| cloning and high-level expression of a chloroperoxidase gene from pseudomonas pyrrocinia in escherichia coli. | a chloroperoxidase gene from pseudomonas pyrrocinia was cloned into escherichia coli using the cosmid vector pjb8. the gene coding for the chloroperoxidase could be localized to a 1.5 kb fragment of dna which was subcloned into the high-copy-number plasmid puc18. in one subclone increased halogenating activity could be found which was 570-fold greater than in p. pyrrocinia. the halogenating enzyme was identified as the chloroperoxidase by sds-polyacrylamide gel electrophoresis. | 1988 | 3049160 |
| purification and characterization of a novel bacterial non-heme chloroperoxidase from pseudomonas pyrrocinia. | the first bacterial chloroperoxidase that is capable of catalyzing the chlorination of indole to 7-chloroindole was detected in pseudomonas pyrrocinia atcc 15958, a bacterium that produces the antifungal antibiotic pyrrolnitrin (wiesner, w., van pée, k.h., and lingens, f. (1986) febs lett. 209, 321-324). here we describe the purification and characterization of this bacterial non-heme chloroperoxidase. the enzyme was purified by deae-cellulose chromatography at different ph values, molecular sie ... | 1988 | 3417677 |
| detection of a new chloroperoxidase in pseudomonas pyrrocinia. | a new chloroperoxidase could be detected in pseudomonas pyrrocinia atcc 15,958, a bacterium that produces the antifungal antibiotic pyrrolnitrin. this enzyme was separated from a ferriprotoporphyrin ix containing bromoperoxidase which was also produced by this bacterium. the enzyme is capable of catalyzing the chorination of indole to 7-chloroindole. this procaryotic chloroperoxidase requires the presence of h2o2 and can also brominate monochlorodimedone, but cannot catalyze its chlorination. th ... | 1986 | 3792552 |
| purification and properties of bromoperoxidase from pseudomonas pyrrocinia. | a bromoperoxidase was purified and partially characterized from pseudomonas pyrrocinia atcc 15958, a bacterium that produces the antifungal antibiotic pyrrolnitrin. the purified enzyme preparation was homogeneous as determined by polyacrylamide gel electrophoresis and ultracentrifugation. the molecular mass of the enzyme was estimated to be 154 kda +/- 3 kda as determined by gel filtration and ultracentrifugation. sodium dodecyl sulfate polyacrylamide gel electrophoresis showed a single band wit ... | 1985 | 4091967 |
| the metabolism of tryptophan and 7-chlorotryptophan in pseudomonas pyrrocinia and pseudomonas aureofaciens. | pseudomonas pyrrocinia atcc 15958 and a mutant strain (acn) of pseudomonas aureofaciens atcc 15926 possess a mechanism for the degradation of the tryptophan side chain. indole, indole-3-carboxylic acid, indole-3-acetic acid and the corresponding compounds chlorinated or brominated at position 7, as well as indole-3-pyruvate and 7-chloroindole-3-pyruvate, were isolated from bacterial cultures. the chlorinated indole derivatives were isolated after the addition of 7-chloro-dl-tryptophan to culture ... | 1983 | 6862384 |
| purification and properties of a non-haem chloroperoxidase from serratia marcescens. | a non-haem chloroperoxidase was isolated from the enteric bacterium serratia marcescens. the enzyme was purified to homogeneity by heat treatment, ammonium sulfate precipitation, ion exchange chromatography, gel filtration and dye-ligand affinity chromatography. native chloroperoxidase has a molecular mass of 58 kda and consists of two identical subunits of 29 kda. whereas chloroperoxidase catalyses only the bromination of monochlorodimedone, indole is chlorinated by this enzyme. chloroperoxidas ... | 1995 | 7607409 |
| cloning of a second non-haem bromoperoxidase gene from streptomyces aureofaciens atcc 10762: sequence analysis, expression in streptomyces lividans and enzyme purification. | the gene for bpo-a1, one of two non-haem bromoperoxidases in the tetracycline and 7-chlorotetracycline producer streptomyces aureofaciens atcc 10762, was cloned in the positive selection vector pij699 and expressed in streptomyces lividans tk64. the cloned bromoperoxidase was over-produced up to 2800-fold by the s. lividans tk64 transformant. by taking advantage of the over-production of bpo-a1 and the heat stability of the enzyme, a new and simple purification procedure was developed. subclonin ... | 1994 | 8012573 |
