Comparative physiology of phenanthrene degradation by two dissimilar pseudomonads isolated from a creosote-contaminated soil

1994 ◽  
Vol 40 (6) ◽  
pp. 432-438 ◽  
Author(s):  
William T. Stringfellow ◽  
Michael D. Aitken
Keyword(s):  
Contaminated Soil ◽  
Genetic Relatedness ◽  
Enrichment Culture ◽  
Acid Methyl Ester ◽  
Pseudomonas Stutzeri ◽  
Maximum Activity ◽  
Resting Cells ◽  
Phenanthrene Degradation ◽  
Naphthoic Acid ◽  
P 16

Two species of bacteria, identified as Pseudomonas stutzeri (P-16) and Pseudomonas saccharophila (P-15) by fatty acid methyl ester analysis, were found in a phenanthrene enrichment culture of a creosote-contaminated soil. The organisms are shown to be physiologically dissimilar, and their genetic relatedness is discussed. Phenanthrene degradation by both organisms followed Michaelis–Menten kinetics, allowing for the determination of half-saturation (Ks) and maximum activity coefficients, using nonlinear regression. Both organisms utilized kinetically similar enzymes for phenanthrene uptake and oxidation, as evidenced by similar Ks coefficients of approximately 0.2 mg/L and temperature optima of 40 °C, but levels of expression differed with different media. Each organism degraded phenanthrene via salicylic acid, but patterns of intermediate metabolism were shown to differ. P-15 excreted 1-hydroxy-2-naphthoic acid during growth on phenanthrene and demonstrated Michaelis–Menten kinetics for the oxidation of 1-hydroxy-2-naphthoic acid by resting cells. P-16 excreted only trace amounts of 1-hydroxy-2-naphthoic acid and demonstrated linear kinetics in response to 1-hydroxy-2-naphthoic acid concentration. P-15 was found to form thick biofilms on phenanthrene crystals and was characterized by a hydrophobic cell surface, whereas P-16 grew mostly in suspension and was hydrophilic. Neither organism produced significant amounts of biosurfactants when grown on phenanthrene. The implications of these findings for the design of systems to remediate contaminated soil are discussed.Key words: phenanthrene, 1-hydroxy-2-naphthoic acid, biodegradation, kinetics, polycyclic aromatic hydrocarbons.

The metabolism of p-fluorophenylacetic acid by a Pseudomonas sp. I. Isolation and identification of intermediates in degradation

1971 ◽  
Vol 17 (5) ◽  
pp. 635-644 ◽  
Author(s):  
D. B. Harper ◽  
E. R. Blakley
Keyword(s):  
Sole Carbon Source ◽  
Culture Medium ◽  
Enrichment Culture ◽  
Culture Technique ◽  
Spectroscopic Techniques ◽  
Resting Cells ◽  
Pseudomonas Sp ◽  
Isolation And Identification ◽  
Hydroxyphenylacetic Acid ◽  
Organic Fluorine

A Pseudomonas sp. capable of growing on p-fluorophenylacetic acid as sole carbon source has been isolated using the enrichment culture technique. All the organic fluorine is released into the culture medium as fluoride ion during growth. A number of fluorinated intermediates have been isolated from the culture medium when resting cells were incubated with the substrate. Using infrared, nuclear magnetic resonance, and mass spectroscopic techniques together with chemical degradative procedures, these have been identified as D(+)-monofluorosuccinic acid, trans-3-fluoro-3-hexenedioic acid, (−)-4-carboxymethyl-4-fluorobutanolide, 4-fluoro-2-hydroxyphenylacetic acid, and 4-fluoro-3-hydroxyphenylacetic acid.

Microbial growth on mercaptosuccinic acid

1968 ◽  
Vol 14 (5) ◽  
pp. 515-523 ◽  
Author(s):  
Mark R. Hall ◽  
Richard S. Berk
Keyword(s):  
Microbial Growth ◽  
Enrichment Culture ◽  
Basal Medium ◽  
Sole Source ◽  
Resting Cells ◽  
Metabolic Fate ◽  
Succinate Oxidation ◽  
Oxidase System ◽  
Mercaptosuccinic Acid ◽  
Inorganic Sulfate

