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Angewandte Chemie (International Ed. in... Sep 2020N,N-dimethyl formamide (DMF) is an extensively used organic solvent but is also a potent pollutant. Certain bacterial species from genera such as Paracoccus,...
N,N-dimethyl formamide (DMF) is an extensively used organic solvent but is also a potent pollutant. Certain bacterial species from genera such as Paracoccus, Pseudomonas, and Alcaligenes have evolved to use DMF as a sole carbon and nitrogen source for growth via degradation by a dimethylformamidase (DMFase). We show that DMFase from Paracoccus sp. strain DMF is a halophilic and thermostable enzyme comprising a multimeric complex of the α β or (α β ) type. One of the three domains of the large subunit and the small subunit are hitherto undescribed protein folds of unknown evolutionary origin. The active site consists of a mononuclear iron coordinated by two Tyr side-chain phenolates and one carboxylate from Glu. The Fe ion in the active site catalyzes the hydrolytic cleavage of the amide bond in DMF. Kinetic characterization reveals that the enzyme shows cooperativity between subunits, and mutagenesis and structural data provide clues to the catalytic mechanism.
Topics: Amidohydrolases; Catalytic Domain; Dimethylformamide; Molecular Structure; Paracoccus; Tyrosine
PubMed: 32452120
DOI: 10.1002/anie.202005332 -
Journal of Clinical Microbiology May 1975The cellular fatty acid composition of 25 clinical isolates of Alcaligenes and Pseudomonas was determined by gas-liquid chromatography (GLC). The GLC fatty acid profiles...
The cellular fatty acid composition of 25 clinical isolates of Alcaligenes and Pseudomonas was determined by gas-liquid chromatography (GLC). The GLC fatty acid profiles of three species of Pseudomonas were markedly different from those of Alcaligenes. The most significant differences were the presence and relative amounts of hydroxy, branched-chain, and cyclopropane fatty acids. One of the major fatty acids in A. faecalis was a 17-carbon cyclopropane (17 delta) acid, whereas a 15-carbon branched-chain acid (13-methyl tetradecanoate) characterized isolates of P. putrefaciens. The determination of these fatty acids by GLC provides a rapid and specific means of distinguishing clinical isolates of Pseudomonas and Alcaligenes.
Topics: Alcaligenes; Bacterial Infections; Chromatography, Gas; Cyclopropanes; Fatty Acids; Humans; Pseudomonas; Species Specificity
PubMed: 1176611
DOI: 10.1128/jcm.1.5.414-419.1975 -
Applied and Environmental Microbiology Mar 1999Two 3-hydroxybenzoate-inducible gentisate 1,2-dioxygenases were purified to homogeneity from Pseudomonas alcaligenes NCIB 9867 (P25X) and Pseudomonas putida NCIB 9869...
Two 3-hydroxybenzoate-inducible gentisate 1,2-dioxygenases were purified to homogeneity from Pseudomonas alcaligenes NCIB 9867 (P25X) and Pseudomonas putida NCIB 9869 (P35X), respectively. The estimated molecular mass of the purified P25X gentisate 1, 2-dioxygenase was 154 kDa, with a subunit mass of 39 kDa. Its structure is deduced to be a tetramer. The pI of this enzyme was established to be 4.8 to 5.0. The subunit mass of P35X gentisate 1, 2-dioxygenase was 41 kDa, and this enzyme was deduced to exist as a dimer, with a native molecular mass of about 82 kDa. The pI of P35X gentisate 1,2-dioxygenase was around 4.6 to 4.8. Both of the gentisate 1,2-dioxygenases exhibited typical saturation kinetics and had apparent Kms of 92 and 143 microM for gentisate, respectively. Broad substrate specificities were exhibited towards alkyl and halogenated gentisate analogs. Both enzymes had similar kinetic turnover characteristics for gentisate, with kcat/Km values of 44.08 x 10(4) s-1 M-1 for the P25X enzyme and 39.34 x 10(4) s-1 M-1 for the P35X enzyme. Higher kcat/Km values were expressed by both enzymes against the substituted gentisates. Significant differences were observed between the N-terminal sequences of the first 23 amino acid residues of the P25X and P35X gentisate 1,2-dioxygenases. The P25X gentisate 1,2-dioxygenase was stable between pH 5.0 and 7.5, with the optimal pH around 8.0. The P35X enzyme showed a pH stability range between 7.0 and 9.0, and the optimum pH was also 8.0. The optimal temperature for both P25X and P35X gentisate 1, 2-dioxygenases was around 50 degrees C, but the P35X enzyme was more heat stable than that from P25X. Both enzymes were strongly stimulated by 0.1 mM Fe2+ but were completely inhibited by the presence of 5 mM Cu2+. Partial inhibition of both enzymes was also observed with 5 mM Mn2+, Zn2+, and EDTA.
