lee, Jehee

    2015-05-01

    Antioxidative defense renders a significant protection against environmental stress in organisms and maintains the correct redox balance in cells, thereby supporting proper immune function. Catalase is an indispensable antioxidant in organisms that detoxifies hydrogen peroxides produced in cellular environments. In this study, we sought to molecularly characterize a homolog of catalase (RfCat), identified from black rockfish (Sebastes schlegelii). RfCat consists of a 1581 bp coding region for a protein of 527 amino acids, with a predicted molecular weight of 60 kD. The protein sequence of RfCat harbored similar domain architecture to known catalases, containing a proximal active site signature and proximal heme ligand signature, and further sharing prominent homology with its teleostan counterparts. As affirmed by multiple sequence alignments, most of the functionally important residues were well conserved in RfCat. Furthermore, our phylogenetic analysis indicates its common vertebrate ancestral origin and a close evolutionary relationship with teleostan catalases. Recombinantly expressed RfCat demonstrated prominent peroxidase activity that varied with different substrate and protein concentrations, and protected against DNA damage. RfCat mRNA was ubiquitously expressed among different tissues examined, as detected by qPCR. In addition, RfCat mRNA expression was modulated in response to pathogenic stress elicited by Streptococcus iniae and poly I:C in blood and spleen tissues. Collectively, our findings indicate that RfCat may play an indispensable role in host response to oxidative stress and maintain a correct redox balance after a pathogen invasion.

  14. Modulatory effect of pineapple peel extract on lipid peroxidation,catalase activity and hepatic biomarker levels in blood plasma of alcoholinduced oxidative stressed rats

    Institute of Scientific and Technical Information of China (English)

    Okafor; OY; Erukainure; OL; Ajiboye; JA; Adejobi; RO; Owolabi; FO; Kosoko; SB

    2011-01-01

    Objective:To investigate the ability of the methanolic extract of pineapple peel to modulate alcohol-induced lipid peroxidation,changes in catalase activities and hepatic biochemical marker levels in blood plasma.Methods:Oxidative stress was induced by oral administration of ethanol(20%w/v) at a dosage of 5 niL/kg bw in rats.After 28 days of treatment,the rats were fasted overnight and sacrificed by cervical dislocation.Blood was collected with a 2 mL syringe by cardiac puncture and was centrifuged at 3000 rpm for 10 min.The plasma was analyzed to evaluate malondialdehyde(MDA),catalase activity,aspartate aminotransferase(AST),alkaline phosphatase(ALP) and alanine aminotransferase(ALT) concentrations.Results:Administration of alcohol caused a drastic increase(87.74%) in MDA level compared with the control.Pineapple peel extract significantly reduced the MDA level by 60.16%at 2.S mL/kg bw.Rats fed alcohol only had the highest catalase activity,treatment with pineapple peel extract at 2.5 mL/kg bw however, reduced the activity.Increased AST,ALP and ALT activities were observed in rats fed alcohol only respectively,treatment with pineapple peel extract drastically reduced their activities. Conclusions:The positive modulation of lipid peroxidation,catalase activities as well as hepatic biomarker levels of blood plasma by the methanolic extract of pineapple peels under alcoholinduced oxidative stress is an indication of its protective ability in the management of alcoholinduced toxicity.

  15. Ultrasensitive fluorescence immunoassay for detection of ochratoxin A using catalase-mediated fluorescence quenching of CdTe QDs

    Science.gov (United States)

    Huang, Xiaolin; Zhan, Shengnan; Xu, Hengyi; Meng, Xianwei; Xiong, Yonghua; Chen, Xiaoyuan

    2016-04-01

    Herein, for the first time we report an improved competitive fluorescent enzyme linked immunosorbent assay (ELISA) for the ultrasensitive detection of ochratoxin A (OTA) by using hydrogen peroxide (H2O2)-induced fluorescence quenching of mercaptopropionic acid-modified CdTe quantum dots (QDs). In this immunoassay, catalase (CAT) was labeled with OTA as a competitive antigen to connect the fluorescence signals of the QDs with the concentration of the target. Through the combinatorial use of H2O2-induced fluorescence quenching of CdTe QDs as a fluorescence signal output and the ultrahigh catalytic activity of CAT to H2O2, our proposed method could be used to perform a dynamic linear detection of OTA ranging from 0.05 pg mL-1 to 10 pg mL-1. The half maximal inhibitory concentration was 0.53 pg mL-1 and the limit of detection was 0.05 pg mL-1. These values were approximately 283- and 300-folds lower than those of horseradish peroxidase (HRP)-based conventional ELISA, respectively. The reported method is accurate, highly reproducible, and specific against other mycotoxins in agricultural products as well. In summary, the developed fluorescence immunoassay based on H2O2-induced fluorescence quenching of CdTe QDs can be used for the rapid and highly sensitive detection of mycotoxins or haptens in food safety monitoring.Herein, for the first time we report an improved competitive fluorescent enzyme linked immunosorbent assay (ELISA) for the ultrasensitive detection of ochratoxin A (OTA) by using hydrogen peroxide (H2O2)-induced fluorescence quenching of mercaptopropionic acid-modified CdTe quantum dots (QDs). In this immunoassay, catalase (CAT) was labeled with OTA as a competitive antigen to connect the fluorescence signals of the QDs with the concentration of the target. Through the combinatorial use of H2O2-induced fluorescence quenching of CdTe QDs as a fluorescence signal output and the ultrahigh catalytic activity of CAT to H2O2, our proposed method could be used to

  16. Metal-based superoxide dismutase and catalase mimics reduce oxidative stress biomarkers and extend life span of Saccharomyces cerevisiae.

    Science.gov (United States)

    Ribeiro, Thales de P; Fonseca, Fernanda L; de Carvalho, Mariana D C; Godinho, Rodrigo M da C; de Almeida, Fernando Pereira; Saint'Pierre, Tatiana D; Rey, Nicolás A; Fernandes, Christiane; Horn, Adolfo; Pereira, Marcos D

    2017-01-15

    Aging is a natural process characterized by several biological changes. In this context, oxidative stress appears as a key factor that leads cells and organisms to severe dysfunctions and diseases. To cope with reactive oxygen species and oxidative-related damage, there has been increased use of superoxide dismutase (SOD)/catalase (CAT) biomimetic compounds. Recently, we have shown that three metal-based compounds {[Fe(HPClNOL)Cl2]NO3, [Cu(HPClNOL)(CH3CN)](ClO4)2 and Mn(HPClNOL)(Cl)2}, harboring in vitro SOD and/or CAT activities, were critical for protection of yeast cells against oxidative stress. In this work, treating Saccharomyces cerevisiae with these SOD/CAT mimics (25.0 µM/1 h), we highlight the pivotal role of these compounds to extend the life span of yeast during chronological aging. Evaluating lipid and protein oxidation of aged cells, it becomes evident that these mimics extend the life expectancy of yeast mainly due to the reduction in oxidative stress biomarkers. In addition, the treatment of yeast cells with these mimics regulated the amounts of lipid droplet occurrence, consistent with the requirement and protection of lipids for cell integrity during aging. Concerning SOD/CAT mimics uptake, using inductively coupled plasma mass spectrometry, we add new evidence that these complexes, besides being bioabsorbed by S. cerevisiae cells, can also affect metal homeostasis. Finally, our work presents a new application for these SOD/CAT mimics, which demonstrate a great potential to be employed as antiaging agents. Taken together, these promising results prompt future studies concerning the relevance of administration of these molecules against the emerging aging-related diseases such as Parkinson's, Alzheimer's and Huntington's.

  17. Growth, toxin production, active oxygen species and catalase activity of Microcystis aeruginosa (Cyanophyceae) exposed to temperature stress.

    Science.gov (United States)

    Giannuzzi, Leda; Krock, Bernd; Minaglia, Melina Celeste Crettaz; Rosso, Lorena; Houghton, Christian; Sedan, Daniela; Malanga, Gabriela; Espinosa, Mariela; Andrinolo, Darío; Hernando, Marcelo

    2016-11-01

    Microcystis are known for their potential ability to synthesize toxins, mainly microcystins (MCs). In order to evaluate the effects of temperature on chlorophyll a (Chl a), growth, physiological responses and toxin production of a native Microcystis aeruginosa, we exposed the cells to low (23°C) and high (29°C) temperature in addition to a 26°C control treatment. Exponential growth rate was significantly higher at 29°C compared to 23°C and control, reaching 0.43, 0.32 and 0.33day(-)(1) respectively. In addition, there was a delay of the start of exponential growth at 23°C. However, the intracellular concentration of Chl a decreased significantly due to temperature change. A significant increase in intracellular ROS was observed in coincidence with the activation of enzymatic antioxidant catalase (CAT) during the first two days of exposure to 23° and 29°C in comparison to the control experiment, decreasing thereafter to nearly initial values. Five MCs were determined by LC-MS/MS analysis. In the experiments, the highest MC concentration, 205fg [Leu(1)] MC-LR.cell(-1) expressed as MC-LR equivalent was measured in the beginning of the experiment and subsequently declined to 160fg.cell(-1) on day 2 and 70fg.cell(-1) on day 4 in cells exposed to 29°C. The same trend was observed for all other MCs except for the least abundant MC-LR which showed a continuous increase during exposure time. Our results suggest a high ability of M. aeruginosa to perceive ROS and to rapidly initiate antioxidant defenses with a differential response on MC production.

  18. Gluconic acid production in bioreactor with immobilized glucose oxidase plus catalase on polymer membrane adjacent to anion-exchange membrane.

    Science.gov (United States)

    Godjevargova, Tzonka; Dayal, Rajeshwar; Turmanova, Sevdalina

    2004-10-20

    Gluconic acid was obtained in the permeate side of the bioreactor with glucose oxidase (GOD) immobilized onto anion-exchange membrane (AEM) of low-density polyethylene grafted with 4-vinylpiridine. The electric resistance of the anion-exchange membranes was increased after the enzyme immobilization on the membrane. The gluconic acid productions were relatively low with the GOD immobilized by any method on the AEM. To increase the enzyme reaction efficiency, GOD was immobilized on membrane of AN copolymer (PAN) adjacent to an anion-exchange membrane in bioreactor. Uses of anion-exchange membrane led to selective removal of the gluconic acid from the glucose solution and reduce the gluconic acid inhibition. The amount of gluconic acid obtained in the permeate side of the bioreactor with the GOD immobilized on the PAN membrane adjacent to the AEM under electrodialysis was about 30 times higher than that obtained with enzyme directly bound to the AEM. The optimal substrate concentration in the feed side was found to be about 1 g/l. Further experiments were carried out with the co-immobilized GOD plus Catalase (CAT) on the PAN membrane adjacent to the AEM to improve the efficiency of the immobilize system. The yield of this process was at least 95%. The storage stability of the co-immobilized GOD and CAT was studied (lost 20% of initial activity for 90 d). The results obtained clearly showed the higher potential of the dual membrane bioreactor with GOD plus CAT bound to ultrafiltration polymer membrane adjacent to the AEM. Storage stability of GOD activity in GOD plus CAT immobilized on PAN//AEM membranes and on AEM.

  19. Enhanced reactive oxygen species scavenging by overproduction of superoxide dismutase and catalase delays postharvest physiological deterioration of cassava storage roots.

    Science.gov (United States)

    Xu, Jia; Duan, Xiaoguang; Yang, Jun; Beeching, John R; Zhang, Peng

    2013-03-01

    Postharvest physiological deterioration (PPD) of cassava (Manihot esculenta) storage roots is the result of a rapid oxidative burst, which leads to discoloration of the vascular tissues due to the oxidation of phenolic compounds. In this study, coexpression of the reactive oxygen species (ROS)-scavenging enzymes copper/zinc superoxide dismutase (MeCu/ZnSOD) and catalase (MeCAT1) in transgenic cassava was used to explore the intrinsic relationship between ROS scavenging and PPD occurrence. Transgenic cassava plants integrated with the expression cassette p54::MeCu/ZnSOD-35S::MeCAT1 were confirmed by Southern-blot analysis. The expression of MeCu/ZnSOD and MeCAT1 was verified by quantitative reverse transcription-polymerase chain reaction and enzymatic activity analysis both in the leaves and storage roots. Under exposure to the ROS-generating reagent methyl viologen or to hydrogen peroxide (H2O2), the transgenic plants showed higher enzymatic activities of SOD and CAT than the wild-type plants. Levels of malondialdehyde, chlorophyll degradation, lipid peroxidation, and H2O2 accumulation were dramatically reduced in the transgenic lines compared with the wild type. After harvest, the storage roots of transgenic cassava lines show a delay in their PPD response of at least 10 d, accompanied by less mitochondrial oxidation and H2O2 accumulation, compared with those of the wild type. We hypothesize that this is due to the combined ectopic expression of Cu/ZnSOD and CAT leading to an improved synergistic ROS-scavenging capacity of the roots. Our study not only sheds light on the mechanism of the PPD process but also develops an effective approach for delaying the occurrence of PPD in cassava.

  20. 3{prime} UTR sequence-specific mRNA-protein complexes and the post-transcriptional regulation of catalase

    Energy Technology Data Exchange (ETDEWEB)

    Reimer, D.L.; Ott, R.N.; Singh, S.M. [Univ. of Western Ontario, London (Canada)

    1994-09-01

    Recently, sequences in the 3{prime} untranslated region (3{prime} UTR) of some genes have been recognized which may play an important role in the post-transcriptional regulation of gene expression. Mutations in this region of the gene are known to cause at least two diseases including myotonic dystrophy and a lysosomal accumulation disease. The mechanism is thought to involve mRNA-protein interactions that affect translation and/or mRNA stability. Reports of this nature are not common and the significance of the often large 3{prime} UTR on the regulation of gene expression remains speculative. Studies on the 3{prime} UTR mRNA-protein interactions in model eukaryotic genes therefore are critical to better understand the molecular mechanisms involved in post-transcriptional gene regulation. Mouse catalase, encoded by Cas-1, was used as a model to characterize the molecular mechanisms of post-transcriptional gene regulation. The 3{prime} UTR (752 bp) of Cas-1 contains three unusual, near repeats [(CA){sub 31}, (U){sub 15} and (TGTGC){sub 7}]. Gel mobility shift assays using {sup 32}P-labelled transcripts which contain these sequences and tissue homogenates from various sources identified mRNA-protein complexes specific to (CA){sub 31} and (U){sub 15} only. In all strains analyzed, a single protein of 69 kDa which was involved in the (CA){sub 31} complex, was observed in most tissues except lung and was localized to the polysomal fraction. Similarly, two proteins involved in the (U){sub 15} complex, 38 and 47 kDa, were observed in all tissues and strains studied. Only the 38 kDa protein was observed in the polysomal fraction. The results argue for a possible role for these 3{prime} UTR mRNA-binding protein complexes in the post-transcriptional regulation of this antioxidant enzyme.

  1. Effects of total dissolved gas supersaturated water on lethality and catalase activity of Chinese sucker (Myxocyprinus asiaticus Bleeker)

    Institute of Scientific and Technical Information of China (English)

    Shi-chao CHEN; Xiao-qing LIU; Wen JIANG; Ke-feng LI; Jun DU; Dan-zhou SHEN; Quan GONG

    2012-01-01

    Total dissolved gas (TDG) supersaturation caused by dam sluicing can result in gas bubble trauma (GBT) in fish and threaten their survival.In the present study,Chinese suckers (Myxocyprinus asiaticus Bleeker) were exposed to TDG supersaturated water at levels ranging from 120% to 145% for 48 h.The median lethal concentration (LC50) and the median lethal time (LT50) were determined to evaluate acute lethal effects on Chinese suckers.The results showed that the LC50 values of 4,6,8,and 10 h were 142%,137%,135%,and 130%,respectively.The LT50 values were 3.2,4.7,7.8,9.2,and 43.4 h,respectively,when TDG supersaturated levels were 145%,140%,135%,130%,and 125%.Furthermore,the biological responses in Chinese suckers were studied by assaying the catalase (CAT) activities in gills and muscles at the supersaturation level of 140% within LT50.The CAT activities in the gills and muscle tissues exhibited a regularity of a decrease after an increase.CAT activities in the muscles were increased significantly at 3/5LT50 (P<0.05) and then came back to the normal level.However,there were no significant differences between the treatment group (TDG level of 140%) and the control group (TDG level of 100%) on CAT activities in the gills before 3/5LT50 (P>0.05),but the activities were significantly lower than the normal level at 4/5LT50 and LT50 (P<0.05).

  2. Combustion products of 1,3-butadiene inhibit catalase activity and induce expression of oxidative DNA damage repair enzymes in human bronchial epithelial cells.

    Science.gov (United States)

    Kennedy, Christopher H; Catallo, W James; Wilson, Vincent L; Mitchell, James B

    2009-10-01

    1,3-Butadiene, an important petrochemical, is commonly burned off when excess amounts need to be destroyed. This combustion process produces butadiene soot (BDS), which is composed of a complex mixture of polycyclic aromatic hydrocarbons in particulates ranging in size from DNA damage in NHBE cells. Thus, our aims were to determine the effect of butadiene soot ethanol extract (BSEE) on both enzyme activity and the expression of proteins involved in the repair of oxidative DNA damage. Catalase was found to be sensitive to BDS as catalase activity was potently diminished in the presence of BSEE. Using Western analysis, both the alpha isoform of human 8-oxoguanine DNA glycosylase (alpha-hOGG1) and human apurinic/apyrimidinic endonuclease (APE-1) were shown to be significantly overexpressed as compared to untreated controls after exposure of NHBE cells to BSEE. Our results indicate that BSEE is capable of effectively inactivating the antioxidant enzyme catalase, presumably via oxidation of protein amino acids. The presence of oxidized biomolecules may partially explain the extranuclear fluorescence that is detected when NHBE cells are treated with an organic extract of BDS. Overexpression of both alpha-hOGG1 and APE-1 proteins following treatment of NHBE cells with BSEE suggests that this mixture causes oxidative DNA damage.

  3. PprM is necessary for up-regulation of katE1, encoding the major catalase of Deinococcus radiodurans, under unstressed culture conditions.