| chloroperoxidase from streptomyces lividans: isolation and characterization of the enzyme and the corresponding gene. | for the first time, a halogenating enzyme which is not known to produce halogenated metabolites has been isolated from a bacterial strain. the gene encoding the nonheme chloroperoxidase (cpo-l) from streptomyces lividans tk64 was cloned, and its gene product was characterized. s. lividans tk64 produced only very small amounts of the enzyme. after cloning of the gene into streptomyces aureofaciens tü24-88, the enzyme was overexpressed up to 3,000-fold. based on the overexpression, a simple purifi ... | 1994 | 8157602 |
| chloroperoxidase-encoding gene from pseudomonas pyrrocinia: sequence, expression in heterologous hosts, and purification of the enzyme. | the nucleotide sequence of a 1.5-kb fragment of pseudomonas pyrrocinia dna containing the chloroperoxidase(cpo)-encoding gene (cpo) and its flanking regions was determined. the cpo codes for a protein of 278 amino acids (aa). the matuĊe enzyme contains no n-terminal methionine, so that the cpo monomer consists of 277 aa with a calculated m(r) of 30,304. expression studies showed that the cpo from p. pyrrocinia is functionally expressed in escherichia coli and streptomyces lividans. based on the ... | 1993 | 8344520 |
| thiocarbamate herbicide-inducible nonheme haloperoxidase of rhodococcus erythropolis ni86/21. | during biodegradation of thiocarbamate herbicides by rhodococcus erythropolis ni86/21, a protein with an m(r) of 30,000 is induced (i. nagy, g. schoofs, f. compernolle, p. proost, j. vanderleyden, and r.de mot, j. bacteriol. 177:676-687, 1995). based on n-terminal sequence data for the protein purified by two-dimensional electrophoresis, the corresponding structural gene, thcf, was cloned and sequenced. the deduced protein sequence of thcf is homologous to those of nonheme haloperoxidases. a par ... | 1997 | 9143122 |
| occurrence of multiple genomovars of burkholderia cepacia in cystic fibrosis patients and proposal of burkholderia multivorans sp. nov. | we performed an integrated genotypic and phenotypic analysis of 128 strains of the genera burkholderia, ralstonia, and pseudomonas in order to study the taxonomic structure of burkholderia cepacia and its relationships with other burkholderia species. our data show that presumed b. cepacia strains isolated from cystic fibrosis patients belong to at least five distinct genomic species, one of which was identified as burkholderia vietnamiensis. this group of five phenotypically similar species is ... | 1997 | 9336927 |
| burkholderia graminis sp. nov., a rhizospheric burkholderia species, and reassessment of [pseudomonas] phenazinium, [pseudomonas] pyrrocinia and [pseudomonas] glathei as burkholderia. | in a survey of soil and wheat or maize rhizoplane bacteria isolated using a medium containing azelaic acid and tryptamine as sole carbon and nitrogen sources, respectively, a large proportion of burkholderia-like bacteria were found. among them, a homogeneous group of strains was identifiable based on phenotypic properties, fatty acid composition, dna-dna hybridizations and 16s rdna sequences. according to molecular data, this group belongs to the genus burkholderia but its weak similarity to pr ... | 1998 | 9731297 |
| antifungal and peroxidative activities of nonheme chloroperoxidase in relation to transgenic plant protection. | nonheme chloroperoxidase (cpo-p) of pseudomonas pyrrocinia catalyzes the oxidation of alkyl acids to peracids by hydrogen peroxide. alkyl peracids possess potent antifungal activity as found with peracetate: 50% killing (ld(50)) of aspergillus flavus occurred at 25 microm compared to 3.0 mm for the hydrogen peroxide substrate. to evaluate whether cpo-p could protect plants from fungal infection, tobacco was transformed with a gene for cpo-p from p. pyrrocinia and assayed for antifungal activity. ... | 2000 | 11052700 |
| [oxidation of indole and its derivatives by heme-independent chloroperoxidases]. | indole, indolylacetic acid, and tryptophan were oxidized by cloroperoxidases isolated from strains of streptomyces lividans and pseudomonas pyrrocinia. indigo (indoxyl), isatin, and anthranilic acid (intermediate products of oxidative degradation of indole and indole derivatives) were extracted from the reaction medium. | 2001 | 11443895 |
| evaluation of peracid formation as the basis for resistance to infection in plants transformed with haloperoxidase. | nonheme haloperoxidase (hpo-p) isolated from pseudomonas pyrrocinia catalyzed the peroxidation of alkyl acids to peracids. among acids tested as substrates, acetic acid was most readily peroxidized. the reaction product peracetate possessed potent antifungal activity: 50% death (ld(50)) of aspergillus flavus occurred at 25 microm peracetate. viability of a. flavus was inhibited by up to 80% by leaf extracts of tobacco plants transformed with the hpo-p gene from p. pyrrocinia compared to viabilit ... | 2002 | 11829632 |
