Search Results (23)

Volatile organic compound pollution caused by toluene has become a global issue. In order to solve this problem, biodegradation of toluene has been applied all over the world. This study investigated the effects of Fe³⁺ on toluene degradation by the Rhodococcus sp. TG-1. The results show that 1 mg L⁻¹ Fe³⁺ increased the degradation rate of 600 mg L⁻¹ toluene from 61.9% to 87.2% at 16 h. The acceleration mechanism of Fe³⁺ was explicated using transmission electron microscope (TEM) and energy-dispersive X-ray spectroscopy (EDX) analyses, coupled plasma optical emission spectroscopy, an enzyme activity assay, and transcriptome analysis. Four genes were detected to be significantly up-regulated under Fe³⁺ induction, suggesting that Fe³⁺ might be implicated in toluene degradation. Meanwhile, Fe³⁺ was a component of the active center of catechol 1,2-dioxygenase (C12O) and significantly improved the enzyme activity of C12O. The mechanism by which Fe³⁺ accelerates toluene degradation was proposed based on the transcription levels of degradation genes and the enzyme activity of C12O. This study provided an improved method for enhancing the degradation effect of toluene and furthered our comprehension of the mechanism of toluene degradation. Full article

Phenolic compounds are an extensive group of natural and anthropogenic organic substances of the aromatic series containing one or more hydroxyl groups. The main sources of phenols entering the environment are waste from metallurgy and coke plants, enterprises of the leather, furniture, and pulp and paper industries, as well as wastewater from the production of phenol–formaldehyde resins, adhesives, plastics, and pesticides. Among this group of compounds, phenol is the most common environmental pollutant. One of the cheapest and most effective ways to combat phenol pollution is biological purification. However, the inability of bacteria to decompose high concentrations of phenol is a significant limitation. Due to the uncoupling of oxidative phosphorylation, phenol concentrations above 1 g/L are toxic and inhibit cell growth. This article presents data on the biodegradative potential of Rhodococcus opacus strain 3D. This strain is capable of decomposing a wide range of toxicants, including phenol. In the present study, cell growth with phenol, growth after rest, growth of immobilized cells before and after rest, phase contrast, and scanning microscopy of immobilized cells on fiber were studied in detail. The free-living and immobilized cells can decompose phenol concentrations up to 1.5 g/L and 2.5 g/L, respectively. The decomposition of the toxicant was catalyzed by the enzymes catechol 1,2-dioxygenase and cis,cis-muconate cycloisomerase. The role of protocatechuate 3,4-dioxygenase in biodegradative processes is discussed. In this work, it is shown that the immobilized cells can be stored for a long time (up to 2 years) without significant loss of their degradation activity. An assessment of the induction of genes potentially involved in this process was taken. Based on our investigation, we can conclude that this strain can be considered an effective destructor that is capable of degrading phenol at high concentrations, increases its biodegradative potential during immobilization, and retains this ability for a long storage time. Therefore, the strain can be used in biotechnology for the purification of aqueous samples at high concentrations from phenolic contamination. Full article

Plant growth-promoting bacteria (PGPB) play a role in stimulating plant growth through mechanisms such as the synthesis of the phytohormone indole-3-acetic acid (IAA). The aims of this study were the characterization of IAA synthesis and degradation by the model aromatic-degrading bacterium Paraburkholderia xenovorans LB400, and its growth promotion of the Nicotiana tabacum plant. Strain LB400 was able to synthesize IAA (measured by HPLC) during growth in the presence of tryptophan and at least one additional carbon source; synthesis of anthranilic acid was also observed. RT-PCR analysis indicates that under these conditions, strain LB400 expressed the ipdC gene, which encodes indole-3-pyruvate decarboxylase, suggesting that IAA biosynthesis proceeds through the indole-3-pyruvate pathway. In addition, strain LB400 degraded IAA and grew on IAA as a sole carbon and energy source. Strain LB400 expressed the iacC and catA genes, which encode the α subunit of the aromatic-ring-hydroxylating dioxygenase in the IAA catabolic pathway and the catechol 1,2-dioxygenase, respectively, which may suggest a peripheral IAA pathway leading to the central catechol pathway. Notably, P. xenovorans LB400 promoted the growth of tobacco seedlings, increasing the number and the length of the roots. In conclusion, this study indicates that the versatile bacterium P. xenovorans LB400 is a PGPB. Full article

