Insight into the mechanism of chlorinated nitroaromatic compounds anaerobic reduction with mackinawite (FeS) nanoparticles.

Anaerobic biotechnology for wastewaters treatment can nowadays be considered as state of the art methods. Nonetheless, this technology exhibits certain inherent limitations when employed for industrial wastewater treatment, encompassing elevated substrate consumption, diminished electron transfer efficiency, and compromised system stability. To address the above issues, increasing interest is being given to the potential of using conductive non-biological materials, e,g., iron sulfide (FeS), as a readily accessible electron donor and electron shuttle in the biological decontamination process. In this study, Mackinawite nanoparticles (FeS NPs) were studied for their ability to serve as electron donors for p-chloronitrobenzene (p-CNB) anaerobic reduction within a coupled system. This coupled system achieved an impressive p-CNB removal efficiency of 78.3 ± 2.9% at a FeS NPs dosage of 1 mg/L, surpassing the efficiencies of 62.1 ± 1.5% of abiotic and 30.6 ± 1.6% of biotic control systems, respectively. Notably, the coupled system exhibited exclusive formation of aniline (AN), indicating the partial dechlorination of p-CNB. The improvements observed in the coupled system were attributed to the increased activity in the electron transport system (ETS), which enhanced the sludge conductivity and nitroaromatic reductases activity. The analysis of equivalent electron donors confirmed that the S2- ions dominated the anaerobic reduction of p-CNB in the coupled system. However, the anaerobic reduction of p-CNB would be adversely inhibited when the FeS NPs dosage exceeded 5 g/L. In a continuous operation, the p-CNB concentration and HRT were optimized as 125 mg/L and 40 h, respectively, resulting in an outstanding p-CNB removal efficiency exceeding 94.0% after 160 days. During the anaerobic reduction process, as contributed by the predominant bacterium of Thiobacillus with a 6.6% relative abundance, a mass of p-chloroaniline (p-CAN) and AN were generated. Additionally, Desulfomonile was emerged with abundances ranging from 0.3 to 0.7%, which was also beneficial for the reduction of p-CNB to AN. The long-term stable performance of the coupled system highlighted that anaerobic technology mediated by FeS NPs has a promising potential for the treatment of wastewater containing chlorinated nitroaromatic compounds, especially without the aid of organic co-substrates.

Geobacter sulfurreducens promoted the biosynthesis of reduced graphene oxide and coupled it for nitrobenzene reduction.

In order to explore an efficient and green method to deal with nitrobenzene (NB) pollutant, reduced graphene oxide (rGO) as an electron shuttle was applied to enhance the extracellular electron transfer (EET) process of Geobacter sulfurreducens, which was a typical electrochemically active bacteria (EAB). In this study, rGO biosynthesis was achieved via the reduction of graphene oxide (GO) by G. sulfurreducens PCA within 3 days. Also, the rGO-PCA combining system completely reduced 50-200 µmol/L of NB to aniline as end product within one day. SEM characterization revealed that PCA cells were partly wrapped by rGO, and therefore the distance of electron transfer between strain PCA and rGO material was reduced. Beside, the ID/IG of GO, rGO, and rGO-PCA combining system were 0.990, 1.293 and 1.31, respectively. Moreover, highest currents were observed in rGO-PCA-NB as 12.950 µA/-12.560 µA at -408 mV/156 mV, attributing to the faster electron transfer efficiency in EET process. Therefore, the NB reduction was mainly due to: (I) direct EET process from G. sulfurreducens PCA to NB; (II) rGO served as electron shuttle and accelerated electron transfer to NB, which was the main degradation pathway. Overall, the biosynthesis of rGO via GO reduction by Geobacter promoted the NB removal process, which provided a facile strategy to alleviate the problematic nitroaromatic pollution in the environment.

Phytotoxicity of nitrobenzene bioaccumulation in rice seedlings: Nitrobenzene inhibits growth, induces oxidative stress, and reduces photosynthetic pigment synthesis.

