Impact analysis of hydraulic residence time and dissolved oxygen on performance efficiency and microbial community in N, N-dimethylformamide wastewater treated by an AnSBR-ASBR.

Suitable operating parameters are one of the key factors to efficient and stable biological wastewater treatment of N, N-dimethylformamide (DMF) wastewater. In this study, an improved AnSBR-ASBR reactor (anaerobic sequencing batch reactor, AnSBR, and aerobic SBR, ASBR, run in series) was used to investigated the effects of operating conditions such as hydraulic residence time (HRT), AnSBR stirring speed and ASBR dissolved oxygen (DO) for DMF wastewater treatment. When HRT decreased from 24 h to 12 h, the average removal rates of COD by the AnSBR were 34.59% and 39.54%, respectively. Meanwhile, the removal rate of NH4+-N by ASBR decreased from 88.38% to 62.81%. The DMF removal rate reached the best at 18 h and the expression of dehydrogenase was the highest in the AnSBR. The abundance of Megasphaera, the dominant sugar-degrading bacteria in the AnSBR, continued to decline due to the decrease of HRT. The relative abundance of Methanobacterium gradually increased to 80.2% with the decrease of HRT and that hydrotrophic methanogenesis dominated the methanogenic process. The HRT decrease promoted butyrate and pyruvate metabolism in anaerobic sludge, but the proportion of glycolysis and methane metabolism decreased. The AnSBR-ASBR reactor had the best operation performance when HRT was 18 h, AnSBR speed was 220 r/min, and ASBR DO content was 3-4 mg/L. This study provided an effective reference for the reasonable selection of operating parameters in the treatment of DMF-containing wastewater by the AnSBR-ASBR.

DMF mineralization and substrate specificity mechanism of Aminobacter ciceronei DMFA1.

N,N-dimethylformamide (DMF) is widely used in various industries, but its direct release into water poses high risks to human beings. Although a lot of DMF-degrading bacteria has been isolated, limited studies focus on the degradation preference among DMF and its analogues. In this study, an efficient DMF mineralization bacterium designated Aminobacter ciceronei DMFA1 was isolated from marine sediment. When exposed to a 0.2% DMF (∼1900 mg/L), strain DMFA1 exhibited a degradation efficiency of 100% within 4 days. The observed growth using formamide as the sole carbon source implied the possible DMF degradation pathway of strain DMFA1. Meanwhile,the strain DMFA1 possesses a broad-spectrum substrate degradation, which could effectively degraded 0.2% N,N-dimethylacetamide (DMAC) and N-methylformamide (NMF). Genomic analysis further confirmed the supposed pathway through annotating the genes encoding N, N-dimethylformamidase (DMFase), formamidase, and formate dehydrogenase. The existence of sole DMFase indicating its substrate specificity controlled the preference of DMAc of strain DMFA1. By integrating multiple sequence alignment, homology modeling and molecular docking, the preference of the DMFase in strain DMFA1 towards DMAc are related to: 1) Mutations in key active site residues; 2) the absence of small subunit; and 3) no energy barrier for substrates entering the active site.

Allyl methyl disulfide (AMDS) prevents N,N-dimethyl formamide-induced liver damage by suppressing oxidative stress and NLRP3 inflammasome activation.

N,N-dimethylformamide (DMF), a widely consumed industrial solvent with persistent characteristics, can induce occupational liver damage and pose threats to the general population due to the enormous DMF-containing industrial efflux and emission from indoor facilities. This study was performed to explore the roles of allyl methyl disulfide (AMDS) in liver damage induced by DMF and the underlying mechanisms. AMDS was found to effectively suppress the elevation in the liver weight/body weight ratio and serum aminotransferase activities, and reduce the mortality of mice induced by DMF. In addition, AMDS abrogated DMF-elicited increases in malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) levels and decreases in glutathione (GSH) levels in mouse livers. The increase in macrophage number, mRNA expression of M1 macrophage biomarkers, and protein expression of key components in the NF-κB pathway and NLRP3 inflammasome induced by DMF exposure were all suppressed by AMDS in mouse livers. Furthermore, AMDS inhibited DMF-induced cell damage and NF-κB activation in cocultured AML12 hepatocytes and J774A.1 macrophages. However, AMDS per se did not significantly affect the protein level and activity of CYP2E1. Collectively, these results demonstrate that AMDS effectively ameliorates DMF-induced acute liver damage possibly by suppressing oxidative stress and inactivating the NF-κB pathway and NLRP3 inflammasome.

Insight into the metabolic pathways of Paracoccus sp. strain DMF: a non-marine halotolerant methylotroph capable of degrading aliphatic amines/amides.