| burkholderia anthina sp. nov. and burkholderia pyrrocinia, two additional burkholderia cepacia complex bacteria, may confound results of new molecular diagnostic tools. | nineteen burkholderia cepacia-like isolates of human and environmental origin could not be assigned to one of the seven currently established genomovars using recently developed molecular diagnostic tools for b. cepacia complex bacteria. various genotypic and phenotypic characteristics were examined. the results of this polyphasic study allowed classification of the 19 isolates as an eighth b. cepacia complex genomovar (burkholderia anthina sp. nov.) and to design tools for its identification in ... | 2002 | 12052570 |
| dna-dna hybridization study of burkholderia species using genomic dna macro-array analysis coupled to reverse genome probing. | the present study was aimed at simplifying procedures to delineate species and identify isolates based on dna-dna reassociation. dna macro-arrays harbouring genomic dna of reference strains of several burkholderia species were produced. labelled genomic dna, hybridized to such an array, allowed multiple relative pairwise comparisons. based on the relative dna-dna relatedness values, a complete data matrix was constructed and the ability of the method to discriminate strains belonging to differen ... | 2003 | 12807195 |
| halogenating enzymes in the biosynthesis of antibiotics. | using blot hybridization, it has been shown that microorganisms producing halogen-containing antibiotics--pseudomonas pyrrocinia, p. aureofaciens acn, p. aureofaciens pa1, p. fluorescens cha0, actinoplanes sp., kitasatasporia sp., sacharothrix aerocolonigenes, actinomadura melliaura, and streptomyces albogriseolus--contain the genes of the halogenating enzymes related to tryptophan-7-halogenase and monodechloroaminopyrrolnitrin halogenase from p. fluorescens bl 915. | 2003 | 14616084 |
| epidemiology and clinical course of burkholderia cepacia complex infections, particularly those caused by different burkholderia cenocepacia strains, among patients attending an italian cystic fibrosis center. | in this study, the epidemiology of burkholderia cepacia complex (bcc) recovered from the sputum of 75 patients attending the genoa cystic fibrosis (cf) center at the gaslini children's hospital (genoa, italy) was investigated, and the clinical course of the cf patients infected with the different species and genomovars of bcc was evaluated. all isolates were analyzed for genomovar status by reca gene polymorphism and subsequently random amplified polymorphic dna fingerprinting. burkholderia ceno ... | 2004 | 15070994 |
| pcr-based identification and characterization of burkholderia cepacia complex bacteria from clinical and environmental sources. | to study the genotypic identification and characterization of the 119 burkholderia cepacia complex (bcc) strains recovered from clinical and environmental sources in japan and thailand. | 2004 | 15482431 |
| polyphasic characterisation of burkholderia cepacia-like isolates leading to the emended description of burkholderia pyrrocinia. | twenty-five burkholderia cepacia-like isolates of human and environmental origin, comprising five different reca rflp types, were examined by using a polyphasic taxonomic approach, including reca gene sequence analysis, 16s rrna gene sequence analysis, dna:dna hybridisation studies, tdna-pcr, fatty acid analysis and biochemical analysis. the results of the present study demonstrated that twenty-three of these strains belong to burkholderia pyrrocinia, a b. cepacia complex species thus far compri ... | 2004 | 15490552 |
| detection of cultured and uncultured burkholderia cepacia complex bacteria naturally occurring in the maize rhizosphere. | the species composition of a burkholderia cepacia complex population naturally occurring in the maize rhizosphere was investigated by using both culture-dependent and culture-independent methods. b. cepacia complex isolates were recovered from maize root slurry on the two selective media pseudomonas cepacia azelaic acid tryptamine (pcat) and trypan blue tetracycline (tb-t) and subjected to identification by a combination of restriction fragment length polymorphism (rflp) analysis and species-spe ... | 2005 | 16232288 |
