Catalase activity and the survival of Pseudomonas putida, a root colonizer, upon treatment with peracetic acid

2001 ◽  
Vol 47 (3) ◽  
pp. 222-228 ◽  
Author(s):  
Anne J Anderson ◽  
Charles D Miller
Keyword(s):  
Hydrogen Peroxide ◽  
Pseudomonas Putida ◽  
Peracetic Acid ◽  
Catalase Activity ◽  
Specific Activity ◽  
Active Oxygen Species ◽  
Oxygen Species ◽  
Deficient Mutant ◽  
Wild Type ◽  
Cell Extracts

Peracetic acid is used as a sterilant in several industrial settings. Cells of a plant-colonizing bacterium, Pseudomonas putida in liquid suspension, were more sensitive to killing by peracetic acid when they lacked a major catalase activity, catalase A. Low doses of peracetic acid induced promoter activity of the gene encoding catalase A and increased total catalase specific activity in cell extracts. Microbes present in native agricultural soils rapidly degraded the active oxygen species present in peracetic acid. The simultaneous release of oxygen was consistent with a role for catalase in degrading the hydrogen peroxide that is part of the peracetic acid-equilibrium mixture. Amendment of sterilized soils with wild-type P. putida restored the rate of degradation of peracetic acid to a higher level than was observed in the soils amended with the catalase A-deficient mutant. The association of the bacteria with the plant roots resulted in protection of the wild-type as well as the catalase-deficient mutant from killing by peracetic acid. No differential recovery of the wild-type and catalase A mutant of P. putida was observed from roots after the growth matrix containing the plants was flushed with peracetic acid.Key words: Pseudomonas putida (Pp), activated oxygen species (AOS), hydrogen peroxide, luciferase, colonization.

Catalase activity in Campylobacter jejuni: comparison of a wild-type strain with an aerotolerant variant

1990 ◽  
Vol 36 (6) ◽  
pp. 449-451 ◽  
Author(s):  
Pamela A. Vercellone ◽  
Robert M. Smibert ◽  
Noel R. Krieg
Keyword(s):  
Campylobacter Jejuni ◽  
Catalase Activity ◽  
Specific Activity ◽  
Wild Type ◽  
Oxygen Tolerance ◽  
Variant Strain ◽  
Cultural Conditions ◽  
Cell Extracts ◽  
Bactericidal Effects

A comparison of Campylobacter jejuni VPI strain H840 (ATCC 29428), which can grow at O2 levels up to 15%, with variant strain MC711-01 (which can grown at O2 levels up to 21–26%) indicated that the specific activity of catalase in crude cell extracts was higher in the variant by a factor of 1.6 to 2.5, depending on cultural conditions. Smaller differences occurred with superoxide dismutase activity, while peroxidase activities were invariably lower in the variant strain. The variant strain was much more resistant than the wild type to the bactericidal effects of H2O2. The results suggest that catalase activity might be one of the factors associated with the greater tolerance of O2 by the variant strain. However, both strains became more susceptible to H2O2 when cultures were initially grown at 6% O2 and then shifted to 21% O2; thus the role of catalase in the oxygen tolerance of C. jejuni is probably minor. Key words: Campylobacter jejuni, catalase, oxygen tolerance.

Characterization of catalase activities in a root-colonizing isolate of Pseudomonas putida

1992 ◽  
Vol 38 (10) ◽  
pp. 1026-1032 ◽  
Author(s):  
J. Katsuwon ◽  
A. J. Anderson
Keyword(s):  
Stationary Phase ◽  
Pseudomonas Putida ◽  
Growth Phase ◽  
Specific Activity ◽  
Active Oxygen Species ◽  
Stationary Growth Phase ◽  
Wild Type ◽  
Ph Optimum ◽  
Chloroform Methanol ◽  
Root Surfaces