Nitrobenzene (NB) has been used in numerous industrial and agricultural fields as an organic compound intermediate. NB has mutagenicity and acute toxicity, and is typically a toxic pollutant in industrial wastewater worldwide. To evaluate its phytotoxicity, we treated rice (Oryza sativa) with different concentrations of NB (0, 5, 25, 50, 75, and 100 mg L-1). NB inhibited growth indices of rice (shoot and root length, fresh shoot and root weight, and dry shoot and root weight) as NB treatment concentrations increased. High concentrations (>25 mg L-1) of NB significantly inhibited rice root and shoot growth; root growth was more susceptible to NB. NB treatment could damage the structure and reduce the activity of rice seedling roots. The result of high performance liquid chromatography (HPLC) indicated that the bioaccumulation of NB in rice seedlings had a dose-dependent effect on the growth inhibition. NB reduced the photosynthetic pigment content and the expression levels of chlorophyll synthesis genes. NB treatment increased active oxygen radicals, electrical conductivity, malondialdehyde (MDA), proline, and soluble sugar contents. The expressions of antioxidant enzyme genes were induced by NB stress, and exhibited a phenomenon of initial increase followed by decrease. When the NB concentration was higher than 50 mg L-1, the gene expression levels decreased rapidly. This study provides insight into the association between exposure to NB and its phytotoxic effects on rice seedlings, and assesses the potential risk of NB bioaccumulation for crops that require a large amount of irrigation water.

Effects of temperature, pH, and salinity on the growth kinetics of Pseudomonas sp. NB-1, a newly isolated cold-tolerant, alkali-resistant, and high-efficiency nitrobenzene-degrading bacterium.

ABSTRACTStrain NB-1, which can efficiently degrade nitrobenzene, was identified as Pseudomonas frederiksbergensis. NB-1 was resistant to cold and alkali with the widest temperature (4-35 °C) and pH (5-11) adaptive range, compared with other reported nitrobenzene-degrading microorganisms. Based on the Haldane-Andrews model, the real maximum specific growth rate µm', specific affinity aA, and inhibition coefficient Ki were used in response surface methodology (RSM) simultaneously for the first time to guide NB-1 to treat nitrobenzene wastewater. According to the RSM model, the environmental factors (temperature, pH, salinity) corresponding to the optimal values of µm', aA, and Ki were determined. By comparing the specific growth rates corresponding to the optimal values of µm', aA, and Ki, respectively, the optimum growth conditions of NB-1 were determined under different nitrobenzene concentrations. The study of µm', aA, and Ki by RSM provided a new approach for a more accurate optimization of biological wastewater treatment conditions.

Construction of complete degradation pathway for nitrobenzene in Escherichia coli.

Nitrobenzene is widely present in industrial wastewater and soil. Biodegradation has become an ideal method to remediate organic pollutants due to its low cost, high efficiency, and absence of secondary pollution. In the present study, 10 exogenous genes that can completely degrade nitrobenzene were introduced into Escherichia coli, and their successful expression in the strain was verified by fluorescence quantitative polymerase chain reaction and proteomic analysis. The results of the degradation experiment showed that the engineered strain could completely degrade 4 mM nitrobenzene within 8 h. The formation of intermediate metabolites was detected, and the final metabolites entered the E. coli tricarboxylic acid cycle smoothly. This process was discovered by isotope tracing method. Results indicated the integrality of the degradation pathway and the complete degradation of nitrobenzene. Finally, further experiments were conducted in soil to verify its degradation ability and showed that the engineered strain could also degrade 1 mM nitrobenzene within 10 h. In this study, engineered bacteria that can completely degrade nitrobenzene have been constructed successfully. The construction of remediation-engineered bacteria by synthetic biology laid the foundation for the industrial application of biological degradation of organic pollutants.

Synergistic effect of bioanode and biocathode on nitrobenzene removal: Microbial community structure and functions.

