SEARCH · PubMed Health

Results for “Rhodococcus”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 415 records · Page 23Linked to original sources

PubMed1990

Structural analysis of the specific capsular polysaccharide of Rhodococcus equi serotype 2.

The specific capsular polysaccharide produced by Rhodococcus equi serotype 2 is a high-molecular-weight acidic polymer composed of D-glucose, D-mannose, D-glucuronic acid and 3-O-[(S)-1-carboxyethyl]-L-rhamnose in equimolar proportions. Structural analysis, employing a combination of chemical and n.m.r. techniques, established that the polysaccharide is composed of linear repeating tetrasaccharide units. (formula; see text) in which the beta-D-mannose residues carry O-acetyl groups at O-2 and O-3 to the extent of 1.7 mol equivalents. Unequivocal determination of the absolute chirality of the 3-O-[(S)-1-carboxyethyl]-alpha-L-rhamnose residues was achieved by chemical correlation with an authentic synthetic sample. The 1H and 13C-n.m.r. resonances of the native and O-deacetylated serotype 2 polysaccharides were fully assigned by homo- and heteronuclear chemical-shift correlation methods.

Carbohydrate Conformation↗

PubMed1994

Structural elucidation of the specific capsular polysaccharide of Rhodococcus equi serotype 7.

Structural analysis of the specific capsular polysaccharide produced by Rhodococcus equi serotype 7 indicated it to be a high-molecular-weight polymer consisting of equal molar amounts of D-galactose, D-mannose, L-rhamnose, and pyruvic acid. By employing a combination of chemical and NMR techniques, it was established that the polysaccharide is composed of the linear repeating trisaccharide units: [formula: see text] -->3)-alpha-D-Galp-(1-->3)-alpha-D-Manp-(1-->3)-alpha-L-Rhap -(1-->, in which the cyclic pyruvic acid acetal groups bridging the O-4 and O-6 positions of the alpha-D-Manp residues have the S-configuration. The 1H and 13C NMR spectra of the native and pyruvic acetal-free polysaccharides were fully assigned.

Bacterial Capsules↗

PubMed1990

Mass spectrometric analysis of O-peracetylated derivatives of 1-monomycoloylglycerol isolated from Corynebacterium pseudotuberculosis, Rhodococcus rhodochrous and R. lentifragmentus.

1-Monomycoloylglycerols from Corynebacterium pseudotuberculosis, Rhodococcus rhodochrous and R. lentifragmentus were reacted with acetic acid ahydride in the presence of pyridine. On infrared spectra the reaction products showed a sharp characteristic absorption of the acetyl ester group at 1235 cm-1; the hydroxyl group absorption (3400 cm-1) was absent. O-Peracetylated monomycoloylglycerols were analyzed by mass spectrometry under electron impact mode. The most common and representative peaks were associated to the following remarkable fragments: (a) peak at m/z 159 represented the backbone of the glycerol unit of peracetylated monomycoloylglycerols; it constituted the diagnostic and base peak of that group of compounds; (b) peak representing the glycerol moiety together with the alpha-subunit of the mycolic acid moiety; it gave the size of the chain length of the hydrocarbon side chain of that mycolic acid; and (c) a series of peaks of acylium ions minus 60 mass units (acetic acid) indicated the size of the chain length of the esterified mycolic acid. Therefore, acetylation of monomycoloylglycerol becomes the derivatized compound suitable for mass spectrometry. Moreover, the derivatives are more thermostable and resistant to pyrolysis. By associating these properties, separation and identification of homologs of monomycoloylglycerols are further expected.

Corynebacterium↗

PubMed1993

Chromatographic and mass spectrometric characterization of 3-O-benzoyl methyl ester derivatives of mycolic acid fractions from Corynebacterium pseudotuberculosis, C. diphtheriae and Rhodococcus rhodochrous.

