Biodegradation Of P-nitrophenol Research Articles

Detoxification of p-nitrophenol (PNP) using Enterococcus gallinarum JT-02 isolated from animal farm waste sludge

Enterococcus gallinarum (JT-02) isolated and identified from the animal farm waste sludge was found to be capable of biodegrading p-nitrophenol (PNP), an organic compound used to manufacture drugs, fungicides, insecticides, dyes, and to darken leather. The intention of this study was to optimize the biodegradation by finding the optimal conditions for the specific strain through single-factor experiments. The bacterial strain was grown in Luria Bertani broth and various parameters were optimized to achieve the prime settings for the p-nitrophenol (PNP) biodegradation. The results indicated that the best setups for the biodegradation by the strain JT-02 was 100 mg/L of PNP; pH 7; 30 °C; 150 rpm in a shaker incubator and 3% (v/v) of inoculum dose. Once the optimal conditions were found, the bacteria were capable of degrading p-nitrophenol (98.21%) in 4 days. Intermediates produced during PNP biodegradation were identified using High Performance Liquid Chromatography (HPLC) analysis and the biodegradation pathway was elucidated. Phytotoxicity studies were carried out with Vigna radiata seeds to confirm the applicability and efficiency of PNP biodegradation.

Sequentially modified carbon felt for enhanced p-nitrophenol biodegradation through direct interspecific electron transfer

The widely applied aromatic nitration in modern industry leads to toxic p-nitrophenol (PNP) in environment. Exploring its efficient degradation routes is of great interests. In this study, a novel four-step sequential modification procedure was developed to increase the specific surface area, functional group, hydrophilicity, and conductivity of carbon felt (CF). The implementation of the modified CF promoted reductive PNP biodegradation, attaining 95.2 ± 0.8% of removal efficiency with less accumulation of highly toxic organic intermediates (e.g., p-aminophenol), compared to carrier-free and CF-packed biosystems. The constructed anaerobic-aerobic process with modified CF in 219-d continuous operation achieved further removal of carbon and nitrogen containing intermediates and partial mineralization of PNP. The modified CF promoted the secretion of extracellular polymeric substances (EPS) and cytochrome c (Cyt c), which were essential components to facilitate direct interspecies electron transfer (DIET). Synergistic relationship was deduced that glucose was converted into volatile fatty acids by fermenters (e.g., Longilinea and Syntrophobacter), which donated electrons to the PNP degraders (e.g., Bacteroidetes_vadinHA17) through DIET channels (CF, Cyt c, EPS) to complete PNP removal. This study proposes a novel strategy using engineered conductive material to enhance the DIET process for efficient and sustainable PNP bioremediation.

Evidence of p-nitrophenol Biodegradation and Study of Genomic Attributes from a Newly Isolated Aquatic Bacterium Pseudomonas Asiatica Strain PNPG3

ABSTRACT A p-nitrophenol (PNP) degrading aquatic bacterial strain PNPG3 was isolated from the Ganges water and was identified as Pseudomonas asiatica based on genome sequence analyses. The optimum catabolic growths for the strain was recorded with 0.5 mM PNP and it could tolerate up to 6 mM PNP. It could carry out biodegradation of PNP through p-benzoquinone (PBQ) and 1, 2, 4-benzenetriol (BT) with concomitant release of nitrite. Genome sequence analysis predicted the presence of all the genes (pdcABC1C2DEFG) responsible for providing the PNP biodegradation phenotype for this strain. Based on homology search, the functional attributes encoded by this gene cluster were predicted to include p-nitrophenol 4-monooxygenase (PdcA), benzoquinone reductase (PdcB), hydroxyquinol 1, 2-dioxygenase (PdcC1), hydroxyquinol 1, 2-dioxygenase large subunit (PdcC2), 4-hydroxymuconic semialdehyde dehydrogenase (PdcD), maleylacetate reductase (PdcE), hydroquinone dioxygenase alpha subunit (PdcF) and putative regulator (PdcG). This is the first report of any representative aquatic strain under Pseudomonas asiatica, having the highest known catabolic PNP utilizing capability from the Ganges water of India to the best of the author’s knowledge, and may find application toward cost-effective bioremediation of PNP-contaminated waterbodies.