Paracoccus sp. strain DMF (P. DMF from henceforth) is a gram-negative heterotroph known to tolerate and utilize high concentrations of N,N-dimethylformamide (DMF). The work presented here elaborates on the metabolic pathways involved in the degradation of C1 compounds, many of which are well-known pollutants and toxic to the environment. Investigations on microbial growth and detection of metabolic intermediates corroborate the outcome of the functional genome analysis. Several classes of C1 compounds, such as methanol, methylated amines, aliphatic amides, and naturally occurring quaternary amines like glycine betaine, were tested as growth substrates. The detailed growth and kinetic parameter analyses reveal that P. DMF can efficiently aerobically degrade trimethylamine (TMA) and grow on quaternary amines such as glycine betaine. The results show that the mechanism for halotolerant adaptation in the presence of glycine betaine is dissimilar from those observed for conventional trehalose-mediated halotolerance in heterotrophic bacteria. In addition, a close genomic survey revealed the presence of a Co(I)-based substrate-specific corrinoid methyltransferase operon, referred to as mtgBC. This demethylation system has been associated with glycine betaine catabolism in anaerobic methanogens and is unknown in denitrifying aerobic heterotrophs. This report on an anoxic-specific demethylation system in an aerobic heterotroph is unique. Our finding exposes the metabolic potential for the degradation of a variety of C1 compounds by P. DMF, making it a novel organism of choice for remediating a wide range of possible environmental contaminants.

Biosynthesis of poly(3-hydroxybutyrate) by N,N-dimethylformamide degrading strain Paracoccus sp. PXZ: A strategy for resource utilization of pollutants.

N,N-dimethylformamide is a toxic chemical solvent, which widely exists in industrial wastewater. Nevertheless, the relevant methods merely achieved non-hazardous treatment of N,N-dimethylformamide. In this study, one efficient N,N-dimethylformamide degrading strain was isolated and developed for pollutant removal coupling with poly(3-hydroxybutyrate) (PHB) accumulation. The functional host was characterized as Paracoccus sp. PXZ, which could consume N,N-dimethylformamide as the nutrient substrate for cell reproduction. Whole-genome sequencing analysis confirmed that PXZ simultaneously possesses the essential genes for poly(3-hydroxybutyrate) synthesis. Subsequently, the approaches of nutrient supplementation and various physicochemical variables to strengthen poly(3-hydroxybutyrate) production were investigated. The optimal biopolymer concentration was 2.74 g·L-1 with a poly(3-hydroxybutyrate) proportion of 61%, showing a yield of 0.29 g-PHB·g-1-fructose. Furthermore, N,N-dimethylformamide served as the special nitrogen matter that could realize a similar poly(3-hydroxybutyrate) accumulation. This study provided a fermentation technology coupling with N,N-dimethylformamide degradation, offering a new strategy for resource utilization of specific pollutants and wastewater treatment.

Metabolic Engineering for the Comprehensive Utilization of N,N-Dimethylformamide-Containing Wastewater.

N, N-dimethylformamide is frequently present in industrial wastewater and is environmentally detrimental. The current study aims to assess the utilization and biodegradation of N, N-dimethylformamide-containing wastewater to lessen the associated environmental load. Results show that addition of wastewater containing N, N-dimethylformamide to Trichoderma reesei fermentation media enhances cellulase production and facilitates cellulose hydrolysis. However, N, N-dimethylformamide is a cellulase enhancer that is not degraded during cellulase production in T. reesei fermentation and is retained in the N, N-dimethylformamide-enhanced cellulase solution. Indeed, the cellulosic sugar solution generated via lignocellulose hydrolysis with N, N-dimethylformamide-enhanced cellulase retains N, N-dimethylformamide. We further identified three core enzyme modules─N, N-dimethylformamidase, dimethylamine dehydrogenase, and methylamine dehydrogenase enzyme─which were inserted into Escherichia coli to develop metabolically engineered strains. These strains degraded N, N-dimethylformamide and produced succinate using N, N-dimethylformamide-enhanced cellulosic sugar as the substrate. The platform described here can be applied to effectively convert waste into valuable bioproducts.

Efficient bioremediation of PAHs-contaminated soils by a methylotrophic enrichment culture.

Bioaugmentation effectively enhances microbial bioremediation of hazardous polycyclic aromatic hydrocarbons (PAHs) from contaminated environments. While screening for pyrene-degrading bacteria from a former manufactured gas plant soil (MGPS), the mixed enrichment culture was found to be more efficient in PAHs biodegradation than the culturable pure strains. Interestingly, analysis of 16S rRNA sequences revealed that the culture was dominated by a previously uncultured member of the family Rhizobiaceae. The culture utilized C1 and other methylotrophic substrates, including dimethylformamide (DMF), which was used as a solvent for supplementing the culture medium with PAHs. In the liquid medium, the culture rapidly degraded phenanthrene, pyrene, and the carcinogenic benzo(a)pyrene (BaP), when provided as the sole carbon source or with DMF as a co-substrate. The efficiency of the culture in the bioremediation of PAHs from the MGPS and a laboratory waste soil (LWS) was evaluated in bench-scale slurry systems. After 28 days, 80% of Σ16 PAHs were efficiently removed from the inoculated MGPS. Notably, the bioaugmentation achieved 90% removal of four-ringed and 60% of highly recalcitrant five- and six-ringed PAHs from the MGPS. Likewise, almost all phenanthrene, pyrene, and 65% BaP were removed from the bioaugmented LWS. This study highlights the application of the methylotrophic enrichment culture dominated by an uncultured bacterium for the efficient bioremediation of PAHs.