Pseudomonas putida, a saprophytic root-colonizing bacterium, produces multiple forms of catalase. Catalase A, which increases in specific activity during growth phase and after treatment with H2O2, is located in the cytoplasm and is inhibited by 3-amino-1,2,4-triazole, EDTA, and cyanide, but not by chloroform–methanol treatment. Catalase B, which is induced by external H2O2 or during stationary phase of growth, is membrane associated and is inhibited by chloroform–methanol, EDTA, and cyanide, but not by aminotriazole. Catalase A has a broad pH optimum, from pH 6.0 to 11.0, with two peaks, at pH 8.0 and 11.0. Catalase B is most active at pH 5.0–11.0. Mutant J-1, generated by ethyl methanesulfonate mutagenesis, lacked catalase A activity in extracts of cells harvested throughout lag to early stationary growth phase in liquid medium. Catalase B was produced by J-1 in stationary phase. Exposure of J-1 to H2O2 caused the production of both catalase A and catalase B. Mutant J-1 was more susceptible to cell death than the wild type upon direct exposure to 2.5 mM H2O2 but survived this treatment after exposure to lower (0.3 mM), nonlethal doses of H2O2. The ability to adapt to H2O2 may be related to the behaviour of J-1 on roots where active oxygen species are produced by root surface enzymes. J-1 colonized root surfaces at wild-type levels and produced catalases A and B after exposure to root surfaces for 12 h. Key words: Pseudomonas putida, catalase, root colonization.

scholarly journals Effects of Combination of Different −10 Hexamers and Downstream Sequences on Stationary-Phase-Specific Sigma Factor ςS-Dependent Transcription in Pseudomonas putida

2000 ◽  
Vol 182 (23) ◽  
pp. 6707-6713 ◽  
Author(s):  
Eve-Ly Ojangu ◽  
Andres Tover ◽  
Riho Teras ◽  
Maia Kivisaar
Keyword(s):  
Stationary Phase ◽  
Pseudomonas Putida ◽  
Sigma Factor ◽  
Sigma Factors ◽  
Deficient Mutant ◽  
Wild Type ◽  
In Vivo Experiments ◽  
Silent Genes ◽  
Rna Polymerase Holoenzyme

ABSTRACT The main sigma factor activating gene expression, necessary in stationary phase and under stress conditions, is ςS. In contrast to other minor sigma factors, RNA polymerase holoenzyme containing ςS (EςS) recognizes a number of promoters which are also recognized by that containing ς70 (Eς70). We have previously shown that transposon Tn4652 can activate silent genes in starvingPseudomonas putida cells by creating fusion promoters during transposition. The sequence of the fusion promoters is similar to the ς70-specific promoter consensus. The −10 hexameric sequence and the sequence downstream from the −10 element differ among these promoters. We found that transcription from the fusion promoters is stationary phase specific. Based on in vivo experiments carried out with wild-type and rpoS-deficient mutant P. putida, the effect of ςS on transcription from the fusion promoters was established only in some of these promoters. The importance of the sequence of the −10 hexamer has been pointed out in several published papers, but there is no information about whether the sequences downstream from the −10 element can affect ςS-dependent transcription. Combination of the −10 hexameric sequences and downstream sequences of different fusion promoters revealed that ςS-specific transcription from these promoters is not determined by the −10 hexameric sequence only. The results obtained in this study indicate that the sequence of the −10 element influences ςS-specific transcription in concert with the sequence downstream from the −10 box.

Lung edema due to hydrogen peroxide is independent of cyclooxygenase products

1984 ◽  
Vol 56 (4) ◽  
pp. 900-905 ◽  
Author(s):  
O. Burghuber ◽  
M. M. Mathias ◽  
I. F. McMurtry ◽  
J. T. Reeves ◽  
N. F. Voelkel
Keyword(s):  
Hydrogen Peroxide ◽  
Glucose Oxidase ◽  
Direct Action ◽  
Active Oxygen Species ◽  
Active Oxygen ◽  
Lung Edema ◽  
Oxygen Species ◽  
Edema Formation ◽  
Prostaglandin Production ◽  
Prostaglandin F2 Alpha