Biodegradation of p-nitrophenol by a member of the genus Brachybacterium, isolated from the river Ganges.

The online version contains supplementary material available at 10.1007/s13205-022-03263-7.

Biodegradation of P-nitro phenol using a novel bacterium Achromobacter denitrifacians isolated from industrial effluent water.

In the present investigation, Achromobacter denitrifacians was isolated from industrial wastewater and used in the degradation of para nitro-phenol. Experiments were made as a function of different carbon sources, organic and inorganic nitrogen sources and metal ions to analyse the removal efficiency of para nitro-phenol present in the industrial wastewater sources. Observations revealed that the rate of phenol biodegradation was significantly affected by pH, temperature of incubation, glucose, peptone and metal ion concentration. The optimal conditions for phenol removal were found to be pH of 7.5, temperature, 35 °C and 0.25 gL-1 supplemented glucose level, 0.25 gL-1 supplemented peptone level, and 0.01 gL-1 zinc ion. The key importance of the present study is the utilization of a native bacterial strain isolated from the industrial effluent water itself having an impending role in the bioremediation process of phenol.

Biodegradation of p-nitrophenol by engineered strain

p-Nitrophenol (PNP) is an important environmental pollutant and can causes significant environmental and health risks. Compared with the traditional methods, biodegradation is a useful one to completely remove the harmful pollutants from the environment. Here, an engineered strain was first constructed by introducing PNP biodegradation pathway via the hydroquinone (HQ) pathway into Escherichia coli. In the engineered strain BL-PNP, PNP was completely degraded to β-ketoadipate and subsequently enter the metabolites of multiple anabolic pathways. The high tolerance and rapid degradation ability to PNP enable the engineered strain to have the potential to degrade toxic substances. The engineered strain created in this study can be used as a functional strain for bioremediation of PNP and potential toxic intermediates, and the method of assembling aromatic hydrocarbons metabolic pathway can be used to eradicate nitroaromatic pollutants in the environment.

Transcriptomic analysis of Burkholderia cenocepacia CEIB S5-2 during methyl parathion degradation.

Methyl parathion (MP) is a highly toxic organophosphorus pesticide associated with water, soil, and air pollution events. The identification and characterization of microorganisms capable of biodegrading pollutants are an important environmental task for bioremediation of pesticide impacted sites. The strain Burkholderia cenocepacia CEIB S5-2 is a bacterium capable of efficiently hydrolyzing MP and biodegrade p-nitrophenol (PNP), the main MP hydrolysis product. Due to the high PNP toxicity over microbial living forms, the reports on bacterial PNP biodegradation are scarce. According to the genomic data, the MP- and PNP-degrading ability observed in B. cenocepacia CEIB S5-2 is related to the presence of the methyl parathion-degrading gene (mpd) and the gene cluster pnpABA'E1E2FDC, which include the genes implicated in the PNP degradation. In this work, the transcriptomic analysis of the strain in the presence of MP revealed the differential expression of 257 genes, including all genes implicated in the PNP degradation, as well as a set of genes related to the sensing of environmental changes, the response to stress, and the degradation of aromatic compounds, such as translational regulators, membrane transporters, efflux pumps, and oxidative stress response genes. These findings suggest that these genes play an important role in the defense against toxic effects derived from the MP and PNP exposure. Therefore, B. cenocepacia CEIB S5-2 has a great potential for application in pesticide bioremediation approaches due to its biodegradation capabilities and the differential expression of genes for resistance to MP and PNP.

