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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 -
BMC Microbiology Mar 2013The agrichemical 4-aminopyridine is used as a bird repellent in crop fields and has an epileptogenic action in a variety of animals, including man and mouse....
BACKGROUND
The agrichemical 4-aminopyridine is used as a bird repellent in crop fields and has an epileptogenic action in a variety of animals, including man and mouse. 4-Aminopyridine is biodegraded in the environment through an unknown mechanism.
RESULTS
A 4-aminopyridine-degrading enrichment culture utilized 4-aminopyridine as a carbon, nitrogen, and energy source, generating 4-amino-3-hydroxypyridine, 3,4-dihydroxypyridine, and formate as intermediates. 4-Amino-3-hydroxypyridine could not be further metabolized and probably accumulated as a dead-end product in the culture. Biodegradability tests and partial sequence analysis of the enrichment culture indicated that 4-aminopyridine was mainly degraded via 3,4-dihydroxypyridine and that the metabolite is probably cleaved by 3-hydroxy-4-pyridone dioxygenase. Seven culturable predominant bacterial strains (strains 4AP-A to 4AP-G) were isolated on nutrient agar plates. Changes in the bacterial populations of 4-aminopyridine, 3,4-dihydroxypyridine, or formate/ammonium chloride enrichment cultures were monitored by denaturing gradient gel electrophoresis (DGGE) profiling of PCR-amplified 16S rRNA gene fragments. Sequence analysis of the 16S rRNA gene fragments derived from predominant DGGE bands indicated that Pseudomonas nitroreducens 4AP-A and Enterobacter sp. 4AP-G were predominant in the three tested enrichment cultures and that the unculturable strains Hyphomicrobium sp. 4AP-Y and Elizabethkingia sp. 4AP-Z were predominant in 4-aminopyridine and formate/ammonium chloride enrichment cultures and in the 3,4-dihydroxypyridine enrichment culture, respectively. Among the culturable strains, strain 4AP-A could utilize 3,4-dihydroxypyridine as a growth substrate. Although we could not isolate strain 4AP-Y on several media, PCR-DGGE analysis and microscopy indicated that the unique bi-polar filamentous bacterial cells gradually became more dominant with increasing 4-aminopyridine concentration in the medium.
CONCLUSIONS
Hyphomicrobium sp. 4AP-Y, P. nitroreducens 4AP-A, and Elizabethkingia sp. 4AP-Z probably play important roles in 4-aminopyridine degradation in crop fields. In the enrichment culture, 3,4-dihydroxypyridine and its metabolites including formate might be shared as growth substrates and maintain the enrichment culture, including these indispensable strains.
Topics: 4-Aminopyridine; Bacteria; Biota; Biotransformation; DNA, Bacterial; DNA, Ribosomal; Environmental Pollutants; Molecular Sequence Data; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Soil Microbiology
PubMed: 23517195
DOI: 10.1186/1471-2180-13-62 -
Journal of Applied Microbiology May 2010To isolate and characterize microbes in the soils containing high contents of phenolics and to dissolve the allelopathic inhibition of plants through microbial...
AIMS
To isolate and characterize microbes in the soils containing high contents of phenolics and to dissolve the allelopathic inhibition of plants through microbial degradation.
METHODS AND RESULTS
Four microbes were isolated from plant soils using a screening medium containing p-coumaric acid as sole carbon source. The isolates were identified by biochemical analysis and sequences of their 16S or 18S rDNA, and designated as Pseudomonas putida 4CD1 from rice (Oryza sativa) soil, Ps. putida 4CD3 from pine (Pinus massoniana) soil, Pseudomonas nitroreducens 4CD2 and Rhodotorula glutinis 4CD4 from bamboo (Bambusa chungii) soil. All isolates degraded 1 g l(-1) of p-coumaric acid by 70-93% in inorganic and by 99% in Luria-Bertani solutions within 48 h. They also effectively degraded ferulic acid, p-hydroxybenzoic acid and p-hydroxybenzaldehyde. The microbes can degrade p-coumaric acid and reverse its inhibition on seed germination and seedling growth in culture solutions and soils. Low pHs inhibited the growth and phenolic degradation of the three bacteria. High temperature inhibited the R. glutinis. Co(2+) completely inhibited the three bacteria, but not the R. glutinis. Cu(2+), Al(3+), Zn(2+), Fe(3+), Mn(2+), Mg(2+) and Ca(2+) had varying degrees of inhibition for each of the bacteria.