    Science.gov (United States)

    Jeong, Sun-Wook; Seo, Ho Seong; Kim, Min-Kyu; Choi, Jong-Il; Lim, Heon-Man; Lim, Sangyong

    2016-06-01

    Deinococcus radiodurans is a poly-extremophilic organism, capable of tolerating a wide variety of different stresses, such as gamma/ultraviolet radiation, desiccation, and oxidative stress. PprM, a cold shock protein homolog, is involved in the radiation resistance of D. radiodurans, but its role in the oxidative stress response has not been investigated. In this study, we investigated the effect of pprM mutation on catalase gene expression. pprM disruption decreased the mRNA and protein levels of KatE1, which is the major catalase in D. radiodurans, under normal culture conditions. A pprM mutant strain (pprM MT) exhibited decreased catalase activity, and its resistance to hydrogen peroxide (H2O2) decreased accordingly compared with that of the wild-type strain. We confirmed that RecG helicase negatively regulates katE1 under normal culture conditions. Among katE1 transcriptional regulators, the positive regulator drRRA was not altered in pprM (-), while the negative regulators perR, dtxR, and recG were activated more than 2.5-fold in pprM MT. These findings suggest that PprM is necessary for KatE1 production under normal culture conditions by down-regulation of katE1 negative regulators.

  4. Reactions of the class II peroxidases, lignin peroxidase and Arthromyces ramosus peroxidase, with hydrogen peroxide. Catalase-like activity, compound III formation, and enzyme inactivation.

    Science.gov (United States)

    Hiner, Alexander N P; Hernández-Ruiz, Josefa; Rodríguez-López, José Neptuno; García-Cánovas, Francisco; Brisset, Nigel C; Smith, Andrew T; Arnao, Marino B; Acosta, Manuel

    2002-07-26

    The reactions of the fungal enzymes Arthromyces ramosus peroxidase (ARP) and Phanerochaete chrysosporium lignin peroxidase (LiP) with hydrogen peroxide (H(2)O(2)) have been studied. Both enzymes exhibited catalase activity with hyperbolic H(2)O(2) concentration dependence (K(m) approximately 8-10 mm, k(cat) approximately 1-3 s(-1)). The catalase and peroxidase activities of LiP were inhibited within 10 min and those of ARP in 1 h. The inactivation constants were calculated using two independent methods; LiP, k(i) approximately 19 x 10(-3) s(-1); ARP, k(i) approximately 1.6 x 10(-3) s(-1). Compound III (oxyperoxidase) was detected as the majority species after the addition of H(2)O(2) to LiP or ARP, and its formation was accompanied by loss of enzyme activity. A reaction scheme is presented which rationalizes the turnover and inactivation of LiP and ARP with H(2)O(2). A similar model is applicable to horseradish peroxidase. The scheme links catalase and compound III forming catalytic pathways and inactivation at the level of the [compound I.H(2)O(2)] complex. Inactivation does not occur from compound III. All peroxidases studied to date are sensitive to inactivation by H(2)O(2), and it is suggested that the model will be generally applicable to peroxidases of the plant, fungal, and prokaryotic superfamily.

  5. Fenton reaction-mediated fluorescence quenching of N-acetyl-L-cysteine-protected gold nanoclusters: analytical applications of hydrogen peroxide, glucose, and catalase detection.

    Science.gov (United States)

    Deng, Hao-Hua; Wu, Gang-Wei; He, Dong; Peng, Hua-Ping; Liu, Ai-Lin; Xia, Xing-Hua; Chen, Wei

    2015-11-21

    Given the importance of hydrogen peroxide (H2O2) in many biological processes and its wide application in various industries, the demand for sensitive, accurate, and economical H2O2 sensors is high. In this study, we used Fenton reaction-stimulated fluorescence quenching of N-acetyl-L-cysteine-protected gold nanoclusters (NAC-AuNCs) as a reporter system for the determination of H2O2. After the experimental conditions were optimized, the sensing platform enabled the analysis of H2O2 with a limit of detection (LOD) as low as 0.027 μM. As the glucose oxidase cascade leads to the generation of H2O2 and catalase catalyzes the decomposition of H2O2, these two biocatalytic procedures can be probed by the Fenton reaction-mediated quenching of NAC-AuNCs. The LOD for glucose was found to be 0.18 μM, and the linear range was 0.39-27.22 μM. The LOD for catalase was 0.002 U mL(-1), and the linear range was 0.01-0.3 U mL(-1). Moreover, the proposed sensing methods were successfully applied for human serum glucose detection and the non-invasive determination of catalase activity in human saliva, demonstrating their great potential for practical applications.

  6. Environmental Lead Exposure, Catalase Gene, and Markers of Antioxidant and Oxidative Stress Relation to Hypertension: An Analysis Based on the EGAT Study

    Directory of Open Access Journals (Sweden)

    Jintana Sirivarasai

    2015-01-01

    Full Text Available Lead has been linked to the development of hypertension via oxidative stress. Catalase plays an important role in the disposal of hydrogen peroxide in erythrocyte and its activity was determined by CAT gene. The aims of this study were to investigate (1 the association between blood levels of antioxidant markers such as catalase, superoxide dismutase, glutathione, glutathione peroxidase, oxidative stress-marker (malondialdehyde, and blood lead level and (2 the influence of genetic polymorphism of CAT gene (rs769217 on change in blood pressure in general population of EGAT study project. This is a cross-sectional study of 332 normotensive, 432 prehypertensive, and 222 hypertensive male subjects. Hypertensive subjects had significantly higher blood lead level (5.28 μg/dL compared to normotensive (4.41 μg/dL and prehypertensive (4.55 μg/dL subjects (P<0.05. These significant findings are also found in MDA levels. Moreover, individuals with TT genotype in hypertensive group had significantly higher blood lead and MDA levels (6.06 μg/dL and 9.67 μmol/L than those with CC genotype (5.32 μg/dL and 8.31 μmol/L, P<0.05. Our findings suggested that decreased blood catalase activity in this polymorphism together with low level lead exposure induced lipid peroxidation may be responsible for hypertension.

  7. CAT基因突变热点区域的PCR扩增方法%Optimization of PCR Amplification Parameters for Mutational Hotspots in the Human Catalase Gene

    Institute of Scientific and Technical Information of China (English)

    李毅; 赵华; 赵红宇; 章锦才

    2009-01-01

    Objective To study the speciality and sensitivity of PCR for mutational hotspots in the human catalase gene. Methods Blood samples were taken from volunteers for genomic DNA preparation. Polymerase chain reaction (PCR) amplification of sixteen gene segments encompassing mutational hotspots in the human catalase was carried out. Touchdown and Hot-start PCR was imple-mented to improve the efficiency of gene amplification. Results High and constant amplification effi-ciency is obtained using touchdown and hot-start PCR. Conclusion The stable and reproducible PCR amplification parameters for mutational hotspots in the human catalase gene were optimized.%目的 探讨过氧化氢酶基因突变热点区域PCR扩增方法 ,提高PCR反应的特异性和灵敏度,有助于快速检测CAT基因相关疾病.方法 从人静脉血液标本提取人血液基因组DNA,设计引物扩增特定的CAT基因片段,联合应用热启动PCR和降落PCR技术.结果 建立了重复性好,分辨率高的PCR反应体系.结论 建立了适用于CAT基因突变热点区域的PCR反应体系,有助于快速检测CAT基因相关痰病.

  8. Marine teleost ortholog of catalase from rock bream (Oplegnathus fasciatus): molecular perspectives from genomic organization to enzymatic behavior with respect to its potent antioxidant properties.

    Science.gov (United States)

    Elvitigala, Don Anushka Sandaruwan; Premachandra, H K A; Whang, Ilson; Priyathilaka, Thanthrige Thiunuwan; Kim, Eunmi; Lim, Bong-Soo; Jung, Hyung-Bok; Yeo, Sang-Yeob; Park, Hae-Chul; Lee, Jehee

    2013-10-01

    Catalases are well known antioxidant enzymes that can mainly dismutate hydrogen peroxide into water and oxygen in order to prevent oxidative stress. The complete genomic DNA (gDNA) sequence of the catalase gene from rock bream (Oplegnathus fasciatus) was identified from our custom-constructed BAC genomic DNA library and designated as RbCat. RbCat consists of 13 exons, separated by 12 introns, within a 13,722-bp gDNA sequence. The complete cDNA sequence (3303 bp) of RbCat is comprised of a 1581-bp coding region, encoding a peptide of 527 amino acids (aa) in length, with a predicted molecular mass of 60 kDa and a theoretical isoelectric point of 8.34. The anticipated promoter region of RbCat contains several transcription factor-binding sites, including sites that bind with immune- and antioxidant-responsive signaling molecules, suggesting its substantial transcriptional regulation. RbCat resembles the typical catalase family signature, i.e., it is composed of the catalase proximal active site motif along with a catalase proximal heme-ligand signature motif and shares great homology with its fish counterparts. According to multiple sequence alignment, functionally important amino acids present in RbCat were thoroughly conserved among its vertebrate counterparts. Phylogenetic analysis revealed that RbCat evolved from a vertebrate origin, and further positioned it in the fish clade. Recombinant RbCat had noticeable peroxidase activity against its substrate, hydrogen peroxide, in a dose-dependent manner. However, it demonstrated substantial peroxidase activity within a broad range of temperatures and pH values. Constitutive RbCat mRNA expression of different magnitudes was detected in a tissue-specific manner, suggesting its diverse role in physiology with respect to the tissue type. Moreover, immune challenge experiments using Edwardsiella tarda and rock bream iridovirus (RBIV) as live pathogens and polyinosinic:polycytidylic acid and lipopolysaccharide as mitogens

  9. 过氧化氢酶活性测定的新方法%A novel method for assaying catalase activity

    Institute of Scientific and Technical Information of China (English)

    刘砚韬; 王振伟; 张伶俐

    2013-01-01

    OBJECTIVE To establish a novel method for determination of catalase activity.METHODS Two-step method was adopted in this research for determination of catalase activity.Firstly,the catalase catalyzed the breakdown of H2O2.Secondly,the peroxidase reacted with residual H2O2 and oxygen donor-TOOS which was oxygenated to a blue quinoneimine dye,then absorbance at 555 nm was determined to calculate the activity.The influence of factors in the reaction,such as chromogenic reagent,substrate concentration,reaction time,enzyme concentration,and sensitivity and stability of chromogenic reagent were compared with other methods.RESULTS Optimal reaction system was composed of pH 7.5,30 ℃ and 16.2 mmol·L-1 of H2O2 as well as TOOS as the chromogenic reagent.CONCLUSION This was a simple,quick and high sensitive method to determine catalase activity and was especially appropriate for assaying catalase activity in some enzymes used in diagnostic kit.%目的 建立一种简单、灵敏和快速检测过氧化氢酶(CAT)活性的方法.方法 采用两步法检测CAT活性.第一步是CAT催化H2O2分解,第二步是剩余的H2O2在CAT作用下,氧供体(TOOS)被氧化成蓝色醌亚胺物质,测定在555 nm处的吸光度,计算CAT活性.对反应的影响因素,如显色剂、底物浓度、反应时间、酶浓度、灵敏度和稳定性等,与其它CAT活性测定方法进行比较分析.结果 从pH7.5、30℃、16.2 mmol·L-1作为最佳反应条件,TOOS作显色试剂.结论 所建方法用于测定CAT活性,具有简单、快速和灵敏度高等特点,尤其适用于诊断试剂盒中CAT活性的检测.

  10. The influence of detorsion of adnexal torsion on melondialdehyde, peroxidase catalase, and catalase of ovarian tissue in rabbits%解除附件扭转对兔卵巢组织MDA、GSH-Px和CAT的影响

    Institute of Scientific and Technical Information of China (English)

    于月新; 李巨; 陈佳; 樊宝剑; 颜宇博

    2013-01-01

    Objective To observe the influence of reperfusion after adnexal torsion (AT) on malondialdehyde (MDA), glutathione peroxidase catalase (GSH-Px) and catalase (CAT) contents of ovarian tissue in rabbits. Methods Forty female Japanese long-eared white rabbits were randomly divided into study group (n=32) and control group (n=8). The left adnexa of rabbits in the study group was clockwise twisted three laps, and then fixed on the left abdominal walls. Adnexal detorsion was then done 24 hours after adnexal torsion in the study group, and then the rabbits were divided into 4 groups (8 each). Both ovaries of each group were removed 24h, 48h, 72h and 96h, respectively, after reperfusion. The rabbits in the control group received sham-operation and both ovaries were removed 96h later. The removed left ovaries were used for biochemical detection of GSH-Px, CAT and MDA. All the right ovaries were used as experimental internal-control. Results The activity of GSH-Px declined significantly 24h to 72h after adnexal detorsion (P0.05). The activity of CAT declined significantly 24h and 48h after adnexal detorsion (P0.05). Conclusions Detorsion after adnexal torsion can affect the degree of oxidative stress injury in rabbits' ovaries as shown by the changes in the activities of GSH-Px, CAT and MDA. With the elongation of detorsion time, ovarian injury will be gradually alleviated.%目的 观察附件扭转(AT)后再灌注对兔卵巢丙二醛(MDA)、谷胱甘肽-过氧化物酶(GSH-Px)、过氧化氢酶(CAT)的影响.方法 雌性成年日本大耳白兔40只,随机分为实验组(n=32)和对照组(n=8).实验组兔将左侧附件按顺时针方向扭转3周后固定于左侧腹壁,24h后解除扭转,再分成4组,每组8只,分别于24、48、72、96h后取双侧卵巢.对照组行假手术,96h后取双侧卵巢.取左侧卵巢检测GSH-Px、CAT活性及MDA含量,以切除的右侧卵巢作为内对照.结果 附件扭转解除后,GSH-Px活力于再灌注24h至72h

  11. 过氧化氢酶与糖尿病的研究进展%Catalase and diabetes mellitus

    Institute of Scientific and Technical Information of China (English)

    赵瑞娥; 刘丽梅

    2008-01-01

    Catalase(CAT)can decompose hydrogen peroxide(H2O2),a product of peroxide dismuted by superoxide dismutase,into oxygen and water,so as to reduce the generation of toxic reactive hydroxyl radicals and prevent damage and dysfunction of islet β cells.Previous studies have shown that two polymorphisms,C-262T and C1167T,of CAT gene were significantly associated with diabetes mellitus,and that their C alleles were risk factors of type 1 diabetes mellitus.Moreover,other ethnical studies demonstrated that both genetic defects and pelymorphisms of the CAT gene could reduce the activity of CAT,increase the concentration of the H2O2 and therefore aggravate the oxidative stress.These data suggested that genetic variation of CAT may be an important pathophysiological mechanism in the development of diabetes and its complications.%过氧化氢酶(CAT)能迅速将超氧化物歧化酶分解过氧化物产生的过氧化氢(H2O2)转化为氧气和水,从而减少更具氧化活性的羟自由基生成,防止氧化应激造成的胰岛β细胞损伤和功能异常.研究发现,CAT基因的两个多态位点C-262T及C1167T均与糖尿病发生显著相关,其C等位基因均为1型糖尿病发病的危险因子.此外,其他种族研究证实,CAT的遗传学缺陷及其基因多态性均可引起CAT活性下降,使机体H2O2浓度增加,加重氧化应激.提示CAT的遗传变异可能是促进糖尿病及其并发症发生和发展的重要病理生理机制之一.

  12. SO4(=) uptake and catalase role in preconditioning after H2O2-induced oxidative stress in human erythrocytes.

    Science.gov (United States)

    Morabito, Rossana; Remigante, Alessia; Di Pietro, Maria Letizia; Giannetto, Antonino; La Spada, Giuseppina; Marino, Angela

    2017-02-01

    Preconditioning (PC) is an adaptive response to a mild and transient oxidative stress, shown for the first time in myocardial cells and not described in erythrocytes so far. The possible adaptation of human erythrocytes to hydrogen peroxide (H2O2)-induced oxidative stress has been here verified by monitoring one of band 3 protein functions, i.e., Cl(-)/HCO3(-) exchange, through rate constant for SO4(=) uptake measurement. With this aim, erythrocytes were exposed to a mild and transient oxidative stress (30 min to either 10 or 100 μM H2O2), followed by a stronger oxidant condition (300- or, alternatively, 600-μM H2O2 treatment). SO4(=) uptake was measured by a turbidimetric method, and the possible role of catalase (CAT, significantly contributing to the anti-oxidant system in erythrocytes) in PC response has been verified by measuring the rate of H2O2 degradation. The preventive exposure of erythrocytes to 10 μM H2O2, and then to 300 μM H2O2, significantly ameliorated the rate constant for SO4(=) uptake with respect to 300 μM H2O2 alone, showing thus an adaptive response to oxidative stress. Our results show that (i) SO4(=) uptake measurement is a suitable model to monitor the effects of a mild and transient oxidative stress in human erythrocytes, (ii) band 3 protein anion exchange capability is retained after 10 μM H2O2 treatment, (iii) PC response induced by the 10 μM H2O2 pretreatment is clearly detected, and (iv) PC response, elicited by low-concentrated H2O2, is mediated by CAT enzyme and does not involve band 3 protein tyrosine phosphorylation pathways. Erythrocyte adaptation to a short-term oxidative stress may serve as a basis for future studies about the impact of more prolonged oxidative events, often associated to aging, drug consumption, chronic alcoholism, hyperglycemia, or neurodegenerative diseases.

  13. Clonal spread of catalase-negative ST5/SCCmec II Staphylococcus aureus carrying the staphylococcal enterotoxin A (sea), staphylococcal enterotoxin b (seb), and toxic shock toxin (tst) virulence genes.

