Pseudomonas Multivorans Research Articles

Purification and properties of a NAD-linked 1,2-propanediol dehydrogenase from propane-grown Pseudomonas fluorescens NRRL B-1244

NAD-dependent 1,2-propanediol dehydrogenase (EC 1.1.1.4) activity was detected in cell-free crude extracts of various propane-grown bacteria. The enzyme activity was much lower in 1-propanol-grown cells than in propane-grown cells of Pseudomonas fluorescens NRRL B-1244, indicating that the enzyme may be inducible by metabolites of propane subterminal oxidation. 1,2-Propanediol dehydrogenase was purified from propane-grown Ps. fluorescens NRRL B-1244. The purified enzyme fraction shows a single-protein band upon acrylamide gel electrophoresis and has a molecular weight of 760,000. It consists of 10 subunits of identical molecular weight (77,600). It oxidizes diols that possess either two adjacent hydroxy groups, or a hydroxy group with an adjacent carbonyl group. Primary and secondary alcohols are not oxidized. The pH and temperature optima for 1,2-propanediol dehydrogenase are 8.5 and 20–25 °C, respectively. The activation energy calculated is 5.76 kcal/mol. 1,2-Propanediol dehydrogenase does not catalyze the reduction of acetol or acetoin in the presence of NADH (reverse reaction). The K m values at 25 °C, pH 7.0, buffer solution for 1,2-propanediol and NAD are 2 × 10 −2 and 9 × 10 −5 m, respectively. The 1,2-propanediol dehydrogenase activity was inhibited by strong thiol reagents, but not by metal-chelating agents. The amino acid composition of the purified enzyme was determined. Antisera prepared against purified 1,2-propanediol dehydrogenase from propane-grown Ps. fluorescens NRRL B-1244 formed homologous precipitin bands with isofunctional enzymes derived from propane-grown Arthrobacter sp. NRRL B-11315, Nocardia paraffinica ATCC 21198, and Mycobacterium sp. P2y, but not from propane-grown Pseudomonas multivorans ATCC 17616 and Brevibacterium sp. ATCC 14649, or 1-propanol-grown Ps. fluorescens NRRL B-1244. Isofunctional enzymes derived from methane-grown methylotrophs also showed different immunological and catalytic properties.

Microcalorimetry of microorganism metabolism of monosaccharides and simple aromatic compounds

AbstractHeat conduction solution enable rapid determination of the heats of aerobic and anaerobic metabolism of substrate by microorganisms. Aliquots of 1.0 ml cell suspension, 5 × 109 cell/ml, were mixed with a few dozen nmol substrate contained in 0.5 ml, under a controlled atmosphere of air, O2, or N2. At these substrate concentration, with adapted microorganisms, metabolism and its heat generation are usually complete within 300 to 600 sec. The raw data yield ΔHapp values. The ΔHapp were determined in the range 0.001 to 0.010% substrate, and extrapolated (limit substrate concentration →0), to yield Δ0H̄, the limiting differential molar heat of metabolism. The Δ0H̄ values express the heat generated when there is rapid metabolism but little new growth, minimal contribution by H+ transfer from metabolites, and maintenance of aerobicity or anaerobicity as specified. Escherichiacoli B/5 was used for aerobic and anaerobic combustion of eight sugars. Pseudomonas multivorans, and an Acinetobacter, strain B‐1, were used for aerobic metabolism of benzene, toluene, naphthalene, and a methylnaphthalene. The larger heats of combustion of the hydrocarbons enable the use of aqueous solutions of hydrocarbons well below their solubility limits. The quotient Δ0H̄/n (n = atoms carbon/molecule substrate) varies from (‐)36 to (‐)67 kcal/mol carbon for the sugars. The most reduced sugar yields the largest exothermic heats. The quotient varies from (‐)27 to (‐)81 kcal/mol carbon for the aromatic hydrocarbons. Comparison of the calorimetric heats of metabolism of those from total aerobic combustion in aquo (where available) give measure of the efficiencies with which the heat contents of the aqueous substrate are used by the bacteria.