A benzoyl group was attached to the 3-hydroxyl group of the methyl ester derivative of corynomycolic acid fraction isolated from Corynebacterium pseudotuberculosis. The infrared spectrum of the 3-O-benzoylated compound displayed a series of characteristic absorptions found at 1110, 1267 and 1603 cm-1 that confirmed the presence of a monosubstituted phenyl grouping. The 1H-NMR spectrum showed peaks representing protons of the aromatic ring at 7.4 ppm and 8.0 ppm. The UV spectrum revealed two absorption maxima: at 190 and 228 nm. The mass spectrum of the 3-O-benzoylated material exhibited the following peaks: (1) a prominent peak at m/z 105 of the benzoyl group that constituted the base peak; (2) peaks of methyl esters representing the alpha-hydrocarbon side chain plus carbon atoms C1 and C2 of the mycolic acid molecule; and (3) peaks of molecular ion minus benzoic acid and/or molecular ion minus benzoxyl group. When subjected to liquid chromatography (LC) on an octadecylsilane-silica gel column the 3-O-benzoylated methyl ester derivatives of the corynomycolic acid fraction were separated into their constituent homologous fractions corresponding to underivatized corynomycolic acids with the chain length C30, C32 and C34. Reversed phase HPLC of saturated and monounsaturated species of 3-O-benzoylated derivatives of the mycolic acid fraction from C. diphtheriae and Rhodococcus rhodochrous led to the separation of the corresponding homologous fractions. Mass spectrometry by electron impact mode identified both series of the homologous materials differing in mass by 28 units.

Chromatography, Liquid↗

PubMed1989

Early events associated with experimental infection of the murine lung with Rhodococcus equi.

Pneumonia due to Rhodococcus equi was induced in the murine lung by deposition of a known dose of organisms. From serial estimations of bacterial numbers in the lungs of inoculated mice, analysis of the cellular composition of bronchoalveolar lavage fluid and morphological examination of the lungs, events in the host-parasite interaction were followed until day 7. Early bacterial clearance from the lung was dose-dependent but was not sustained. A proportion of the inoculated R. equi was susceptible to the early nonspecific phagocytic cell response, and the contribution of neutrophils to bacterial clearance appeared largely limited to the first 24 hours. A substantial fraction of the organisms survived in the alveoli, probably within macrophages. The contribution phagocytes make to resistance against R. equi is similar to that which prevails in infection with Listeria monocytogenes.

Actinomycetales Infections↗

PubMed1991

Crystallization of a photosensitive nitrile hydratase from Rhodococcus sp. N-771.

A photosensitive nitrile hydratase from Rhodococcus sp. N-771 has been crystallized in two different crystal forms in its inactive form. One crystal form belongs to an orthorhombic space group P2(1)2(1)2 with unit cell dimensions of a = 117.4 A, b = 145.7 A and c = 52.1 A, and the other form belongs to a hexagonal space group P6(3)22 with unit cell dimensions of a = 110.2 A and c = 412.1 A.

Crystallization↗

PubMed1989

Synthesis of 9 alpha-hydroxysteroids by a Rhodococcus sp.

9 alpha-Hydroxylation of delta 5-3 beta-hydroxysteroids (of androstane, pregnane, 24-nor- and 21,24-bisnorcholane groups) was carried out by a Rhodococcus sp., isolated from a petroleum-containing soil sample. A large number of the investigated steroids was transformed into 9 alpha-hydroxy-delta 4-3-ketones in satisfactory yields (50-90%) at high initial concentrations of the substrates (0.5-5.0 g/L). The influence of some structural features of the steroid molecule on the progress and effectiveness of the microbial transformation was also shown.

Androstanes↗

PubMed1991

Accumulation of a poly(hydroxyalkanoate) copolymer containing primarily 3-hydroxyvalerate from simple carbohydrate substrates by Rhodococcus sp. NCIMB 40126.

A number of taxonomically-related bacteria have been identified which accumulate poly(hydroxyalkanoate) (PHA) copolymers containing primarily 3-hydroxyvalerate (3HV) monomer units from a range of unrelated single carbon sources. One of these, Rhodococcus sp. NCIMB 40126, was further investigated and shown to produce a copolymer containing 75 mol% 3HV and 25 mol% 3-hydroxybutyrate (3HB) from glucose as sole carbon source. Polyesters containing both 3HV and 3HB monomer units, together with 4-hydroxybutyrate (4HB), 5-hydroxyvalerate (5HV) or 3-hydroxyhexanoate (3HHx), were also produced by this organism from certain accumulation substrates. With valeric acid as substrate, almost pure (99 mol% 3HV) poly(3-hydroxyvalerate) was produced. N.m.r. analysis confirmed the composition of these polyesters. The thermal properties and molecular weight of the copolymer produced from glucose were comparable to those of PHB produced by Alcaligenes eutrophus.