Carbon-neutral treatment of N, N-dimethylformamide-containing industrial wastewater by anaerobic membrane bioreactor (AnMBR): Bio-energy recovery and CO2 emission reduction.

High-strength industrial wastewater containing approximately 2000 mg/L of N, N-dimethylformamide (DMF) was treated by the anaerobic membrane bioreactor (AnMBR) during a long-term operation with the concept of carbon neutrality in this study. Bio-methane was recovered as bio-energy or bio-resource from DMF-containing wastewater along with the CO2 emission reduction. The results are clear evidence of the feasibility of carbon-neutral treatment of DMF-containing wastewater by the AnMBR. With an effective degradation under the organic loading rate of 6.53 COD kg/m3/d at the HRT of 12 h, the AnMBR completely covered the energy consumption during long-term operation by saving electricity of 4.16 kWh/m3 compared with the conventional activated sludge process. The CO2 emission of the AnMBR was just 1.06 kg/m3, remarkably reducing 1.45 kg/m3 of CO2. The treatment of DMF-containing wastewater by the AnMBR perfectly realized the goal of carbon neutrality, and was considered as an alternative to the conventional activated sludge process.

Exposure of mouse oocytes to N,N-dimethylformamide impairs mitochondrial functions and reduces oocyte quality.

N,N-dimethylformamide (DMF) is a widely-used solvent for the synthesis of synthetic fibers such as polyacrylonitrile fiber, and can also be used to make medicine. Although this organic solvent has multipurpose applications, its biological toxicity cannot be ignored and its impact on mammalian reproduction remains largely unexplored. Our study found that DMF exposure inhibited oocyte maturation and fertilization ability. Transcriptomic analysis indicated that DMF exposure changed the expression of genes and transposable elements in oocytes. Subcellular structure examination found that DMF exposure caused mitochondrial dysfunction, abnormal aggregation of mitochondria and decreased mitochondrial membrane potential in mouse oocytes. Its exposure also caused abnormal distribution of Golgi apparatus and endoplasmic reticulum which formed large number of clusters. In addition, oxidative stress occurs in oocytes exposed to DMF, which was manifested by an increase in the level of reactive oxygen species. We found that DMF exposure induced disordered spindle and chromosomes abnormality. Meanwhile, we examined various histone modification levels in oocytes exposed to DMF and found that DMF exposure reduced H3K9me3, H3K9ac, H3K27ac, and H4K16ac levels in mouse oocytes. Moreover, DMF-treated oocytes failed to form pronuclei after fusion with normal sperm. Collectively, DMF exposure caused mitochondrial damage, oxidative stress, spindle assembly and chromosome arrangement disorder, leading to oocyte maturation arrest and fertilization failure.

Temperature and solvent exposure response of three fatty acid peroxygenase enzymes for application in industrial enzyme processes.

Free fatty acids (FFAs) are a useful feedstock for a range of industrial chemical synthesis applications. However, efficiently converting FFAs to molecules for biofuel and other high-value chemicals requires more efficient and cost-effective catalysts. Cytochrome P450 fatty acid peroxygenases (CYP152) have a unique chemistry that allows use of the peroxide shunt pathway for biochemical conversion of FFAs. Known CYP152s are heat labile, however, requiring characterization of more thermotolerant versions for use in industrial applications. A fatty acid peroxygenase from Bacillus methanolicus (CYP152K6) was shown here to have a higher optimal reaction temperature than OleT (CYP152L1). CYP152K6 was stable up to 50 °C and showed great stability in 3% acetone and dimethylformamide. Stability in solvents helps the enzyme's substrates remain soluble in solution for more efficient catalysis, and heat stability allows enzymes to remain active longer during industrial processes.

A 2-Tyr-1-carboxylate Mononuclear Iron Center Forms the Active Site of a Paracoccus Dimethylformamidase.

