RESUMO
BACKGROUND: Bioaugmentation is considered a sustainable and cost-effective methodology to recover contaminated environments, but its outcome is highly variable. Predation is a key top-down control mechanism affecting inoculum establishment, however, its effects on this process have received little attention. This study focused on the impact of trophic interactions on bioaugmentation success in two soils with different pollution exposure histories. We inoculated a 13C-labelled pollutant-degrading consortium in these soils and tracked the fate of the labelled biomass through stable isotope probing (SIP) of DNA. We identified active bacterial and eukaryotic inoculum-biomass consumers through amplicon sequencing of 16S rRNA and 18S rRNA genes coupled to a novel enrichment factor calculation. RESULTS: Inoculation effectively increased PAH removal in the short-term, but not in the long-term polluted soil. A decrease in the relative abundance of the inoculated genera was observed already on day 15 in the long-term polluted soil, while growth of these genera was observed in the short-term polluted soil, indicating establishment of the inoculum. In both soils, eukaryotic genera dominated as early incorporators of 13C-labelled biomass, while bacteria incorporated the labelled biomass at the end of the incubation period, probably through cross-feeding. We also found different successional patterns between the two soils. In the short-term polluted soil, Cercozoa and Fungi genera predominated as early incorporators, whereas Ciliophora, Ochrophyta and Amoebozoa were the predominant genera in the long-term polluted soil. CONCLUSION: Our results showed differences in the inoculum establishment and predator community responses, affecting bioaugmentation efficiency. This highlights the need to further study predation effects on inoculum survival to increase the applicability of inoculation-based technologies. Video Abstract.
Assuntos
Bactérias , Biodegradação Ambiental , RNA Ribossômico 16S , RNA Ribossômico 18S , Microbiologia do Solo , Poluentes do Solo , Solo , Solo/química , Poluentes do Solo/metabolismo , RNA Ribossômico 16S/genética , RNA Ribossômico 18S/genética , Bactérias/metabolismo , Bactérias/classificação , Bactérias/genética , Biomassa , Isótopos de Carbono/metabolismo , Cadeia Alimentar , Hidrocarbonetos Policíclicos Aromáticos/metabolismo , Marcação por IsótopoRESUMO
Ocean oil pollution has a large impact on the environment and the health of living organisms. Bioremediation cleaning strategies are promising eco-friendly alternatives for tackling this problem. Previously, we designed and reported a hydrocarbon (HC) degrading microbial consortium of four marine strains belonging to the species Alloalcanivorax xenomutans, Halopseudomonas aestusnigri, Paenarthrobacter sp., and Pseudomonas aeruginosa. However, the knowledge about the metabolic potential of this bacterial consortium for HC bioremediation is not yet well understood. Here, we analyzed the complete genomes of these marine bacterial strains accompanied by a phylogenetic reconstruction along with 138 bacterial strains. Synteny between complete genomes of the same species or genus, revealed high conservation among strains of the same species, covering over 91% of their genomic sequences. Functional predictions highlighted a high abundance of genes related to HC degradation, which may result in functional redundancy within the consortium; however, unique and complete gene clusters linked to aromatic degradation were found in the four genomes, suggesting substrate specialization. Pangenome gain and loss analysis of genes involved in HC degradation provided insights into the evolutionary history of these capabilities, shedding light on the acquisition and loss of relevant genes related to alkane and aromatic degradation. Our work, including comparative genomic analyses, identification of secondary metabolites, and prediction of HC-degrading genes, enhances our understanding of the functional diversity and ecological roles of these marine bacteria in crude oil-contaminated marine environments and contributes to the applied knowledge of bioremediation.