Phenol and p-nitrophenol biodegradations by acclimated activated sludge: Influence of operational conditions on biodegradation kinetics and responding microbial communities

This study aims to evaluate the influence of operational factors (i.e. substrate concentration, activated sludge concentration and acclimation concentration) and the microbial communities of activated sludge on the aerobic biodegradation of phenol and p-nitrophenol (PNP). During the biodegradation process, the phenol or PNP biodegradation and activated sludge growth kinetics were determined and the interaction effects of the operational factors on the kinetics were assessed. The phenol biodegradation kinetics was well described by zero-order model (R2 > 0.813). At low phenol concentration (< 200 mg/L), the biodegradation rate can be enhanced by increasing activated sludge concentration and acclimation concentration. From the specific oxygen uptake rate (SOUR) analysis, lower SOUR values for PNP relative to phenol suggests higher toxicity of PNP, with inhibition detected above 50 mg/L for both phenol and PNP. In view of the inadequacy of zero-order equation for PNP biodegradation, the kinetic data was fitted into Haldane equation that incorporate the inhibitory effect. From the results, the value of critical PNP concentration was observed to increase at higher acclimation concentration, indicating the attenuation of inhibitory effects on PNP-acclimated activated sludge. For microbial growth, only the factor of acclimation concentration greatly affected the growth kinetics, in which the Ks/Ki ratio was greatly reduced when the acclimation concentration was doubled. Lastly, potential degrading strains of Acinetobacter sp. USM, Rhodococcus sp. USM1 and Caballeronia sp. USM2, were successfully isolated from the acclimated activated sludge. From the results of dioxygenase genes amplification, the phenol degradation mechanism was identified following ortho fission pathway.

Coupled biodegradation of p-nitrophenol and p-aminophenol in bioelectrochemical system: Mechanism and microbial functional diversity

Biodegradation mechanisms and microbial functional diversity during coupled p-nitrophenol (PNP) and p-aminophenol (PAP) degradation were studied in a bioelectrochemical system. PNP in the biocathode and PAP in the bioanode were almost completely removed within 28hr and 68hr respectively. The degradation followed the steps including hydrating hydroxyalkylation, dehydrogenating carbonylation, and hydrolating ring cleavage, etc. Metagemomic analysis based on the KEGG and eggNOG database annotations revealed the microbial composition and functional genes/enzymes related to phenol degradation in the system. The predominant bacteria genera were Lautropia, Pandoraea, Thiobacillus, Ignavibacterium, Truepera and Hyphomicrobium. The recognized biodegradation genes/enzymes related to pollutant degradation were as follows: pmo, hbd, & ppo for phenol degradation, nzba, amie, & badh for aromatic degradation, and CYP & p450 for xenobiotics degradation, etc. The co-occurrence of ARGs (antibiotic resistant genes), such as adeF, MexJ, ErmF, PDC-93 and Escherichia_coli_mdfA, etc., were annotated in CARD database during the biodegradation process. The Proteobacteria & Actinobacteria phylum was the primary host of both the biodegradation genes & ARGs in this system. The microbial functional diversity ensured the effective biodegradation of the phenol pollutants in the bioelectrochemical system.

Statistical optimization of immobilization of activated sludge in PVA/alginate cryogel beads using response surface methodology for p-nitrophenol biodegradation

This study focuses on the modelling and optimization of process variables for the immobilization of activated sludge in polyvinyl alcohol (PVA)/alginate cryogel beads using response surface methodology (RSM). The results from analysis of variance showed that the models were reliable and significant in predicting the biodegradation rate and beads breakage. The interaction effects of the five independent variables, namely number of freezing-thawing cycle, bead size as well as PVA, alginate and CaCl2 concentrations, were statistically analyzed using central composite design (CCD). Alginate concentration was classified by models for contributing the most significant effect on both responses. The interactions between alginate concentration with CaCl2 concentration, alginate concentration with number of freezing-thawing cycle and alginate concentration with bead size significantly affected the performances of cryogel beads on p-nitrophenol (PNP) biodegradation rate and beads breakage. The predicted results were in conformity with the experimental runs, in which the maximum PNP biodegradation rate of 7.4 mg/L h and minimum breakage of 0 % could be achieved using 8.0 wt % of PVA, 1.411 wt % of sodium alginate, 3.012 wt % of CaCl2, 3.659 mm of bead and 3 cycles for freezing-thawing process. At low initial PNP concentration of 100 mg/L, the PVA/alginate-AS cryogel beads were reusable with no significant loss in PNP biodegradation efficiency for 20 consecutive biodegradation cycles.