Active oxygen species can cause lung injury. Although a direct action on endothelial cells is proposed, the possibility exists that they might cause injury via mediators. We considered that active oxygen species would stimulate the generation of cyclooxygenase metabolites, which then alter pulmonary vasoreactivity and cause edema. We chemically produced hydrogen peroxide by adding glucose oxidase to a plasma- and cell-free, but beta-D-glucose-containing, solution, which perfused isolated rat lungs. Addition of glucose oxidase to the perfusate caused a marked decrease in pulmonary vasoreactivity, accompanied by an increase in the concentrations of prostacyclin, thromboxane A2, and prostaglandin F2 alpha. Pretreatment with catalase, a specific scavenger of hydrogen peroxide, preserved pulmonary vasoreactivity, inhibited the increase of the concentration of the measured prostaglandins, and prevented edema formation. Indomethacin effectively blocked lung prostaglandin production but neither prevented the decrease in vasoreactivity nor inhibited edema formation. From these data we conclude that hydrogen peroxide impaired pulmonary vasoreactivity and subsequently caused edema. Despite the fact that hydrogen peroxide stimulated lung prostaglandin production, cyclooxygenase-derived products neither caused the decrease in vasoreactivity nor the development of edema.

scholarly journals NADPH Oxidase-Mediated Reactive Oxygen Species Production: Subcellular Localization and Reassessment of Its Role in Plant Defense

2009 ◽  
Vol 22 (7) ◽  
pp. 868-881 ◽  
Author(s):  
Jeannine Lherminier ◽  
Taline Elmayan ◽  
Jérôme Fromentin ◽  
Khadija Tantaoui Elaraqui ◽  
Simona Vesa ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Hydrogen Peroxide ◽  
Plasma Membrane ◽  
Nadph Oxidase ◽  
Subcellular Localization ◽  
Acquired Resistance ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Wild Type ◽  
Hydrogen Peroxide Production

Chemiluminescence detection of reactive oxygen species (ROS) triggered in tobacco BY-2 cells by the fungal elicitor cryptogein was previously demonstrated to be abolished in cells transformed with an antisense construct of the plasma membrane NADPH oxidase, NtrbohD. Here, using electron microscopy, it has been confirmed that the first hydrogen peroxide production occurring a few minutes after challenge of tobacco cells with cryptogein is plasma membrane located and NtrbohD mediated. Furthermore, the presence of NtrbohD in detergent-resistant membrane fractions could be associated with the presence of NtrbohD-mediated hydrogen peroxide patches along the plasma membrane. Comparison of the subcellular localization of ROS in wild-type tobacco and in plants transformed with antisense constructs of NtrbohD revealed that this enzyme is also responsible for the hydrogen peroxide production occurring at the plasma membrane after infiltration of tobacco leaves with cryptogein. Finally, the reactivity of wild-type and transformed plants to the elicitor and their resistance against the pathogenic oomycete Phytophthora parasitica were examined. NtrbohD-mediated hydrogen peroxide production does not seem determinant for either hypersensitive response development or the establishment of acquired resistance but it is most likely involved in the signaling pathways associated with the protection of the plant cell.

scholarly journals Activation of hepatocyte protein kinase C by redox-cycling quinones

1989 ◽  
Vol 260 (2) ◽  
pp. 499-507 ◽  
Author(s):  
G E Kass ◽  
S K Duddy ◽  
S Orrenius
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Kinase Activity ◽  
Specific Activity ◽  
Active Oxygen Species ◽  
Redox Cycling ◽  
Active Oxygen ◽  
Oxygen Species ◽  
Cytosolic Protein ◽  
Kinase C