This study aimed to reveal the synergistic effect of bioanode and biocathode on nitrobenzene (NB) removal with different microbial community structures and functions. Single-chamber bioelectrochemical reactors were constructed and operated with different initial concentrations of NB and glucose as the substrate. With the synergistic effect of biocathode and bioanode, NB was completely removed within 8 h at a kinetic rate constant of 0.8256 h-1, and high conversion rate from NB to AN (92%) was achieved within 18 h. The kinetic rate constant of NB removal was linearly correlated with the maximum current density and total coulombs (R2 > 0.95). Increase of glucose and NB concentrations had significantly positive and negative effects, respectively, on the NB removal kinetics (R2 > 0.97 and R2 > 0.93, respectively). Geobacter sp. and Enterococcus sp. dominated in the bioanode and biocathode, respectively. The presence of Klebsiella pneumoniae in the bioanode was beneficial for Geobacter species to produce electricity and to alleviate the NB inhibition. As one of the dominant species at the biocathode, Methanobacterium formicicum has the ability of nitroaromatics degradation according to KEGG analysis, which played a crucial role for NB reduction. Fermentative bacteria converted glucose into volatile fatty acids or H2, to provide energy sources to other species (e.g., Geobacter sulfurreducens and Methanobacterium formicicum). The information from this study is useful to optimize the bioelectrocatalytic system for nitroaromatic compound removal.

Unveiling the chemotactic response and mechanism of Shewanella oneidensis MR-1 to nitrobenzene.

Bioreduction by electroactive bacteria (EAB) is considered as a potential and cost-effective approach for the removal of nitroaromatic compounds (NACs). However, little is known about how the widespread EAB sense and respond to slightly soluble NACs in aquatic environments. Here, the chemotactic behaviors of Shewanella oneidensis MR-1, a model EAB, toward several NACs were examined and their underlying molecular mechanism was elucidated. S. oneidensis MR-1 was found to exhibit a strong chemotactic response to nitrobenzene (NB), but not to other selected NACs under aerobic conditions. To sense NB, this bacterium requires both the histidine kinase (CheA-3)-involved chemotactic signal transduction pathway and an inner-membrane c-type cytochrome CymA. Such a chemotactic response is mediated by an energy taxis mechanism. Additionally, external riboflavin was shown to greatly enhance the Shewanella taxis toward NB, implying a feasible way to increase the bioavailability of NACs. The present study deepens our understanding of the role of microbial chemotaxis in the removal of NACs and provides more options for the bioremediation of NAC-contaminated sites.

A Recently Assembled Degradation Pathway for 2,3-Dichloronitrobenzene in Diaphorobacter sp. Strain JS3051.

Diaphorobacter sp. strain JS3051 utilizes 2,3-dichloronitrobenzene (23DCNB), a toxic anthropogenic compound, as the sole carbon, nitrogen, and energy source for growth, but the metabolic pathway and its origins are unknown. Here, we establish that a gene cluster (dcb), encoding a Nag-like dioxygenase, is responsible for the initial oxidation of the 23DCNB molecule. The 2,3-dichloronitrobenzene dioxygenase system (DcbAaAbAcAd) catalyzes conversion of 23DCNB to 3,4-dichlorocatechol (34DCC). Site-directed mutagenesis studies indicated that residue 204 of DcbAc is crucial for the substrate specificity of 23DCNB dioxygenase. The presence of glutamic acid at position 204 of 23DCNB dioxygenase is unique among Nag-like dioxygenases. Genetic, biochemical, and structural evidence indicate that the 23DCNB dioxygenase is more closely related to 2-nitrotoluene dioxygenase from Acidovorax sp. strain JS42 than to the 34DCNB dioxygenase from Diaphorobacter sp. strain JS3050, which was isolated from the same site as strain JS3051. A gene cluster (dcc) encoding the enzymes for 34DCC catabolism, homologous to a clc operon in Pseudomonas knackmussii strain B13, is also on the chromosome at a distance of 2.5 Mb from the dcb genes. Heterologously expressed DccA catalyzed ring cleavage of 34DCC with high affinity and catalytic efficiency. This work not only establishes the molecular mechanism for 23DCNB mineralization, but also enhances the understanding of the recent evolution of the catabolic pathways for nitroarenes. IMPORTANCE Because anthropogenic nitroaromatic compounds have entered the biosphere relatively recently, exploration of the recently evolved catabolic pathways can provide clues for adaptive evolutionary mechanisms in bacteria. The concept that nitroarene dioxygenases shared a common ancestor with naphthalene dioxygenase is well established. But their phylogeny and how they evolved in response to novel nitroaromatic compounds are largely unknown. Elucidation of the molecular basis for 23DCNB degradation revealed that the catabolic pathways of two DCNB isomers in different isolates from the same site were derived from different recent origins. Integrating structural models of catalytic subunits and enzymatic activities data provided new insight about how recently modified enzymes were selected depending on the structure of new substrates. This study enhances understanding and prediction of adaptive evolution of catabolic pathways in bacteria in response to new chemicals.