    Science.gov (United States)

    Lee, Hae Kyung; Kim, Jung-Beom; Kim, Hyunjung; Jekarl, Dong Wook; Kim, Yang Ree; Yu, Jin Kyung; Park, Yeon-Joon

    2014-01-01

    17 catalase-negative methicillin-resistant Staphylococcus aureus (MRSA) isolates were recovered from respiratory specimens of patients at a 700-bed hospital in Korea. The goal of this study was to determine the molecular characteristics of catalase-negative MRSA strains in Korea for the first time. Characteristics that we explored included kat A gene mutation sequence, sequence type, staphylococcal cassette chromosome (SCC) mec subtype classification, and toxin gene profiles. All 17 isolates showed similar pulsed field gel electrophoresis (PFGE) pattern. Four mutations were identified in the kat A gene of a representative catalase-negative MRSA strain: A602G causing a histidine 201 to arginine change, A695T causing a glutamic acid 232 to valine change, T778A causing a tryptophan 260 to arginine change, and G1438A causing a glycine 480 to serine change. Previous studies suggest that the A695T and T778A mutations may have strong effects on the catalase activity of catalase-negative MRSA. The sequence type (ST) and SCCmec type of this isolate were ST 5 and SCCmec type II, respectively. All 17 isolates harbored toxic shock toxin (tst), staphylococcal enterotoxin A (sea), and staphylococcal enterotoxin B (seb) virulence genes. The mortality rate of the present study was 11.8%, suggesting that the clinical relevance of catalase-negative MRSA requires further study in the future.

  14. Gaharu Leaf Extract Water Reduce MDA and 8-OHdG Levels and Increase Activities SOD and Catalase in Wistar Rats Provided Maximum Physical Activity

    Directory of Open Access Journals (Sweden)

    I Made Oka Adi Parwata

    2016-09-01

    Full Text Available Background: Oxidative stress occurs due to an imbalance of the number of free radicals by the number of endogenous antioxidant produced by the body i.e. Superoxide Dismutase (SOD, Gluthathione Peroxidase (GPx, and Catalase. The imbalance between the number of free radicals and antioxidants can be overcome with the endogenous antioxidant intake that exogenous oxidative stress can be reduced. One of exogenous antioxidants is natural Gaharu leaf water extract. Objective: This research focus on the effect of Gaharu leaf water extract in reducing MDA and 8-OHdG and increase the activity of SOD and Catalase. Methods: This study was an experimental with post only controls group design. Experiment was divided  into 5 groups of wistar rats, each consisting of 5 animals, i.e. negative control group without extract [K (-], treatment 1 treated 50 mg/kg BW/day of the extract (T1, treatment 2 treated 100 mg/kg BW/day of the extract (T2, treatment 3 treated 200 mg/ kg BW/day of the extract (T3, and positive control group [K (+] treated with vitamin Cat a dose 50 mg/kg BW/day. All groups treated for 10 weeks. Every day, before treatment, each group was given a maximum swimming activity for 1.5 hours for 10 weeks. ELISA was used to measure MDA, 8-OHdG, SOD, and Catalase activities. Result: The research results showed that treatment of extract of  leaves of Gaharu with an higher dose from 50 mg/kg BW up to 200 mg/ kg BW significantly decline (p <0.05 levels of MDA with the average ranging from 6.37±0.23, 5,56±0.27 and 4.32±0.27, 8-OHdG with a mean of 1.64±0.11, 1.26±0.46, and 1.09±0.17. On the other hand the treatment also increase SOD activity with less ranging from 12.15±1.04, 15.70±2.02, and 18.84±1.51, and Catalase ranging from 6,68±0.63, 8.20±1.14 and 9.29±0,79 in the blood of Wistar rats were given a maximum activity compared to the negative control group. This is probably higher phenol compounds (bioflavonoids quantity content of the extract

  15. Eukaryotic extracellular catalase-peroxidase from Magnaporthe grisea - Biophysical/chemical characterization of the first representative from a novel phytopathogenic KatG group.

    Science.gov (United States)

    Zámocký, Marcel; Droghetti, Enrica; Bellei, Marzia; Gasselhuber, Bernhard; Pabst, Martin; Furtmüller, Paul G; Battistuzzi, Gianantonio; Smulevich, Giulietta; Obinger, Christian

    2012-03-01

    All phytopathogenic fungi have two catalase-peroxidase paralogues located either intracellularly (KatG1) or extracellularly (KatG2). Here, for the first time a secreted bifunctional, homodimeric catalase-peroxidase (KatG2 from the rice blast fungus Magnaporthe grisea) has been produced heterologously with almost 100% heme occupancy and comprehensively investigated by using a broad set of methods including UV-Vis, ECD and resonance Raman spectroscopy (RR), thin-layer spectroelectrochemistry, mass spectrometry, steady-state & presteady-state spectroscopy. RR spectroscopy reveals that MagKatG2 shows a unique mixed-spin state, non-planar heme b, and a proximal histidine with pronounced imidazolate character. At pH 7.0 and 25 °C, the standard reduction potential E°' of the Fe(III)/Fe(II) couple for the high-spin native protein was found to fall in the range typical for the KatG family. Binding of cyanide was relatively slow at pH 7.0 and 25 °C and with a K(d) value significantly higher than for the intracellular counterpart. Demonstrated by mass spectrometry MagKatG2 has the typical Trp118-Tyr251-Met277 adduct that is essential for its predominantly catalase activity at the unique acidic pH optimum. In addition, MagKatG2 acts as a versatile peroxidase using both one- and two-electron donors. Based on these data, structure-function relationships of extracellular eukaryotic KatGs are discussed with respect to intracellular KatGs and possible role(s) in host-pathogen interaction.

  16. Immobilization of catalase on electrospun PVA/PA6-Cu(II) nanofibrous membrane for the development of efficient and reusable enzyme membrane reactor.

    Science.gov (United States)

    Feng, Quan; Zhao, Yong; Wei, Anfang; Li, Changlong; Wei, Qufu; Fong, Hao

    2014-09-02

    In this study, a mat/membrane consisting of overlaid PVA/PA6-Cu(II) composite nanofibers was prepared via the electrospinning technique followed by coordination/chelation with Cu(II) ions; an enzyme of catalase (CAT) was then immobilized onto the PVA/PA6-Cu(II) nanofibrous membrane. The amount of immobilized catalase reached a high value of 64 ± 4.6 mg/g, while the kinetic parameters (Vmax and Km) of enzyme were 3774 μmol/mg·min and 41.13 mM, respectively. Furthermore, the thermal stability and storage stability of immobilized catalase were improved significantly. Thereafter, a plug-flow type of immobilized enzyme membrane reactor (IEMR) was assembled from the PVA/PA6-Cu(II)-CAT membrane. With the increase of operational pressure from 0.02 to 0.2 MPa, the flux value of IEMR increased from 0.20 ± 0.02 to 0.76 ± 0.04 L/m(2)·min, whereas the conversion ratio of H2O2 decreased slightly from 92 ± 2.5% to 87 ± 2.1%. After 5 repeating cycles, the production capacity of IEMR was merely decreased from 0.144 ± 0.006 to 0.102 ± 0.004 mol/m(2)·min. These results indicated that the assembled IEMR possessed high productivity and excellent reusability, suggesting that the IEMR based on electrospun PVA/PA6-Cu(II) nanofibrous membrane might have great potential for various applications, particularly those related to environmental protection.

  17. Enhanced killing of penicillin-treated gram-positive cocci by human granulocytes: role of bacterial autolysins, catalase, and granulocyte oxidative pathways.

    Science.gov (United States)

    Isturiz, R; Metcalf, J A; Root, R K

    1985-01-01

    Staphylococci pretreated with subminimal inhibitory concentrations (subMIC) of cell-wall active antibiotics exhibit increased susceptibility to killing by human polymorphonuclear leukocytes (PMNs), even when phagosome information is impaired by the mold metabolite, cytochalasin B. To investigate the role of specific bacterial factors in the process, studies were carried out with organisms lacking catalase (streptococci) or cell-wall autolytic enzymes and compared to findings with Staphylococcus aureus 502A. Neutrophil factors were studied using inhibitors, oxygen radical scavengers, myeloperoxidase (MPO)-deficient PMNs, or PMNs from a patient with chronic granulomatous disease (CGD). Documentation of the enhanced susceptibility of the streptococcal strains to killing by PMNs following subMIC penicillin pretreatment required the use of cytochalasin B. Enhancement of killing occurred independent of the presence or absence of bacterial autolysins or catalase. SubMIC penicillin pretreatment of S. pneumoniae R36A specifically promoted the susceptibility of these organisms to killing by myeloperoxidase (MPO)-mediated mechanisms (enhancement lost using MPO-deficient or azide-treated cells). Factors other than MPO or toxic oxygen products generated by the PMN respiratory burst are responsible for enhanced killing of penicillin-pretreated S. aureus 502A (enhancement preserved using MPO-deficient, azide-treated, or chronic granulomatous disease patient cells). These studies define methods to study the interaction of antimicrobial agents and PMNs in the killing of microorganisms. They also demonstrate that penicillin treatment can change the susceptibility of gram-positive cocci to the action of specific PMN microbicidal mechanisms. The mechanism of the enhancement appears to be bacterial strain-dependent and not predictable by bacterial autolysin or catalase activity.

  18. Effect on post-cryopreserved semen characteristics of Holstein bulls of adding combinations of vitamin C and either catalase or reduced glutathione to Tris extender.

    Science.gov (United States)

    Eidan, Sajeda M

    2016-04-01

    This study was undertaken to investigate the influence of adding combinations of vitamin C to Tris extender with either catalase or reduced glutathione on post-cryopreserved semen characteristics of Holstein bulls for different preservation periods (cooling at 5°C, 48 h, 1, 2 and 3 months post cryopreservation, PC). Seven Holstein bulls of 2.5-3 years of age were used in this experiment. Semen was collected via artificial vagina in one ejaculate per bull per week for the 7 week experimental period. Pooled semen was equally divided into three treatments using Tris extender. Combinations of vitamin C (2.5mM) were added with either catalase (100 IU/ml, T2) or reduced glutathione (2mM, T3) to Tris extender and comparisons in response were made with the control group (Tris extender, T1). Individual sperm motility (IM), viability (V), plasma membrane integrity (PMI), and acrosome integrity (AI) were assessed during all periods of the study along with Malondialdehyde (MDA) concentrations and freezing ability. The IM was greater (P ≤ 0.01) in the T2 as compared with the T1 group at all periods of the study. Furthermore, the IM were greater (P ≤ 0.01) in the T3 as compared with the T1 group at the 48 h time period and at 3 months PC. The V, PMI and AI were greater (P ≤ 0.01) in T2 and T3 as compared with the T1 group at all the experimental periods. The MDA was greater (P ≤ 0.01) in the T2 as compared with the T1 group at 3 months PC. In conclusion, there was improved semen quality if semen of Holstein bulls was collected and stored in combinations of vitamin C with either catalase (T2) or reduced glutathione (T3) being added to Tris extender.

  19. Purification and immobilization of a thermoalkaliphilic catalase from Thermoascus aurantiacus%一种嗜热嗜碱过氧化氢酶的分离纯化和固定化

    Institute of Scientific and Technical Information of China (English)

    柯尊柱; 张朝晖; 陈小龙

    2011-01-01

    After treating with (NH4)2SO4 fractional precipitation and DEAE-Sepharose anion-exchange column, a thermoalkaliphilic catalase from Thermoascus aurantiacus was purified. The catalase identified by SDS-PAGE was composed of two identical sub-units and had a molecular weight of 1. 9×105.The catalase was highly active over a temperature range from 25℃ to 751 and a pH range from 7 to 13, having the optimum activity at 75℃ and pH 12. These results demonstrated that the catalase was thermoalkaliphilic. The Km value for the purified catalase was 40. 74 mmol ? L-1 and the k2 value was 1. 59X104 μmol ? (min ? Mg protein)"1. The catalase was immobilized onto chitosan beads by covalent binding. The immobilized catalase showed higher pH stability and thermostability than the free catalase. It retained about 55% of its initial activity after 13 repeated cycles during the total 130 min reaction time. The properties mentioned above showed that this catalase had a potential application in removing residual hydrogen peroxide from bleaching streams in industrial bleaching processes.%引言过氧化氢酶(catalase,简称CAT,EC 1.11.1.6)是一种高效催化分解过氧化氢的酶,广泛存在于好氧微生物和动植物体内.在纺织印染工业、造纸工业、废水处理、纸浆、牛奶保鲜以及临床分析等方面有着广泛的应用[1-3].其中嗜热嗜碱过氧化氢酶由于其独特的耐热耐碱性质,已有多篇文献报道该酶在处理纺织业漂白废水中残留H2O2的应用研究[4-8].

  20. KatG, the Bifunctional Catalase of Xanthomonas citri subsp. citri, Responds to Hydrogen Peroxide and Contributes to Epiphytic Survival on Citrus Leaves

    Science.gov (United States)

    Tondo, María Laura; Delprato, María Laura; Kraiselburd, Ivana; Fernández Zenoff, María Verónica; Farías, María Eugenia; Orellano, Elena G.

    2016-01-01

    Xanthomonas citri subsp. citri (Xcc) is the bacterium responsible for citrus canker. This bacterium is exposed to reactive oxygen species (ROS) at different points during its life cycle, including those normally produced by aerobic respiration or upon exposition to ultraviolet (UV) radiation. Moreover, ROS are key components of the host immune response. Among enzymatic ROS-detoxifying mechanisms, catalases eliminate H2O2, avoiding the potential damage caused by this specie. Xcc genome includes four catalase genes. In this work, we studied the physiological role of KatG, the only bifunctional catalase of Xcc, through the construction and characterization of a modified strain (XcckatG), carrying an insertional mutation in the katG gene. First, we evaluated the involvement of KatG in the bacterial adaptive response to H2O2. XcckatG cultures exhibited lower catalase activity than those of the wild-type strain, and this activity was not induced upon treatment with sub-lethal doses of H2O2. Moreover, the KatG-deficient mutant exhibited decreased tolerance to H2O2 toxicity compared to wild-type cells and accumulated high intracellular levels of peroxides upon exposure to sub-lethal concentrations of H2O2. To further study the role of KatG in Xcc physiology, we evaluated bacterial survival upon exposure to UV-A or UV-B radiation. In both conditions, XcckatG showed a high mortality in comparison to Xcc wild-type. Finally, we studied the development of bacterial biofilms. While structured biofilms were observed for the Xcc wild-type, the development of these structures was impaired for XcckatG. Based on these results, we demonstrated that KatG is responsible for Xcc adaptive response to H2O2 and a key component of the bacterial response to oxidative stress. Moreover, this enzyme plays an important role during Xcc epiphytic survival, being essential for biofilm formation and UV resistance. PMID:26990197

  1. Propagation of human iPS cells in alginate-based microcapsules prepared using reactions catalyzed by horseradish peroxidase and catalase.

    Science.gov (United States)

    Ashida, Tomoaki; Sakai, Shinji; Taya, Masahito

    2016-09-01

    Cell encapsulation has been investigated as a bioproduction system in the biomedical and pharmaceutical fields. We encaps-ulated human induced pluripotent stem (hiPS) cells in duplex microcapsules prepared from an alginate derivative possessing phenolic hydroxyl moieties, in a single-step procedure based on two competing enzymatic reactions catalyzed by horseradish peroxidase (HRP) and catalase. The encapsulated cells maintained 91.4% viability and proliferated to fill the microcapsules following 19 days of culture. Encapsulated hiPS cells showed pluripotency comparable to that of unencapsulated cells during the cultures, as demonstrated by the expression of the SSEA-4 marker.

  2. KatG, the Bifunctional Catalase of Xanthomonas citri subsp. citri, Responds to Hydrogen Peroxide and Contributes to Epiphytic Survival on Citrus Leaves.

    Directory of Open Access Journals (Sweden)

    María Laura Tondo

    Full Text Available Xanthomonas citri subsp. citri (Xcc is the bacterium responsible for citrus canker. This bacterium is exposed to reactive oxygen species (ROS at different points during its life cycle, including those normally produced by aerobic respiration or upon exposition to ultraviolet (UV radiation. Moreover, ROS are key components of the host immune response. Among enzymatic ROS-detoxifying mechanisms, catalases eliminate H2O2, avoiding the potential damage caused by this specie. Xcc genome includes four catalase genes. In this work, we studied the physiological role of KatG, the only bifunctional catalase of Xcc, through the construction and characterization of a modified strain (XcckatG, carrying an insertional mutation in the katG gene. First, we evaluated the involvement of KatG in the bacterial adaptive response to H2O2. XcckatG cultures exhibited lower catalase activity than those of the wild-type strain, and this activity was not induced upon treatment with sub-lethal doses of H2O2. Moreover, the KatG-deficient mutant exhibited decreased tolerance to H2O2 toxicity compared to wild-type cells and accumulated high intracellular levels of peroxides upon exposure to sub-lethal concentrations of H2O2. To further study the role of KatG in Xcc physiology, we evaluated bacterial survival upon exposure to UV-A or UV-B radiation. In both conditions, XcckatG showed a high mortality in comparison to Xcc wild-type. Finally, we studied the development of bacterial biofilms. While structured biofilms were observed for the Xcc wild-type, the development of these structures was impaired for XcckatG. Based on these results, we demonstrated that KatG is responsible for Xcc adaptive response to H2O2 and a key component of the bacterial response to oxidative stress. Moreover, this enzyme plays an important role during Xcc epiphytic survival, being essential for biofilm formation and UV resistance.

  3. Changing of Bacteria Catalase Activity Under the Influence of Electro-Magnetic Radiation on a Frequency of Nitric Oxide Absorption and Radiation Molecular Spectrum

    Directory of Open Access Journals (Sweden)

    G.M. Shub

    2009-09-01

    Full Text Available The dynamics of catalase activity degree changing in Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa is described under the influence of electro-magnetic radiation on a frequency of nitric oxide absorption and radiation molecular spectrum. The panoramic spectrometric measuring complex, developed in Central Scientific Research Institute of measuring equipment Public corporation, Saratov, was used while carrying out the research. Electromagnetic vibrations of extremely high frequencies were stimulated in this complex imitating the structure of nitric oxide absorption and radiation molecular spectrum. The growth of activity of the mentioned enzyme of the strains under research was detected. The most significant changes were observed under 60-minutes exposure.

  4. Identification of Scedosporium boydii catalase A1 gene, a reactive oxygen species detoxification factor highly expressed in response to oxidative stress and phagocytic cells.