The effects of quinone-generated active oxygen species on rat hepatocyte protein kinase C were investigated. The specific activity of cytosolic protein kinase C was increased 2-3-fold in hepatocytes incubated with the redox-cycling quinones, menadione, duroquinone or 2,3-dimethoxy-1,4-naphthoquinone, without alterations in particulate protein kinase C specific activity or Ca2+- and lipid-independent kinase activities. Redox-cycling quinones did not stimulate translocation of protein kinase C; however, activated protein kinase C was redistributed from cytosol to the particulate fraction when quinone-treated hepatocytes were exposed to 12-O-tetradecanoylphorbol 13-acetate (TPA). Quinone treatment did not alter cytosolic phorbol 12,13-dibutyrate (PDBu) binding capacity, and the cytosol of both control and quinone-treated hepatocytes exhibited a Kd for PDBu binding of 2 nM. Quinone-mediated activation of cytosolic protein kinase C was reversed by incubation with 10 mM-beta-mercaptoethanol, dithiothreitol or GSH, at 4 degrees C for 24 h. Furthermore, protein kinase C specific activity in control cytosol incubated in air increased by over 100% within 3 h; this increase was reversed by thiol-reducing agents. Similarly, incubation of partially-purified rat brain protein kinase C in air, or with low concentrations of GSSG in the presence of GSH, resulted in a 2-2.5-fold increase in Ca2+- and lipid-dependent kinase activity. In contrast with the effects of the redox-cycling quinones, when hepatocytes were treated with the thiol agents N-ethylmaleimide (NEM), p-benzoquinone (pBQ) or p-chloromercuribenzoic acid (pCMB), the cytosolic Ca2+- and lipid-dependent kinase activity was significantly inhibited, but the particulate-associated protein kinase C activity was unaffected. The Ca2+- and lipid-independent kinase activity of both the cytosolic and particulate fractions was significantly stimulated by NEM, but was unaffected by pBQ and pCMB. These results show that hepatocyte cytosolic protein kinase C is activated to a high-Vmax form by quinone-generated active oxygen species, and this effect is due to a reduction-sensitive modification of the thiol/disulphide status of protein kinase C.

scholarly journals A Helicobacter hepaticus catalase mutant is hypersensitive to oxidative stress and suffers increased DNA damage

2007 ◽  
Vol 56 (4) ◽  
pp. 557-562 ◽  
Author(s):  
Yang Hong ◽  
Ge Wang ◽  
Robert J. Maier
Keyword(s):  
Oxidative Stress ◽  
Dna Damage ◽  
Catalase Activity ◽  
Insertional Mutagenesis ◽  
Specific Activity ◽  
Bacterial Species ◽  
Helicobacter Hepaticus ◽  
Wild Type ◽  
Crude Cell Extract ◽  
H Pylori

Catalase (KatA) is known to play an important role in oxidative stress resistance in many bacterial species and a homologue exists in Helicobacter hepaticus, a member of the enterohepatic Helicobacter species. Here, a katA mutant was constructed by insertional mutagenesis and its oxidative stress phenotype was investigated. Catalase activity was readily detected [196 units (mg protein crude cell extract)−1] in the wild-type, whereas the mutant strain was deficient in, but not devoid of, activity. In contrast, Helicobacter pylori katA strains lack detectable catalase activity and wild-type H. pylori generally contains higher specific activity than H. hepaticus. Wild-type H. hepaticus cells tolerated 6 % O2 for growth, whilst the katA mutant could not survive at this oxygen level. Even at the optimal O2 level, the growth of the H. hepaticus katA strain was severely inhibited, which is also in contrast to H. pylori katA strains. Wild-type H. hepaticus cells withstood exposure to 100 mM H2O2 but the katA mutant cells were killed by the same treatment. Wild-type cells suffered no significant DNA damage by H2O2 treatment (100 mM for 6 min), whilst the same treatment resulted in severe DNA fragmentation in the katA mutant. Thus H. hepaticus KatA plays an important role as an antioxidant protein.

scholarly journals KatG Is the Primary Detoxifier of Hydrogen Peroxide Produced by Aerobic Metabolism in Bradyrhizobium japonicum