Design, Synthesis, and Evaluation of o-(Biphenyl-3-ylmethoxy)nitrophenyl Derivatives as PD-1/PD-L1 Inhibitors with Potent Anticancer Efficacy In Vivo.

Two series of novel o-(biphenyl-3-ylmethoxy)nitrophenyl compounds (A1-31 and B1-17) were designed as programmed cell death protein 1 (PD-1)/PD-ligand 1 (PD-L1) inhibitors. All compounds showed significant inhibitory activity with IC50 values ranging from 2.7 to 87.4 nM except compound A17, and compound B2 displayed the best activity. Further experiments showed that B2 bound to the PD-L1 protein without obvious toxicity in Lewis lung carcinoma (LLC) cells. Furthermore, B2 significantly promoted interferon-gamma secretion in a dose-dependent manner in vitro and in vivo. Especially, B2 exhibited potent in vivo anticancer efficacy in an LLC-bearing allograft mouse model at a low dose of 5 mg/kg, which was more active than BMS-1018 (tumor growth inhibition rate: 48.5% vs 17.8%). A panel of immunohistochemistry and flow cytometry assays demonstrated that B2 effectively counteracted PD-1-induced immunosuppression in the tumor microenvironment, thereby triggering antitumor immunity. These results indicate that B2 is a promising PD-1/PD-L1 inhibitor worthy of further development.

Intracellular Hybrid Biosystem in a Protozoan to Trigger Visible-Light-Driven Photocatalysis.

Incorporating artificial photosensitizers with microorganisms has recently been recognized as an effective way to convert light energy into chemical energy. However, the incorporated biosystem is usually constructed in an extracellular manner and is vulnerable to the external environment. Here, we develop an intracellular hybrid biosystem in a higher organism protozoa Tetrahymena pyriformis, in which the in vivo synthesized CdS nanoparticles trigger photoreduction of nitrobenzene into aniline under visible-light irradiation. Integrating a photosensitizer CdS into T. pyriformis enables the photosensitizer CdS, inherent nitroreductase, and the cytoplasmic reductive substance in T. pyriformis to synergistically engage in the photocatalysis process, generating a greatly enhanced aniline yield with a 40-fold increment. Moreover, building an intracellular hybrid biosystem in mutant T. pyriformis could even grant it new capability of reducing nitrobenzene into aniline under visible-light irradiation. Such an intracellular hybrid biosystem paves a new way to functionalize higher organisms and diversify light energy conversion.

Amino acid derivative reactivity assay-organic solvent reaction system: A novel alternative test for skin sensitization capable of assessing highly hydrophobic substances.

The amino acid derivative reactivity assay (ADRA) is an in chemico alternative to animal testing that focuses on protein binding. The ADRA is a skin sensitization test that solves problems associated with the direct peptide reactivity assay. However, when utilizing the ADRA to evaluate highly hydrophobic substances with octanol/water partition coefficients (logKow) of >6, the test substances may not dissolve in the reaction solution, which can prevent the accurate assessment of skin sensitization. Therefore, we developed the ADRA-organic solvent (ADRA-OS) reaction system, which is a novel skin sensitization test that enables the assessment of highly hydrophobic substances with a logKow of >6. We discovered that the organic solvent ratio, the triethylamine concentration, and the ethylenediaminetetraacetic acid disodium salt dihydrate concentration participate in reactions with the nucleophile N-(2-(1-naphthyl)acetyl)-l-cysteine (NAC) and sensitizers that are used in ADRA and in stabilizing NAC. Thus, we determined the optimal reaction composition of the ADRA-OS according to L9 (33 ) orthogonal array experiments. Using this test, we assessed 14 types of highly hydrophobic substances. When we compared the results with ADRA, we found that ADRA-OS reaction system has high solubility for highly hydrophobic substances and that it has a high predictive capacity (sensitivity: 63%, specificity: 100%, accuracy: 79%). The implication of the results is that the novel ADRA-OS reaction system should provide a useful method for assessing the skin sensitization of highly hydrophobic substances with a logKow of >6.