CONCLUSIONS
Phenolics in plant culture solutions and soils can be decomposed through application of soil microbes in laboratory or controlled conditions. However, modification of growth conditions is more important for acidic and ions-contaminated media.
SIGNIFICANCE AND IMPACT OF THE STUDY
The four microbes were first isolated and characterized from the soils of bamboo, rice or pine. This study provides some evidence and methods for microbial control of phenolic allelochemicals.
Topics: Coumaric Acids; Germination; Hydrogen-Ion Concentration; Metals; Oryza; Phenols; Pheromones; Pseudomonas; RNA, Ribosomal, 16S; RNA, Ribosomal, 18S; Rhodotorula; Sasa; Soil Microbiology; Temperature
PubMed: 19912433
DOI: 10.1111/j.1365-2672.2009.04589.x -
International Journal of Molecular... Nov 2022The B12-producing strains DSM 1650 and sp. CCUG 2519 (both formerly ), with the most distributed pathway among bacteria for exogenous choline/betaine utilization, are...
The B12-producing strains DSM 1650 and sp. CCUG 2519 (both formerly ), with the most distributed pathway among bacteria for exogenous choline/betaine utilization, are promising recombinant hosts for the endogenous production of B12 precursor betaine by direct methylation of bioavailable glycine or non-proteinogenic -alanine. Two plasmid-based de novo betaine pathways, distinguished by their enzymes, have provided an expression of the genes encoding for -methyltransferases of the halotolerant cyanobacterium or plant to synthesize the internal glycine betaine or -alanine betaine, respectively. These betaines equally allowed the recombinant pseudomonads to grow effectively and to synthesize a high level of cobalamin, as well as to increase their protective properties against abiotic stresses to a degree comparable with the supplementation of an exogenous betaine. Both de novo betaine pathways significantly enforced the protection of bacterial cells against lowering temperature to 15 °C and increasing salinity to 400 mM of NaCl. However, the expression of the single plant-derived gene for the -alanine-specific -methyltransferase additionally increased the effectiveness of exogenous glycine betaine almost twofold on cobalamin biosynthesis, probably due to the ' ability to use two independent pathways, their own choline/betaine pathway and the plant -alanine betaine biosynthetic pathway.
Topics: Betaine; Choline; Pseudomonas; Stress, Physiological; Methyltransferases; beta-Alanine; Vitamin B 12
PubMed: 36430408
DOI: 10.3390/ijms232213934 -
Heliyon Jun 2023Polystyrene is a plastic that leads to environmental pollution. In particular, expanded polystyrene is very light and takes up much space, causing additional...
OBJECTIVES
Polystyrene is a plastic that leads to environmental pollution. In particular, expanded polystyrene is very light and takes up much space, causing additional environmental problems. The aim of this study was to isolate new symbiotic bacteria which degraded polystyrene from mealworms.
METHODS
The population of polystyrene degrading bacteria was increased by enrichment culture of intestinal bacteria from mealworms with polystyrene as a sole carbon source. The degradation activity of isolated bacteria was evaluated by morphological change of micro-polystyrene particles and the surface change of polystyrene films.
RESULTS
Eight isolated species (, , , , , , , and ) were identified that degrade polystyrene.
CONCLUSION
Bacterial identification shows that a broad spectrum of bacteria decomposing polystyrene coexists in the intestinal tract of mealworms.
PubMed: 37426801
DOI: 10.1016/j.heliyon.2023.e17352 -
Bioscience, Biotechnology, and... 2012Vanillin is one of the most valuable compounds in the flavoring and fragrance industries, and many attempts to produce natural vanillin have been made in recent years....