Assuntos
Biodegradação Ambiental , Genoma Bacteriano , Genômica , Hidrocarbonetos , Filogenia , Hidrocarbonetos/metabolismo , Genômica/métodos , Consórcios Microbianos/genética , Bactérias/genética , Bactérias/metabolismo , Bactérias/classificação , Água do Mar/microbiologiaRESUMO
In vitro plant cultures are able to remove and metabolise xenobiotics, making them promising tools for decontamination strategies. In this work, we evaluated Brassica napus hairy roots (HRs) to tolerate and remove high concentrations of the azo dye Naphthol Blue-Black (NBB). Experiments were performed using both growing and resting culture systems at different pHs. Reuse of HRs biomass was evaluated in successive decolourisation cycles. Proteomics was applied to understand the molecular responses likely to be involved in the tolerance and removal of NBB. The HRs tolerated up to 480 µg mL-1 NBB, and 100 % removal was achieved at 180 µg mL-1 NBB after 10 days using both culture systems. Interestingly, the HRs are robust enough to be reused, showing 55-60 % removal even after three reuse cycles. The highest dye removal rates were achieved during the first 2 days of incubation, as initial removal is mainly driven by passive processes. Active mechanisms are triggered later by regulating the expression of proteins with different biological functions, mainly those related to xenobiotic metabolism, such as hydrolytic and redox enzymes. These results suggest that B. napus HRs are a robust tool that could make a significant contribution to textile wastewater treatment.
Assuntos
Biodegradação Ambiental , Brassica napus , Raízes de Plantas , Proteômica , Brassica napus/metabolismo , Raízes de Plantas/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Corantes/metabolismo , Corantes/química , Compostos Azo/metabolismo , Compostos Azo/química , Poluentes Químicos da Água/metabolismoRESUMO
Bacteria can synthesize a broad spectrum of multifunctional polysaccharides including extracellular polysaccharides (EPS). Bacterial EPS can be utilized in the food, pharmaceutical, and biomedical areas owing to their physical and rheological properties in addition to generally presenting low toxicity. From an ecological viewpoint, EPS are biodegradable and environment compatible, offering several advantages over synthetic compounds. This study investigated the EPS produced by Klebsiella oxytoca (KO-EPS) by chemically characterizing and evaluating its properties. The monosaccharide components of the KO-EPS were determined by HPLC coupled with a refractive index detector and GC-MS. The KO-EPS was then analyzed by methylation analysis, FT-IR and NMR spectroscopy to give a potential primary structure. KO-EPS demonstrated the ability to stabilize hydrophilic emulsions with various hydrophobic compounds, including hydrocarbons and vegetable and mineral oils. In terms of iron chelation capacity, the KO-EPS could sequester 41.9 % and 34.1 % of the most common iron states, Fe2+ and Fe3+, respectively. Moreover, KO-EPS exhibited an improvement in the viscosity of aqueous dispersion, being proportional to the increase in its concentration and presenting a non-Newtonian pseudoplastic flow behavior. KO-EPS also did not present a cytotoxic effect indicating that the KO-EPS could have potential applications as a natural thickener, bioemulsifier, and bioremediation agent.
Assuntos
Biodegradação Ambiental , Emulsões , Klebsiella oxytoca , Polissacarídeos Bacterianos , Reologia , Klebsiella oxytoca/metabolismo , Polissacarídeos Bacterianos/química , Polissacarídeos Bacterianos/biossíntese , Emulsificantes/química , Emulsificantes/metabolismo , Biotecnologia/métodos , Viscosidade , Interações Hidrofóbicas e HidrofílicasRESUMO
Cildáñez stream (in Matanza-Riachuelo basin, Buenos Aires) is one of the most polluted watercourses of Argentina, containing a mixed contamination from agricultural and industrial wastes. The application of water bioremediation processes for this kind of effluent will require microorganisms with a high tolerance to contamination. In this sense, obtaining higher contaminant-resistant microalgae lines is widely desired. In this study, adaptive laboratory evolution (ALE) and random mutagenesis were used to obtain Chlorella vulgaris LMPA-40 strains adapted to grow in polluted water from the Cildáñez stream. The ALE process was performed by 22 successive subcultures under selective pressure (Cildáñez wastewater alone or with the addition of phenol or H2O2) while random mutagenesis was performed with UV-C radiation at 275nm. Not all the cell lines obtained after ALE could adapt enough to overcome the stress caused by the Cildáñez wastewater, indicating that the process is quite random and depends on the stressor used. The best results were obtained for the Cildáñez wastewater adapted cells (Cild 3 strain) that were more resistant than the original strain. The concentration of protein, Chlorophyll A, Chlorophyll B, and carotenoids in the Cild 3 ALE evolved strain was higher than that of the control strain. However, this strain exhibited half of the lipid content compared to the same control strain. Interestingly, these alterations and the acquired tolerance may be reversed over time during storage. These findings suggest that the acquisition of novel cell lines could not be permanent, a fact that must be considered for future trials.