DNA-templated silver nanoclusters as an efficient catalyst for reduction of nitrobenzene derivatives: a systematic study.

Nitrobenzene compounds are highly toxic pollutants with good stability, and they have a major negative impact on both human health and the ecological environment. Herein, it was found for the first time that fluorescent DNA-silver nanoclusters (DNA-AgNCs) can catalyze the reduction of toxic and harmful nitro compounds into less toxic amino compounds with excellent tolerance to high temperature and organic solvents. In this study, the reduction of p-nitrophenol (4-NP) as a model was systematically investigated, followed by expending the substrate to disclose the versatility of this reaction. This report not only expanded the conditions for utilizing catalytic reduction conditions of DNA-AgNCs as an efficient catalyst in the control of hazardous chemicals but also widened the substrate range of DNA-AgNCs reduction, providing a new angle for the application of noble metal nanoclusters.

Modeling the Bioactivation and Subsequent Reactivity of Drugs.

Electrophilically reactive drug metabolites are implicated in many adverse drug reactions. In this mechanism-termed bioactivation-metabolic enzymes convert drugs into reactive metabolites that often conjugate to nucleophilic sites within biological macromolecules like proteins. Toxic metabolite-product adducts induce severe immune responses that can cause sometimes fatal disorders, most commonly in the form of liver injury, blood dyscrasia, or the dermatologic conditions toxic epidermal necrolysis and Stevens-Johnson syndrome. This study models four of the most common metabolic transformations that result in bioactivation: quinone formation, epoxidation, thiophene sulfur-oxidation, and nitroaromatic reduction, by synthesizing models of metabolism and reactivity. First, the metabolism models predict the formation probabilities of all possible metabolites among the pathways studied. Second, the exact structures of these metabolites are enumerated. Third, using these structures, the reactivity model predicts the reactivity of each metabolite. Finally, a feedfoward neural network converts the metabolism and reactivity predictions to a bioactivation prediction for each possible metabolite. These bioactivation predictions represent the joint probability that a metabolite forms and that this metabolite subsequently conjugates to protein or glutathione. Among molecules bioactivated by these pathways, we predicted the correct pathway with an AUC accuracy of 89.98%. Furthermore, the model predicts whether molecules will be bioactivated, distinguishing bioactivated and nonbioactivated molecules with 81.06% AUC. We applied this algorithm to withdrawn drugs. The known bioactivation pathways of alclofenac and benzbromarone were identified by the algorithm, and high probability bioactivation pathways not yet confirmed were identified for safrazine, zimelidine, and astemizole. This bioactivation model-the first of its kind that jointly considers both metabolism and reactivity-enables drug candidates to be quickly evaluated for a toxicity risk that often evades detection during preclinical trials. The XenoSite bioactivation model is available at http://swami.wustl.edu/xenosite/p/bioactivation.

A Nag-like dioxygenase initiates 3,4-dichloronitrobenzene degradation via 4,5-dichlorocatechol in Diaphorobacter sp. strain JS3050.

The chemical synthesis intermediate 3,4-dichloronitrobenzene (3,4-DCNB) is an environmental pollutant. Diaphorobacter sp. strain JS3050 utilizes 3,4-DCNB as a sole source of carbon, nitrogen and energy. However, the molecular determinants of its catabolism are poorly understood. Here, the complete genome of strain JS3050 was sequenced and key genes were expressed heterologously to establish the details of its degradation pathway. A chromosome-encoded three-component nitroarene dioxygenase (DcnAaAbAcAd) converted 3,4-DCNB stoichiometrically to 4,5-dichlorocatechol, which was transformed to 3,4-dichloromuconate by a plasmid-borne ring-cleavage chlorocatechol 1,2-dioxygenase (DcnC). On the chromosome, there are also genes encoding enzymes (DcnDEF) responsible for the subsequent transformation of 3,4-dichloromuconate to ß-ketoadipic acid. The fact that the genes responsible for the catabolic pathway are separately located on plasmid and chromosome indicates that recent assembly and ongoing evolution of the genes encoding the pathway is likely. The regiospecificity of 4,5-dichlorocatechol formation from 3,4-DCNB by DcnAaAbAcAd represents a sophisticated evolution of the nitroarene dioxygenase that avoids misrouting of toxic intermediates. The findings enhance the understanding of microbial catabolic diversity during adaptive evolution in response to xenobiotics released into the environment.