Glucose↗

PubMed1992

Rhodococcus equi infection in patients with AIDS.

Rhodococcus equi is an emerging opportunistic pathogen of HIV-I infected patients. It is an aerobic, Gram-positive coryneform bacterium which acts as a facultative intracellular micro-organism, multiplying in the phagosome of macrophages. Eighteen cases of R. equi infection in HIV-I positive patients have now been reported. Sixteen of these had pneumonia, of which 12 had cavitating lung lesions. A history of contact with farm animals, which are the primary hosts of R. equi, was found in only three patients. There was a delay in establishing a definite diagnosis in most cases as this depended upon the isolation of R. equi from sputum, bronchoalveolar lavage fluid, or blood. Treatment included surgical resection in five patients and erythromycin with a second antibiotic in 13 cases, but II of the 18 patients died from the infection. In this report we describe our experience of R. equi pneumonia in two AIDS patients and review the published cases of the disease in man.

Acquired Immunodeficiency Syndrome↗

PubMed1989

The effect of immunosuppression on resistance to Rhodococcus equi in mice.

Rhodococcus equi, a natural pathogen of horses, produces lesions in mice following experimental infection. The effect of various immunosuppressing agents on the sequential development of these lesions has been assessed by measuring the growth of R. equi following intravenous or intranasal challenge and by histological examination. Cyclophosphamide treatment of mice, challenged intranasally, resulted in the development of lesions not unlike that seen in experimental and natural infection in foals. Cortisone acetate also impaired bacterial clearance from the lungs and affected the accumulation of mononuclear cells at infective foci. Most of the agents chosen to impair macrophage function failed to affect the resistance of mice to R. equi. Carbon, carrageenan and silica failed to alter significantly the growth kinetics of R. equi. Dextran sulphate depressed the rate of pulmonary clearance of organisms and affected the ability of animals to eliminate R. equi following rechallenge. Overall, these results support other evidence that cell mediated immunity is involved in host resistance to R. equi and that activated macrophages play a role in acquired immunity to this organism.

Actinomycetales Infections↗

PubMed1992

An assay to quantitate the binding of Rhodococcus equi to macrophages.

A Rhodococcus equi radiobinding assay has been developed using organisms labeled with 3H-uracil. These labeled organisms resemble their unlabeled counterparts with respect to colony morphology, viability, and buoyant density. Bacteria routinely incorporate between 5 x 10(-3) and 5 x 10(-2) counts per minute per colony forming unit (cfu) which in this assay allows the detection of fewer than 0.2 cfu per macrophage. Once incorporated, greater than 90% of the label remains bacterial associated for at least 4 h postlabeling. The majority of the label is trichloroacetic acid precipitable, partitions into the aqueous phase following phenol/chloroform extraction and is ethanol precipitable. RNAse treatment of the ethanol precipitate abolishes label trichloroacetic acid precipitation. This radiolabeling technique has been used to quantitate the attachment of R. equi to both murine peritoneal and equine alveolar macrophages adherent to 13 mm glass coverslips. R. equi binding is dose dependent, saturable, and specific to macrophages. Further, binding is enhanced in the presence of fresh serum. Inhibition of radiolabeled bacterial binding can be obtained by competition with cold R. equi. This radiolabeled binding assay represents a crucial step in identifying the receptors on macrophages involved in the recognition of R. equi and may help to provide information on how macrophages recognize intracellular bacteria in general.

Animals↗

PubMed1991

Cloning and characterization of an amidase gene from Rhodococcus species N-774 and its expression in Escherichia coli.