Topics: Amino Acid Sequence; Biodegradation, Environmental; Dioxygenases; Gentisates; Hydrogen-Ion Concentration; Kinetics; Molecular Sequence Data; Oxygenases; Pseudomonas; Pseudomonas putida; Substrate Specificity; Temperature; Water Microbiology
PubMed: 10049846
DOI: 10.1128/AEM.65.3.946-950.1999 -
Journal of Clinical Microbiology Oct 1984This study evaluated the ability of the Rapid NFT system (API System SA, Montalieu-Vercieu, France) to accurately identify 262 clinically isolated, gram-negative,...
This study evaluated the ability of the Rapid NFT system (API System SA, Montalieu-Vercieu, France) to accurately identify 262 clinically isolated, gram-negative, nonfermentative rods without additional tests. Identifications were classified as correct; low discrimination, with a spectrum of two or more possibilities (additional tests necessary for accurate identification); and incorrect. Correct identification rates were analyzed in two categories: (i) correct to species or biotype for all organism groups except Alcaligenes faecalis-odorans, Moraxella, Pseudomonas testosteroni-alcaligenes-pseudoalcaligenes, and Acinetobacter calcoaceticus biotype haemolyticus-alcaligenes (in this category, the latter four genus-biotype group identifications were taken as correct) and (ii) correct to species or biotype in all cases, including the above four groups. In category i, 87.4% of the strains were correctly identified, with 4.2% low discrimination and 8.4% incorrect. When the criteria of category ii were used, 71.8% of the strains were correctly identified, with 19.9% low discrimination. The Rapid NFT system provided excellent species identification of Pseudomonas and Flavobacterium spp., Bordetella bronchiseptica, and Achromobacter xylosoxidans strains. Within Acinetobacter calcoaceticus, differentiation between biotypes anitratus and lwoffi was satisfactory, but the system did not differentiate between biotypes haemolyticus and alcaligenes. Species resolution within the genera Moraxella and Alcaligenes was incomplete. All Alcaligenes faecalis strains were misidentified and accounted for 50% of misidentifications with the Rapid NFT system; however, these results may reflect taxonomic differences rather than true misidentifications. The Rapid NFT system is easy to inoculate and interpret and represents a worthwhile advance in the identification of gram-negative, nonfermentative rods.
Topics: Bacteria; DNA, Bacterial; Fermentation; Reagent Kits, Diagnostic; Tryptophanase
PubMed: 6490857
DOI: 10.1128/jcm.20.4.730-734.1984 -
Applied and Environmental Microbiology Dec 2005The presence of tetracycline resistance (Tc(r)) genes and class I integrons (in-1), and their ability to cotransfer were investigated in Tc(r) gram-negative (185...
The presence of tetracycline resistance (Tc(r)) genes and class I integrons (in-1), and their ability to cotransfer were investigated in Tc(r) gram-negative (185 strains) and gram-positive (72 strains) bacteria from Danish farmland and pigsties. The isolates belonged to the groups or species Escherichia coli, Enterobacter spp., Arthrobacter spp., Alcaligenes spp., Pseudomonas spp., and Corynebacterium glutamicum. The 257 isolates were screened for in-1. Eighty-one of the gram-negative isolates were also screened for the Tc(r) genes tet(A), tet(B), and tet(C), and all (n = 72) gram-positive isolates were screened for tet(33). Fourteen (7%) of the soil isolates and eleven (25%) of the pigsty isolates contained in-1. All isolates that contained tet genes also contained in-1, except one gram-negative isolate from a pigsty that contained tet(B). All gram-positive isolates with in-1 also contained tet(33). No isolates contained more than one tet gene. The in-1-positive isolates were tested for resistance to selected antimicrobial agents and showed resistance to three to nine drugs. Filter-mating experiments showed cotransfer of Tc(r) and class I integrons from soil isolates to Escherichia coli and/or Pseudomonas putida. We conclude that soil bacteria in close contact to manure or pigsty environment may thus have an important role in horizontal spread of resistance. Use of tetracyclines in food animal production may increase not only Tc(r) but also multidrug resistance (caused by the presence tet genes and in-1) in bacteria.