Using ultrasonic treated sludge to accelerate pyridine and p-nitrophenol biodegradation

Aerobic biomass (acclimated activated sludge) was treated by ultrasound and used to accelerate biodegradations of pyridine and p-nitrophenol (PNP), which begin with mono-oxygenation reactions that need an internal electron donor. Ultrasound treatment disrupted the biomass and produced more soluble and (especially) colloidal organic material. Compared with untreated biomass, pyridine- and PNP-biodegradation rates increased when treated biomass provided electron donor. Pyridine- and PNP-biodegradation rates increased by 10% and 20%, respectively, over the control experiments when supernatants from treated biomass were added into the medium. For accelerating pyridine- and PNP removal rates, adding supernatants of treated biomass was equivalent to adding succinate of 0.35 mmol/L and 0.21 mmol/L, respectively. The rates were increased by 63% for both substrates when the entire treated biomass was added. Colloidal solids in the treated biomass contained most of biodegradable electron donor able to stimulate initial monooxygenations of pyridine and PNP. Adding treated biomass together with untreated biomass gave a synergistic impact on enhancing pyridine and PNP biodegradation, because the treated biomass supplied extra donor, while the untreated biomass maintained a high level of active biomass.

Aerobic biodegradation of p-nitrophenol in a nitrifying sludge bioreactor: System performance, sludge property and microbial community shift

The system performance, sludge property and microbial community shift were evaluated in a nitrifying sludge (NS) bioreactor for simultaneous treating p-Nitrophenol (PNP) and high ammonia wastewater. After long-term acclimation for 80 days, the removal efficiencies of PNP and NH4+-N reached to 99.9% and 99.5%, respectively. Meanwhile, the effluent PNP gradually decreased from 7.9 to 0.1 mg/L by acclimation of sludge. The particle size of NS increased from 115.2 μm to 226.3 μm accompanied by the decreased zeta potential as a self-protection strategy. The presence of PNP exposure altered the effluent soluble microbial products (SMP) fluorescent components and molecular composition. The increase in the relative abundance of Thauera, Nitrospiraceae and Nitrosomonas indicated the nitrification and denitrification capacities of NS increased, which maybe the PNP cometabolic biodegradation effect. Moreover, Ignavibacteria and Aeromonas were responsible as the dominant bacteria for degrading PNP in the nitrifying system.

Carboxymethyl sago pulp/chitosan hydrogel as an immobilization medium for activated sludge for p‐nitrophenol biodegradation

ABSTRACTIn this study, carboxymethyl sago pulp (CMSP) derived from sago waste was successfully crosslinked with ferric ions in the presence of chitosan forming a novel immobilization matrix for p‐nitrophenol (PNP)‐acclimated activated sludge. On the basis of the shortest biodegradation time of PNP, the optimized operational conditions of immobilization were found to be: 7 w/v% CMSP, 9 g L−1 of activated sludge, 0.1 M ferric ion, and 15 min of crosslinking time. Observable inhibited PNP biodegradation was exhibited by the suspended activated sludge at the initial PNP concentration of 400 mg L−1. In contrast, complete mineralization was achieved by the CMSP/chitosan‐immobilized activated sludge (CMSP/Ch‐AS) beads. The results revealed the important role of CMSP/Ch hydrogel in protecting activated sludge from the toxicity of PNP. The CMSP/Ch‐AS beads could be reused consecutively up to three and two cycles, respectively, for the biodegradation of PNP at 200 and 400 mg L−1, with the attainment of more than 99% of PNP removal at each cycle. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019, 136, 47531.