Bound detergent molecules in bacterial reaction centers facilitate detection of tetryl explosive.

Bacterial reaction centers (BRC) from Rhodobacter sphaeroides were found to accelerate, about 100-fold, the reaction between tetryl (2,4,6-trinitrophenylmethylnitramine) explosive and n-lauryl-N-N-dimethylamine-N-oxide (LDAO) that results in the formation of picric acid-like product with characteristic UV-VIS absorption spectrum with peaks at 345 and 415 nm. Moreover, this product also affects the spectra of BRC cofactors in the NIR spectral region and stabilizes the conformational changes associated with slow charge recombination. The evolution of the NIR absorption changes correlated with the kinetics of the product formation. Comparison between the wild-type and the R26 carotenoid-less strain indicates that tetryl-LDAO reaction is roughly five times faster for R26, which allows for identifying the carotenoid binding site as the optimal reaction site. Another, less-defined reaction site is located in the BRC's hydrophobic cavity. These effects are highly selective for tetryl and not observed for several other widespread nitric explosives; slowed-down charge recombination allows for distinguishing between tetryl and QB-site herbicides. The current limit of detection is in the ppb range or ~ 100 nM. Details of the molecular mechanisms of the reactions and perspectives of using these effects in bioassays or biosensors for explosives detection are also discussed.

Influence of plant radial oxygen loss in constructed wetland combined with microbial fuel cell on nitrobenzene removal from aqueous solution.

This study investigated the effects of radial oxygen loss (ROL) of three different plants on nitrobenzene (NB) wastewater treatment and bioelectricity generation performance in constructed wetland-microbial fuel cell (CW-MFC). ROL and root biomass from wetland plants showed positive effects on NB wastewater compared to unplanted CW-MFC. Scirpus validus exhibited higher tolerance to NB than Typha orientalis and Iris pseudacorus at 20-200 mg/L NB. As NB concentration reached 200 mg/L, the CW-MFC with Scirpus validus had relatively high DO (2.57 ±â€¯0.17 mg/L) and root biomass (16.42 ±â€¯0.18 g/m2), which resulted in the highest power density and voltage (19.5 mW/m2, 590 mV) as well as NB removal efficiency (93.9 %) among four reactors. High-throughput sequencing results suggested that electrochemically active bacteria (EAB) (e.g., Geobacter, Ferruginibacter) and dominant NB-degrading bacteria (e.g., Comamonas, Pseudomonas) could be enhanced by wetland plants, especially in CW-MFC with Scirpus validus. Therefore, Scirpus validus was a good option for simultaneously treating NB wastewater and producing bioelectricity.

Reduction of nitroarenes by magnetically recoverable nitroreductase immobilized on Fe3O4 nanoparticles.

Enzymes as catalysts have attracted significant attention due to their excellent specificity and incomparable efficiency, but their practical application is limited because these catalysts are difficult to separate and recover. A magnetically recoverable biocatalyst has been effectively prepared through the immobilization of a nitroreductase (oxygen-insensitive, purified from Enterobacter cloacae) onto the Fe3O4 nanoparticles. The magnetic nanoparticles (MNPs) were synthesized by a coprecipitation method in an aqueous system. The surfaces of the MNPs were modified with sodium silicate and chloroacetic acid (CAA). Using 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) through a covalent binding, nitroreductase was loaded onto the modified magnetic carriers through covalent coupling, and thus, a magnetically recoverable biocatalyst was prepared. The free and immobilized nitroreductase activity was also investigated by the reduction of p-nitrobenzonitrile using nicotinamide adenine dinucleotide phosphate (NAPDH) as a cofactor. The activity of the immobilized enzyme was able to maintain 83.23% of that of the free enzyme. The prepared enzyme can easily reduce substituted nitrobenzene to substituted aniline at room temperature and atmospheric pressure, and the yield is up to 60.9%. Most importantly, the loaded nitroreductase carriers can be easily separated and recycled from the reaction system using an externally applied magnetic field. The magnetically recoverable biocatalyst can be recycled and reused 7 times while maintaining high activities and the activity of the magnetic catalyst can be maintained at more than 85.0% of that of the previous cycle. This research solves the recovery problem encountered in industrial applications of biocatalysts and presents a clean and green method of preparing substituted aniline.