In recent years, pharmaceuticals have emerged as pollutants due to their incomplete degradation in sewage treatment plants and their ability to cause physiological problems in humans even at low doses. Understanding the environmental fate of pharmaceutical pollutants and the mechanisms involved in their degradation is crucial for developing strategies to mitigate their impact on ecosystems and human health. In this study, the degradation of pharmaceutical compound ibuprofen was achieved by employing two bacterial strains, Achromobacter spanius strain S11 and Achromobacter piechaudii S18, previously isolated from contaminated water. These strains were capable of degrading ibuprofen as their sole carbon source. The study aimed to identify intermediate metabolites, determine the enzymes involved, and detect specific genes related to ibuprofen degradation. Different concentrations of ibuprofen, temperatures, and pH levels were tested. Both A. spanius S11 and A. piechaudii S18 successfully degraded ibuprofen. A. spanius S11 showed a degradation efficiency of 91.18% after only 72 h and reached 95.7% after 144 h, while A. piechaudii S18 exhibited degradation efficiencies of 72.39% and 73.01% after three and seven days, respectively. The LC-MS technique was used to identify biodegradation metabolites produced by A. spanius S11. The results indicated that the first step was hydroxylation followed by oxidation via the combination of monooxygenases that catalyze the C-H hydroxylation and dehydrogenases. Furthermore, the detection of intermediate metabolites of trihydroxyibuprofen suggested that the biodegradation of ibuprofen by A. spanius S11 can occur through multiple mechanisms. The highest enzyme activities were recorded for catechol 1,2-dioxygenase, 4.230 ± 0.026 U/mg, followed by laccase, 2.001 ± 0.215 U/mg. This study demonstrates the potential of Achromobacter strains, particularly A. spanius (S11), in degrading ibuprofen. These findings provide insights into the ibuprofen degradation process, intermediate metabolites, and relevant genes. Full article

During adaptation to waters that are rich in xenobiotics, biological systems pass through multiple stages. The first one is related to the restructuring of communities, pronounced destruction of the structure, and multiplication of active biodegradants. The purpose of the present research was to describe the microbiome restructuring that occurs during the adaptation stage in landfill leachate treatment. In a model SBR (sequencing batch reactor), a 21-day purification process of landfill leachate was simulated. Wastewater was fed in increasing concentrations. When undiluted leachate entered, the activated sludge structure disintegrated (Sludge Volume Index—4.6 mL/g). The Chemical Oxygen Demand and ammonium nitrogen concentration remained at high values in the influent (2321.11 mgO₂/L and 573.20 mg/L, respectively). A significant amount of free-swimming cells was found, and the number of aerobic heterotrophs and bacteria of the genera Pseudomonas and Acinetobacter increased by up to 125 times. The Azoarcus-Thauera cluster (27%) and Pseudomonas spp. (16%) were registered as the main bacterial groups in the activated sludge. In the changed structure of the microbial community, Gammaproteobacteria, family Rhizobiaceae, class Saccharimonadia were predominantly represented. Among the suspended bacteria, Microbactericeae and Burkholderiaceae, which are known for their ability to degrade xenobiotics, were present in larger quantities. The enzymological analysis demonstrated that the ortho-pathway of cleavage of aromatic structures was active in the community. The described changes in the leachate-purifying microbial community appear to be destructive at the technological level. At the microbiological level, however, trends of initial adaptation were clearly outlined, which, if continued, could provide a highly efficient biodegradation community. Full article

