The mycelium-associated Streptomyces reticuli catalase-peroxidase, its gene and regulation by FurS (original) (raw)
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Protein Science, 2009
True catalases are tyrosine-liganded, usually tetrameric, hemoproteins with subunit sizes of approximately 55-84 kDa. Recently characterized hemoproteins with a catalase-related structure, yet lacking in catalatic activity, include the 40-43 kDa allene oxide synthases of marine invertebrates and cyanobacteria. Herein, we describe the 1.8 A X-ray crystal structure of a 33 kDa subunit hemoprotein from Mycobacterium avium ssp. paratuberculosis (annotated as MAP-2744c), that retains the core elements of the catalase fold and exhibits an organic peroxide-dependent peroxidase activity. MAP-2744c exhibits negligible catalatic activity, weak peroxidatic activity using hydrogen peroxide (20/s) and strong peroxidase activity (approximately 300/s) using organic hydroperoxides as co-substrate. Key amino acid differences significantly impact prosthetic group conformation and placement and confer a distinct activity to this prototypical member of a group of conserved bacterial "minicatalases". Its structural features and the result of the enzyme assays support a role for MAP-2744c and its close homologues in mitigating challenge by a variety of reactive oxygen species.
European Journal of Biochemistry, 2000
Streptomyces reticuli produces a heme-containing homodimeric enzyme (160 kDa), the catalase-peroxidase CpeB, which is processed to the enzyme CpeC during prolonged growth. CpeC contains four subunits of 60 kDa each that do not include the C-terminal portion of the progenitor subunits. A genetically engineered cpeB gene encodes a truncated subunit lacking 195 of the C-terminal amino acids; four of these subunits assemble to form the enzyme CpeD. Heme binds most strongly in CpeB, least in CpeD. The catalaseperoxidase CpeB and its apo-form (obtained after extraction of heme) catalyze the peroxidation of Mn(II) to Mn(III), independent of the presence or absence of the heme inhibitor KCN. CpeC and CpeD, in contrast, do not exhibit manganese-peroxidase activity. The data show for the first time that a bacterial catalase-peroxidase has a heme-independent manganeseperoxidase activity, which depends on the presence of the C-terminal domain.
Frankia Symbiosis, 2003
A monofunctional catalase and a bifunctional catalase-peroxidase were revealed by activity staining of nondenaturing PAGE in Frankia strain R43. Both enzymes were shown to be cytoplasmatic, growth regulated and expressed mainly during the stationary growth phase. Nevertheless, low levels of constitutive expression could also be detected during the early stages of growth. Immunoblot analyses using a polyclonal antibody raised against a catalase-peroxidase purified from Streptomyces reticuli showed a band of 83.2 kDa, with the same growth dependent pattern as obtained by the non-denaturing PAGE analyses. Induction studies revealed that both enzymes were strongly induced by raising the intracellular concentration of H 2 O 2 with paraquat, but not with exogenous H 2 O 2 . In addition, no acquisition of tolerance to exogenous H 2 O 2 was observed whatever the pretreatment of the inocula, i.e. despite the expression level of both hydroperoxidases.
Biochimica Et Biophysica Acta-protein Structure and Molecular Enzymology, 2001
The catalase-peroxidase encoded by katG of Mycobacterium tuberculosis is a more effective activator of the antibiotic isoniazid than is the equivalent enzyme from Escherichia coli. The environment of the heme iron was investigated using X-ray absorption spectroscopy to determine if differences in this region were associated with the differences in reactivity. The variation in the distal side Fe^ligand distances between the two enzymes was the same within experimental error indicating that it was not the heme iron environment that produced the differences in reactivity. Analysis of variants of the E. coli catalase-peroxidase containing changes in active site residues Arg102 and His106 revealed small differences in Fe^water ligand distance including a shorter distance for the His106Tyr variant. The Arg102Leu variant was 5-coordinate, but His106Cys and Arg102Cys variants showed no changes within experimental error. These results are compared with those reported for other peroxidases. ß 2001 Elsevier Science B.V. All rights reserved.