Rhodococcus wratislaviensis strain 9: An efficient p-nitrophenol degrader with a great potential for bioremediation

A Gram-positive bacterium, Rhodococcus wratislaviensis strain 9, was isolated from groundwater contaminated with nitrophenolics and trichloroethene following enrichment culture technique. The cells of strain 9 grown on LB broth (uninduced) degraded 720 μM p-nitrophenol (PNP) within 12 h, and utilized as a source of carbon and energy. Orthogonal experimental design analysis to determine optimal conditions for biodegradation of PNP showed that pH had a significant positive effect (P ≤ .05) on bacterial degradation of PNP, while glucose, di- and tri-nitrophenols exhibited significant negative effect. Cell-free extracts obtained from PNP-grown culture that contained 20 μg mL−1 protein degraded 90% of 720 μM PNP within 5 h of incubation. Two-dimensional protein analysis revealed differential expression of the oxygenase component of PNP monooxygenase and an elongation factor Tu in PNP-grown cells, but not in those grown on glucose. The strain 9 remediated laboratory wastewater containing 900 μM PNP efficiently within 14 h, indicating its great potential in bioremediation of PNP-contaminated waters.

Spatial and temporal variability in the potential of river water biofilms to degrade p-nitrophenol

In order to predict the fate of chemicals in the environment, a range of regulatory tests are performed with microbial inocula collected from environmental compartments to investigate the potential for biodegradation. The abundance and distribution of microbes in the environment is affected by a range of variables, hence diversity and biomass of inocula used in biodegradation tests can be highly variable in space and time. The use of artificial or natural biofilms in regulatory tests could enable more consistent microbial communities be used as inocula, in order to increase test consistency. We investigated spatial and temporal variation in composition, biomass and chemical biodegradation potential of bacterial biofilms formed in river water. Sampling time and sampling location impacted the capacity of biofilms to degrade p-nitrophenol (PNP). Biofilm bacterial community structure varied across sampling times, but was not affected by sampling location. Degradation of PNP was associated with increased relative abundance of Pseudomonas syringae. Partitioning of the bacterial metacommunity into core and satellite taxa revealed that the P. syringae could be either a satellite or core member of the community across sampling times, but this had no impact on PNP degradation. Quantitative PCR analysis of the pnpA gene showed that it was present in all samples irrespective of their ability to degrade PNP. River biofilms showed seasonal variation in biomass, microbial community composition and PNP biodegradation potential, which resulted in inconsistent biodegradation test results. We discuss the results in the context of the mechanisms underlying variation in regulatory chemical degradation tests.

Cloning and expression of vgb gene in Bacillus cereus, improve phenol and p-nitrophenol biodegradation

In this work, the vgb gene from Vitrocilla stercoraria was used to genetically modify a Bacillus cereus strain isolated from pulp and paper wastewater effluent. The gene was cloned in a multicopy plasmid (pUB110) or uni-copy gene using a chromosome integrative vector (pTrpBG1). B. cereus and its recombinant strains were used for phenol and p-nitrophenol biodegradation using aerobic or micro-aerobic conditions and two different temperatures (i.e. 37 and 25 °C). Complete (100%) phenol degradation was obtained for the strain where the multicopy of vgb gene was present, 98% for the strain where uni-copy gene was present and 45% for wild type strain for the same experimental conditions (i.e. 37 °C and aerobic condition). For p-nitrophenol degradation at the same conditions, the strain with the multi-copy vgb gene was capable to achieve 50% of biodegradation, ∼100% biodegradation was obtained using the uni-copy strain and ∼24% for wild type strain. When the micro-aerobic condition was tested, the biodegradation yield showed a significant decreased. The biodegradation trend observed for aerobic was similar for micro-aerobic assessments: the modified strains showed higher degradation rates when compared with wild type strain. For all experimental conditions, the highest p-nitrophenol degradation was observed using the strain with uni-copy of vgb gene. Besides the increase of biodegradative capability of the strain, insertion of the vgb gene was observed able to modify other morphological characteristics such as avoiding the typical flake formation in the B. cereus culture. In both cases, the modification seems to be related with the enhancement of oxygen supply to the cells generated by the vgb gene insertion. The application of the genetically modified microorganism (GMM) to the biodegradation of pollutants in contaminated water possesses high potential as an environmentally friendly technology to facing this emergent problem.