2004 ◽  
Vol 186 (23) ◽  
pp. 7874-7880 ◽  
Author(s):  
Heather R. Panek ◽  
Mark R. O'Brian
Keyword(s):  
Catalase Activity ◽  
Bradyrhizobium Japonicum ◽  
Aerobic Metabolism ◽  
Short Exposure ◽  
Wild Type ◽  
Gene Encoding ◽  
Whole Cells ◽  
Cell Extracts ◽  
Genes Encoding ◽  
High Concentrations

ABSTRACT Bacteria are exposed to reactive oxygen species from the environment and from those generated by aerobic metabolism. Catalases are heme proteins that detoxify H2O2, and many bacteria contain more than one catalase enzyme. Also, the nonheme peroxidase alkyl hydroperoxide reductase (Ahp) is the major scavenger of endogenous H2O2 in Escherichia coli. Here, we show that aerobically grown Bradyrhizobium japonicum cells express a single catalase activity. Four genes encoding putative catalases in the B. japonicum genome were identified, including a katG homolog encoding a catalase-peroxidase. Deletion of the katG gene resulted in loss of catalase activity in cell extracts and of exogenous H2O2 consumption by whole cells. The katG strain had a severe aerobic growth phenotype but showed improved growth in the absence of O2. By contrast, a B. japonicum ahpCD mutant grew well aerobically and consumed H2O2 at wild-type rates. A heme-deficient hemA mutant expressed about one-third of the KatG activity as the wild type but grew well aerobically and scavenged low concentrations of exogenous H2O2. However, cells of the hemA strain were deficient in consumption of high concentrations of H2O2 and were very sensitive to killing by short exposure to H2O2. In addition, KatG activity did not decrease as a result of mutation of the gene encoding the transcriptional activator OxyR. We conclude that aerobic metabolism produces toxic levels of H2O2 in B. japonicum, which is detoxified primarily by KatG. Furthermore, the katG level sufficient for detoxification does not require OxyR.

scholarly journals Contribution of Mn-Cofactored Superoxide Dismutase (SodA) to the Virulence of Streptococcus agalactiae

2001 ◽  
Vol 69 (8) ◽  
pp. 5098-5106 ◽  
Author(s):  
Claire Poyart ◽  
Elisabeth Pellegrini ◽  
Olivier Gaillot ◽  
Claire Boumaila ◽  
Marina Baptista ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Superoxide Dismutase ◽  
Streptococcus Agalactiae ◽  
Specific Activity ◽  
Infection Model ◽  
Mouse Bone Marrow ◽  
Group B Streptococci ◽  
Bacterial Killing ◽  
Cell Extracts

ABSTRACT Superoxide dismutases convert superoxide anions to molecular oxygen and hydrogen peroxide, which, in turn, is metabolized by catalases and/or peroxidases. These enzymes constitute one of the major defense mechanisms of cells against oxidative stress and hence play a role in the pathogenesis of certain bacteria. We previously demonstrated that group B streptococci (GBS) possess a single Mn-cofactored superoxide dismutase (SodA). To analyze the role of this enzyme in the pathogenicity of GBS, we constructed a sodA-disrupted mutant of Streptococcus agalactiae NEM316 by allelic exchange. This mutant was subsequently cis complemented by integration into the chromosome of pAT113/Sp harboring the wild-typesodA gene. The SOD specific activity detected by gel analysis in cell extracts confirmed that active SODs were present in the parental and complemented strains but absent in thesodA mutant. The growth rates of these strains in standing cultures were comparable, but the sodA mutant was extremely susceptible to the oxidative stress generated by addition of paraquat or hydrogen peroxide to the culture medium and exhibited a higher mutation frequency in the presence of rifampin. In mouse bone marrow-derived macrophages, the sodA mutant showed an increased susceptibility to bacterial killing by macrophages. In a mouse infection model, after intravenous injection the survival of thesodA mutant in the blood and the brain was markedly reduced in comparison to that of the parental and complemented strains whereas only minor effects on survival in the liver and the spleen were observed. These results suggest that SodA plays a role in GBS pathogenesis.

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