Assuntos
Chlorella vulgaris , Chlorella vulgaris/genética , Águas Residuárias/microbiologia , Argentina , Biodegradação Ambiental , Evolução Molecular Direcionada , Mutagênese , Clorofila A , Clorofila/análise , Peróxido de Hidrogênio/farmacologiaRESUMO
The enzyme PETase fromIdeonella sakaiensis (IsPETase) strain 201-F6 can catalyze the hydrolysis of polyethylene terephthalate (PET), mainly converting it into mono(2-hydroxyethyl) terephthalic acid (MHET). In this study, we used quantum mechanics/molecular mechanics (QM/MM) simulations to explore the molecular details of the catalytic reaction mechanism of IsPETase in the formation of MHET. The QM region was described with AM1d/PhoT and M06-2X/6-31+G(d,p) potential. QM/MM simulations unveil the complete enzymatic PET hydrolysis mechanism and identify two possible reaction pathways for acylation and deacylation steps. The barrier obtained at M06-2X/6-31+G(d,p)/MM potential for the deacylation step corresponds to 20.4 kcal/mol, aligning with the experimental value of 18 kcal/mol. Our findings indicate that deacylation is the rate-limiting step of the process. Furthermore, per-residue interaction energy contributions revealed unfavorable contributions to the transition state of amino acids located at positions 200-230, suggesting potential sites for targeted mutations. These results can contribute to the development of more active and selective enzymes for PET depolymerization.
Assuntos
Polietilenotereftalatos , Teoria Quântica , Polietilenotereftalatos/química , Polietilenotereftalatos/metabolismo , Simulação de Dinâmica Molecular , Burkholderiales/enzimologia , Burkholderiales/metabolismo , Hidrólise , Biodegradação Ambiental , Biocatálise , AcilaçãoRESUMO
Environmental contamination by chromium represents a serious public health problem. Therefore, it is crucial to develop and optimize remediation technologies to reduce its concentration in the environment. The aims of this study were to evaluate the uptake of chromium by live and complete microbial mats in experimental mesocosms under different pH and salinity conditions to understand how these factors affect the microphytobenthic community and, consequently, how chromium removal process is influenced. Microbial mats from the estuarine environment were exposed to 15 mg Cr/L under different pH (2, 4, and 8) and salinity (2, 15, and 33) conditions. Salinity, redox potential, and pH were measured throughout the trial in solutions and in microbial mats, while total Cr determinations were performed at the end of the assay. The results demonstrated that the removal efficiency of Cr by microbial mats was significantly improved in solutions at pH 2, remaining unaffected by variations in salinity. Notably, both cyanobacteria and diatoms showed remarkable resistance to Cr exposure under all conditions tested, highlighting their exceptional adaptability. Microbial mats have proved to be effective filters for reducing the concentration of chromium in aqueous solutions with varying pH and salinity levels.