Investigation on the regulation of aniline utilization in Pseudomonas multivorans strain An 1

Pseudomonas multivorans strain An 1 was isolated from forest soil after enrichment in a medium containing 0.1% aniline as the sole source of carbon and energy. Increasing aniline concentrations increasingly inhibited bacterial growth. At pH 7, aniline concentrations greater than 16mM were toxic enough to completely arrest growth. The optimal pH for growth on aniline was 6.5. On binary mixtures of aniline and additional carbon sources, diauxic growth was observed. The addition carbon sources caused various degrees of repression of the aniline catabolizing enzyme system. The fastest induction of this system occurred at pH 4, suggesting that protonization of the aniline molecule is crucial.

Comparative studies on the metabolism of aniline and chloroanilines by Pseudomonas multivorans strain An 1

Pseudomonas multivorans strain An 1 used aniline but not chloroanilines as the sole source of carbon and energy for growth. The aniline-adapted cells, however, were able to oxygenate chloroanilines. Relative oxygenation rates for aniline, 2-chloroaniline, 3-chloroaniline, 4-chloroaniline, and 3,4-dichloroaniline were 100, 46, 66, 20, and 3%, respectively.

Emendation of the Description of Pseudomonas cepacia Burkholder (Synonyms: Pseudomonas multivorans Stainer et al., Pseudomonas kingae Jonsson; EO-1 Group)

It has been reported that the names Pseudomonas multivorans, P. kingae, and P. cepacia are synonymous and that P. cepacia has priority. We have confirmed this synonymy by subjecting a larger number of strains (15 each) of P. multivorans and P. kingae to a wider variety of morphological, nutritional, physiological, and biochemical tests than employed in other studies. A total of 127 tests performed on each strain showed that the two groups of strains were 94% similar, differing only in the following nine characters: growth at 41 C; pigmentation; growth on cetrimide agar; nitrate reduction; acid from sucrose and maltose on O-F medium; utilization of nicotinate and n-dodecane as sole carbon sources; and susceptibility to nalidixic acid. Salient properties for both groups of strains are nutritional versatility and accumulation of poly-β-hydroxybutyrate as a cellular reserve material.

Purification and characterization of the two 6-phosphogluconate dehydrogenase species from Pseudomonas multivorans.

The two species of 6-phosphogluconate dehydrogenase (EC 1.1.1.43) from Pseudomonas multivorans were resolved from extracts of gluconate-grown bacteria and purified to homogeneity. Each enzyme comprised between 0.1 and 0.2% of the total cellular protein. Separation of the two enzymes, one which is specific for nicotinamide adenine dinucleotide phosphate and the other which is active with nicotinamide adenine dinucleotide or nicotinamide adenine dinucleotide phosphate was facilitated by the marked difference in their respective isoelectric points, which were at pH 5.0 and 6.9. Comparison of the subunit compositions of the two enzymes indicated that they do not share common peptide chains. The enzyme active with nicotinamide adenine dinucleotide was composed of two subunits of about 40,000 molecular weight, and the nicotinamide adenine dinucleotide phosphate-specific enzyme was composed of two subunits of about 60,000 molecular weight. Immunological studies indicated that the two enzymes do not share common antigenic determinants. Reduced nicotinamide adenine dinucleotide phosphate strongly inhibited the 6-phosphogluconate dehydrogenase active with nicotinamide adenine dinucleotide by decreasing its affinity for 6-phosphogluconate. Guanosine-5'-triphosphate had a similar influence on the nicotinamide adenine dinucleotide phosphate-specific 6-phosphogluconate dehydrogenase. These results in conjunction with other data indicating that reduced nicotinamide adenine dinucleotide phosphate stimulates the conversion of 6-phosphogluconate to pyruvate by crude bacterial extracts suggest that in P. multivorans, the relative distribution of 6-phosphogluconate into the pentose phosphate and Entner-Doudoroff pathways might be determined by the intracellular concentrations of reduced nicotinamide adenine dinucleotide phosphate and purine nucleotides.