Topics: Alcaligenes; Animals; Anti-Bacterial Agents; Arthrobacter; DNA Primers; Integrons; Manure; Microbial Sensitivity Tests; Polymerase Chain Reaction; Pseudomonas; Soil Microbiology; Swine; Tetracycline Resistance
PubMed: 16332771
DOI: 10.1128/AEM.71.12.7941-7947.2005 -
BMC Infectious Diseases Aug 2006Purified water for pharmaceutical purposes must be free of microbial contamination and pyrogens. Even with the additional sanitary and disinfecting treatments applied to...
BACKGROUND
Purified water for pharmaceutical purposes must be free of microbial contamination and pyrogens. Even with the additional sanitary and disinfecting treatments applied to the system (sequential operational stages), Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas alcaligenes, Pseudomonas picketti, Flavobacterium aureum, Acinetobacter lowffi and Pseudomonas diminuta were isolated and identified from a thirteen-stage purification system. To evaluate the efficacy of the chemical agents used in the disinfecting process along with those used to adjust chemical characteristics of the system, over the identified bacteria, the kinetic parameter of killing time (D-value) necessary to inactivate 90% of the initial bioburden (decimal reduction time) was experimentally determined.
METHODS
Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas alcaligenes, Pseudomonas picketti, Flavobacterium aureum, Acinetobacter lowffi and Pseudomonas diminuta were called in house (wild) bacteria. Pseudomonas diminuta ATCC 11568, Pseudomonas alcaligenes INCQS , Pseudomonas aeruginosa ATCC 15442, Pseudomonas fluorescens ATCC 3178, Pseudomonas picketti ATCC 5031, Bacillus subtilis ATCC 937 and Escherichia coli ATCC 25922 were used as 'standard' bacteria to evaluate resistance at 25 degrees C against either 0.5% citric acid, 0.5% hydrochloric acid, 70% ethanol, 0.5% sodium bisulfite, 0.4% sodium hydroxide, 0.5% sodium hypochlorite, or a mixture of 2.2% hydrogen peroxide (H2O2) and 0.45% peracetic acid.
RESULTS
The efficacy of the sanitizers varied with concentration and contact time to reduce decimal logarithmic (log10) population (n cycles). To kill 90% of the initial population (or one log10 cycle), the necessary time (D-value) was for P. aeruginosa into: (i) 0.5% citric acid, D = 3.8 min; (ii) 0.5% hydrochloric acid, D = 6.9 min; (iii) 70% ethanol, D = 9.7 min; (iv) 0.5% sodium bisulfite, D = 5.3 min; (v) 0.4% sodium hydroxide, D = 14.2 min; (vi) 0.5% sodium hypochlorite, D = 7.9 min; (vii) mixture of hydrogen peroxide (2.2%) plus peracetic acid (0.45%), D = 5.5 min.
CONCLUSION
The contact time of 180 min of the system with the mixture of H2O2+ peracetic acid, a total theoretical reduction of 6 log10 cycles was attained in the water purified storage tank and distribution loop. The contact time between the water purification system (WPS) and the sanitary agents should be reviewed to reach sufficient bioburden reduction (over 6 log10).
Topics: Disinfectants; Drug Industry; Gram-Negative Bacteria; Water Purification
PubMed: 16914053
DOI: 10.1186/1471-2334-6-131 -
Biotechnology Reports (Amsterdam,... Dec 2021The mechanisms of tolerance to heavy metals used by some microorganisms identified by bioprospection processes are useful for the development and implementation of...