Using enrichment culture technics a species of Alcaligenes (M1) was isolated from soil which was able to utilize mercaptosuccinic acid (MS) as a sole source of carbon, sulfur, and energy. Growth on a MS–salts basal medium was not significantly enhanced by single supplements of B-vitamins or by yeast extract. Comparative studies on succinate and MS oxidation by Alcaligenes and Pseudomonas aeruginosa indicated that MS was an inhibitor of succinate oxidation by resting cells of both microorganisms when they were grown in a medium lacking mercaptosuccinic acid such as a succinate–salts basal. However, when M1 was grown on MS, it was able to oxidize both succinate and MS, thereby indicating that the MS oxidase system was inducible. In addition, the MS oxidase system in cell-free extracts of M1 was relatively insensitive to 10–30 μmoles of malonate, whereas the succinoxidase system was inhibited 66% by 30 μmoles of the inhibitor. Cell-free extracts of succinate-grown cells of P. aeruginosa were unable to oxidize MS, indicating that the inactivity of resting cells was not due to a permease problem. Investigation of the metabolic fate of the sulfur moiety of MS by growing cells of M1 indicated that all of the available sulfur was liberated as inorganic sulfate, while no free sulfide was detected. Thiosulfate sulfurtransferase (rhodanese) was detected in extracts from cells grown both with and without mercaptosuccinic acid. However, growth in the MS medium enhanced the production of rhodanese approximately 40%. In addition, thiosulfate oxidase activity was also detected in resting cells and cell-free extracts prepared from MS-grown cells, but not from cells grown without mercaptosuccinic acid.

scholarly journals Biofilm Interaction Mapping and Analysis (BIMA): A tool for deconstructing interspecific interactions in co-culture biofilms

2021 ◽  
Author(s):  
Suzanne M Kosina ◽  
Peter Rademacher ◽  
Kelly M Wetmore ◽  
Markus de Raad ◽  
Marcin Zemla ◽  
...  
Keyword(s):  
Contaminated Soil ◽  
Interspecific Interactions ◽  
Pseudomonas Stutzeri ◽  
Interspecific Interaction ◽  
Deletion Mutants ◽  
Bacterial Strains ◽  
Chromium Vi ◽  
Pseudomonas Species ◽  
Aquaculture Management ◽  
Culture Growth

Pseudomonas species are ubiquitous in nature and include numerous medically, agriculturally and technologically beneficial strains of which the interspecific interactions are of great interest for biotechnologies. Specifically, co-cultures containing Pseudomonas stutzeri have been used for bioremediation, biocontrol, aquaculture management and wastewater denitrification. Furthermore, the use of P. stutzeri biofilms, in combination with consortia based approaches, may offer advantages for these processes. Understanding the interspecific interaction within biofilm co-cultures or consortia provides a means for improvement of current technologies. However, the investigation of biofilm based consortia has been limited. We present an adaptable and scalable method for the analysis of macroscopic interactions (colony morphology, inhibition and invasion) between colony forming bacterial strains using an automated printing method followed by analysis of the genes and metabolites involved in the interactions. Using Biofilm Interaction Mapping and Analysis (BIMA), these interactions were investigated between P. stutzeri strain RCH2, a denitrifier isolated from chromium (VI) contaminated soil, and thirteen other species of pseudomonas isolated from non-contaminated soil. The metabolites and genes associated with both active co-culture growth and inhibitory growth were investigated using mass spectrometry based metabolomics and mutant fitness profiling of a DNA-barcoded mutant library. One interaction partner, Pseudomonas fluorescens N1B4 was selected for mutant fitness profiling; with this approach four genes of importance were identified and the effects on interactions were evaluated with deletion mutants and metabolomics.

Characteristics of phenanthrene-degrading bacteria isolated from soils contaminated with polycyclic aromatic hydrocarbons

1998 ◽  
Vol 44 (8) ◽  
pp. 743-752 ◽  
Author(s):  
Michael D Aitken ◽  
William T Stringfellow ◽  
Robert D Nagel ◽  
Chikoma Kazunga ◽  
Shu-Hwa Chen
Keyword(s):  
Polycyclic Aromatic Hydrocarbons ◽  
Aromatic Hydrocarbons ◽  
Contaminated Soils ◽  
Acid Methyl Ester ◽  
Aqueous Solubility ◽  
Bacterial Strains ◽  
Phenanthrene Degradation ◽  
Acid Methyl ◽  
Degrading Bacteria ◽  
Polycyclic Aromatic