Purification and characterization of the Pseudomonas multivorans glucose-6-phosphate dehydrogenase active with nicotinamide adenine dinucleotide.

The Pseudomonas multivorans glucose-6-phosphate dehydrogenase (EC 1.1.1.49) active with nicotinamide adenine dinucleotide, which is inhibitable by adenosine-5'-triphosphate, was purified approximately 1,000-fold from extracts of glucose-grown bacteria, and characterized with respect to subunit composition, response to different inhibitory ligands, and certain other properties. The enzyme was found to be an oligomer composed of four subunits of about 60,000 molecular weight. Reduced nicotinamide adenine dinucleotide phosphate, but not reduced nicotinamide adenine dinucleotide, was found to be a potent inhibitor of its activity. The range of concentrations of reduced nicotinamide adenine dinucleotide phosphate over which inhibition occurred was about 100-fold lower than that for adenosine-5'-triphosphate. The data suggest that reduced nicotinamide adenine dinucleotide phosphate may play an important role in regulation of hexose phosphate metabolism in P. multivorans. Antisera prepared against the purified enzyme strongly inhibited its activity, but failed to inhibit the activity of the nicotinamide adenine dinucleotide phosphate-specific glucose-6-phosphate dehydrogenase which is also present in extracts of this bacterium. Immunodiffusion experiments confirmed the results of the enzyme inhibition studies, and failed to support the idea that the two glucose-6-phosphate dehydrogenase species from P. multivorans represent different oligomeric forms of the same protein.

The fatty acids of Pseudomonas multivorans (Pseudomonas cepacia) and Pseudomonas kingii.

SUMMARY: The fatty acid compositions of three strains of Pseudomonas multivorans and three of P. kingii were determined by gas-liquid chromatography. The major fatty acids identified were 16:0, 16:1, 18:1, 3-OH 14:0, 3-OH 16:0 and cyclopropane acids 17 Δ and 19 Δ. The fatty acid compositions of these strains were similar to the acids identified in sixteen clinical isolates of the P. multivorans (P. cepacia)/P. kingii group’. These data support the current view that P. multivorans and P. kingii are identical species.

Multiple forms of Pseudomonas multivorans glucose-6-phosphate and 6-phosphogluconate dehydrogenases: differences in size, pyridine nucleotide specificity, and susceptibility to inhibition by adenosine 5'-triphosphate.

Two major species of glucose-6-phosphate dehydrogenase (EC 1.1.1.49) differing in size, pyridine nucleotide specificity, and susceptibility to inhibition by adenosine 5'-triphosphate (ATP) were detected in extracts of Pseudomonas multivorans (which has recently been shown to be synonymous with the species Pseudomonas cepacia) ATCC 17616. The large species (molecular weight ca. 230,000) was active with nicotinamide adenine dinucleotide (NAD) or nicotinamide adenine dinucleotide phosphate (NADP) and was markedly inhibited by ATP, which decreased its affinity for glucose-6-phosphate and for pyridine nucleotides. This form of the enzyme exhibited homotropic effects for glucose-6-phosphate. The small species (molecular weight ca. 96,000) was active with NADP but not with NAD, was not inhibited by ATP, and exhibited no homotropic effects for glucose-6-phosphate. Under certain conditions multiplicity of 6-phosphogluconate dehydrogenase (EC 1.1.1.43) activities was also noted. One form of the enzyme (80,000 molecular weight) was active with either NAD or NADP and was inhibited by ATP, which decreased its affinity for 6-phosphogluconate. The other form (120,000 molecular weight) was highly specific for NADP and was not susceptible to inhibition by ATP. Neither form of the enzyme exhibited homotropic effects for 6-phosphogluconate. The possible relationships between the different species of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase are discussed.