The quantity of industrially polluted waters is increasing everywhere, of which a significant part is occupied by a number of mono- and poly-aromatic compounds. Toxins enter the soil, sewage, and clean water by mixing with or seeping into them from industrial wastewater. By using 18S RNA and ITS sequences, the Penicillium commune AL5 strain that was isolated from Antarctic soil was identified. This study is dedicated to exploring its capacity to metabolize hazardous aromatic compounds. The strain showed very good potential in the degradation of hydroxylated monophenols and possessed exceptional abilities in terms of resorcinol degradation. The strain’s ability to metabolize 0.3 g/L of p-cresol at 10 °C is notable. The strain is also capable of metabolizing LMW PAHs (naphthalene, anthracene, and phenanthrene) and eliminating all three tested compounds under 23 °C, respectively, 77.5%, 93.8%, and 75.1%. At 10 °C, the process slowed down, but the degradation of naphthalene continued to be over 50%. The quantity of PAH and a few significant intermediary metabolites were determined using GC–MS analysis. Sequencing of the enzymes phenol hydroxylase and catechol 1,2-dioxygenase revealed a close association with the genes and proteins in some fungal strains that can degrade the aromatic compounds examined thus far. Full article

Halogenated organic compounds (HOCs) pose a serious problem for the environment. Many are highly toxic and accumulate both in soil and in organisms. Their biological transformation takes place by dehalogenation, in which the halogen substituents are detached from the carbon in the organic compound by enzymes produced by microorganisms. This increases the compounds’ water solubility and bioavailability, reduces toxicity, and allows the resulting compound to become more susceptible to biodegradation. The microbial halogen cycle in soil is an important part of global dehalogenation processes. The aim of the study was to examine the potential of microbial communities inhabiting natural and anthropogenically modified environments to carry out the dehalogenation process. The potential of microorganisms was assessed by analyzing the metagenomes from a natural environment (forest soils) and from environments subjected to anthropopression (agricultural soil and sludge from wastewater treatment plants). Thirteen genes encoding enzymes with dehalogenase activity were identified in the metagenomes of both environments, among which, 2-haloacid dehalogenase and catechol 2,3-dioxygenase were the most abundant genes. Comparative analysis, based on comparing taxonomy, identified genes, total halogens content and content of DDT derivatives, demonstrated the ability of microorganisms to transform HOCs in both environments, indicating the presence of these compounds in the environment for a long period of time and the adaptive need to develop mechanisms for their detoxification. Metagenome analyses and comparative analyses indicate the genetic potential of microorganisms of both environments to carry out dehalogenation processes, including dehalogenation of anthropogenic HOCs. Full article

Phenol poses a threat as one of the most important industrial environmental pollutants that must be removed before disposal. Biodegradation is a cost-effective and environmentally friendly approach for phenol removal. This work aimed at studying phenol degradation by Pseudarthrobacter phenanthrenivorans Sphe3 cells and also, investigating the pathway used by the bacterium for phenol catabolism. Moreover, alginate-immobilized Sphe3 cells were studied in terms of phenol degradation efficiency compared to free cells. Sphe3 was found to be capable of growing in the presence of phenol as the sole source of carbon and energy, at concentrations up to 1500 mg/L. According to qPCR findings, both pathways of ortho- and meta-cleavage of catechol are active, however, enzymatic assays and intermediate products identification support the predominance of the ortho-metabolic pathway for phenol degradation. Alginate-entrapped Sphe3 cells completely degraded 1000 mg/L phenol after 192 h, even though phenol catabolism proceeds slower in the first 24 h compared to free cells. Immobilized Sphe3 cells retain phenol-degrading capacity even after 30 days of storage and also can be reused for at least five cycles retaining more than 75% of the original phenol-catabolizing capacity. Full article

Phenol is an important pollutant widely discharged as a component of hydrocarbon fuels, but its degradation in cold regions is challenging due to the harsh environmental conditions. To date, there is little information available concerning the capability for phenol biodegradation by indigenous Antarctic bacteria. In this study, enzyme activities and genes encoding phenol degradative enzymes identified using whole genome sequencing (WGS) were investigated to determine the pathway(s) of phenol degradation of Arthrobacter sp. strains AQ5-05 and AQ5-06, originally isolated from Antarctica. Complete phenol degradative genes involved only in the ortho-cleavage were detected in both strains. This was validated using assays of the enzymes catechol 1,2-dioxygenase and catechol 2,3-dioxygenase, which indicated the activity of only catechol 1,2-dioxygenase in both strains, in agreement with the results from the WGS. Both strains were psychrotolerant with the optimum temperature for phenol degradation, being between 10 and 15 °C. This study suggests the potential use of cold-adapted bacteria in the bioremediation of phenol pollution in cold environments. Full article