    Science.gov (United States)

    Mina, Sara; Staerck, Cindy; d'Almeida, Sènan M; Marot, Agnès; Delneste, Yves; Calenda, Alphonse; Tabiasco, Julie; Bouchara, Jean-Philippe; Fleury, Maxime J J

    2015-12-01

    Scedosporium boydii is an opportunistic filamentous fungus which may be responsible for a large variety of infections in both immunocompetent and immunocompromised individuals. This fungus belongs to the Scedosporium apiospermum species complex which usually ranks second among the filamentous fungi colonizing the airways of patients with cystic fibrosis (CF). Species of the S. apiospermum complex are able to chronically colonize the CF airways suggesting pathogenic mechanisms allowing persistence and growth of these fungi in the respiratory tract. Few putative virulence factors have been purified and characterized so far in the S. apiospermum complex including a cytosolic Cu,Zn-superoxide dismutase (SOD) and a monofunctional catalase (catalase A1). Upon microbial infection, host phagocytes release reactive oxygen species (ROS), such as hydrogen peroxide, as part of the antimicrobial response. Catalases are known to protect pathogens against ROS by degradation of the hydrogen peroxide. Here, we identified the S. boydii catalase A1 gene (CATA1) and investigated its expression in response to the environmental conditions encountered in the CF airways and to the oxidative stress. Results showed that S. boydii CATA1 gene expression is not affected by hypoxia, hypercapnia or pH changes. In contrast, CATA1 gene was overexpressed in response to a chemically induced oxidative stress with a relative gene expression 37-fold higher in the presence of 250 μM H(2)O(2), 20-fold higher with 250 μM menadione and 5-fold higher with 2 mM paraquat. Moreover, S. boydii CATA1 gene expression progressively increased upon exposure to activated THP-1-derived macrophages, reaching a maximum after 12 h (26 fold). Activated HL60-derived neutrophils and activated human peripheral blood neutrophils more rapidly induced S. boydii CATA1 gene overexpression, a maximum gene expression level being reached at 75 min (17 fold) and 60 min (15 fold), respectively. In contrast expression of the gene

  5. 细菌过氧化氢酶基因的克隆方法研究%An Effective Method for Cloning Catalase Gene from Bacteria

    Institute of Scientific and Technical Information of China (English)

    李充璧; 李琛; 李赛男

    2012-01-01

    [Objective] To study a method for cloning the active catalase from bacteria. [Method] E. coli W3110 was cultivated and genomic DNA was extracted and then was partially digested with restrictive enzyme Sau3AI. Genomic DNA fragments over 1.5 kb were separated and purified. Vector pUC18 after digested with BamH I was linked to the different DNA fragements partially digested with Sau3AI by ligases T4. And then the new ligated products were transformed into the E. coli competent DH5a. The transformers were incubated ananaer obically on brain heart infusion (BHI)containing tannic acid,then were kept aerobically for a further 24 h. The catalase-positive clones were screened. [ Result] Through PCR identification and restriction enzyme digestion analysis,the recombinant plasmid Pucl8-cat was constrcted successfully. The catalase gene cloned was correct in conforming to the theory. [Conclusion] This method lays a foundation for rapidly and simply cloning an active catalase.%[目的]研究细菌过氧化酶基因的克隆方法.[方法]培养扩增E.coli W3110,提取其基因组DNA,采用限制性内切酶Sau3A I对其进行部分酶切,分离纯化1.5 kb以上的DNA片段.将经BamH I酶切的大肠杆菌质粒pUC18与经Sau3A I部分酶切的不同片段用T4连接酶进行连接,并转化大肠杆菌DH5α.通过BHI-单宁酸培养介质,筛选出含有活性过氧化氢酶基因的重组子.[结果]经酶切和PCR鉴定,证实所克隆的过氧化氢酶基因是正确的,与理论相符.[结论]该研究结果为快速简便地克隆筛选活性细菌过氧化氢酶基因奠定了基础.

  6. KatG, the Bifunctional Catalase of Xanthomonas citri subsp. citri, Responds to Hydrogen Peroxide and Contributes to Epiphytic Survival on Citrus Leaves.

    Science.gov (United States)

    Tondo, María Laura; Delprato, María Laura; Kraiselburd, Ivana; Fernández Zenoff, María Verónica; Farías, María Eugenia; Orellano, Elena G

    2016-01-01

    Xanthomonas citri subsp. citri (Xcc) is the bacterium responsible for citrus canker. This bacterium is exposed to reactive oxygen species (ROS) at different points during its life cycle, including those normally produced by aerobic respiration or upon exposition to ultraviolet (UV) radiation. Moreover, ROS are key components of the host immune response. Among enzymatic ROS-detoxifying mechanisms, catalases eliminate H2O2, avoiding the potential damage caused by this specie. Xcc genome includes four catalase genes. In this work, we studied the physiological role of KatG, the only bifunctional catalase of Xcc, through the construction and characterization of a modified strain (XcckatG), carrying an insertional mutation in the katG gene. First, we evaluated the involvement of KatG in the bacterial adaptive response to H2O2. XcckatG cultures exhibited lower catalase activity than those of the wild-type strain, and this activity was not induced upon treatment with sub-lethal doses of H2O2. Moreover, the KatG-deficient mutant exhibited decreased tolerance to H2O2 toxicity compared to wild-type cells and accumulated high intracellular levels of peroxides upon exposure to sub-lethal concentrations of H2O2. To further study the role of KatG in Xcc physiology, we evaluated bacterial survival upon exposure to UV-A or UV-B radiation. In both conditions, XcckatG showed a high mortality in comparison to Xcc wild-type. Finally, we studied the development of bacterial biofilms. While structured biofilms were observed for the Xcc wild-type, the development of these structures was impaired for XcckatG. Based on these results, we demonstrated that KatG is responsible for Xcc adaptive response to H2O2 and a key component of the bacterial response to oxidative stress. Moreover, this enzyme plays an important role during Xcc epiphytic survival, being essential for biofilm formation and UV resistance.

  7. Caribbean yellow band disease compromises the activity of catalase and glutathione S-transferase in the reef-building coral Orbicella faveolata exposed to anthracene.

    Science.gov (United States)

    Montilla, Luis Miguel; Ramos, Ruth; García, Elia; Cróquer, Aldo

    2016-05-03

    Healthy and diseased corals are threatened by different anthropogenic sources, such as pollution, a problem expected to become more severe in the near future. Despite the fact that coastal pollution and coral diseases might represent a serious threat to coral reef health, there is a paucity of controlled experiments showing whether the response of diseased and healthy corals to xenobiotics differs. In this study, we exposed healthy and Caribbean yellow band disease (CYBD)-affected Orbicella faveolata colonies to 3 sublethal concentrations of anthracene to test if enzymatic responses to this hydrocarbon were compromised in CYBD-affected tissues. For this, a 2-factorial fully orthogonal design was used in a controlled laboratory bioassay, using tissue condition (2 levels: apparently healthy and diseased) and pollutant concentration (4 levels: experimental control, 10, 30 and 100 ppb concentration) as fixed factors. A permutation-based ANOVA (PERMANOVA) was used to test the effects of condition and concentration on the specific activity of 3 enzymatic biomarkers: catalase, glutathione S-transferase, and glutathione peroxidase. We found a significant interaction between the concentration of anthracene and the colony condition for catalase (Pseudo-F = 3.84, df = 3, p anthracene in CYBD-affected tissues was compromised, as the activity of these enzymes decreased 3- to 4-fold compared to healthy tissues. These results suggest that under a potential scenario of increasing hydrocarbon coastal pollution, colonies of O. faveolata affected with CYBD might become more vulnerable to the deleterious effects of chemical pollution.

  8. Effects of catalase, 2% chlorhexidine gel and 1% sodium hypochlorite on the microtensile bond strength of teeth bleached with 35% hydrogen peroxide

    Directory of Open Access Journals (Sweden)

    Ricardo Ferreira

    2011-07-01

    Full Text Available Introduction and objective: Dental bleaching is an effective, relatively simple and noninvasive technique. The aim of this study was to evaluate the effects of catalase, 2% chlorhexidine gel, and 1% sodium hypochlorite on the microtensile bond strength to enamel of bovine teeth submitted to internal and external bleaching with 35% hydrogen peroxide. Material and methods: Sixty bovine incisors were used. They had their debris removed, washed in tap water and stored frozen. The samples were divided into five experimental groups according to the treatment applied after bleaching (n = 12: 1 − control/no bleaching (C; 2 − catalase (CA; 3 − 2% chlorhexidine gel (CG; 4 − 1% sodium hypochlorite (SH; 5 − distilled water (DW. For microtensile test, samples were prepared into blocks of enamel/resin, which were sectioned to obtain hourglass-like specimens. Bond strength was calculated in MPa and data analyzed statistically by Anova (p < 0.05. Results: Microtensile bond strength means decreased in comparison to control group, but no statistically significant difference between groups was found. Conclusion: The substances used after dental bleaching did not result in statistically significant microtensile bond strength means of the tested groups.

  9. Fluctuations in peroxidase and catalase activities of resistant and susceptible black gram (Vigna mungo (L.) Hepper) genotypes elicited by Bemisia tabaci (Gennadius) feeding.

    Science.gov (United States)

    Taggar, Gaurav Kumar; Gill, Ranjit Singh; Gupta, Anil Kumar; Sandhu, Jeet Singh

    2012-10-01

    Whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleryrodidae), is a serious pest of black gram, (Vigna mungo (L.) Hepper), an important legume pulse crop grown in north India. This research investigated the potential role of selected plant oxidative enzymes in resistance/susceptibility to whitefly in nine black gram genotypes. Oxidative enzyme activity was estimated spectrophotometrically from leaf samples collected at 30 and 50 d after sowing (DAS) from whitefly infested and uninfested plants. The enzymes showed different activity levels at different times after the infestation. The results indicated that in general, whitefly infestation increased the activities of peroxidase and decreased the catalase activity. Resistant genotypes NDU 5-7 and KU 99-20 recorded higher peroxidase and catalase activities at 30 and 50 DAS under whitefly-stress conditions as compared with non-stressed plants. The results suggest that the enhanced activities of the enzymes may contribute to bioprotection of black gram plants against B. tabaci infestation. The potential mechanisms to explain the correlation of resistance to whitefly in black gram genotypes with higher activities of oxidative enzymes are also discussed.

  10. Peroxiredoxin-glutaredoxin and catalase promote resistance of nontypeable Haemophilus influenzae 86-028NP to oxidants and survival within neutrophil extracellular traps.

    Science.gov (United States)

    Juneau, Richard A; Pang, Bing; Armbruster, Chelsie E; Murrah, Kyle A; Perez, Antonia C; Swords, W Edward

    2015-01-01

    Nontypeable Haemophilus influenzae (NTHI) is a common commensal and opportunistic pathogen of the human airways. For example, NTHI is a leading cause of otitis media and is the most common cause of airway infections associated with chronic obstructive pulmonary disease (COPD). These infections are often chronic/recurrent in nature and involve bacterial persistence within biofilm communities that are highly resistant to host clearance. Our previous work has shown that NTHI within biofilms has increased expression of factors associated with oxidative stress responses. The goal of this study was to define the roles of catalase (encoded by hktE) and a bifunctional peroxiredoxin-glutaredoxin (encoded by pdgX) in resistance of NTHI to oxidants and persistence in vivo. Isogenic NTHI strain 86-028NP mutants lacking hktE and pdgX had increased susceptibility to peroxide. Moreover, these strains had persistence defects in the chinchilla infection model for otitis media, as well as in a murine model for COPD. Additional work showed that pdgX and hktE were important determinants of NTHI survival within neutrophil extracellular traps (NETs), which we have shown to be an integral part of NTHI biofilms in vivo. Based on these data, we conclude that catalase and peroxiredoxin-glutaredoxin are determinants of bacterial persistence during chronic/recurrent NTHI infections that promote bacterial survival within NETs.

  11. Degradation of direct yellow 9 by electro-Fenton: process study and optimization and, monitoring of treated water toxicity using catalase.

    Science.gov (United States)

    Kourdali, Sidali; Badis, Abdelmalek; Boucherit, Ahmed

    2014-12-01

    The present study was undertaken to investigate the degradation and removal of direct yellow 9 (DY9) by the electro-Fenton (EF) process in batch reactor using iron and stainless steel electrodes. DY9 removal decreased with the increase in pH (3 to 8) and increased with the increase in current intensity (0.05 to 0.2A) and [H2O2] (0 to 0.5gL(-1), but not with high doses which led to low rates of DY9 removal and OH(∙) uptake). The regression quadratic models describing DY9 degradation yield "R (percent)" and electrical energy consumption "EEC (kWhkg(-1))" were validated by the analysis of variance (ANOVA) and were both noted to fit well with the experimental data. The R(2) correlation coefficients (0.995, 0.978), those adjusted coefficients (0.986, 0.939), and F values (110.7, 24.9) obtained for the responses validated the efficiency of model. The results revealed that among several other parameters, EEC depended essentially on the degradation yield. The eco-toxicity tests showed a positive correlation between catalase activity and DY9 concentration, and catalase could be qualitatively identified to assess the effect of dye and its by-products generated during the EF process.

  12. 过氧化氢酶与酵母菌共固定的研究%Co-immobilizing of Catalase and Yeast

    Institute of Scientific and Technical Information of China (English)

    胡杨; 雷女孝; 王征; 谢达平

    2001-01-01

    采用聚阳离子复合物,用于过氧化氢酶与酵母细胞的共固定化,可防止使用过程中酶的渗漏,改善了供氧效果和酵母的增殖。固定化酶的最适反应pH值范围为6~8。当聚阳离子浓度达150 mmol/L时,固定化酶活力回收可达68.6%,在室温条件下,5 d时酶活力下降58%。这种酶和酵母细胞的共固定颗粒,在1 d时间内使颗粒内酵母细胞增殖至50亿/g。%Catalase and yeast were immobilized together by polycation complex.Catalase leakage was overcome in the doing so. Oxygen supply and yeast multiplication were improved. The optimum pH of the immobilizing was 6~8.The activity recovery of the immobilized enzyme reached 68% as polycation concentration was kept at 150 mmol/L.At room temperature,the activity of enzyme decreased about 58% in the 5th day.In the co-immobilizing particle yeast cell increased up to 5×109 /g in one day.

  13. High level extracellular production of a recombinant alkaline catalase in E. coli BL21 under ethanol stress and its application in hydrogen peroxide removal after cotton fabrics bleaching.

    Science.gov (United States)

    Yu, Zhenxiao; Zheng, Hongchen; Zhao, Xingya; Li, Shufang; Xu, Jianyong; Song, Hui

    2016-08-01

    The effects of induction parameters, osmolytes and ethanol stress on the productivity of the recombinant alkaline catalase (KatA) in Escherichia coli BL21 (pET26b-KatA) were investigated. The yield of soluble KatA was significantly enhanced by 2% ethanol stress. And a certain amount of Triton X-100 supplementation could markedly improved extracellular ratio of KatA. A total soluble catalase activity of 78,762U/mL with the extracellular ratio of 92.5% was achieved by fed-batch fermentation in a 10L fermentor, which was the highest yield so far. The purified KatA showed high stability at 50°C and pH 6-10. Application of KatA for elimination of H2O2 after cotton fabrics bleaching led to less consumption of water, steam and electric power by 25%, 12% and 16.7% respectively without productivity and quality losing of cotton fabrics. Thus, the recombinant KatA is a promising candidate for industrial production and applications.

  14. Cu/Zn superoxide dismutase and catalase activities in Pinus mugo needles growing at elevated stands in the mountains, and their photochemical efficiency of PSII.

    Science.gov (United States)

    Miszalski, Zbigniew; Libik, Marta; Surówka, Ewa; Niewiadomska, Ewa

    2005-08-01

    Pinus mugo needles were sampled at different altitudes (1420, 1590 and 1920 m a.s.l.) to analyse levels of oxidative stress and changes in maximum photochemical efficiency of PSII. Polyacrylamide gel electrophoresis demonstrated that almost all superoxide dismutase activity represented Cu/Zn superoxide dismutase, and only 4-6% represents Mn superoxide dismutase. In extracts from plants sampled at 1590 and 1920 m a.s.l., lower activity of Cu/Zn superoxide dismutase was found. Comparing these data with immunoblots, it can be concluded that the differences in superoxide dismutase activity was related to protein amount. In needles from higher altitudes, a decrease in catalase activity was detected, as opposed to the protein amount, which was higher in needles from the higher stands. Considering the decrease in catalase and Cu/Zn superoxide dismutase activities in needles collected at 1590 and 1920 m a.s.l., we suggest that higher levels of oxidative stress may induce changes in photochemical efficiency of PSII.

  15. COMPARED ANALYSIS OF CATALASE AND PEROXIDASE ACTIVITY IN CELLULOLYTIC FUNGUS TRICHODERMA REESEI GROWN ON MEDIUM WITH DIFFERENT CONCENTRATIONS OF GRINDED WHEAT AND BARLEY STRAWS

    Directory of Open Access Journals (Sweden)

    Mihaela Cristica

    2010-09-01

    Full Text Available The purpose of this study was to assess the evolution of catalase and peroxidase activity in Trichoderma reesei grown on medium containing grinded wheat and barley straws. Carbon source of cultivation medium - glucose was replaced by various concentrations of grinded wheat and barley straws, finally resulting three experimental variants as follows: V1 = 20 g/l, V2 = 30 g/l, V3 = 40 g/l. ĂŽn addition to these variants a control sample was added in which composition remainded unchanged. The catalase activity was determined by spectrophotometric Sinha method (Artenie et al., 2008 while peroxidase activity was assesed using the o-dianisidine method (Cojocaru, 2009. Enzymatic determinations were carried out at 7 and 14 days from inoculation, in both fungus mycelium and culture liquid. The enzymatic assay showed significant differences between determinations intervals and work variants. Enzyme activity is influenced by the age of fungus and by the different nature of the substrate used.