Nucleic acid enzyme studies of nonfermentative gram-negative bacteria using thin-layer chromatography.

A rapid and easy to perform procedure for determining the nucleic acid enzyme reactions of intact bacterial cells was developed. Overnight organism growth on triple sugar agar was tested for nucleoside phosphotransferase, nucleosidase, and nucleotidase activity. Reaction products were detected by means of thin-layer chromatography and fluorescence. Characteristic patterns were seen with certain strains of nonfermentative gram-negative bacteria, which indicated that these tests could aid in classification and identification of E0-1 group, Pseudomonas multivorans, P. cepacia, P. maltophilia, certain other Pseudomonas species, and Herellea vaginicola.

Nucleic Acid Enzyme Studies of Nonfermentative Gram-Negative Bacteria Using Thin-Layer Chromatography1

A rapid and easy to perform procedure for determining the nucleic acid enzyme reactions of intact bacterial cells was developed. Overnight organism growth on triple sugar agar was tested for nucleoside phosphotransferase, nucleosidase, and nucleotidase activity. Reaction products were detected by means of thin-layer chromatography and fluorescence. Characteristic patterns were seen with certain strains of nonfermentative gram-negative bacteria, which indicated that these tests could aid in classification and identification of E0-1 group, Pseudomonas multivorans, P. cepacia, P. maltophilia, certain other Pseudomonas species, and Herellea vaginicola.

WOUND INFECTION WITH PSEUDOMONAS MULTIVORANS: A WATER-BORNE CONTAMINANT OF DISINFECTANT SOLUTIONS

Over a period of 5 weeks, Pseudomonas multivorans was isolated from infected operation wounds of nine patients in a hospital. In some cases the organism seemed to be a wound contaminant but in others it behaved as a pathogen. It was isolated from bottles of a 1 in 30 dilution of' Savlon' (chlorhexidine 0·05%, cetrimide 0·5%) in the hospital, and from samples of the piped water supply in the hospital and outside.

Characterization of saccharolytic nonfermentative bacteria associated with man.

Features of 378 clinical isolates of saccharolytic, nonfermentative Gram-negative rods and 20 reference strains were examined. All but four of the clinical strains were assigned to recognized taxa, namely Acinetobacter, Chromobacterium, Flavobacterium, Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas maltophilia, Pseudomonas multivorans, Pseudomonas putida, Pseudomonas stutzeri, and Xanthomonas.

Early intermediates in the degradation of alpha-conidendrin by a Pseudomonas multivorans.

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTEarly intermediates in the degradation of α-conidendrin by a Pseudomonas multivoransAnne Toms and John Mathew WoodCite this: Biochemistry 1970, 9, 4, 733–740Publication Date (Print):February 1, 1970Publication History Published online1 May 2002Published inissue 1 February 1970https://doi.org/10.1021/bi00806a005RIGHTS & PERMISSIONSArticle Views32Altmetric-Citations10LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit PDF (567 KB) Get e-Alerts Get e-Alerts

Evidence for the presence of fimbriae (pili) on vibrio species.

SUMMARY: Three strains of Vibrio were found to agglutinate both guinea-pig and human red cells. The vibrios differed in haemagglutination tests from fim-briated enterobacteria by showing only partial inhibition by mannose and by agglutinating guinea-pig and human red cells equally well. Electron microscopy of the vibrios revealed filamentous appendages which resembled the common fimbriae of Escherichia coli in having average diameters between 6 and 10 mμ and in being more numerous on organisms from liquid media than from solid media. The fimbriae on vibrios also had some characteristics of F fimbriae of enterobacteria. In comparative studies fimbriae which resembled common fimbriae of E. coli were found on Pseudomonas multi-vorans and Aeromonas liquefaciens.