Long-term performance and stability of a continuous granular airlift reactor treating a high-strength wastewater containing a mixture of aromatic compounds

Continuous feeding operation of an airlift reactor and its inoculation with mature aerobic granules allowed the successful treatment of a mixture of aromatic compounds (p-nitrophenol, o-cresol and phenol). Complete biodegradation of p-nitrophenol, o-cresol, phenol and their metabolic intermediates was achieved at an organic loading rate of 0.61g COD L−1d−1. Stable granulation was obtained throughout the long-term operation (400 days) achieving an average granule size of 2.0±1mm and a sludge volumetric index of 26±1mLg−1 TSS. The identified genera in the aerobic granular biomass were heterotrophic bacteria able to consume aromatic compounds. Therefore, the continuous feeding regimen and the exposure of aerobic granules to a mixture of aromatic compounds make possible to obtain good granulation and high removal efficiency.

Experimental study and kinetic modeling of cometabolic degradation of phenol and p-nitrophenol by loofa-immobilized Ralstonia eutropha

In the present study, phenol-adapted cells of Ralstonia eutropha were used to degrade p-nitrophenol (PNP) in the presence of phenol. PNP at initial concentrations ranging from 5 to 15 mg/L was degraded almost completely by free cells of R. eutropha. The use of loofa-immobilized cells increased the complete removal of PNP up to 30 mg/L. Kinetic data for PNP biodegradation by immobilized cells of R. eutropha best fitted the Haldane model. The kinetic parameters were ks = 0.0006 (mg PNP/mg biomass.h), Ks = 8.83 (mg/L) and Ki = 30.77 (mg/L). The degradation pathways of PNP through the metabolites, 4-nitro-catechol (4-NC) and hydroquinone (HQ), were investigated using HPLC.

BIOREMEDIATION CONTAINING WASTEWATER BY OF P-NITROPHENOL AEROBIC GRANULES

Aerobic granulation is a novel biotechnique for the treatment of coloured wastewater containing toxic xenobiotics in sequencing batch reactor (SBR). The aerobic granules were investigated as a potential biomaterial for biodegradation of p-nitrophenol. The removal efficiency of p-nitrophenol was 93 % after 65 days of operation at 8 hr hydraulic retention time (HRT). The aerobic granules were cultivated at a p-nitrophenol loading rate of 0.9 kg m-3 day-1, with sodium acetate as co-substrate to accelerate the growth of p-nitrophenol degrading biomass. SBR was inoculated with activated sludge that was initially conditioned as a batch culture for a period of 30 days by supplying p-nitrophenol. Optimal pH of 9 was maintained throughout the experimental setup. This study clearly demonstrates that aerobic granules are efficient to degrade up to 300 mgL-1 of p-nitrophenol to tolerate the toxicity in high-strength wastewater.

Simultaneous nitritation and p-nitrophenol removal using aerobic granular biomass in a continuous airlift reactor

The chemical and petrochemical industries produce wastewaters containing ammonium and phenolic compounds. Biological treatment of these wastewaters could be problematic due to the possible inhibitory effects exerted by phenolic compounds. The feasibility of performing simultaneous nitritation and p-nitrophenol (PNP) biodegradation using a continuous aerobic granular reactor was evaluated. A nitrifying granular sludge was bioaugmented with a PNP-degrading floccular sludge, while PNP was progressively added to the feed containing a high ammonium concentration. Nitritation was sustained throughout the operational period with ca. 85% of ammonium oxidation and less than 0.3% of nitrate in the effluent. PNP biodegradation was unstable and the oxygen limiting condition was found to be the main explanation for this unsteadiness. An increase in dissolved oxygen concentration from 2.0 to 4.5 mg O2 L(-1) significantly enhanced PNP removal, achieving total elimination. Acinetobacter genus and ammonia-oxidising bacteria were the predominant bacteria species in the granular biomass.