Assuntos
Cromo , Salinidade , Poluentes Químicos da Água , Cromo/análise , Concentração de Íons de Hidrogênio , Poluentes Químicos da Água/análise , Cianobactérias , Diatomáceas , Biodegradação AmbientalRESUMO
Plastics derived from fossil fuels are used ubiquitously owing to their exceptional physicochemical characteristics. However, the extensive and short-term use of plastics has caused environmental challenges. The biotechnological plastic conversion can help address the challenges related to plastic pollution, offering sustainable alternatives that can operate using bioeconomic concepts and promote socioeconomic benefits. In this context, using soil from a plastic-contaminated landfill, two consortia were established (ConsPlastic-A and -B) displaying versatility in developing and consuming polyethylene or polyethylene terephthalate as the carbon source of nutrition. The ConsPlastic-A and -B metagenomic sequencing, taxonomic profiling, and the reconstruction of 79 draft bacterial genomes significantly expanded the knowledge of plastic-degrading microorganisms and enzymes, disclosing novel taxonomic groups associated with polymer degradation. The microbial consortium was utilized to obtain a novel Pseudomonas putida strain (BR4), presenting a striking metabolic arsenal for aromatic compound degradation and assimilation, confirmed by genomic analyses. The BR4 displays the inherent capacity to degrade polyethylene terephthalate (PET) and produce polyhydroxybutyrate (PHB) containing hydroxyvalerate (HV) units that contribute to enhanced copolymer properties, such as increased flexibility and resistance to breakage, compared with pure PHB. Therefore, BR4 is a promising strain for developing a bioconsolidated plastic depolymerization and upcycling process. Collectively, our study provides insights that may extend beyond the artificial ecosystems established during our experiments and supports future strategies for effectively decomposing and valorizing plastic waste. Furthermore, the functional genomic analysis described herein serves as a valuable guide for elucidating the genetic potential of microbial communities and microorganisms in plastic deconstruction and upcycling.
Assuntos
Biodegradação Ambiental , Microbiota , Plásticos , Plásticos/metabolismo , Microbiologia do Solo , Polietilenotereftalatos/metabolismo , Poluentes do Solo/metabolismo , Polímeros/metabolismo , Bactérias/metabolismo , Bactérias/genética , Plásticos Biodegradáveis/metabolismo , Consórcios Microbianos , Pseudomonas putida/metabolismo , Pseudomonas putida/genéticaRESUMO
The present study explored the potential of leaf litter as a source of fungi able to produce ligninolytic enzymes for the biodegradation of anthraquinone dyes. Within the colonies isolated from the leaf litter, only three colonies of two species Trametes were selected based on the detection of oxidation and decolorization halos in Petri dishes with PDA (potato-dextrose-agar) + Guaicol and PDA + RBBR (Remazol Brilliant Blue R). The identification of the colonies was done through sequencing of the ITS region. The enzymatic activity of Lac (lacase), MnP (manganês peroxidase) and LiP (lignina peroxidase) was analyzed by spectrophotometry during fermentation in PD+RBBR imedium. Isolates A1SSI01 and A1SSI02 were identified as Trametes flavida, while A5SS01 was identified as Trametes sp. Laccase showed the highest enzymatic activity, reaching 452.13 IU.L-1 (A1SSI01, 0.05% RBBR) after 96h. Isolate A1SSI02 reached the highest percentage of decolorization, achieving 89.28% in seven days. The results imply that these Trametes isolates can be highly effective in waste treatment systems containing toxic anthraquinone dyes. Keywords: laccase, peroxidases, basidiomycete, litter and biodecolorization.
Assuntos
Biodegradação Ambiental , Lacase , Peroxidases , Folhas de Planta , Trametes , Folhas de Planta/química , Folhas de Planta/microbiologia , Trametes/enzimologia , Peroxidases/metabolismo , Lacase/metabolismo , Florestas , Antraquinonas/metabolismo , Corantes , Lignina/metabolismo , BrasilRESUMO
Bioemulsifiers are compounds produced by microorganisms that reduce the interfacial forces between hydrophobic substances and water. Due to their potential in the pharmaceutical and food industries and their efficiency in oil spill remediation, they have been the subject of study in the scientific community while being safe, biodegradable, and sustainable compared to synthetic options. These biomolecules have high molecular weight and polymeric structures, distinguishing them from traditional biosurfactants. Emulsan, a bioemulsifier exopolysaccharide, is produced by Acinetobacter strains and is highly efficient in forming stable emulsions. Its low toxicity and high potential as an emulsifying agent promote its application in pharmaceutical and food industries as a drug-delivery vehicle and emulsion stabilizer. Due to the high environmental impact of oil spills, bioemulsifiers have great potential for environmental applications, such as bioremediation. This unique feature gives them a distinct mechanism of action in forming emulsions, resulting in minimal environmental impact. A better understanding of these aspects can improve the use of bioemulsifiers and environmental remediation in various industries. This review will discuss the production and characterization of Emulsan, focusing on recent advancements in cultivation conditions, purification techniques, compound identification, and ecotoxicity.