Molecular cloning, sequence analysis and expression of the gene for catalase‐peroxidase (<i>cpeA</i>) from the photosynthetic bacterium <i>Rhodobacter capsulatus</i> B10

Forkl, Hubert; Vandekerckhove, Joël; Drews, Gerhart; Tadros, Monier H.

doi:10.1111/j.1432-1033.1993.tb17918.x

Cited by 37 publications

(23 citation statements)

References 42 publications

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“…Among the others, L. pneumophila CuZnSOD is also periplasmic (52), but the catalase-peroxidases of C. crescentus (51) and R. capsulatus (18,19,27,29) have not been subjected to localization studies and are not predicted to contain a signal peptide (PSORT Prediction, http://psort.nibb.ac.jp/form.html; Signal P V1.1, http://www.cbs.dtu.dk/services/signalP/). Our studies raise the question of whether a periplasmic location is a sine qua non for the stationary-phase function.…”

Section: Discussionmentioning

confidence: 99%

Catalase-Peroxidases of Legionella pneumophila : Cloning of the katA Gene and Studies of KatA Function

Bandyopadhyay

Steinman

2000

J Bacteriol

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Legionella pneumophila, the causative organism of Legionnaires' pneumonia, contains two enzymes with catalatic and peroxidatic activity, KatA and KatB. To address the issue of redundant, overlapping, or discrete in vivo functions of highly homologous catalase-peroxidases, the gene for katA was cloned and its function was studied in L. pneumophila and Escherichia coli and compared with prior studies of katB in this laboratory. katA is induced during exponential growth and is the predominant peroxidase in stationary phase. When katA is inactivated, L. pneumophila is more sensitive to exogenous hydrogen peroxide and less virulent in the THP-1 macrophage cell line, similar to katB. Catalatic-peroxidatic activity with different peroxidatic cosubstrates is comparable for KatA and KatB, but KatA is five times more active towards dianisidine. In contrast with these examples of redundant or overlapping function, stationary-phase survival is decreased by 100-to 10,000-fold when katA is inactivated, while no change from wild type is seen for the katB null. The principal clue for understanding this discrete in vivo function was the demonstration that KatA is periplasmic and KatB is cytosolic. This stationary-phase phenotype suggests that targets sensitive to hydrogen peroxide are present outside the cytosol in stationary phase or that the peroxidatic activity of KatA is critical for stationary-phase redox reactions in the periplasm, perhaps disulfide bond formation. Since starvation-induced stationary phase is a prerequisite to acquisition of virulence by L. pneumophila, further studies on the function and regulation of katA in stationary phase may give insights on the mechanisms of infectivity of this pathogen.

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Section: Discussionmentioning

confidence: 99%

Catalase-Peroxidases of Legionella pneumophila : Cloning of the katA Gene and Studies of KatA Function

Bandyopadhyay

Steinman

2000

J Bacteriol

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“…This may be due to the steric hindrance that does not provide access to the heme prosthetic group of the enzyme (62). Secondly, comparison of the conserved motifs in the KatB amino acid sequence revealed that the heme-binding ligands and active-site residues are highly conserved with the true catalases, whereas the conserved active site and domains of peroxidases that are present in bacterial catalase-peroxidases (19,73) were not identified in the B. fragilis catalase. The restricted peroxidatic activity associated with B. fragilis KatB is probably due to the nonspecific catalysis that occurs in typical catalases toward certain types of organic electron donor compounds (62).…”

Section: Fragilismentioning

confidence: 99%

Biochemical and genetic analyses of a catalase from the anaerobic bacterium Bacteroides fragilis

Rocha

Smith

1995

J Bacteriol

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A single catalase enzyme was produced by the anaerobic bacterium Bacteroides fragilis when cultures at late log phase were shifted to aerobic conditions. In anaerobic conditions, catalase activity was detected in stationary-phase cultures, indicating that not only oxygen exposure but also starvation may affect the production of this antioxidant enzyme. The purified enzyme showed a peroxidatic activity when pyrogallol was used as an electron donor. It is a hemoprotein containing one heme molecule per holomer and has an estimated molecular weight of 124,000 to 130,000. The catalase gene was cloned by screening a B. fragilis library for complementation of catalase activity in an Escherichia coli catalase mutant (katE katG) strain. The cloned gene, designated katB, encoded a catalase enzyme with electrophoretic mobility identical to that of the purified protein from the B. fragilis parental strain. The nucleotide sequence of katB revealed a 1,461-bp open reading frame for a protein with 486 amino acids and a predicted molecular weight of 55,905. This result was very close to the 60,000 Da determined by denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified catalase and indicates that the native enzyme is composed of two identical subunits. The N-terminal amino acid sequence of the purified catalase obtained by Edman degradation confirmed that it is a product of katB. The amino acid sequence of KatB showed high similarity to Haemophilus influenzae HktE (71.6% identity, 66% nucleotide identity), as well as to gram-positive bacterial and mammalian catalases. No similarities to bacterial catalase-peroxidase-type enzymes were found. The active-site residues, proximal and distal hemebinding ligands, and NADPH-binding residues of the bovine liver catalase-type enzyme were highly conserved in B. fragilis KatB.

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“…Little is known about the conditions prevailing in the bacterial cells following the establishment of respiration, but oxygen-centered derivatives such as superoxide and peroxides are expected to be produced by reduced intermediates of the electron transport chain (36). In line with this assumption, a number of protective antioxidant proteins are known to be present in Rhodobacter species, including two peroxidases (12) and an oxygen-responsive thioredoxin which is essential for both aerobic and anaerobic growth (28). We have recently identified a single SOD (SOD Rc ) in R. capsulatus cells cultured under various growth regimes (8).…”

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confidence: 97%

The Single Superoxide Dismutase of Rhodobacter capsulatus Is a Cambialistic, Manganese-Containing Enzyme

et al. 2003

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The phototrophic bacterium Rhodobacter capsulatus contains a single, oxygen-responsive superoxide dismutase (SOD Rc ) homologous to iron-containing superoxide dismutase enzymes. Recombinant SOD Rc , however, displayed higher activity after refolding with Mn 2؉ , especially when the pH of the assay mixture was raised. SOD Rc isolated from Rhodobacter cells also preferentially contains manganese, but metal discrimination depends on the culture conditions, with iron fractions increasing from 7% in aerobic cultures up to 40% in photosynthetic cultures. Therefore, SOD Rc behaves as a Mn-containing dismutase with cambialistic properties.

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Molecular cloning, sequence analysis and expression of the gene for catalase‐peroxidase (cpeA) from the photosynthetic bacterium Rhodobacter capsulatus B10

Cited by 37 publications

References 42 publications

Catalase-Peroxidases of Legionella pneumophila : Cloning of the katA Gene and Studies of KatA Function

Catalase-Peroxidases of Legionella pneumophila : Cloning of the katA Gene and Studies of KatA Function

Biochemical and genetic analyses of a catalase from the anaerobic bacterium Bacteroides fragilis

The Single Superoxide Dismutase of Rhodobacter capsulatus Is a Cambialistic, Manganese-Containing Enzyme

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