Vanillin is one of the most valuable compounds in the flavoring and fragrance industries, and many attempts to produce natural vanillin have been made in recent years. Isoeugenol monooxygenase (Iem) converts the phenylpropanoid compound isoeugenol to vanillin. In Pseudomonas nitroreducens Jin1, the positive regulatory protein IemR is divergently expressed from Iem, and the promoter region is located between the genes. In this study, we investigated the transcriptional regulation of iem in Escherichia coli. We focused on inducers and regulatory protein IemR. Transcription of iem was found to be dependent on the amounts of isoeugenol and IemR. Isoeugenol was found to be the best inducer of iem, followed by trans-anethole, which induced iem to 58% of the transcription level observed for isoeugenol. Overproduction of IemR in E. coli significantly increased the transcription of iem, up to 96-fold, even in the absence of isoeugenol, as compared to basally expressed IemR. Results of this study indicate that the transcription of iem iss dependent on the type of inducers and on IemR. They should contribute to the development of bioengineering strategies for increased production of vanillin through high-level expression of the isoeugenol monooxygenase gene in microorganisms.
Topics: Bacterial Proteins; Base Sequence; Escherichia coli; Eugenol; Gene Expression Regulation, Bacterial; Mixed Function Oxygenases; Molecular Sequence Data; Pseudomonas; Transcription, Genetic
PubMed: 23047101
DOI: 10.1271/bbb.120375 -
AoB PLANTS Apr 2019Gametophytes of the fern develop into either hermaphrodites or males. As hermaphrodites develop, they secrete antheridiogen, or A, into the environment, inducing male...
Gametophytes of the fern develop into either hermaphrodites or males. As hermaphrodites develop, they secrete antheridiogen, or A, into the environment, inducing male development in undifferentiated gametophytes. Hermaphrodites are composed of archegonia, antheridia, rhizoids and a notch meristem, while males consist of antheridia and rhizoids. Much of the research on sexual and morphological development concerns gametophytes grown in sterile environments. Using biochemical and molecular techniques we identify a soil bacterium and explore its effects on sexual and rhizoid development. Hermaphrodite and male gametophytes were exposed to this bacterium and the effects on sexual development, rhizoid length and rhizoid number were explored. The bacterium was identified as a pseudomonad, . Gametophytes grown in the presence of the pseudomonad were more likely to develop into hermaphrodites across all gametophyte densities. Across all gametophyte sizes, hermaphrodites had rhizoids that were 2.95× longer in the presence of the pseudomonad while males had rhizoids that were 2.72× longer in the presence of the pseudomonad. Both hermaphrodite and male gametophytes developed fewer rhizoids in the presence of the pseudomonad. Control hermaphrodites produced 1.23× more rhizoids across all gametophyte sizes. For male gametophytes grown in the absence of the pseudomonad, the rate of increase in the number of rhizoids was greater with increasing size in the control than the rate of increase in males grown in the presence of the pseudomonad. The pseudomonad may be acting on gametophyte sexual development via several potential mechanisms: degradation of A, changes in nutrient availability or phytohormone production. The pseudomonad may also increase rhizoid number through production of phytohormones or changes in nutrient availability.
PubMed: 31019671
DOI: 10.1093/aobpla/plz012 -
Journal of Inorganic Biochemistry Nov 2018A strategy for elucidating sequence determinants of function in the class of cytochrome P450 (CYP) enzymes that catalyze the first steps of terpene metabolism in wild...
A strategy for elucidating sequence determinants of function in the class of cytochrome P450 (CYP) enzymes that catalyze the first steps of terpene metabolism in wild microbiomes is described. Wild organisms that can use camphor, terpineol, pinene and limonene were isolated from soils rich in coniferous waste. Cell free extracts and growth beers were analyzed by gas chromatography/mass spectrometry to identify primary oxidative metabolites. For one organism, Pseudomonas nitroreducens TPJM, a cytochrome P450 (CYP108B1) isolated from cell free extracts was demonstrated to catalyze the oxidation of α-terpineol in assays combining the native ferredoxin and putidaredoxin reductase, and the resulting oxidation products identified by gas chromatography/mass spectrometry. Shotgun sequencing of PnTPJM identified four candidate P450 genes, including an apparently fragmentary gene with a high degree of homology with the known enzyme CYP108 (P450).
Topics: Bacterial Proteins; Cytochrome P-450 Enzyme System; Pseudomonas; Soil Microbiology; Terpenes
PubMed: 30170307
DOI: 10.1016/j.jinorgbio.2018.08.006 -
Scientific Reports May 2019Bacteria in the environment play a major role in the degradation of widely used man-made recalcitrant organic compounds. Pseudomonas nitroreducens TX1 is of special...