| microbial conversion of major ginsenoside rb(1) to pharmaceutically active minor ginsenoside rd. | more than seventy strains of aerobic bacteria showing beta-glucosidase activity were isolated from a ginseng field, using a newly designed esculin-r2a agar, and identified by their 16s rrna gene sequences. of these microorganisms, twelve strains could convert the major ginsenoside, rb(1), to the pharmaceutically active minor ginsenoside rd. three strains, burkholderia pyrrocinia gp16, bacillus megaterium gp27 and sphingomonas echinoides gp50, were phylogenetically studied, and observed to be mos ... | 2005 | 16273039 |
| metabolic profiling of burkholderia cenocepacia, burkholderia ambifaria, and burkholderia pyrrocinia isolates from maize rhizosphere. | burkholderia cenocepacia, burkholderia ambifaria, and burkholderia pyrrocinia are the burkholderia cepacia complex (bcc) species most frequently associated with roots of crop plants. to investigate the ecophysiological diversity of these species, metabolic profiling of maize rhizosphere isolates was carried out by means of the biolog system, using gn2 and sfn2 plates and different parameters related to optical density (od). the metabolic profiles produced by the sfn2 and gn2 plates were identica ... | 2005 | 16328653 |
| distribution and genomic location of active insertion sequences in the burkholderia cepacia complex. | this study aimed firstly to establish the distribution and copy number within the burkholderia cepacia complex of three insertion sequences (is402, is407 and is1416) that possess the ability to activate transcription and hence influence gene expression. a second aim was to map the genomic insertion sites of one of the active insertion sequences (is407) to establish putative links between insertion site and downstream gene activation. the resulting data revealed that all three insertion sequences ... | 2006 | 16388024 |
| differentiation of species combined into the burkholderia cepacia complex and related taxa on the basis of their fatty acid patterns. | using the established commercial system sherlock (midi, inc.), cellular fatty acid methyl ester analysis for differentiation among burkholderia cepacia complex species was proven. the identification key based on the diagnostic fatty acids is able to discern phenotypically related ralstonia pickettii and pandoraea spp. and further distinguish burkholderia pyrrocinia, burkholderia ambifaria, and burkholderia vietnamiensis. | 2006 | 16517920 |
| effect of agricultural management regime on burkholderia community structure in soil. | the main objective of this study was to determine the burkholderia community structure associated with areas under different agricultural management and to evaluate to which extent this community structure is affected by changes in agricultural management. two fields with distinct soil history (arable land and permanent grassland) were exposed to three agricultural management regimes (crop rotation, maize monoculture, and grassland). by using a culture-independent approach, based on a burkholder ... | 2006 | 16897309 |
| diversity and distribution of burkholderia cepacia complex in the rhizosphere of rice and maize. | a survey of burkholderia cepacia complex (bcc) species was conducted in agricultural fields within hangzhou, china. out of the 251 bacterial isolates recovered on the selective media from the rhizosphere of rice and maize, 112 of them were assigned to bcc by pcr assays. the species composition of the bcc isolates was analyzed by a combination of reca-restriction fragment length polymorphism assays, species-specific pcr tests and reca gene sequencing. the results revealed that the majority belong ... | 2007 | 17233735 |
| molecular characterization of burkholderia strains isolated from rice cultivars (oryza sativa l.) for species identification and phylogenetic grouping. | the genus burkholderia consists of extremely versatile bacteria that occupy diverse niches and are commonly encountered in the rhizosphere of crop plants. in this study, we characterized three plant growth promoting strains assigned as burkholderia sp. using biochemical and molecular characterization. the burkholderia spp. strains cbmb40, cbpb-him, and cbpb-hod were characterized using biochemical tests, biolog carbon substrate utilization, fatty acid methyl ester analysis, analysis of reca gene ... | 2008 | 18600039 |