The mechanisms of tolerance to heavy metals used by some microorganisms identified by bioprospection processes are useful for the development and implementation of bioremediation strategies for contaminated environments with high toxic load caused by heavy metals. A total of seven native microbial isolates were obtained from wastewater bodies from an industrial zone in the municipality of Girardota, Antioquia, Colombia. Subsequently, they were selected to evaluate their lead tolerance capacity at different concentrations. In addition, some parameters were determined, such as the capacity to produce exopolysaccharides and their biosorption to understand potential mechanisms associated to lead tolerance. According to the biocehemical test (Vitek) and the molecular analysis of sequences of 16S rDNA, bacterial were identified as , and . We determined that the seven isolates had the capacity to tolerate concentrations higher than 50 mg/ml of lead, and that the concentration and exposure time (40 h) to this metal significantly affect the spp. isolates. Statistically significant differences were detected ( < 0.05) in the production of the exopolysaccharide (EPS) among the isolates. (P16) was the strain with the maximum absorbance exopolysaccharide measured. We evidenced that (P14) and (P20) have 80% capacity to biosorber lead using live mass (minimum range from 80.9% to 87%). It is suggested that the tolerance to lead exhibited by the environmental isolates of spp. can be attributed to the production of exopolysaccharides and biosorption, which are protection factors for its survival in contaminated places. Finally, it was determined that the adsorption measured from dead biomass was significant ( < 0.05) from 40 h of exposure to metal (Average 182.2 ± 7). We generated new knowledge about the potential use of the spp. genus to bioremediate affluent contaminated with heavy metals.
PubMed: 34765463
DOI: 10.1016/j.btre.2021.e00685 -
The Journal of General and Applied... 2012Polyhydroxyalkanoates (PHAs) accumulating bacteria were isolated under various selective conditions such as pH, salt concentrations and types of heavy metal. Fifty...
Polyhydroxyalkanoates (PHAs) accumulating bacteria were isolated under various selective conditions such as pH, salt concentrations and types of heavy metal. Fifty strains of bacterial isolates were found to belong to Bacillus, Proteus, Pseudomonas, Aeromonas, Alcaligenes and Chromobacterium, based on phenotypical features and genotypic investigation. Only twenty five bacterial isolates were selected and observed for the production of PHAs. Interestingly, bacteria belonging to Firmucutes Bacillus sp. produced a high amount of PHAs. The maximum PHAs were accumulated by B. licheniformis PHA 007 at 68.80% of dry cell weight (DCW). Pseudomonas sp., Aeromonas sp., Alcaligenes sp. and Chromobacterium sp. were recorded to produce a moderate amount of PHAs, varying from 10.00-44.32% of DCW. The enzymatic activity was preliminarily analyzed by the ratio of the clear zone diameter to colony diameter. Bacillus gave the highest ratio of hydrolysis zone which corresponds to the highest hydrolytic enzyme activities. Bacillus licheniformis PHA 007 had the highest lipase and protease activity at 2.1 and 5.1, respectively. However, the highest amylase activity was observed in Bacillus sp. PHA 023 at 1.4. Determination of metabolic characteristics was also investigated to check for their ability to consume a wide range of substrates. Bacillus, Aeromonas sp. and Alcaligenes sp. had great ability to utilize a variety of substrates. To decrease high PHA cost, different sources of cheap substrates were tested for the production of PHAs. Bacillus cereus PHA 008 gave the maximal yield of PHA production (64.09% of DCW) when cultivated in anaerobically treated POME. In addition, the accumulation of PHA copolymers such as 3-hydroxyvalerate and 3-hydroxyhexanoate was also observed in Bacillus and Pseudomomas sp. strain 012 and 045, respectively. Eight of the nine isolates accumulated a significant amount of PHAs when inexpensive carbon sources were used as substrates. Here it varied from 1.69% of DCW by B. licheniformis PHA 007 to 64.09% of DCW by B. cereus PHA 008.
Topics: Animals; Bacteria; Bacterial Typing Techniques; DNA, Bacterial; DNA, Ribosomal; Environmental Microbiology; Feces; Food Microbiology; Humans; Molecular Sequence Data; Polyhydroxyalkanoates; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Sewage
PubMed: 22878735
DOI: 10.2323/jgam.58.173 -
PloS One 2024Dysbiotic biliary bacterial profile is reported in cancer patients and is associated with survival and comorbidities, raising the question of its effect on the influence...
BACKGROUND
Dysbiotic biliary bacterial profile is reported in cancer patients and is associated with survival and comorbidities, raising the question of its effect on the influence of anticancer drugs and, recently, the suggestion of perichemotherapy antibiotics in pancreatic cancer patients colonized by the Escherichia coli and Klebsiella pneumoniae.