For investigation of an unknown open reading frame which is present upstream of the nitrile hydratase (NHase) gene from Rhodococcus sp. N-774, a longer DNA fragment covering the entire gene was cloned in Escherichia coli. Nucleotide sequencing and detailed subcloning experiments predicted a single open reading frame consisting of 521 amino acid residues of Mr 54,671. The amino acid sequence, especially its NH2-terminal portion, showed significant homology with those of indoleacetamide hydrolases from Pseudomonas savastanoi and Agrobacterium tumefaciens, and acetamidase from Aspergillus nidulans. The 521-amino acid coding region was therefore expressed by use of the E. coli lac promoter in E. coli, and was found to direct a considerable amidase activity. This amidase hydrolyzed propionamide efficiently, and also hydrolyzed, at a lower efficiency, acetamide, acrylamide and indoleacetamide. These data clearly show that the unknown open reading frame present upstream of the NHase coding region encodes an amidase. Because the TAG translational stop codon of the amidase is located only 75 base pairs apart from the ATG start codon of the alpha-subunit of NHase, these genes are probably translated in a polycistronic manner.

Amidohydrolases↗

PubMed1995

Bacterial steroid monooxygenase catalyzing the Baeyer-Villiger oxidation of C21-ketosteroids from Rhodococcus rhodochrous: the isolation and characterization.

Steroid monooxygenase from Rhodococcus rhodochrous, isolated in homogeneity with a high yield, catalyzes Baeyer-Villiger oxidation of progesterone to produce testosterone acetate with the stoichiometric consumptions of NADPH and molecular oxygen. It is a flavoenzyme with the molecular size of 60 kDa in the monomeric form and the isoelectric point of 4.9. The absorption spectrum has the maxima at 278, 376, and 439 nm and the shoulders at 360 and 465 nm, indicating a strong hypsochromic shift (blue-shift) of the absorption peak in the visible wavelength region. The prosthetic group of the enzyme was identified to be FAD, and the Kd value was estimated to be 0.95 microM. The enzyme catalyzed only the oxidative esterification of progesterone, 11 alpha- and 11 beta-hydroxyprogesterone and not the oxidative lactonization of androstenedione. Km for progesterone was 100 microM, for NADPH was 3.3 microM, and the turnover number was 185 min-1. Kd values for progesterone, 11 alpha-hydroxyprogesterone, deoxycorticosterone, and androstenedione were 110, 130, 2000, and 450 microM, respectively. The optimum pH of the reaction was about 8.5. The reaction was inhibited competitively by 17 alpha-hydroxyprogesterone and androstenedione. Amino terminal sequences of the enzymes from the bacterium and also from fungus, Cylindrocarpon radiocicola were considerably different, and the potential flavin-binding site could be detected on the amino-terminal region of the fungus enzyme but not on that of the bacterial enzyme. Western blotting analyses of the two steroid monooxygenases resulted that mouse antiserum raised for each enzyme reacted only with the antigenic enzyme protein but did not show the cross-reactions. It is clarified that bacterial steroid monooxygenase is distinctly different from the fungal enzyme in the molecular and enzymic properties.

Amino Acid Sequence↗

PubMed1994

Purification and characterization of a novel carbonyl reductase isolated from Rhodococcus erythropolis.

During growth on n-tetradecane a novel NADH-dependent carbonyl reductase is induced in the Gram-positive bacterium Rhodococcus erythropolis (Peters, P., Zelinski, T. and Kula, M.R. (1992) Appl. Microbiol. Biotechnol. 38, 334-340). The enzyme has been purified to homogeneity using fractional pH precipitation, anion exchange chromatography and affinity chromatography. The isoelectric point of the oxidoreductase is 4.4. The apparent molecular mass of the native enzyme is 161 kDa, that of the subunits 40 kDa as determined by SDS gel electrophoresis. A tetrameric structure of the carbonyl reductase is consistent with these results. Important biochemical data concerning the application of the reductase are: a broad pH-optimum, temperature optimum at 40 degrees C and stability at room temperature for more than 5 days. The oxidoreductase accepted as substrate aliphatic and aromatic ketones, keto esters (esters of keto carboxylic acids) and halogenated carbonyl compounds and reduced them to the corresponding hydroxyl compounds with (S)-configuration with more than 98% enantiomeric excess. The NAD(+)-dependent oxidation of primary alcohols was not catalyzed by the carbonyl reductase, whereas secondary alcohols and hydroxy acid esters were oxidized to the corresponding carbonyl compounds at about 10-fold slower reaction rates compared to the reduction.