Despite the well-described abundance of phenol-degrading bacteria, knowledge concerning their degradation abilities under suboptimal conditions is still very limited and needs to be expanded. Therefore, this work aimed to study the growth and degradation potential of Stenotrophomonas maltophilia KB2 and Pseudomonas moorei KB4 strains toward phenol under suboptimal temperatures, pH, and salinity in connection with the activity of catechol dioxygenases, fatty acid profiling, and membrane permeability. The methodology used included: batch culture of bacteria in minimal medium supplemented with phenol (300 mg/L), isolating and measuring the activity of catechol 1,2- and 2,3-dioxygenases, calculating kinetic parameters, chromatographic analysis of fatty acid methyl esters (FAMEs) and determining the membrane permeability. It was established that the time of phenol utilisation by both strains under high temperatures (39 and 40 °C) proceeded 10 h; however, at the lowest temperature (10 °C), it was extended to 72 h. P. moorei KB4 was more sensitive to pH (6.5 and 8.5) than S. maltophilia KB2 and degraded phenol 5–6 h longer. Salinity also influenced the time of phenol removal. S. maltophilia KB2 degraded phenol in the presence of 2.5% NaCl within 28 h, while P. moorei KB4 during 72 h. The ability of bacteria to degrade phenol in suboptimal conditions was coupled with a relatively high activity of catechol 1,2- and/or 2,3-dioxygenases. FAME profiling and membrane permeability measurements indicated crucial alterations in bacterial membrane properties during phenol degradation leading predominantly to an increase in fatty acid saturation and membrane permeability. The obtained results offer hope for the potential use of both strains in environmental microbiology and biotechnology applications. Full article

Biotechnologies based on microbial species capable of destroying harmful pollutants are a successful way to solve some of the most important problems associated with a clean environment. The subject of investigation is the Antarctic fungal strain Aspergillus glaucus AL1. The culturing of the examined strain was performed with 70 mg of wet mycelium being inoculated in a Czapek Dox liquid medium containing naphthalene, anthracene, or phenanthrene (0.3 g/L) as the sole carbon source. Progressively decreasing naphthalene and anthracene concentrations were monitored in the culture medium until the 15th day of the cultivation of A. glaucus AL1. The degradation was determined through gas chromatography–mass spectrometry. Both decreased by 66% and 44%, respectively, for this period. The GC-MS analyses were applied to identify salicylic acid, catechol, and ketoadipic acid as intermediates in the naphthalene degradation. The intermediates identified in anthracene catabolism are 2-hydroxy-1-naphthoic acid, o-phthalic acid, and protocatechuic acid. The enzyme activities for phenol 2-monooxygenase (1.14.13.7) and catechol 1,2-dioxygenase (1.13.11.1) were established. A gene encoding an enzyme with catechol 1,2-dioxygenase activity was identified and sequenced (GeneBank Ac. No KM360483). The recent study provides original data on the potential of an ascomycete’s fungal strain A. glaucus strain AL 1 to degrade naphthalene and anthracene. Full article

Heavy metal co-contamination in crude oil-polluted environments may inhibit microbial bioremediation of hydrocarbons. The model heavy metal-resistant bacterium Cupriavidus metallidurans CH34 possesses cadmium and mercury resistance, as well as genes related to the catabolism of hazardous BTEX aromatic hydrocarbons. The aims of this study were to analyze the aromatic catabolic potential of C. metallidurans CH34 and to determine the functionality of the predicted benzene catabolic pathway and the influence of cadmium and mercury on benzene degradation. Three chromosome-encoded bacterial multicomponent monooxygenases (BMMs) are involved in benzene catabolic pathways. Growth assessment, intermediates identification, and gene expression analysis indicate the functionality of the benzene catabolic pathway. Strain CH34 degraded benzene via phenol and 2-hydroxymuconic semialdehyde. Transcriptional analyses revealed a transition from the expression of catechol 2,3-dioxygenase (tomB) in the early exponential phase to catechol 1,2-dioxygenase (catA1 and catA2) in the late exponential phase. The minimum inhibitory concentration to Hg (II) and Cd (II) was significantly lower in the presence of benzene, demonstrating the effect of co-contamination on bacterial growth. Notably, this study showed that C. metallidurans CH34 degraded benzene in the presence of Hg (II) or Cd (II). Full article