Ten bacterial strains were isolated from seven contaminated soils by enrichment with phenanthrene as the sole carbon source. These isolates and another phenanthrene-degrading strain were examined for various characteristics related to phenanthrene degradation and their ability to metabolize 12 other polycyclic aromatic hydrocarbons (PAH), ranging in size from two to five rings, after growth in the presence of phenanthrene. Fatty acid methyl ester analysis indicated that at least five genera (Agrobacterium, Bacillus, Burkholderia, Pseudomonas, and Sphingomonas) and at least three species of Pseudomonas were represented in this collection. All of the strains oxidized phenanthrene according to Michaelis-Menten kinetics, with half-saturation coefficients well below the aqueous solubility of phenanthrene in all cases. All but one of the strains oxidized 1-hydroxy-2-naphthoate following growth on phenanthrene, and all oxidized at least one downstream intermediate from either or both of the known phenanthrene degradation pathways. All of the isolates could metabolize (oxidize, mineralize, or remove from solution) a broad range of PAH, although the exact range and extent of metabolism for a given substrate were unique to the particular isolate. Benz[a]anthracene, chrysene, and benzo[a]pyrene were each mineralized by eight of the strains, while pyrene was not mineralized by any. Pyrene was, however, removed from solution by all of the isolates, and the presence of at least one significant metabolite from pyrene was observed by radiochromatography for the five strains in which such metabolites were sought. Our results support earlier indications that the mineralization of pyrene by bacteria may require unique metabolic capabilities that do not appear to overlap with the determinants for mineralization of phenanthrene or other high molecular weight PAH.Key words: kinetics, polycyclic aromatic hydrocarbons, phenanthrene, mineralization, benzo[a]pyrene.

scholarly journals Biodegradation of cis-Dichloroethene as the Sole Carbon Source by a β-Proteobacterium

2002 ◽  
Vol 68 (6) ◽  
pp. 2726-2730 ◽  
Author(s):  
Nicholas V. Coleman ◽  
Timothy E. Mattes ◽  
James M. Gossett ◽  
Jim C. Spain
Keyword(s):  
Enrichment Culture ◽  
Growth Yield ◽  
Utilization Rate ◽  
Specific Substrate ◽  
Resting Cells ◽  
Aerobic Bacterium ◽  
16S Ribosomal Dna ◽  
Epoxidation Reaction ◽  
Specific Substrate Utilization Rate ◽  
Ribosomal Dna Sequence

ABSTRACT An aerobic bacterium capable of growth on cis-dichloroethene (cDCE) as a sole carbon and energy source was isolated by enrichment culture. The 16S ribosomal DNA sequence of the isolate (strain JS666) had 97.9% identity to the sequence from Polaromonas vacuolata, indicating that the isolate was a β-proteobacterium. At 20°C, strain JS666 grew on cDCE with a minimum doubling time of 73 ± 7 h and a growth yield of 6.1 g of protein/mol of cDCE. Chloride analysis indicated that complete dechlorination of cDCE occurred during growth. The half-velocity constant for cDCE transformation was 1.6 ± 0.2 μM, and the maximum specific substrate utilization rate ranged from 12.6 to 16.8 nmol/min/mg of protein. Resting cells grown on cDCE could transform cDCE, ethene, vinyl chloride, trans-dichloroethene, trichloroethene, and 1,2-dichloroethane. Epoxyethane was produced from ethene by cDCE-grown cells, suggesting that an epoxidation reaction is the first step in cDCE degradation.

scholarly journals Metabolism of Thioamides by Ralstonia pickettii TA

2006 ◽  
Vol 72 (12) ◽  
pp. 7468-7476 ◽  
Author(s):  
Anthony G. Dodge ◽  
Jack E. Richman ◽  
Gilbert Johnson ◽  
Lawrence P. Wackett
Keyword(s):  
Nitrogen Source ◽  
Enrichment Culture ◽  
Carbon Sources ◽  
Resting Cells ◽  
Sole Nitrogen Source ◽  
Antituberculosis Drug ◽  
Ralstonia Pickettii ◽  
Dead End ◽  
Spontaneous Reaction ◽  
Related Compounds

ABSTRACT Information on bacterial thioamide metabolism has focused on transformation of the antituberculosis drug ethionamide and related compounds by Mycobacterium tuberculosis. To study this metabolism more generally, a bacterium that grew using thioacetamide as the sole nitrogen source was isolated via enrichment culture. The bacterium was identified as Ralstonia pickettii and designated strain TA. Cells grown on thioacetamide also transformed other thioamide compounds. Transformation of the thioamides tested was dependent on oxygen. During thioamide degradation, sulfur was detected in the medium at the oxidation level of sulfite, further suggesting an oxygenase mechanism. R. pickettii TA did not grow on thiobenzamide as a nitrogen source, but resting cells converted thiobenzamide to benzamide, with thiobenzamide S-oxide and benzonitrile detected as intermediates. Thioacetamide S-oxide was detected as an intermediate during thioacetamide degradation, but the only accumulating metabolite of thioacetamide was identified as 3,5-dimethyl-1,2,4-thiadiazole, a compound shown to derive from spontaneous reaction of thioacetamide and oxygenated thioacetamide species. This dead-end metabolite accounted for only ca. 12% of the metabolized thioacetamide. Neither acetonitrile nor acetamide was detected during thioacetamide degradation, but R. pickettii grew on both compounds as nitrogen and carbon sources. It is proposed that R. pickettii TA degrades thioamides via a mechanism involving consecutive oxygenations of the thioamide sulfur atom.

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