| matrix-assisted laser desorption ionisation-time-of of-flight mass spectrometry of intact cells allows rapid identification of burkholderia cepacia complex. | the present study examined the potential of intact-cell matrix-assisted laser desorption ionisation-time-of-flight mass spectrometry (maldi-tof ms) for a rapid identification of burkholderia cepacia complex (bcc) bacteria using an applied biosystems 4700 proteomics analyser. two software packages were used to analyse mass profiles based on densitometric curves and peak positions. the 75 strains examined, represented the nine established bcc species and some commonly misidentified species, closel ... | 2008 | 18627778 |
| production of the antifungal compound pyrrolnitrin is quorum sensing-regulated in members of the burkholderia cepacia complex. | members of the genus burkholderia are known for their ability to suppress soil-borne fungal pathogens by the production of various antibiotic compounds. in this study we investigated the role of n-acylhomoserine lactone (ahl)-dependent quorum sensing (qs) in the expression of antifungal traits. using a quorum quenching approach, that is, by heterologous expression of the bacillus sp. aiia lactonase, we show that expression of antifungal activities is ahl-dependent in the large majority of the in ... | 2009 | 19220396 |
| polymorphisms within the prnd and pltc genes from pyrrolnitrin and pyoluteorin-producing pseudomonas and burkholderia spp. | abstract pyrrolnitrin (prn) and pyoluteorin (plt) are broad-spectrum antibiotics produced by several strains of pseudomonas and burkholderia species. both antibiotics play an important role in the suppression of multiple plant pathogenic fungi. primers were developed from conserved sequences and amplified prnd and pltc fragments from 18 pseudomonas and four burkholderia spp. of worldwide origin that produce either prn or plt or both. subsequent rflp (restriction fragment length polymorphisms) an ... | 2003 | 19719693 |
| inactivation of burkholderia cepacia complex phage ks9 gp41 identifies the phage repressor and generates lytic virions. | the burkholderia cepacia complex (bcc) is made up of at least 17 species of gram-negative opportunistic bacterial pathogens that cause fatal infections in patients with cystic fibrosis and chronic granulomatous disease. ks9 (vb_bcens_ks9), one of a number of temperate phages isolated from bcc species, is a prophage of burkholderia pyrrocinia lmg 21824. transmission electron micrographs indicate that ks9 belongs to the family siphoviridae and exhibits the b1 morphotype. the 39,896-bp ks9 genome, ... | 2010 | 19939932 |
| the ternary complex of prnb (the second enzyme in the pyrrolnitrin biosynthesis pathway), tryptophan, and cyanide yields new mechanistic insights into the indolamine dioxygenase superfamily. | pyrrolnitrin (3-chloro-4-(2'-nitro-3'-chlorophenyl)pyrrole) is a broad-spectrum antifungal compound isolated from pseudomonas pyrrocinia. four enzymes (prna, prnb, prnc, and prnd) are required for pyrrolnitrin biosynthesis from tryptophan. prnb rearranges the indole ring of 7-cl-l-tryptophan and eliminates the carboxylate group. prnb shows robust activity in vivo, but in vitro activity for prnb under defined conditions remains undetected. the structure of prnb establishes that the enzyme belongs ... | 2010 | 20421301 |
| five novel acid-tolerant oligotrophic thiosulfate-metabolizing chemolithotrophic acid mine drainage strains affiliated with the genus burkholderia of betaproteobacteria and identification of two novel soxb gene homologues. | five acid-tolerant thiosulfate-metabolizing bacteria were isolated from acid mine drainage samples from garubathan, india. 16s rrna gene analysis revealed that the strains were affiliated with the genus burkholderia of the class of betaproteobacteria. comparative 16s rrna gene sequence analyses indicated that the strains designated as gah1 and gah2 produced a separate phylogenetic branch having burkholderia pyrrocinia atcc 51958(t) (96-98%) as the closest relative. strains gah4 and burkholderia ... | 2011 | 21349327 |
| suppression of bacterial wilt and fusarium wilt by a burkholderia nodosa strain isolated from kalimantan soils, indonesia. | a trial was conducted to suppress bacterial wilt of tomato (bwt) caused by ralstonia solanacearum using biocontrol agents (bcas) isolated from soils in kalimantan, indonesia. five isolates were selected from 270 isolates as better performing bcas through screening four times using a pumice medium. the isolates selected were identified as burkholderia nodosa, burkholderia sacchari, burkholderia pyrrocinia and burkholderia terricola according to 16s rdna sequences, fatty acid composition and carbo ... | 2008 | 21558699 |