  16. Effect of the Ascorbic Acid, Pyridoxine and Hydrogen Peroxide Treatments on Germination, Catalase Activity, Protein and Malondialdehyde Content of Three Oil Seeds

    Directory of Open Access Journals (Sweden)

    Aria DOLATABADIAN

    2008-08-01

    Full Text Available Oil seed production has an important role in human nutrition and industry. Success in oil plant cultivation is related to seed production with high viability and rapid germination, because these seeds rapidly loose their viability by fats oxidation. Thus, in this work we studied the effects of ascorbic acid, pyridoxine and hydrogen peroxide solutions on germination quantitative traits, catalase activity, protein and malondialdehyde content of three old oil seeds (sunflower, rape seed and safflower. The results showed that ascorbic acid and pyridoxine stimulated significantly the sunflower and rape seed germination. These vitamins, however, didn't have any effect on safflower germination. Hydrogen peroxide strongly increased safflower germination. Ascorbic acid and pyridoxine decreased catalase activity in sunflower and rape seed, whereas hydrogen peroxide increased it. Ascorbic acid and pyridoxine prevented protein degradation and lipid peroxidation in germinated seeds. Consequently, we understand that ascorbic acid and pyridoxine can increase sunflower and rape seed germination and stimulate rate of growth. Also safflower germination increased due to germination inhibitor oxidation by hydrogen peroxide. In conclusion, this report shows that oil seeds treated with ascorbic acid, pyridoxine and hydrogen peroxide remarkably increase the capacity of germination. We suggest that treatments with such substances can improve the old oil seed germination during storage.

  17. THE ANALYSIS OF CATALASE AND PEROOXIDASE ACTIVITY IN SAPROPHYTIC FUNGUS RHIZOPUS NIGRICANS GROWN ON MEDIUM WHITH DIFERENT CONCENTRATION OF GRINDED CORN CARYOPSIS

    Directory of Open Access Journals (Sweden)

    Tamara Barbaneagra

    2010-09-01

    Full Text Available The purpose of this study was to assay catalase and peroxidase activity in the saprophytic fungus Rhizopus nigricans, grown on mediums containing grinded corn caryopsis, which, in our experiments have replaced carbon source – sucrose in composition of liquid culture medium Czapeck Dox, resulting in the final three experimental variants: V1 = 20 g/l, V2 = 30 g/l, V3 = 40 g/l, while the control variant composition remained unchanged. Measurements were made at three time intervals: 5 days and 10 days and 15 days after inoculation, using fungus mycelium and culture liquid. Determination of catalase activity was performed using Sinha method (Artenie Vl., et al., 2008, and determination of peroxidase was carried out on the basis of ortho-dianisidine method (Cojocaru D.C., 2009. The results showed significant deferens in dynamic of enzyme activity depending on the concentration of carbon source introduced into the medium and age of the fungus.

  18. Isolation,purification and characterization of Alocasia macrorrhiza catalase%海芋过氧化氢酶的分离纯化及其性质

    Institute of Scientific and Technical Information of China (English)

    李思莹; 黄卓烈; 郑华章; 黄丹丹

    2015-01-01

    为了探索海芋( Alocasia macrorrhiza)过氧化氢酶的分子结构和酶学性质,提取海芋新鲜叶片、叶柄和块茎的粗酶进行活力比较和同工酶分析。用Sephadex G 200凝胶过滤层析法将粗酶分离纯化,以PAGE电泳对酶纯度鉴定。结果表明:海芋过氧化氢酶较单一,没有同工酶。经层析法进行分离获得电泳纯的过氧化氢酶,其活力回收率为40�83%,纯化倍数为8�88倍。十二烷基硫酸钠聚丙烯酰氨凝胶电泳( SDS PAGE)分析结果发现:该酶由5条肽链组成,全酶相对分子质量为2�6211×105。酶学性质研究结果表明:该酶最适反应温度40℃,最适pH为7�5。在25~45℃和pH 6�0~9�0下,该酶能保持较强的催化活力,稳定性较好。在最适条件下,该酶的Km 值为16�43 mmol/L,对底物的亲和力较好。甲醇、乙醇和异丙醇对此酶活力有显著的抑制作用。%In order to characterize catalase, crude enzyme was extracted from leaves, petioles, tuber of Alocasia macrorrhiza. The enzyme activities and isozymes were compared and analyzed among the organs. Sephadex G 200 gel chromatography was used to purify the enzyme. Polyacrylamide gel electrophoresis (PAGE) was used to identify the enzyme purity. There was no isozyme of catalase in Alocasia macrorrhiza. The activity recovery was 40�83% and the purification multiple was 8�88 folds. It was indicated by SDS⁃PAGE that the catalase in Alocasia macrorrhiza was composed of 5 polypeptides. The molecular weight of this catalase was 2�621 1×105. The optimal reaction temperature was 40 ℃ and the optimal pH was 7�5. Under the optimal conditions, the Km of this catalase was 16�43 mmol/L. The enzyme activity was inhibited by methanol,ethanol and isopropanol.

  19. Atividade da catalase no pulmão, rim e intestino delgado não isquemiado de ratos após reperfusão intestinal Catalase activity in lung, kidney and small bowel non-ischemic in rats after intestinal reperfusion

    Directory of Open Access Journals (Sweden)

    Camila de Oliveira Ferro

    2010-02-01

    Full Text Available OBJETIVO: Avaliar a atividade catalase, após lesão por isquemia e reperfusão intestinal e estudar as alterações deste antioxidante em órgãos situados à distância do insulto inicial. MÉTODOS: Utilizaram-se 18 ratos do tipo Wistar, aleatoriamente distribuídos em três grupos. 1-Controle, 2-Simulação e 3-Isquemia/Reperfusão. Neste último, realizou-se isquemia no íleo, por 60 minutos, seguida de reperfusão por 30 minutos. No grupo 2 efetuou-se apenas uma laparotomia. Foram retirados, de todos os animais, segmentos do intestino com e sem reperfusão, além do pulmão e rim direitos para exame com microscopia óptica. A atividade da catalase foi aferida em espectrofotômetro ajustado para 240 nm. Utilizaram-se os testes estatísticos Mann e Whitney e Kruskal Wallis. RESULTADOS: Observou-se aumento significante (p OBJECTIVE: This study aimed to assess the catalase activity after ischemia and reperfusion and to study the changes of this antioxidant in organs located far from the initial insult. METHODS: Eighteen Wistar rats were randomly divided into three groups. 1-Control, 2-Simulation and 3-Ischemia and Reperrfusion. In the latter it was done an ischemia of the ileum for 60 minutes followed by reperfusion for 30 minutes. In group 2 only laparotomy was performed. From all animals it was taken segments of the reperfused and non reperfused intestine, as well of the right kidney and lung to be evaluated under light microscopy. Catalase activity was measured in spectrophotometer with a wavelength set to 240 nm. It was used Mann Whitney and Kruskal Wallis statistical tests. RESULTS: There was a significant increase (p <0.05 in the catalase activity not only at small bowel ischemic and non-ischemic segments but also at lungs. However the enzymatic activity decreases in the kidney. In all organs studied at reperfusion group it was found a slight villi derangement, mild congestion and infiltration with inflammatory cells, and areas of

  20. Acidic pH conditions induce dissociation of the haem from the protein and destabilise the catalase isolated from Aspergillus terreus.

    Science.gov (United States)

    Vatsyayan, Preety; Goswami, Pranab

    2011-02-01

    The stability (half-life, t(½)) of the large catalase (CAT) isolated from Aspergillus terreus was decreased under acidic conditions (maximum t(½) approximately 8.5 months at pH ≤ 6) versus alkaline conditions (t(½) approximately 15 months at pH 8-12). Acidic conditions induce the dissociation of haem from CAT, as revealed from a reduction in the Soret peak intensity at 405 nm and an increase in the peak current at Fe(3+)/Fe(2+) redox potentials. This increase in current is attributed to the facile electron transfer from the free haem generated on the electrode surface as a result of its disintegration from the insulating protein matrix. The haem isolated from CAT at acidic condition was reconstituted with apo-CAT at alkaline denaturing conditions to regenerate the CAT activity.

  1. Infective Endocarditis: Identification of Catalase-Negative, Gram-Positive Cocci from Blood Cultures by Partial 16S rRNA Gene Analysis and by Vitek 2 Examination

    DEFF Research Database (Denmark)

    Abdul-Redha, Rawaa Jalil; Kemp, Michael; Bangsborg, Jette M;

    2010-01-01

    Streptococci, enterococci and Streptococcus-like bacteria are frequent etiologic agents of infective endocarditis and correct species identification can be a laboratory challenge. Viridans streptococci (VS) not seldomly cause contamination of blood cultures. Vitek 2 and partial sequencing of the 16......S rRNA gene were applied in order to compare the results of both methods. STRAINS ORIGINATED FROM TWO GROUPS OF PATIENTS: 149 strains from patients with infective endocarditis and 181 strains assessed as blood culture contaminants. Of the 330 strains, based on partial 16S rRNA gene sequencing...... results, 251 (76%) were VS strains, 10 (3%) were pyogenic streptococcal strains, 54 (16%) were E. faecalis strains and 15 (5%) strains belonged to a group of miscellaneous catalase-negative, Gram-positive cocci. Among VS strains, respectively, 220 (87,6%) and 31 (12,3%) obtained agreeing and non...

  2. A fungal catalase reacts selectively with the 13S fatty acid hydroperoxide products of the adjacent lipoxygenase gene and exhibits 13S-hydroperoxide-dependent peroxidase activity.

    Science.gov (United States)

    Teder, Tarvi; Boeglin, William E; Schneider, Claus; Brash, Alan R

    2017-03-28

    The genome of the fungal plant pathogen Fusarium graminearum harbors six catalases, one of which has the sequence characteristics of a fatty acid peroxide-metabolizing catalase. We cloned and expressed this hemoprotein (designated as Fg-cat) along with its immediate neighbor, a 13S-lipoxygenase (cf. Brodhun et al., PloS One, e64919, 2013) that we considered might supply a fatty acid hydroperoxide substrate. Indeed, Fg-cat reacts abruptly with the 13S-hydroperoxide of linoleic acid (13S-HPODE) with an initial rate of 700-1300s(-1). By comparison there was no reaction with 9R- or 9S-HPODEs and extremely weak reaction with 13R-HPODE (~0.5% of the rate with 13S-HPODE). Although we considered Fg-cat as a candidate for the allene oxide synthase of the jasmonate pathway in fungi, the main product formed from 13S-HPODE was identified by UV, MS, and NMR as 9-oxo-10E-12,13-cis-epoxy-octadecenoic acid (with no traces of AOS activity). The corresponding analog is formed from the 13S-hydroperoxide of α-linolenic acid along with novel diepoxy-ketones and two C13 aldehyde derivatives, the reaction mechanisms of which are proposed. In a peroxidase assay monitoring the oxidation of ABTS, Fg-cat exhibited robust activity (kcat 550s(-1)) using the 13S-hydroperoxy-C18 fatty acids as the oxidizing co-substrate. There was no detectable peroxidase activity using the corresponding 9S-hydroperoxides, nor with t-butyl hydroperoxide, and very weak activity with H2O2 or cumene hydroperoxide at micromolar concentrations of Fg-cat. Fg-cat and the associated lipoxygenase gene are present together in fungal genera Fusarium, Metarhizium and Fonsecaea and appear to constitute a partnership for oxidations in fungal metabolism or defense.

  3. Effect of Yerbimat herbicide on lipid peroxidation, catalase activity, and histological damage in gills and liver of the freshwater fish Goodea atripinnis.

    Science.gov (United States)

    Ortiz-Ordoñez, Esperanza; Uría-Galicia, Esther; Ruiz-Picos, Ricardo Arturo; Duran, Angela Georgina Sánchez; Trejo, Yoseline Hernández; Sedeño-Díaz, Jacinto Elías; López-López, Eugenia

    2011-10-01

    The use of herbicides for agricultural and aquatic weed control has increased worldwide. These substances are potentially toxic pollutants because they induce the production of reactive oxygen species for biological systems and exert oxidative stress in nontarget organisms living in the treated aquatic systems. Recent evidence suggests differences in the toxicity of glyphosate in the form of an active ingredient compared to the toxicity of glyphosate in combination with surfactants, such as those found in commercial formulations. In Mexico, one of the most widely used glyphosate-based herbicides is Yerbimat, which has agricultural as well as aquatic weed control applications. However, there are no aquatic toxicity data, particularly regarding native fish. Therefore, we determined the acute toxicity of commercial-formulation Yerbimat in a static bioassay at 96 h (LC(50)). We also determined its toxicity at 96 h in sublethal concentrations to assess the lipid peroxidation levels (LPX), catalase activity, hepatic glycogen content, and histological damage in the liver and gills of the fish Goodea atripinnis associated with chronic exposure (75 days). The LC(50) was 38.95 ± 0.33 mg/L. The results of the short-term exposure study indicate that Yerbimat can potentially induce oxidative stress in G. atripinnis, because LPX was increased in the gills and liver. Catalase activity was reduced in the gills but increased in the liver, whereas hepatic glycogen was depleted. Chronic exposure was associated with histopathological damage in the gills and liver, some of which was irreversible. Yerbimat represents a potential risk for aquatic biota; therefore, we recommend that its application be carefully considered.

  4. Soluble expression of Catalase/GST fusion protein of helicobacter pylori and thrombin cleavage of GST tag on column%幽门螺杆菌Catalas/GST融合蛋白的可溶性表达及凝血酶柱上切割GST标签

    Institute of Scientific and Technical Information of China (English)

    李妍; 姜茵; 奚月; 何殿殿; 宁云山

    2012-01-01

    目的 利用GST融合基因表达系统表达幽门螺杆菌Catalase/GST融合蛋白,并利用凝血酶柱上切割GST标签.方法 将重组表达质粒Catalase/pGEX-4T-1转化大肠杆菌BL21 (DE3)感受态中并用IPTG进行诱导表达,菌体经反复冻融、溶菌酶裂解及超声破菌进行裂解.采用谷胱甘肽琼脂糖树脂Glutathione Sepharose 4B对Catalase/GST融合蛋白可溶性表达上清进行纯化,并同时利用凝血酶柱上切割GST标签,用鼠抗Catalase抗体对纯化产物进行Western blot鉴定.结果 高效表达出相对分子质量约85 kDa的Catalase/GST融合蛋白,以部分可溶性的形式表达,凝血酶柱上成功地切割了GST标签,Catalase蛋白能被鼠抗Catalase单克隆抗体识别.结论 成功表达了Catalase/GST融合蛋白并柱上切割了GST标签,为深入研究Catalase的功能奠定了基础.%OBJECTIVE To express Catalase/GST fusion protein by GST gene expression system and cleave GST-tag on column using thrombin. METHODS The recombinant expression plasmid Catalase/pGEX4T-l was transformed into E. Coli BL21 (DE3) component cell and induced by IPTG. The bacterial sediment was lysed by repeating freezing and thawing, lysozyme lysis and ultrasonication. Catalase/GST fusion protein was purified by Glutathione Sepharose 4B and cleaved the GST-tag on column using thrombin. Purified Catalase was identified by anti-Catalase monoclonal antibody (mAb) using Western blot. RESULTS The fusion Catalase/GST was partly expressed in soluble form with relative molecular mass of 85kDa. Thrombin cleaved the GST tag on column and Catalase protein was recognized by mouse anti-Catalase mAb. CONCLUSION The recombinant Catalase/GST is successfully expressed and GST-tag is cleaved on column, which lays a foundation for further study on function of Catalase protein.

  5. 过氧化氢酶基因多态性与疾病相关性的研究%The research of association of catalase gene polymorphisms with diseases

    Institute of Scientific and Technical Information of China (English)

    涂茜; 单可人; 官志忠; 任锡麟

    2010-01-01

    Catalase is one of three enzymes within the important huamn antioxidant enzyme system, which also includes superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). They coordinate to reduce the production of active oxygen free radicals , to prevent lipid peroxidation and its intermediate metabolites from damaging human body, and to maintain oxidative-antioxidative balance. Catalase gene polymorphisms,and its haplotypes may be related to enzyme activity and transcriptional activity, and thus affect the sensitivity to some human diseases in different individuals. The study reviewed the structure and function of catalase, and gene polymorphism and its association with human diseases.%过氧化氢酶(catalase,CAT)是人体内很重要的一种抗氧化酶,它与超氧化物歧化酶(superoxide dismutase,SOD),谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)共同构成人体重要的抗氧化酶系统,三者协同,减少了活性氧自由基的产生,防止脂质过氧化及其中间代谢产物对机体的损害,维持体内氧化-抗氧化平衡.过氧化氢酶的基因多态性可能与该酶的活性及转录活性有关,从而影响个体间对疾病的易感性不同.

  6. Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry Analysis of Gram-Positive, Catalase-Negative Cocci Not Belonging to the Streptococcus or Enterococcus Genus and Benefits of Database Extension

    DEFF Research Database (Denmark)

    Christensen, Jens Jørgen; Dargis, Rimtas; Hammer, Monja;

    2012-01-01

    Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry with a Bruker Daltonics microflex LT system was applied to 90 well-characterized catalase-negative, Gram-positive cocci not belonging to the streptococci or enterococci. Biotyper version 2.0.43.1 software was...

  7. Effect of lead (Pb) exposure on the activity of superoxide dismutase and catalase in battery manufacturing workers (BMW) of Western Maharashtra (India) with reference to heme biosynthesis.