Facilitated bioreduction of nitrobenzene by lignite acting as low-cost and efficient electron shuttle.

The searching for efficient and economical redox mediators to promote the treatment of wastewater containing recalcitrant organic compounds is greatly needed. In this study, the redox mediator activities of four different lignite samples to facilitate the bioreduction of nitrobenzene by Shewanella oneidensis MR-1 were tested for the first time. The initial nitrobenzene reduction rate was increased by 40.4%-90.3% in the presence of 50 mg/L of different lignite samples. Lignite collected from Xinjiang (XJL) having more oxygenated groups performed better in enhancing nitrobenzene bioreduction. The stimulating effects increased with the increase of lignite dosage (0-200 mg/L) and the decrease of lignite particle size (150-0.1 µm). However, the pristine XJL samples with assorted sizes of particles exhibited better stimulating effects than size-fractionated ones, implying that different-sized XJL particles might have synergetic effects on the bioreduction process. When humic acid or iron was removed from XJL, its promoting effects were decreased, demonstrating the crucial roles of both components in lignite-enhanced nitrobenzene bioreduction. Nitric acid treatment could form more oxygenated moieties on lignite surface, which played vital roles in promoting nitrobenzene bioreduction. The initial nitrobenzene bioreduction rate in the presence of HNO3-treated XJL was 80.8% higher than that obtained with pristine XJL. This study proposed an effective and readily available redox mediator that could be applied to promote the bioreduction of recalcitrant electrophilic pollutants.

Optimization ofS/Fe ratio for enhanced nitrobenzene biological removal in anaerobicSystem amended withSulfide-modified nanoscale zerovalent iron.

Anaerobic reduction of nitrobenzene (NB) can be efficiently enhanced bySupplementing withSulfide-modified nanoscale zerovalent iron (S-nZVI). In thisStudy,S/Fe ratio ofS-nZVI was further optimized for enhancing biological NB removal in anaerobicSystem amended withS-nZVI and inoculated by anaerobicSludge. The results indicated that the performance andStability of the coupled anaerobicSystem for NB reduction and aniline formation were remarkably improved byS-nZVI atS/Fe molar ratio of 0.3 (0.3S-nZVI). TheSecretion of extracellular polymericSubstances (EPS), transformation of volatile fatty acids (VFAs), yield of methane and activity ofSeveral key enzymes could be efficiently improved by 0.3S-nZVI. Furthermore,Species related to NB reduction, fermentation, electroactivity and methanogenesis could be enriched in 0.3S-nZVI coupled anaerobicSystem, with remarkable improvement in the biodiversity observed. ThisStudy demonstrated thatSulfidation would be a promising method to improve the performance of nZVI in coupled anaerobicSystems for the removal of recalcitrant nitroaromatic compounds from wastewater.

Enhancing nitrobenzene biodegradation in aquatic systems: Feasibility of using plain soil as an inoculant and effects of adding ascorbic acid and peptone.

Nitrobenzene (NB) is recalcitrant to microbial biodegradation due to the electron-deficient character of the nitro group (NO2-). Prior work has found that the reductant could enhance NB biodegradation by providing excess electron donors. However, the existing theory couldn't explain the increase-and-decrease pattern of the NB biodegradation rate with an increase in a reductant concentration. Our results suggest that the reductant affects NB biodegradation by two mechanisms: the available electron donors and the stimulation or inhibition of biomass growth, which are linked by a pseudo-first-order reaction kinetics. In addition, the results showed that directly inoculating the plain soil into the aquatic system and then allowing the synergistic effect of the organic reductant (ascorbic acid) and the substrate (peptone) enhance NB biodegradation. Employing the new method, 200 mg L-1 NB was transformed in 72 h. GC-MS analysis detected two novel intermediate metabolites, indicating that NB was degraded into aniline and further transformed into acetanilide and 9-octadecenamide before its mineralization. This study sheds light on how to exploit the synergistic effects of the availability of excess electron donors and biomass growth by controlling the reductant and a substrate in the right concentration range (e.g., ascorbic acid < 0.8 mgL-1 + peptone).