It is shown that bacteria Bradyrhizobium japonicum 273 were capable of degrading phenol at moderate concentrations either in a free cell culture or by immobilized cells on granulated activated carbon particles. The amount of degraded phenol was greater in an immobilized cell preparation than in a free culture. The application of a constant electric field during cultivation led to enhanced phenol biodegradation in a free culture and in immobilized cells on granulated activated carbon. The highest phenol removal efficiency was observed for an anode potential of 1.0 V/S.H.E. The effect was better pronounced in a free culture. The enzyme activities of free cells for phenol oxidation and benzene ring cleavage were very sensitive to the anode potential in the first two steps of the metabolic pathway of phenol biodegradation catalyzed by phenol hydroxylase—catechol-1,2-dioxygenase and catechol-2,3-dioxygenase. It was observed that at an anode potential of 0.8 V/S.H.E., the meta-pathway of cleavage of the benzene ring catalyzed by catechol-2,3-dioxygenase became competitive with the ortho-pathway, catalyzed by catechol-1,2-dioxygenase. The obtained results showed that the positive effect of constant electric field on phenol biodegradation was rather due to electric stimulation of enzyme activity than electrochemical anode oxidation. Full article

Microbiota from Alpine forest soils are key players in carbon cycling, which can be greatly affected by climate change. The aim of this study was to evaluate the degradation potential of culturable bacterial strains isolated from an alpine deciduous forest site. Fifty-five strains were studied with regard to their phylogenetic position, growth temperature range and degradation potential for organic compounds (microtiter scale screening for lignin sulfonic acid, catechol, phenol, bisphenol A) at low (5 °C) and moderate (20 °C) temperature. Additionally, the presence of putative catabolic genes (catechol-1,2-dioxygenase, multicomponent phenol hydroxylase, protocatechuate-3,4-dioxygenase) involved in the degradation of these organic compounds was determined through PCR. The results show the importance of the Proteobacteria phylum as its representatives did show good capabilities for biodegradation and good growth at −5 °C. Overall, 82% of strains were able to use at least one of the tested organic compounds as their sole carbon source. The presence of putative catabolic genes could be shown over a broad range of strains and in relation to their degradation abilities. Subsequently performed gene sequencing indicated horizontal gene transfer for catechol-1,2-dioxygenase and protocatechuate-3,4-dioxygenase. The results show the great benefit of combining molecular and culture-based techniques. Full article

Within the broad group of Fe non-heme oxidases, our attention was focused on the catechol 1,2- and 2,3-dioxygenases, which catalyze the oxidative cleavage of aromatic rings. A large group of Fe complexes with N/O ligands, ranging from N₃ to N₂O₂S, was developed to mimic the activity of these enzymes. The Fe complexes discussed in this work can mimic the intradiol/extradiol catechol dioxygenase reaction mechanism. Electronic effects of the substituents in the ligand affect the Lewis acidity of the Fe center, increasing the ability to activate dioxygen and enhancing the catalytic activity of the discussed biomimetic complexes. The ligand architecture, the geometric isomers of the complexes, and the substituent steric effects significantly affect the ability to bind the substrate in a monodentate and bidentate manner. The substrate binding mode determines the preferred mechanism and, consequently, the main conversion products. The preferred mechanism of action can also be affected by the solvents and their ability to form the stable complexes with the Fe center. The electrostatic interactions of micellar media, similar to SDS, also control the intradiol/extradiol mechanisms of the catechol conversion by discussed biomimetics. Full article