Bacteria in the environment play a major role in the degradation of widely used man-made recalcitrant organic compounds. Pseudomonas nitroreducens TX1 is of special interest because of its high efficiency to remove nonionic ethoxylated surfactants. In this study, a novel approach was demonstrated by a bacterial enzyme involved in the formation of radicals to attack ethoxylated surfactants. The dihydrolipoamide dehydrogenase was purified from the crude extract of strain TX1 by using octylphenol polyethoxylate (OPEO) as substrate. The extent of removal of OPEOs during the degradation process was conducted by purified recombinant enzyme from E. coli BL21 (DE3) in the presence of the excess of metal mixtures (Mn, Mg, Zn, and Cu). The metabolites and the degradation rates were analyzed and determined by liquid chromatography-mass spectrometry. The enzyme was demonstrated to form Fenton reagent in the presence of an excess of metals. Under this in vitro condition, it was shown to be able to shorten the ethoxylate chains of OPEO. After 2 hours of reaction, the products obtained from the degradation experiment revealed a prominent ion peak at m/z = 493.3, namely the ethoxylate chain unit is 6 (OPEO) compared to OPEO (m/z = 625.3), the main undegraded surfactant in the no enzyme control. It revealed that the concentration of OPEO and OPEO decreased by 90% and 40% after 4 hours, respectively. The disappearance rates for the OPEO homologs correlated to the length of the exothylate chains, suggesting it is not a specific enzymatic reaction which cleaves one unit by unit from the end of the ethoxylate chain. The results indicate the diverse and novel strategy by bacteria to catabolize organic compounds by using existing housekeeping enzyme(s).
Topics: Bacterial Proteins; Chromatography, Liquid; Copper; Dihydrolipoamide Dehydrogenase; Escherichia coli; Magnesium; Manganese; Mass Spectrometry; Phenols; Pseudomonas; Surface-Active Agents; Zinc
PubMed: 31048711
DOI: 10.1038/s41598-019-43266-8 -
Bioscience, Biotechnology, and... 2013The isoeugenol monooxygenase (iem) gene from Pseudomonas nitroreducens Jin1, responsible for the conversion of isoeugenol to vanillin, was cloned and overexpressed in...
The isoeugenol monooxygenase (iem) gene from Pseudomonas nitroreducens Jin1, responsible for the conversion of isoeugenol to vanillin, was cloned and overexpressed in Escherichia coli. The purified Iem had a predicted molecular mass of 54 kDa. The V(max), K(M), and k(cat) values for it, using isoeugenol as substrate, were 4.2 µmol vanillin min(-1) mg(-1) of protein, 120 µM, and 3.8 s(-1), respectively. Maximum substrate turnover for Iem occurred at 30 °C and pH 9.0. An (18)Oxygen-labeling experiment revealed that oxidative cleavage of isoeugenol by Iem was catalyzed via a monooxygenation reaction, and that incorporation of the oxygen atom from O(2) into vanillin was preferable to incorporation from water. While the catalytic activity of Iem, as prepared, did not require the addition of any organic or metal cofactor, ICP-MS analysis showed 0.7 mol of iron per mol of Iem. Moreover site-directed mutagenesis of Iem of four conserved histidine residues individually, His(167), His(218), His(282), and His(471), which appear to be involved in ligand bonding with Fe(2+), resulted in a loss of activity. Enzyme activity was not appreciably influenced by preincubation of Iem with high concentrations of chelators, suggesting that iron is tightly bound. Iem has an iron-mediated mechanism that is widely spread among the three domains of life.
Topics: Bacterial Proteins; Benzaldehydes; Biotransformation; Cloning, Molecular; Escherichia coli; Eugenol; Gene Expression; Histidine; Hydrogen-Ion Concentration; Iron; Iron Chelating Agents; Kinetics; Mixed Function Oxygenases; Molecular Weight; Mutagenesis, Site-Directed; Oxygen; Oxygen Isotopes; Pseudomonas; Recombinant Proteins
PubMed: 23391906
DOI: 10.1271/bbb.120715