OBJECTIVE
In this study, we investigated the microbial communities that colonize tumours and which bacteria could aid in diagnosing pancreatic and biliary cancer and managing bile-colonized patients.
METHODS
A retrospective study on positive bile cultures of 145 Italian patients who underwent cholangiopancreatography with PC and EPC cancer hospitalized from January 2006 to December 2020 in a QA-certified academic surgical unit were investigated for aerobic/facultative-anaerobic bacteria and fungal organisms.
RESULTS
We found that among Gram-negative bacteria, Escherichia coli and Pseudomonas spp were the most frequent in the EPC group, while Escherichia coli, Klebsiella spp, and Pseudomonas spp were the most frequent in the PC group. Enterococcus spp was the most frequent Gram-positive bacteria in both groups. Comparing the EPC and PC, we found a significant presence of patients with greater age in the PC compared to the EPC group. Regarding Candida spp, we found no significant but greater rate in the PC group compared to the EPC group (11.7% vs 1.96%). We found that Alcaligenes faecalis was the most frequent bacteria in EPC than the PC group, among Gram-negative bacterial species.
CONCLUSIONS
Age differences in gut microbiota composition may affect biliary habitats in our cancer population, especially in patients with pancreatic cancer. Alcaligenes faecalis isolated in the culture of bile samples could represent potential microbial markers for a restricted follow-up to early diagnosis of extra-pancreatic cancer. Finally, the prevalence of Candida spp in pancreatic cancer seems to trigger new aspects about debate about the role of fungal microbiota into their relationship with pancreatic cancer.
Topics: Humans; Bile; Retrospective Studies; Bacteria; Anti-Bacterial Agents; Gram-Negative Bacteria; Biliary Tract Neoplasms; Candida; Escherichia coli; Pancreatic Neoplasms; Microbial Sensitivity Tests
PubMed: 38381746
DOI: 10.1371/journal.pone.0294049 -
Microbiology (Reading, England) Aug 1997Pseudomonas alcaligenes NCIB 9867 (strain P25X), which grows on 2,5-xylenol and harbours the plasmid RP4, was mated with a plasmid-free derivative of Pseudomonas putida... (Comparative Study)
Comparative Study
Pseudomonas alcaligenes NCIB 9867 (strain P25X), which grows on 2,5-xylenol and harbours the plasmid RP4, was mated with a plasmid-free derivative of Pseudomonas putida NCIB 9869, strain RA713, which cannot grow on 2,5-xylenol. Some RA713 transconjugants, initially selected on 2,5-xylenol, were found to carry RP4 plasmids that had acquired additional fragments (designated Xin) which ranged in size from 2 kb to approximately 26 kb instability of DNA inserts in RP4::Xin hybrid plasmids was observed. The smallest insert present in a stable RP4::Xin6 hybrid plasmid, termed Xin6, yielded multiple bands when it was used as a probe with digested P25X chromosomal DNA. Sequence analysis of Xin6 led to the discovery of an open reading frame with homology to the maturases of group II introns. The Xin6 insert also exhibited several features characteristic of a group II intron. These included the presence of the consensus sequence GUGYG at the 5' and and RAY at the 3' end of the intron. RNA secondary structure modelling of Xin6 also revealed the presence of perfectly conserved domains V and VI. Differences were detected in the Xin6 hybridization profiles of several P25X catabolic mutants that have lost the ability to grow on 2,5-xylenol. In these mutants the loss of 2,5-xylenol degradative ability could be due to genome rearrangements mediated by sequences related to the Xin6 group II intron. This is the first reported group II intron isolated from Pseudomonas spp. and the first time that the mobility of a bacterial group II intron has been demonstrated.
Topics: Amino Acid Sequence; Base Sequence; Biodegradation, Environmental; Chimera; Cloning, Molecular; Computer Simulation; Conjugation, Genetic; Consensus Sequence; Genes, Bacterial; Introns; Models, Molecular; Molecular Sequence Data; Mutagenesis, Insertional; Nucleic Acid Conformation; Plasmids; Pseudomonas; RNA, Bacterial; Recombination, Genetic; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Xylenes
PubMed: 9274037
DOI: 10.1099/00221287-143-8-2833