Revised sequence and activity of Bacillus stearothermophilus catalase I (formerly peroxidase)
Journal of Fermentation and Bioengineering, 1992
The revised nucleotide sequence of the perA gene, encoding catalase I (formerly peroxidase) of Bacillus stearothermophilus, is presented. The deduced amino acid sequence was significantly similar to those of Escherichia coli catalase HPI, Salmonella typhimurium catalase HPI, and yeast cytochrome c peroxidase. Catalase I was purified and the ratio of the catalatie to peroxidatic activity was measured and found to be 654 to 1.
Biochimie, 2000
Bifunctional catalase-peroxidases are the least understood type of peroxidases. A high-level expression in Escherichia coli of a fully active recombinant form of a catalase-peroxidase (KatG) from the cyanobacterium Anacystis nidulans (Synechococcus PCC 6301) is reported. Since both physical and kinetic characterization revealed its identity with the wild-type protein, the large quantities of recombinant KatG allowed the examination of both the spectral characteristics and the reactivity of its redox intermediates by using the multi-mixing stopped-flow technique. The homodimeric acidic protein (pI = 4.6) contained high catalase activity (apparent K m = 4.8 mM and apparent k cat = 8850 s-1). Cyanide is shown to be an effective inhibitor of the catalase reaction. The second-order rate constant for cyanide binding to the ferric protein is (6.9 ± 0.2) × 10 5 M-1 s-1 at pH 7.0 and 15°C and the dissociation constant of the cyanide complex is 17 µM. Because of the overwhelming catalase activity, peroxoacetic acid has been used for compound I formation. The apparent second-order rate constant for formation of compound I from the ferric enzyme and peroxoacetic acid is (1.3 ± 0.3) × 10 4 M-1 s-1 at pH 7.0 and 15°C. The spectrum of compound I is characterized by about 40% hypochromicity, a Soret region at 406 nm, and isosbestic points between the native enzyme and compound I at 355 and 428 nm. Rate constants for reduction of KatG compound I by o-dianisidine, pyrogallol, aniline and isoniazid are shown to be (7.3 ± 0.4) × 10 6 M-1 s-1 , (5.4 ± 0.3) × 10 5 M-1 s-1 , (1.6 ± 0.3) × 10 5 M-1 s-1 and (4.3 ± 0.2) × 10 4 M-1 s-1 , respectively. The redox intermediate formed upon reduction of compound I did not exhibit the classical red-shifted peroxidase compound II spectrum which characterizes the presence of a ferryl oxygen species. Its spectral features indicate that the single oxidizing equivalent in KatG compound II is contained on an amino acid which is not electronically coupled to the heme.
FEMS Microbiology Letters, 2002
The catalase gene katA of Staphylococcus xylosus was cloned. It encodes a protein of 494 amino acids with a molecular mass of 56.9 kDa, closely related to monofunctional catalases. A katA mutant still showed a relatively high catalase activity demonstrating that S. xylosus possesses more than one enzyme. By Southern blot analysis using a katA probe, a second genetic locus distinct from katA was detected that probably contained the additional catalase gene. To analyse katA expression, a transcriptional fusion of the katA promoter region to a promoterless L-galactosidase gene was integrated into the genome of S. xylosus. katA expression is induced upon entry into stationary phase, by oxygen and hydrogen peroxide. Iron and manganese depletion induced katA transcription. Comparing the resistance of S. xylosus wild-type and the katA mutant strain to hydrogen peroxide clearly showed that KatA is essential for S. xylosus to cope with hydrogen peroxide stress. Therefore, S. xylosus has at least two differentially expressed catalases.
A catalase from Streptomyces coelicolor A3(2)
Microbiology, 1995
Catalase was purified from the Gram-positive bacterium Streptomyces coelicolor A3(2) in a three-step purification procedure comprising (NH,J,SO, fractionation, Phenyl-Sepharose chromatography and Mono Q chromatography. The purification of catalase, as judged by the final specific activity of 110000 U mg-l, was 250-fold with a 35% yield. The native protein was a homotetramer with a subunit Mr 55000. N-terminal and internal peptide sequence analyses showed that there was a high degree of sequence similarity between the 5. coelicolor catalase and other microbial and mammalian catalases. Southern blot analysis indicated that there was a single catalase gene in 5. coelicolor. The specific activity of catalase throughout the growth of batch cultures was investigated and elevated catalase activity was found in stationary-phase cells.