    Science.gov (United States)

    Patil, Arun J; Bhagwat, Vinod R; Patil, Jyotsna A; Dongre, Nilima N; Ambekar, Jeevan G; Jailkhani, Rama; Das, Kusal K

    2006-12-01

    The aim of this study was to estimate the activity of superoxide dismutase (SOD) and catalase in erythrocytes and malondialdehyde (MDA) in plasma of battery manufacturing workers (BMW) of Western Maharashtra (India) who were occupationally exposed to lead (Pb) over a long period of time (about 15 years). This study was also aimed to determine the Pb intoxication resulted in a disturbance of heme biosynthesis in BMW group. The blood Pb level of BMW group (n = 28) was found to be in the range of 25.8 - 78.0 microg/dL (mean + SD, 53.63 + 16.98) whereas in Pb unexposed control group (n = 35) the range was 2.8 - 22.0 microg/dL (mean + SD, 12.52 + 4.08). The blood level (Pb-B) and urinary lead level (Pb-U) were significantly increased in BMW group as compared to unexposed control. Though activated d- aminolevulinic acid dehydratase (ALAD) activities in BMW group did not show any significant change when compared to control group but activated / non activated erythrocyte - ALAD activities in BMW group showed a significant increase. Erythrocyte- zinc protoporphyrin (ZPP), urinary daminolevulinic acid (ALA-U) and porphobilinogen (PBG-U) of BMW groups elevated significantly as compared to control. A positive correlation (r = 0.66, p BMW group but no such significant correlation (r = 0.02, p> 1.0) were observed in control group. Hematological study revealed a significant decrease of hemoglobin concentration, packed cell volume (%) and other blood indices and a significant increase of total leucocytes count in BMW group in comparison to control group. The serum MDA content was significantly increased (p BMW group as compared to control group. A positive correlation (r = 0.45, p BMW group (Pb-B range 25.8 - 78.0 microg / dL) but such significant correlation did not notice in control group (Pb-B range 2.8 - 22.0 microg / dL). The study clearly showed an adverse effect of heme biosynthesis and imbalance of pro-oxidant / antioxidant status in lead exposed battery manufacturing

  8. Haemoprotein b-590 (Escherichia coli), a reducible catalase and peroxidase: evidence for its close relationship to hydroperoxidase I and a 'cytochrome a1b' preparation.

    Science.gov (United States)

    Poole, R K; Baines, B S; Appleby, C A

    1986-06-01

    A reducible hydroperoxidase, haemoprotein b-590, has been purified 16-fold from a soluble fraction of Escherichia coli K12, grown anaerobically with glycerol and fumarate. The Mr of the native protein, determined by gel filtration, was 331,000 although a minor, smaller species with a Mr of 188,000 was also detected; both had catalase activities. Based on the subunit Mr, determined from SDS gel electrophoresis to be 75,000, the above species are tentatively identified as tetramers and dimers, respectively. The isoelectric point of both species was 4.4. The absorption spectrum of the isolated haemoprotein is typical of ferric, high-spin haem. The A405/A280 ratio never exceeded 0.27, a value half of that obtained for E. coli hydroperoxidase I. On reduction with dithionite, the gamma, beta, and alpha bands were at 441, 559 and 590 nm respectively, the alpha-band being unusually distinct. Treatment of the reduced form with CO gave a sharp prominent gamma-band at 426 nm and caused significant shifts of the alpha and beta bands to shorter (574 and 545 nm) wavelengths. The pyridine haemochrome spectra showed the haem to be protohaem IX; the spectra were featureless between 580 and 630 nm, thus excluding the presence of haem a. However, some features of the difference spectra of the haemoprotein were reminiscent of cytochrome a1, notably the maxima in reduced minus oxidized spectra at 444 and 593 nm and the peaks and troughs in CO difference spectra at 426 and 446 nm respectively. The haemoprotein had high catalase activity: Vmax was 2.3 X 10(6) mol H2O2 (mol haem)-1 min-1 and the Km was 11 mM. At 10 mM-H2O2 the first order rate constant was 0.3 X 10(7) M-1 s-1. The haemoprotein was also a peroxidase with o-dianisidine or 2,3',6-trichloroindophenol as substrates; for the latter substrate, the Km was 0.18 mM. It is concluded that haemoprotein b-590 strongly resembles the hydroperoxidase I purified by Claiborne & Fridovich (Journal of Biological Chemistry 254, 4245-4252, 1979

  9. 以Fe(Ⅲ)改性胶原纤维为载体固定过氧化氢酶%Immobilization of catalase on Fe (Ⅲ) modified collagen fiber

    Institute of Scientific and Technical Information of China (English)

    陈爽; 宋娜; 廖学品; 石碧

    2011-01-01

    将胶原纤维用三价铁改性后作为载体,通过戊二醛的交联作用将过氧化氢酶固定在该载体上.制备的固定化过氧化氢酶蛋白固载量为16.7 mg/g,酶活收率为35%.研究了固定化酶与自由酶的最适pH、最适温度、热稳定性、贮存稳定性及操作稳定性.结果表明:过氧化氢酶经此法固定化后,最适pH及最适温度与自由酶相同,分别为pH 7.0和25℃;但固定化酶的热稳定性显著提高,在75℃保存5 h后,仍能保留30%的活力,而自由酶则完全失活;固定化酶在室温下保存12 d后,酶活力仍保持在88%以上,而自由酶在此条件下则完全失活;此外,固定化过氧化氢酶还表现出了良好的操作稳定性,在室温下连续反应26次后,相对活力为57%.该研究表明胶原纤维可作为固定化过氧化%Fe (III) modified collagen fibers were used to immobilize catalase through the cross-linking of glutaraldehyde. The loading amount of catalase on the supporting matrix was 16.7 mg/g, and 35% enzymatic activity was remained. A series of experiments were conducted on free and immobilized catalase in order to investigate their optimal pH and temperature, and the thermal, storage and operation stability. Results suggest that the free and immobilized catalase prefer similar pH and temperature condition, which were pH 7.0 and 25 ℃. It should be noted that the thermal stability of catalase was considerably improved after immobilization owing to the fact that the enzyme kept 30% of relative activity after incubation at 75 ℃ for 5 h. On the contrary, the free catalase was completely inactive. As for the storage stability, the immobilized catalase kept 88% of relative activity after stored at room temperature for 12 days while the free one was completely inactive under the same conditions. Moreover, the immobilized catalase preserved 57% of relative activity after being reused 26 times, exhibiting excellent operation stability. Consequently

  10. Catalase-like activity of bovine met-hemoglobin: interaction with the pseudo-catalytic peroxidation of anthracene traces in aqueous medium.

    Science.gov (United States)

    Paco, Laveille; Galarneau, Anne; Drone, Jullien; Fajula, François; Bailly, Carole; Pulvin, Sylviane; Thomas, Daniel

    2009-10-01

    Hemoglobin is a member of the hemoprotein superfamily whose main role is to transport O(2) in vertebrate organisms. It has two known promiscuous enzymatic activities, peroxidase and oxygenase. Here we show for the first time that bovine hemoglobin also presents a catalase-like activity characterized by a V(max )of 344 microM/min, a K(M )of 24 mM and a k(cat) equal to 115/min. For high anthracene and hemoglobin concentrations and low hydrogen peroxide concentrations, this activity inhibits the expected oxidation of anthracene, which occurs through a peroxidase-like mechanism. Anthracene belongs to the polycyclic aromatic hydrocarbon (PAH) family whose members are carcinogenic and persistent pollutants found in industrial waste waters. Our results show that anthracene oxidation by hemoglobin and hydrogen peroxide follows a typical bi-bi ping-pong mechanism with a V(max) equal to 0.250 microM/min, K(M(H2O2) )of 80 microM, K(M(ANT)) of 1.1 microM and k(cat) of 0.17/min. The oxidation of anthracene is shown to be pseudo-catalytic because an excess of hemoglobin and hydrogen peroxide is required to make PAH completely disappear. Thus, bovine hemoglobin presents, in different degrees, all the catalytic activities of the hemoprotein group, which makes it a very interesting protein for biotechnological processes and one with which structure-activity relationships can be studied.

  11. Nickel in Soil Modifies Sensitivity to Diazinon Measured by the Activity of Acetylcholinesterase, Catalase, and Glutathione S-Transferase in Earthworm Eisenia fetida

    Directory of Open Access Journals (Sweden)

    Agnieszka Zawisza-Raszka

    2013-01-01

    Full Text Available Nickel in typical soils is present in a very low concentration, but in the contaminated soils it occurs in locally elevated concentrations. The aim of this study was to examine the effect of nickel in the concentrations of 300 (very high, close to LOEC for reproduction and 900 (extremely high, close to LOEC for mortality mg/kg dry soil on the life history and acetylcholinesterase, catalase, and glutathione S-transferase activities in earthworm Eisenia fetida and to establish how nickel modifies the sensitivity to organophosphorous pesticide—diazinon. Cocoons production and juveniles’ number were significantly lower only in groups exposed to Ni in the concentration of 900 mg/kg dry soil for two months. Diazinon administration diminished the AChE activity in the GI tract and in the body wall. The interaction between diazinon and nickel was observed, and, in consequence, the AChE activity after the pesticide treatment was similar to controls in worms preexposed to nickel. Both pesticide administration and exposure to nickel caused an increase in the GST activity in examined organs and CAT activity in body wall. Both biometric and development data and simple enzymatic analysis, especially the AChE and GST, show a Ni pretreatment effect on the subsequent susceptibility to pesticide.

  12. Specific amplification of gene encoding N-terminal region of catalase-peroxidase protein (KatG-N) for diagnosis of disseminated MAC disease in HIV patients.

    Science.gov (United States)

    Latawa, Romica; Singh, Krishna Kumar; Wanchu, Ajay; Sethi, Sunil; Sharma, Kusum; Sharma, Aman; Laal, Suman; Verma, Indu

    2014-10-01

    Disseminated Mycobacterium avium-intracellulare complex (MAC) infection is considered as severe complication of advanced HIV/AIDS disease. Currently available various laboratory investigations have not only limited ability to discriminate between MAC infection and tuberculosis but are also laborious and time consuming. The aim of this study was, therefore, to design a molecular-based strategy for specific detection of MAC and its differentiation from Mycobacterium tuberculosis (M. tb) isolated from the blood specimens of HIV patients. A simple PCR was developed based on the amplification of 120-bp katG-N gene corresponding to the first 40 amino acids of N-terminal catalase-peroxidase (KatG) protein of Mycobacterium avium that shows only ~13% sequence homology by clustal W alignment to N-terminal region of M. tb KatG protein. This assay allowed the accurate and rapid detection of MAC bacteremia, distinguishing it from M. tb in a single PCR reaction without any need for sequencing or hybridization protocol to be performed thereafter. This study produced enough evidence that a significant proportion of Indian HIV patients have disseminated MAC bacteremia, suggesting the utility of M. avium katG-N gene PCR for early detection of MAC disease in HIV patients.

  13. Plasmonic Enzyme-Linked Immunosorbent Assay Using Nanospherical Brushes as a Catalase Container for Colorimetric Detection of Ultralow Concentrations of Listeria monocytogenes.

    Science.gov (United States)

    Chen, Rui; Huang, Xiaolin; Xu, Hengyi; Xiong, Yonghua; Li, Yanbin

    2015-12-30

    Plasmonic enzyme-linked immunosorbent assay (pELISA) based on catalase (CAT)-mediated gold nanoparticle growth exhibits ultrahigh sensitivity for detecting disease-related biomarkers using sandwich formats. However, the limit of detection (LOD) of this strategy for Listeria monocytogenes is only around 10(3) CFU/mL, which considerably exceeds the amount of L. monocytogenes commonly present in food products (container" to increase enzyme loading for enhancing the detection signal. Under optimal conditions, the proposed pELISA exhibits good specificity and excellent sensitivity for L. monocytogenes with a LOD of 8 × 10(1) CFU/mL in 0.01 M phosphate-buffered saline, via a reaction that can be discriminated by the naked eye. The LOD obtained by this method was 2 and 5 orders of magnitude lower than that of conventional CAT-based pELISA and horseradish peroxidase (HRP)-based conventional ELISA, respectively. Coupled with large-volume immunomagnetic separation, the LOD for L. monocytogenes-spiked lettuce samples reached 8 × 10(1) CFU/g. The improved pELISA also exhibited a great potential in detecting a single cell of L. monocytogenes in 100 μL of solution.

  14. Ferric haem forms of Mycobacterium tuberculosis catalase-peroxidase probed by EPR spectroscopy: Their stability and interplay with pH.

    Science.gov (United States)

    Svistunenko, Dimitri A; Worrall, Jonathan A R; Chugh, Snehpriya B; Haigh, Sarah C; Ghiladi, Reza A; Nicholls, Peter

    2012-06-01

    Low temperature EPR spectroscopy was used to characterise Mycobacterium tuberculosis catalase-peroxidase in its resting ferric haem state. Several high spin ferric haem forms and no low spin forms were found in the enzyme samples frozen in methanol on dry ice. The EPR spectra depended not only on the pH but also on the buffer type. As a general trend, the higher the pH, the greater the 'rhombic' fraction of the high spin ferric haem that was observed. The rhombic form was characterised by well separated two lines in the g = 6 region whereas in the 'axial' form the two lines overlap. This pH dependence of the equilibrium of axial and rhombic ferric haem forms is also seen in rapidly freeze-quenched samples. Different high spin ferric haem forms were monitored during a 3 week storage of the enzyme at 4 °C. For some forms, extremal dependences, i.e. those progressing via maxima or minima over storage time, were found. This indicates that the mechanism of the time-dependent transition from one high spin ferric haem form to another must be more complex than a simple single site oxidation.

  15. Modulation of the Activities of Catalase, Cu-Zn, Mn Superoxide Dismutase, and Glutathione Peroxidase in Adipocyte from Ovariectomised Female Rats with Metabolic Syndrome

    Directory of Open Access Journals (Sweden)

    Rebeca Cambray Guerra

    2014-01-01

    Full Text Available The aim of this study was to evaluate the association between estrogen removal, antioxidant enzymes, and oxidative stress generated by obesity in a MS female rat model. Thirty two female Wistar rats were divided into 4 groups: Control (C, MS, MS ovariectomized (Ovx, and MS Ovx plus estradiol (E2. MS was induced by administering 30% sucrose to drinking water for 24 weeks. After sacrifice, intra-abdominal fat was dissected; adipocytes were isolated and lipid peroxidation, non-enzymatic antioxidant capacity, and the activities of Cu-Zn and Mn superoxide dismutase (SOD, catalase (CAT, and glutathione peroxidase (GPx were determined. There were no significant differences in the activities of Cu-Zn, Mn SOD, CAT, and GPx between the C and MS groups, but in the MS Ovx group there was a statistically significant decrease in the activities of these enzymes when compared to MS and MS Ovx+E2. The increased lipid peroxidation and nonenzymatic antioxidant capacity found in MS Ovx was significantly decreased when compared to MS and MS Ovx+E2. In conclusion, the removal of E2 by ovariectomy decreases the activity of the antioxidant enzymes in the intra-abdominal tissue of MS female rats; this is reflected by increased lipid peroxidation and decreased nonenzymatic antioxidant capacity.

  16. Sesamin modulates tyrosine hydroxylase, superoxide dismutase, catalase, inducible NO synthase and interleukin-6 expression in dopaminergic cells under MPP+-induced oxidative stress.

    Science.gov (United States)

    Lahaie-Collins, Vicky; Bournival, Julie; Plouffe, Marilyn; Carange, Julie; Martinoli, Maria-Grazia

    2008-01-01

    Oxidative stress is regarded as a mediator of nerve cell death in several neurodegenerative disorders, such as Parkinson's disease. Sesamin, a lignan mainly found in sesame oil, is currently under study for its anti-oxidative and possible neuroprotective properties. We used 1-methyl-4-phenyl-pyridine (MPP(+)) ion, the active metabolite of the potent parkinsonism-causing toxin 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine, to produce oxidative stress and neurodegeneration in neuronal PC12 cells, which express dopamine, as well as neurofilaments. Our results show that picomolar doses of sesamin protected neuronal PC12 cells from MPP(+)-induced cellular death, as revealed by colorimetric measurements and production of reactive oxygen species. We also demonstrated that sesamin acted by rescuing tyrosine hydroxylase levels from MPP(+)-induced depletion. Sesamin, however, did not modulate dopamine transporter levels, and estrogen receptor-alpha and -beta protein expression. By examining several parameters of cell distress, we found that sesamin also elicited a strong increase in superoxide dismutase activity as well as protein expression and decreased catalase activity and the MPP(+) stimulated inducible nitric oxide synthase protein expression, in neuronal PC12 cells. Finally, sesamin possessed significant anti-inflammatory properties, as disclosed by its potential to reduce MPP(+)-induced interleukin-6 mRNA levels in microglia. From these studies, we determined the importance of the lignan sesamin as a neuroprotective molecule and its possible role in complementary and/or preventive therapies of neurodegenerative diseases.

  17. Sesamin Modulates Tyrosine Hydroxylase, Superoxide Dismutase, Catalase, Inducible No Synthase and Interleukin-6 Expression in Dopaminergic Cells Under Mpp+-Induced Oxidative Stress

    Directory of Open Access Journals (Sweden)

    Vicky Lahaie-Collins

    2008-01-01

    Full Text Available Oxidative stress is regarded as a mediator of nerve cell death in several neurodegenerative disorders, such as Parkinson's disease. Sesamin, a lignan mainly found in sesame oil, is currently under study for its anti-oxidative and possible neuroprotective properties. We used 1-methyl-4-phenyl-pyridine (MPP+ ion, the active metabolite of the potent parkinsonism-causing toxin 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine, to produce oxidative stress and neurodegeneration in neuronal PC12 cells, which express dopamine, as well as neurofilaments. Our results show that picomolar doses of sesamin protected neuronal PC12 cells from MPP+-induced cellular death, as revealed by colorimetric measurements and production of reactive oxygen species. We also demonstrated that sesamin acted by rescuing tyrosine hydroxylase levels from MPP+-induced depletion. Sesamin, however, did not modulate dopamine transporter levels, and estrogen receptor-alpha and -beta protein expression. By examining several parameters of cell distress, we found that sesamin also elicited a strong increase in superoxide dismutase activity as well as protein expression and decreased catalase activity and the MPP+ stimulated inducible nitric oxide synthase protein expression, in neuronal PC12 cells. Finally, sesamin possessed significant anti-inflammatory properties, as disclosed by its potential to reduce MPP+-induced interleukin-6 mRNA levels in microglia. From these studies, we determined the importance of the lignan sesamin as a neuroprotective molecule and its possible role in complementary and/or preventive therapies of neurodegenerative diseases.