Alcohol Oxidoreductases↗

PubMed1992

Cloning of DNA from a Rhodococcus strain conferring the ability to decolorize sulfonated azo dyes.

Azo dyes are recalcitrant pollutants. Two sulfonated azo dyes, Orange II and Amido black, are effectively decolorized by certain nocardioform strains of the genus Rhodococcus. A mutant of one of these strains was isolated which had lost azo-dye decolorizing ability and the strain was used to clone DNA conferring this ability, by screening a BclI library constructed from DNA of a decolorizing strain. The relevant genetic information was located on a 6.3-kb fragment of DNA.

Amido Black↗

PubMed1992

Identification, cloning and sequence analysis of the poly(3-hydroxyalkanoic acid) synthase gene of the gram-positive bacterium Rhodococcus ruber.

The first polyhydroxyalkanoic acid (PHA) synthase gene (phbCRr) of a Gram-positive bacterium was cloned from a genomic library of Rhodococcus ruber in the broad-host-range plasmid vector pRK404. The hybrid plasmid harboring phbCRr allowed the expression of polyhydroxybutyric acid (PHB) synthase activity and restored the ability of PHB synthesis in a PHB-negative mutant of Alcaligenes eutrophus. Nucleotide sequence analysis of phbCRr revealed an open reading frame of 1686 bp starting with the rare codon TTG and encoding a protein of relative molecular mass 61,371. The deduced amino acid sequence of phbCRr exhibited homologies to the primary structures of the PHA synthases of A. eutrophus and Pseudomonas oleovorans. Preparation of PHA granules by discontinuous density gradient centrifugation of crude cellular extracts revealed four major bands in an SDS polyacrylamide gel. A Mr 61,000 protein was identified as the PHA synthase of R. ruber by N-terminal amino acid sequence determination.

Acyltransferases↗

PubMed1992

Sequence of a Rhodococcus gene encoding a protein with extensive homology to the mammalian propionyl-CoA carboxylase beta chain.

Sequence analysis of a 2-kilobase DNA fragment from Rhodococcus sp. NI86/21 revealed an open reading frame encoding a 476-amino-acid protein with striking homology to the rat and human propionyl-CoA carboxylase beta subunits. The nucleotide sequence of a corresponding prokaryotic gene has not yet been reported. Upstream, the C-terminal part of a putative beta-ketoacyl synthase was identified.

3-Oxoacyl-(Acyl-Carrier-Protein) Synthase↗

PubMed1995

The evolutionary relationship of biphenyl dioxygenase from gram-positive Rhodococcus globerulus P6 to multicomponent dioxygenases from gram-negative bacteria.

The Gram+ bacterium Rhodococcus globerulus P6 (RgP6) catabolizes a range of polychlorinated biphenyl (PCB) congeners, thus being of interest in bioelimination processes for PCB. The first step in the pathway, a dioxygenase attack of one of the biphenyl (BP) rings, is catalyzed by biphenyl dioxygenase (BDO). In this study, the nucleotide (nt) sequences of the four clustered cistrons, bphA1A2A3A4, encoding the subunits of BDO and forming part of the bph operon of RgP6 for BP degradation, were determined. A conserved motif proposed to bind a Rieske-type [2Fe-2S] cluster was identified in the deduced amino acid (aa) sequence of both the a subunit of the terminal oxygenase (BphA1) and ferredoxin (BphA3). The ferredoxin reductase subunit (BphA4) contains conserved sites for FAD and NADH binding. Deduced aa sequences of the BDO subunits shared homologies to multicomponent aromatic ring-hydroxylating dioxygenases from Gram- microorganisms. Stronger identity was found to toluene dioxygenase (TDO) of Pseudomonas putida F1 than to other BDO. Aa sequence comparisons suggest that BP degradation genes of RgP6 may have originated in Gram- microorganisms, probably Pseudomonas, and subsequently transferred to this Gram+ bacterium.

Amino Acid Sequence↗