  18. Lipid peroxidation as pathway of aluminium cytotoxicity in human skin fibroblast cultures: prevention by superoxide dismutase+catalase and vitamins E and C.

    Science.gov (United States)

    Anane, R; Creppy, E E

    2001-09-01

    Lipid peroxidation is one of the main manifestations of oxidative damage and has been found to play an important role in the toxicity and carcinogenicity of many xenobiotics. In the present study, we investigated the possible induction of lipid peroxidation by aluminium in human foreskin fibroblast cultures by assaying the malondialdehyde (MDA) produced inside the cells. The MDA-thiobarbituric acid (TBA) adduct was assayed by HPLC using fluorometric quantification after extraction in n-butanol. Lactate dehydrogenase (LDH) release was used as a marker of aluminium toxicity. MDA production was significantly increased after 24 h incubation with aluminium and paralleled LDH release. Superoxide dismutase (SOD)+catalase and vitamins C and E added in the culture medium as oxygen radical and free radical scavengers were efficient in preventing MDA production by aluminium, indicating that oxidative processes are one of the main pathways whereby this metal induces cytotoxicity. The latter is also largely prevented, thus confirming the link between oxidative stress induced by aluminium and its cytotoxicity in human skin fibroblasts.

  19. Glutathione S-transferase, catalase, superoxide dismutase, glutathione peroxidase, and lipid peroxidation as biomarkers of oxidative stress in snails: A review

    Directory of Open Access Journals (Sweden)

    J Bhagat

    2016-10-01

    Full Text Available Antioxidant defense plays a crucial role in the response of an organism to pollutants. Several processes stimulate the production of free radicals or deplete the antioxidant defense, which if not regulated properly, may cause oxidative stress in the organisms, leading to damage in DNA, proteins or lipids. Free radicals are also beneficial as it plays an important role in defense against infectious agents, and signal transduction. Hence a delicate balance between antioxidants and free radicals is required. Oxidative stress biomarkers are very useful in disease etiology and environmental toxicological studies. The increase in anthropogenic activities and environmental awareness has resulted in an explosive increase of research in the field of oxidative stress. Snails are excellent organisms for environmental biomonitoring and contribute a major proportion of the invertebrate biomass. In our article, we have summarized the research carried out using glutathione S-transferase (GST, catalase (CAT, superoxide dismutase (SOD, glutathione peroxidase (GPx, and lipid peroxidation (LPO in snails exposed to various toxicants and their implication in the environmental monitoring programs. In the end, we have discussed different factors affecting the variations in oxidative biomarkers response for a better understanding of the phenomenon.

  20. RNAi-mediated knockdown of catalase causes cell cycle arrest in SL-1 cells and results in low survival rate of Spodoptera litura (Fabricius.

    Directory of Open Access Journals (Sweden)

    Haiming Zhao

    Full Text Available Deregulated reactive oxygen species (ROS production can lead to the disruption of structural and functional integrity of cells as a consequence of reactive interaction between ROS and various biological components. Catalase (CAT is a common enzyme existing in nearly all organisms exposed to oxygen, which decomposes harmful hydrogen peroxide, into water and oxygen. In this study, the full length sequence that encodes CAT-like protein from Spodoptera litura named siltCAT (GenBank accession number: JQ_663444 was cloned and characterized. Amino acid sequence alignment showed siltCAT shared relatively high conservation with other insect, especially the conserved residues which defined heme and NADPH orientation. Expression pattern analysis showed that siltCAT mRNA was mainly expressed in the fat body, midgut, cuticle and malpighian tube, and as well as over last instar larvae, pupa and adult stages. RNA interference was used to silence CAT gene in SL-1 cells and the fourth-instar stage of S. litura larvae respectively. Our results provided evidence that CAT knockdown induced ROS generation, cell cycle arrest and apoptosis in SL-1 cells. It also confirmed the decrease in survival rate because of increased ROS production in experimental groups injected with double-stranded RNA of CAT (dsCAT. This study implied that ROS scavenging by CAT is important for S. litura survival.

  1. Brain catalase in the streptozotocin-rat model of sporadic Alzheimer's disease treated with the iron chelator-monoamine oxidase inhibitor, M30.

    Science.gov (United States)

    Sofic, E; Salkovic-Petrisic, M; Tahirovic, I; Sapcanin, A; Mandel, S; Youdim, M; Riederer, P

    2015-04-01

    Low intracerebroventricular (icv) doses of streptozotocin (STZ) produce regionally specific brain neurochemical changes in rats that are similar to those found in the brain of patients with sporadic Alzheimer's disease (sAD). Since oxidative stress is thought to be one of the major pathologic processes in sAD, catalase (CAT) activity was estimated in the regional brain tissue of animals treated intracerebroventricularly with STZ and the multitarget iron chelator, antioxidant and MAO-inhibitor M30 [5-(N-methyl-N-propargylaminomethyl)-8-hydroxyquinoline]. Five-day oral pre-treatment of adult male Wistar rats with 10 mg/kg/day M30 dose was followed by a single injection of STZ (1 mg/kg, icv). CAT activity was measured colorimetrically in the hippocampus (HPC), brain stem (BS) and cerebellum (CB) of the control, STZ-, M30- and STZ + M30-treated rats, respectively, 4 weeks after the STZ treatment. STZ-treated rats demonstrated significantly lower CAT activity in all three brain regions in comparison to the controls (p iron chelators such as M30 might also have beneficial effects in this non-transgenic sAD model.

  2. Disruption of AtWNK8 Enhances Tolerance of Arabidopsis to Salt and Osmotic Stresses via Modulating Proline Content and Activities of Catalase and Peroxidase

    Directory of Open Access Journals (Sweden)

    Hong Liao

    2013-03-01

    Full Text Available With no lysine kinases (WNKs play important roles in plant growth and development. However, its role in salt and osmotic stress tolerance is unclear. Here, we report that AtWNK8 is mainly expressed in primary root, hypocotyl, stamen and pistil and is induced by NaCl and sorbitol treatment. Compared to the wild-type, the T-DNA knock-out wnk8 mutant was more tolerant to severe salinity and osmotic stresses, as indicated by 27% and 198% more fresh weight in the NaCl and sorbitol treatment, respectively. The wnk8 mutant also accumulated 1.43-fold more proline than the wild-type in the sorbitol treatment. Under NaCl and sorbitol stresses, catalase (CAT activity in wnk8 mutant was 1.92- and 3.7-times of that in Col-0, respectively. Similarly, under salt and osmotic stress conditions, peroxidase (POD activities in wnk8 mutant were 1.81- and 1.58-times of that in Col-0, respectively. Taken together, we revealed that maintaining higher CAT and POD activities might be one of the reasons that the disruption of AtWNK8 enhances the tolerance to salt stress, and accumulating more proline and higher activities of CAT and POD might result in the higher tolerance of WNK8 to osmotic stress.

  3. Catalase and superoxide dismutase activities and the total protein content of protocorm-like bodies of Dendrobium sonia-28 subjected to vitrification.

    Science.gov (United States)

    Poobathy, Ranjetta; Sinniah, Uma Rani; Xavier, Rathinam; Subramaniam, Sreeramanan

    2013-07-01

    Dendrobium sonia-28 is an important ornamental orchid in the Malaysian flower industry. However, the genus faces both low germination rates and the risk of producing heterozygous progenies. Cryopreservation is currently the favoured long-term storage method for orchids with propagation problems. Vitrification, a frequently used cryopreservation technique, involves the application of pretreatments and cryoprotectants to protect and recover explants during and after storage in liquid nitrogen. However, cryopreservation may cause osmotic injuries and toxicity to cryopreserved explants from the use of highly concentrated additives, and cellular injuries from thawing, devitrification and ice formation. Reactive oxygen species (ROS), occurring during dehydration and cryopreservation, may also cause membrane damage. Plants possess efficient antioxidant systems such as the superoxide dismutase (SOD) and catalase (CAT) enzymes to scavenge ROS during low temperature stress. In this study, protocorm-like bodies (PLBs) of Dendrobium sonia-28 were assayed for the total protein content, and both SOD and CAT activities, at each stage of a vitrification exercise to observe for deleterious stages in the protocol. The results indicated that cryopreserved PLBs of Dendrobium sonia-28 underwent excessive post-thawing oxidative stress due to decreased levels of the CAT enzyme at the post-thawing recovery stage, which contributed to the poor survival rates of the cryopreserved PLBs.

  4. Paraquat Resistance in Leaf Discs of PSAG12-IPT Modified Gerbera Is Related to the Activities of Superoxide Dismutase, Catalase, and Dehydroascorbate Reductase

    Institute of Scientific and Technical Information of China (English)

    LAI Qi-xian; BAO Zhi-yi; ZHU Zhu-jun; MAO Bi-zeng; QIAN Qiong-qiu

    2007-01-01

    In this paper, using in vitro leaf disc culture system, the changes of contents of chlorophylls, carotenoids, soluble protein, thiobarbituric acid reactive substance (TBARS), and activities of antioxidant enzymes were investigated during the incubation of leaf discs of PSAG12-IPT modified gerbera in 0, 25, 50 μmol L-1 paraquat (PQ) under continuous light intensity of 130 μmol m-2 s-1, compared with the control plant (wild type). The results showed that PQ treatment significantly decreased the contents of chlorophylls, carotenoids, and soluble protein, therefore, promoted leaf senescence. However, the decreases in the leaf discs of modified gerbera were considerably smaller. The activities of superoxide dismutase (SOD), catalase (CAT), and dehydroascorbate reductase (DHAR) were significantly increased by PQ treatment and with the increasing of PQ concentration, particularly in the modified plants. The activities of ascorbate peroxidase (APX) and guaiacol peroxidase (GPX) could not be detected in the leaf discs of PQ treatments, which suggested that they were labile to the oxidative stress induced by PQ. As a product of lipid peroxidation, TBARS significantly increased in content with the increase of PQ concentration, while its concentration in the modified plants was significantly lower than that of control plants. Therefore, it could be concluded that the chimeric gene PSAG12-IPT transformed gerbera leaves had higher antioxidative potential, thus causing the delay of senescence under oxidative stress induced by PQ.

  5. The link between antioxidant enzymes catalase and glutathione S-transferase and physiological condition of a control population of terrestrial isopod (Porcellio scaber).

    Science.gov (United States)

    Jemec, Anita; Lešer, Vladka; Drobne, Damjana

    2012-05-01

    The aim of this work was to investigate if the activities of catalase and glutathione S-transferase in a control population of terrestrial isopods (Porcellio scaber) are correlated with the physiological condition of the isopods. For this purpose, the activities of these enzymes were analysed in isopods from a stock population and in parallel, the physiological condition of the same specimens was assessed using a histological approach based on epithelial thickness and lipid droplets. We found a correlation between antioxidant enzymes and the physiological condition of the isopods. This implies that these enzymes could be used as predictive indicators of the physiological condition in a stock population before comprehensive toxicological studies are conducted and also in control group after the experiment. When a control group is found to be very heterogeneous in terms of physiological condition, the experiment should be repeated with a larger number of experimental animals. The findings of this study will contribute to more accurate experimental design of toxicity tests when using biomarkers. This should encourage other researchers to increase their effort to know the physiological state of their test organisms.

  6. Genome-wide Screening of Regulators of Catalase Expression: ROLE OF A TRANSCRIPTION COMPLEX AND HISTONE AND tRNA MODIFICATION COMPLEXES ON ADAPTATION TO STRESS.

    Science.gov (United States)

    García, Patricia; Encinar Del Dedo, Javier; Ayté, José; Hidalgo, Elena

    2016-01-01

    In response to environmental cues, the mitogen-activated protein kinase Sty1-driven signaling cascade activates hundreds of genes to induce a robust anti-stress cellular response in fission yeast. Thus, upon stress imposition Sty1 transiently accumulates in the nucleus where it up-regulates transcription through the Atf1 transcription factor. Several regulators of transcription and translation have been identified as important to mount an integral response to oxidative stress, such as the Spt-Ada-Gcn5-acetyl transferase or Elongator complexes, respectively. With the aim of identifying new regulators of this massive gene expression program, we have used a GFP-based protein reporter and screened a fission yeast deletion collection using flow cytometry. We find that the levels of catalase fused to GFP, both before and after a threat of peroxides, are altered in hundreds of strains lacking components of chromatin modifiers, transcription complexes, and modulators of translation. Thus, the transcription elongation complex Paf1, the histone methylase Set1-COMPASS, and the translation-related Trm112 dimers are all involved in full expression of Ctt1-GFP and in wild-type tolerance to peroxides.

  7. Manganese superoxide dismutase (SOD2/MnSOD)/catalase and SOD2/GPx1 ratios as biomarkers for tumor progression and metastasis in prostate, colon, and lung cancer.

    Science.gov (United States)

    Miar, Ana; Hevia, David; Muñoz-Cimadevilla, Henar; Astudillo, Aurora; Velasco, Julio; Sainz, Rosa M; Mayo, Juan C

    2015-08-01

    The role of manganese-dependent superoxide dismutase (SOD2/MnSOD) during tumor progression has been studied for several decades with controversial results. While SOD2 downregulation was initially associated with tumor initiation and was proposed as a tumor suppressor gene, recent studies have reported that SOD2 might favor tumor progression and dissemination. To our knowledge this is the first time that changes in SOD2 expression in three different types of tumors, i.e., prostate, lung, and colon cancer, are studied by analyzing both SOD2 mRNA and protein levels in a total of 246 patients' samples. In prostate samples, SOD2 protein levels were also increased, especially in middle stage tumors. In the case of colon and lung tumors both mRNA and protein SOD2 levels were increased in malignant tissues compared to those in nontumor samples. More importantly, all metastases analyzed showed increased levels of SOD2 when compared to those of normal primary tissue and healthy adjacent tissue. Together, these results suggest that a common redox imbalance in these three types of tumor occurs at intermediate stages which then might favor migration and invasion, leading to a more aggressive cancer type. Consequently, the ratios SOD2/catalase and SOD2/Gpx1 could be considered as potential markers during progression from tumor growth to metastasis.

  8. Amperometric cholesterol biosensor based on the direct electrochemistry of cholesterol oxidase and catalase on a graphene/ionic liquid-modified glassy carbon electrode.

    Science.gov (United States)

    Gholivand, Mohammad Bagher; Khodadadian, Mehdi

    2014-03-15

    Cholesterol oxidase (ChOx) and catalase (CAT) were co-immobilized on a graphene/ionic liquid-modified glassy carbon electrode (GR-IL/GCE) to develop a highly sensitive amperometric cholesterol biosensor. The H2O2 generated during the enzymatic reaction of ChOx with cholesterol could be reduced electrocatalytically by immobilized CAT to obtain a sensitive amperometric response to cholesterol. The direct electron transfer between enzymes and electrode surface was investigated by cyclic voltammetry. Both enzymes showed well-defined redox peaks with quasi-reversible behaviors. An excellent sensitivity of 4.163 mA mM(-1)cm(-2), a response time less than 6s, and a linear range of 0.25-215 μM (R(2)>0.99) have been observed for cholesterol determination using the proposed biosensor. The apparent Michaelis-Menten constant (KM(app)) was calculated to be 2.32 mM. The bienzymatic cholesterol biosensor showed good reproducibility (RSDsanalytical performance for the determination of free cholesterol in human serum samples.

  9. Anti-inflammatory effects of Lactobacillus casei BL23 producing or not a manganese-dependant catalase on DSS-induced colitis in mice

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    Corthier Gérard

    2007-07-01

    Full Text Available Abstract Background Human immune cells generate large amounts of reactive oxygen species (ROS throughout the respiratory burst that occurs during inflammation. In inflammatory bowel diseases, a sustained and abnormal activation of the immune system results in oxidative stress in the digestive tract and in a loss of intestinal homeostasis. We previously showed that the heterologous production of the Lactobacillus plantarum ATCC14431 manganese-dependant catalase (MnKat in Lb. casei BL23 successfully enhances its survival when exposed to oxidative stress. In this study, we evaluated the preventive effects of this antioxidative Lb. casei strain in a murine model of dextran sodium sulfate (DSS-induced moderate colitis. Results Either Lb. casei BL23 MnKat- or MnKat+ was administered daily to mice treated with DSS for 10 days. In contrast to control mice treated with PBS for which DSS induced bleeding diarrhea and mucosal lesions, mice treated with both Lb. casei strains presented a significant (p Conclusion No contribution of MnKat to the protective effect from epithelial damage has been observed in the tested conditions. In contrast, these results confirm the high interest of Lb. casei as an anti-inflammatory probiotic strain.

  10. Role of Burkholderia pseudomallei Sigma N2 in Amino Acids Utilization and in Regulation of Catalase E Expression at the Transcriptional Level

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    Duong Thi Hong Diep

    2015-01-01

    Full Text Available Burkholderia pseudomallei is the causative agent of melioidosis. The complete genome sequences of this pathogen have been revealed, which explain some pathogenic mechanisms. In various hostile conditions, for example, during nitrogen and amino acid starvation, bacteria can utilize alternative sigma factors such as RpoS and RpoN to modulate genes expression for their adaptation and survival. In this study, we demonstrate that mutagenesis of rpoN2, which lies on chromosome 2 of B. pseudomallei and encodes a homologue of the sigma factor RpoN, did not alter nitrogen and amino acid utilization of the bacterium. However, introduction of B. pseudomallei rpoN2 into E. coli strain deficient for rpoN restored the ability to utilize amino acids. Moreover, comparative partial proteomic analysis of the B. pseudomallei wild type and its rpoN2 isogenic mutant was performed to elucidate its amino acids utilization property which was comparable to its function found in the complementation assay. By contrast, the rpoN2 mutant exhibited decreased katE expression at the transcriptional and translational levels. Our finding indicates that B. pseudomallei RpoN2 is involved in a specific function in the regulation of catalase E expression.

  11. 电泳鉴定脱墨浆生产系统过氧化氢酶的研究%A PRELIMINARY STUDY ON APPLYING ELECTOPHORESIS TO IDENTIFY CATALASE IN DIP PRODUCTION SYSTEMS

    Institute of Scientific and Technical Information of China (English)

    刘蓓; 王海毅

    2008-01-01

    This paper discussed the experimental conditions of polyacrylamide gel electrophoresis (PAGE) to identify catalase in deinked pulp (DIP) production systems and analyzed the isozymes of catalase. The results indicated that the sample size of repulping stage was 3 μL; the sample size of the first concentration stage was 5 μL; the sample size of the second concentration stage was 10 μL; the isolation gel concentration was 9% and the concentration gel concentration was 4%. According to results of identification, there were 6 enzyme bands in production systems and each section had different enzyme bands. Repulping stage had the largest number of catalase isozymes species and there were 4 bands in the gel. The first concentration stage had three bands, two of which were unique ones. The second concentration only contained one catalase band what each stages had with the weakest enzyme activity. By analyzing this phenomenon, the main reason for this was that microorganism affects the kinds of catalase. According to the electrophoresis mobility, catalase molecular size ranking was C1>C2>C3>C4>C5>C6>C7. All of them were less than the liver catalse's molecular weight in DIP system.%研究了聚丙烯酰胺凝胶电泳(polyacrylamide gel electrophoresis,简称PAGE)鉴定脱墨浆(DIP)生产系统中过氧化氢酶的实验条件,并对各工段过氧化氢酶同工酶进行了分析.实验结果表明,在碎浆工段点样量3 μL,一段浓缩点样量5 μL,二段浓缩点样量10 μL,浓缩胶质量分数4%,分离胶质量分数9%条件下才能得到较高质量的电泳结果.生产系统中共检测出6条酶带,各工段出现的酶带的数量不同,种类也不同,仅一条酶带在3个工段均出现,进一步探讨了此现象产生的原因,并对6种同工酶的相对分子质量进行了排序.

  12. 小鼠组织中过氧化氢酶的活性与年龄的关系%Age-related changes in catalase activity in different tissues of mice

    Institute of Scientific and Technical Information of China (English)

    孔德胜; 王晓然; 李文君; 陈永春; 姚敏; 高媛

    2012-01-01

    The age-related changes in catalase activity were observed in mice liver, kidney, lung, heart, spleen, stomach and brain. The catalase activity in these tissues in mice aged of 1-month, 4-month and 18-month was determined by Permanganate Titration. The results showed that the activity of catalase varied in different tissues, in descending order as follows: liver > kidney > lung > heart, spleen, stomach > brain. The catalase in mice lung, heart, spleen, stomach and brain showed an age-related increase in the activity from 1-month to 4-month,and an age-related decrease from 4-month to 18-month. In liver and kidney, there was no significant age-associated change in the catalase activity from 1-month to 4-month, while the activity of catalase declined with increasing age from 4-month to 18-month. The age-related change of catalase activity in mice liver, kidney, lung, heart, spleen, stomach and brain suggested that the decrease of catalase activity was closely correlated with the aging process.%为观察小鼠组织中过氧化氢酶的活性与年龄的关系,采用高锰酸钾滴定法测定不同年龄(1、4、18月龄)小鼠肝、肾、肺、心、脾、胃、脑组织中过氧化氢酶的活性.结果显示:小鼠过氧化氢酶在不同组织中活性不同,活性高低顺序基本表现为:肝>肾>肺>心、脾、胃>脑;小鼠肺、心、脾、胃、脑各组织中过氧化氢酶的活性在1~4月龄间随年龄增加而增加,在4~18月龄间随年龄增加而降低;小鼠肝、肾组织中过氧化氢酶的活性在1~4月龄间与年龄相关性不显著,在4~18月龄间随年龄增加而降低.结果表明,小鼠肝、肾、肺、心、脾、胃、脑等组织中过氧化氢酶的活性随年龄变化而变化,机体过氧化氢酶活性的降低与机体衰老密切相关.

  13. Effect of Lead (Pb Exposure on the Activity of Superoxide Dismutase and Catalase in Battery Manufacturing Workers (BMW of Western Maharashtra (India with Reference to Heme biosynthesis

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    Kusal K. Das

    2006-12-01

    Full Text Available The aim of this study was to estimate the activity of superoxide dismutase (SOD and catalase in erythrocytes and malondialdehyde (MDA in plasma of battery manufacturing workers (BMW of Western Maharashtra (India who were occupationally exposed to lead (Pb over a long period of time (about 15 years. This study was also aimed to determine the Pb intoxication resulted in a disturbance of heme biosynthesis in BMW group. The blood Pb level of BMW group (n = 28 was found to be in the range of 25.8 – 78.0 μg/dL (mean + SD, 53.63 + 16.98 whereas in Pb unexposed control group (n = 35 the range was 2.8 – 22.0 μg/dL (mean + SD, 12.52 + 4.08. The blood level (Pb-B and urinary lead level (Pb-U were significantly increased in BMW group as compared to unexposed control. Though activated d- aminolevulinic acid dehydratase (ALAD activities in BMW group did not show any significant change when compared to control group but activated / non activated erythrocyte – ALAD activities in BMW group showed a significant increase. Erythrocyte- zinc protoporphyrin (ZPP, urinary daminolevulinic acid (ALA-U and porphobilinogen (PBG-U of BMW groups elevated significantly as compared to control. A positive correlation (r = 0.66, p 1.0 were observed in control group. Hematological study revealed a significant decrease of hemoglobin concentration, packed cell volume (% and other blood indices and a significant increase of total leucocytes count in BMW group in comparison to control group. The serum MDA content was significantly increased (p< 0.001 and the activities of antioxidant enzymes such as erythrocyte- SOD (p< 0.001 and erythrocytecatalase (p< 0.001 were significantly reduced in BMW group as compared to control group. A positive correlation (r = 0.45, p<0.02 between Pb-B and serum MDA level was observed in BMW group (Pb-B range 25.8 – 78.0 μg / dL but such significant correlation did not notice in

  14. Hinokitiol Exerts Anticancer Activity through Downregulation of MMPs 9/2 and Enhancement of Catalase and SOD Enzymes: In Vivo Augmentation of Lung Histoarchitecture

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    Chien-Hsun Huang

    2015-09-01

    Full Text Available Melanoma is extremely resistant to chemotherapy and the death rate is increasing hastily worldwide. Extracellular matrix promotes the migration and invasion of tumor cells through the production of matrix metalloproteinase (MMP-2 and -9. Evidence has shown that natural dietary antioxidants are capable of inhibiting cancer cell growth. Our recent studies showed that hinokitiol, a natural bioactive compound, inhibited vascular smooth muscle cell proliferation and platelets aggregation. The present study is to investigate the anticancer efficacy of hinokitiol against B16-F10 melanoma cells via modulating tumor invasion factors MMPs, antioxidant enzymes in vitro. An in vivo mice model of histological investigation was performed to study the patterns of elastic and collagen fibers. Hinokitiol inhibited the expression and activity of MMPs-2 and -9 in B16-F10 melanoma cells, as measured by western blotting and gelatin zymography, respectively. An observed increase in protein expression of MMPs 2/9 in melanoma cells was significantly inhibited by hinokitiol. Notably, hinokitiol (1–5 μM increased the activities of antioxidant enzymes catalase (CAT and superoxide dismutase (SOD from the reduction in melanoma cells. Also, hinokitiol (2–10 µM concentration dependently reduced in vitro Fenton reaction induced hydroxyl radical (OH· formation. An in vivo study showed that hinokitiol treatment increased elastic fibers (EF, collagens dispersion, and improved alveolar alterations in the lungs of B16/F10 injected mice. Overall, our findings propose that hinokitiol may be a potent anticancer candidate through down regulation of MMPs 9/2, reduction of OH· production and enhancement of antioxidant enzymes SOD and CAT.

  15. Polysaccharide-Rich Fraction of Noni Fruit (Morinda citrifolia L.) as Doxorubicin Co-Chemotherapy: Evaluation of Catalase, Macrophages, and TCD8+ Lymphocytes.

    Science.gov (United States)

    Sasmito, Ediati; Hertiani, Triana; Novlita Renggani, Tiya; Jaya Laksana, Brata

    2015-01-01

    Noni fruit (Morinda citrifolia L.) has been acknowledged for its cytotoxic and immunostimulatory activity. Our previous results on the immunomodulatory effect of a noni juice polysaccharide-rich fraction encouraged this research to evaluate the potency of the polysaccharide-rich fraction as co-chemotherapy with doxorubicin (DOX) administration. Macrophage activity (MA) was evaluated with the latex bead method. The phagocytic index (PI) was measured as the number of latex beads ingested by 100 macrophages, while the phagocytosis ratio (PR) was indicated by the percentage of macrophages that ingested three or more latex beads. The CEC was evaluated by using a commercial assay kit, while CD8+ T lymphocyte proliferation was evaluated using a flowcytometry method following in vivo administration. Thirty male Wistar rats were divided into five groups (n = 6 each). The control group received DOX via i.p. at a concentration of 4.67 mg/kg BW on days 1 and 4; four treatment groups received PF p.o. at a concentration of 25; 50; 100; 200 mg/kg BW daily, respectively, and additionally DOX i.p. 4.67 mg/kg BW (days 1 and 4) for 7 days. The phagocytic activity was not affected significantly by PF administration compared to the Dox control, but PF administration at a dose of 25 and 50 mg/kg BW has been proven to increase TCD8+ cell proliferation in combination with DOX. The catalase concentration, on the other hand, significantly decreased following PF administration at a dose of 100 mg/kg BW. The results suggest that the polysaccharide-rich fraction of noni juice might induce immunomodulatory effects via TCD8+ activation, have antioxidant activity, and thus might be a potential candidate to be used as an adjuvant to DOX chemotherapy.

  16. Molecular cloning, characterization, and the response of Cu/Zn superoxide dismutase and catalase to PBDE-47 and -209 from the freshwater bivalve Anodonta woodiana.

    Science.gov (United States)

    Xia, Xichao; Huang, Chuanfeng; Zhang, Dongxian; Zhang, Yi; Xue, Shipeng; Wang, Xiying; Zhang, Qingyuan; Guo, Lianghong

    2016-04-01

    Polybrominated diphenyl ethers-47 (PBDE-47) and -209 are significant components of total PBDEs in water and can catalyze the production of reactive oxygen species (ROS) in the organisms. Anti-oxidant enzymes play an important role in scavenging the high level of ROS. In the current study, two full-length cDNAs of Cu/Zn superoxide dismutase (CuZnSODs) and catalase (CAT) were isolated from freshwater bivalve Anodonta woodiana by rapid amplification of cDNA ends approach and respectively named as AwSOD and AwCAT. The nucleotide sequence of AwSOD cDNA had an open reading frame (ORF) of 465 bp encoding a polypeptide of 155 amino acids in which signature 1 GKHGFHVHEFGDNT and signature 2 GNAGARSACGVI of SODs were observed. Deduced amino acid sequence of AwSOD showed a significant similarity with that of CuZnSODs. AwCAT had an ORF 1536 bp encoding a polypeptide of 512 amino acids which contains a conserved catalytic site motif, and a proximal heme-ligand signature motif of CATs. The time-course expressions of AwSOD and AwCAT in hepatopancreas were measured by quantitative real-time PCR. Expressions of AwSOD and AwCAT showed a significant up-regulation in groups at a low concentration treatment of PBDE-47, a biphasic pattern in groups with a high concentration treatment. Administration of PBDE-209 could result in an up-regulation of AwSOD and AwCAT expressions with time- and dose-dependent matter. These results indicate that up-regulations of AwSOD and AwCAT expression of hepatopancreas of freshwater bivalve A. woodiana contribute to eliminate oxidative stress derived from PBDE-47 and -209 treated.

  17. Roles of catalase (CAT) and ascorbate peroxidase (APX) genes in stress response of eggplant (Solanum melongena L.) against Cu(+2) and Zn(+2) heavy metal stresses.

    Science.gov (United States)

    Soydam-Aydın, Semra; Büyük, İlker; Cansaran-Duman, Demet; Aras, Sümer

    2015-12-01

    Eggplant (Solanum melongena L.) is a good source of minerals and vitamins and this feature makes its value comparable with tomato which is economically the most important vegetable worldwide. Due to its common usage as food and in medicines, eggplant cultivation has a growing reputation worldwide. But genetic yield potential of an eggplant variety is not always attained, and it is limited by some factors such as heavy metal contaminated soils in today's world. Today, one of the main objectives of plant stress biology and agricultural biotechnology areas is to find the genes involved in antioxidant stress response and engineering the key genes to improve the plant resistance mechanisms. In this regard, the current study was conducted to gain an idea on the roles of catalase (CAT) and ascorbate peroxidase (APX) genes in defense mechanism of eggplant (S. melongena L., Pala-49 (Turkish cultivar)) treated with different concentrations of Cu(+2) and Zn(+2). For this aim, the steady-state messenger RNA (mRNA) levels of CAT and APX genes were determined by quantitative real-time PCR (qRT-PCR) in stressed eggplants. The results of the current study showed that different concentrations of Cu(+2) and Zn(+2) stresses altered the mRNA levels of CAT and APX genes in eggplants compared to the untreated control samples. When the mRNA levels of both genes were compared, it was observed that CAT gene was more active than APX gene in eggplant samples subjected to Cu(+2) contamination. The current study highlights the importance of CAT and APX genes in response to Cu(+2) and Zn(+2) heavy metal stresses in eggplant and gives an important knowledge about this complex interaction.

  18. Host genetic variations in glutathione-S-transferases, superoxide dismutases and catalase genes influence susceptibility to malaria infection in an Indian population.

    Science.gov (United States)

    Fernandes, Rayzel C; Hasan, Marriyah; Gupta, Himanshu; Geetha, K; Rai, Padmalatha S; Hande, Manjunath H; D'Souza, Sydney C; Adhikari, Prabha; Brand, Angela; Satyamoorthy, Kapaettu

    2015-06-01

    Antioxidant enzymes can contribute to disease susceptibility or determine response to therapy in individuals with malaria. Genetic variations due to polymorphisms in host genes encoding antioxidant enzymes such as glutathione S-transferases-theta, mu, pi (GSTT, GSTM, GSTP), superoxide dismutases (SOD) and catalase (CAT), may therefore, influence inter-individual response to malaria pathology and propensity of infection caused by Plasmodium vivax (Pv) and Plasmodium falciparum (Pf). Therefore, using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing, we investigated the association of deletions of GSTT1 and GSTM1, single nucleotide polymorphisms (SNPs) of GSTP1 (rs1695), SOD1 (rs2234694), SOD2 (rs4880, rs1141718), SOD3 (rs2536512) and CAT (rs1001179) in individuals infected with Pf (n = 100) and Pv (n = 100) against healthy controls (n = 150). Our data suggest a significant role for GSTM1 deletions in complicated Pv (p = 0.0007) malaria with ODDs ratio 3.8 [with 95 % confidence interval (CI) 1.9-7.4]. The results also indicated that polymorphisms present in GSTP1, SOD1 and CAT genes may be associated with malaria susceptibility (p < 0.05), whereas SOD3 polymorphism may play a role in malarial resistance (p < 0.05). In addition, we observed significant SNP-SNP interactions with synergistic genetic effects in SOD2, SOD3 and CAT genes for Pv and in SOD2 and SOD3 genes for Pf. In conclusion, our results provide convincing evidence for a relationship between polymorphisms in host antioxidant enzymes and susceptibility to malaria infection.

  19. Enhanced Reactive Oxygen Species Scavenging by Overproduction of Superoxide Dismutase and Catalase Delays Postharvest Physiological Deterioration of Cassava Storage Roots1[C][W][OA

    Science.gov (United States)

    Xu, Jia; Duan, Xiaoguang; Yang, Jun; Beeching, John R.; Zhang, Peng

    2013-01-01

    Postharvest physiological deterioration (PPD) of cassava (Manihot esculenta) storage roots is the result of a rapid oxidative burst, which leads to discoloration of the vascular tissues due to the oxidation of phenolic compounds. In this study, coexpression of the reactive oxygen species (ROS)-scavenging enzymes copper/zinc superoxide dismutase (MeCu/ZnSOD) and catalase (MeCAT1) in transgenic cassava was used to explore the intrinsic relationship between ROS scavenging and PPD occurrence. Transgenic cassava plants integrated with the expression cassette p54::MeCu/ZnSOD-35S::MeCAT1 were confirmed by Southern-blot analysis. The expression of MeCu/ZnSOD and MeCAT1 was verified by quantitative reverse transcription-polymerase chain reaction and enzymatic activity analysis both in the leaves and storage roots. Under exposure to the ROS-generating reagent methyl viologen or to hydrogen peroxide (H2O2), the transgenic plants showed higher enzymatic activities of SOD and CAT than the wild-type plants. Levels of malondialdehyde, chlorophyll degradation, lipid peroxidation, and H2O2 accumulation were dramatically reduced in the transgenic lines compared with the wild type. After harvest, the storage roots of transgenic cassava lines show a delay in their PPD response of at least 10 d, accompanied by less mitochondrial oxidation and H2O2 accumulation, compared with those of the wild type. We hypothesize that this is due to the combined ectopic expression of Cu/ZnSOD and CAT leading to an improved synergistic ROS-scavenging capacity of the roots. Our study not only sheds light on the mechanism of the PPD process but also develops an effective approach for delaying the occurrence of PPD in cassava. PMID:23344905

  20. CATALASA, PEROXIDASA Y POLIFENOLOXIDASA EN PITAYA AMARILLA (Acanthocereus pitajaya: MADURACIÓN Y SENESCENCIA Catalase, Peroxidase and Polyphenoloxidase from Pitaya Amarilla (Acanthocereus pitajaya Fruits: Ripening and Senescense

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    CARLOS EDUARDO NARVÁEZ CUENCA

    Full Text Available Se evaluó la relación entre algunos síntomas de deterioro y la actividad de enzimas vinculadas tanto con el pardeamiento como con el sistema antioxidante en frutos de pitaya amarilla (Acanthocereus pitajaya cosechados en su madurez fisiológica y almacenados durante 15 días a 24 °C y 85% de humedad relativa. En los frutos enteros se evaluaron la intensidad respiratoria y el color externo; en la corteza se determinaron la actividad de