N,N-dimethyl formamide (DMF) is an extensively used organic solvent but is also a potent pollutant. Certain bacterial species from genera such as Paracoccus, Pseudomonas, and Alcaligenes have evolved to use DMF as a sole carbon and nitrogen source for growth via degradation by a dimethylformamidase (DMFase). We show that DMFase from Paracoccus sp. strain DMF is a halophilic and thermostable enzyme comprising a multimeric complex of the α2 ß2 or (α2 ß2 )2 type. One of the three domains of the large subunit and the small subunit are hitherto undescribed protein folds of unknown evolutionary origin. The active site consists of a mononuclear iron coordinated by two Tyr side-chain phenolates and one carboxylate from Glu. The Fe3+ ion in the active site catalyzes the hydrolytic cleavage of the amide bond in DMF. Kinetic characterization reveals that the enzyme shows cooperativity between subunits, and mutagenesis and structural data provide clues to the catalytic mechanism.

A comparative long-term operation using up-flow anaerobic sludge blanket (UASB) and anaerobic membrane bioreactor (AnMBR) for the upgrading of anaerobic treatment of N, N-dimethylformamide-containing wastewater.

Synthetic industrial wastewater containing approximately 2000 mg/L N, N-dimethylformamide (DMF) was treated using a lab-scale anaerobic sludge blanket (UASB) and an anaerobic membrane bioreactor (AnMBR) in this study. The inoculum consisted of two sources of sludge: Co-culture of anaerobic digested sludge (ADS) with DMF-hydrolyzing activated sludge (DAS) for the AnMBR, and co-culture of anaerobic granular sludge (AGS) with DAS for the UASB. Effective DMF methanogenic degradation of nearly 100% removal was achieved in both reactors on the first day. Both reactors obtained excellent DMF removal efficiency and high methane production under a low organic loading rate (OLR) of around 3-4 g COD/L/d. However, excessive elevation of OLR significantly limited DMF hydrolysis. When OLR exceeded 6 g COD/L/d, the removal efficiency and methane production in both reactors dramatically dropped. Despite their different forms and shapes, the ADS and AGS both provide methanogens which are responsible for methanogenesis. The UASB tolerated a higher OLR while the AnMBR was limited by membrane fouling due to the increased sludge concentration. However, the AnMBR obtained high-quality effluent without suspended solid. Whether DMF can be effectively degraded depends on DAS, in which abundant DMF-hydrolyzing bacteria (DHB) provide sufficient quantities of the hydrolytic enzyme for effective hydrolysis of DMF. However, these DHB were facultative and were also identified as denitrifying bacteria which require nitrate as the electron acceptor or otherwise survive under the aerobic condition. They gradually decayed rather than proliferated and were outcompeted by methanogens. Therefore, it is conceivable that a slight dosage of nitrate would enrich the abundance of DHB in both the UASB and the AnMBR, and provide a sufficient quantity of enzymes for the DMF hydrolysis. The cultivation of the anaerobic DMF-degrading granular sludge using the UASB is considered an upgraded solution to the effective treatment of DMF-containing wastewater.

The essential role of CYP2E1 in metabolism and hepatotoxicity of N,N-dimethylformamide using a novel Cyp2e1 knockout mouse model and a population study.

N,N-Dimethylformamide (DMF) is a widespread contaminant of leather factories and their surrounding environment. There is a lack of direct in vivo evidence supporting CYP2E1 as a primary enzyme responsible for DMF metabolism and hepatotoxicity. In this study, a novel Cyp2e1 knockout (KO) mouse model was generated and used to assess whether DMF metabolism and hepatotoxicity is CYP2E1 dependent using an acute toxicity protocol with a single dose of 1500 mg DMF/kg. An epidemiological study in 698 DMF-exposed workers and 188 non-DMF-exposed controls was conducted to investigate the associations between functional polymorphisms of CYP2E1 (rs6413432/rs2031920) and DMF metabolite (N-methylcarbmoylated-hemoglobin [NMHb]). We successfully established Cyp2e1 KO mice with evidence from DNA sequence analysis, which showed 1-bp insertion at 65 bp (C) site of Cyp2e1 Exon 1. In addition, western blot and in vivo pharmacokinetic study also showed a complete absence of CYP2E1 protein and a 92% and 88% reduction in CYP2E1 activity among males and females, respectively. DMF metabolism as evidenced by increased blood NMHb, and hepatotoxicity as evidenced by elevated liver/body weight ratio, activity of liver enzymes and massive liver necrosis were detected in wild-type (WT) mice but were completely abrogated in KO mice, strongly supporting a CYP2E1-dependent pattern of DMF metabolism and hepatotoxicity. Moreover, variant allele of CYP2E1-rs6413432 was also significantly associated with higher NMHb levels in DMF-exposed workers (P = 0.045). The increase of glucose-regulated protein 94 detected in WT mice but not in KO mice suggested CYP2E1-dependent endoplasmic reticulum stress may be a key mechanism underlying DMF-induced hepatotoxicity.

Capacity of an in vitro rat skin model to predict human dermal absorption: Influences of aging and anatomical site.

In addition to inhalation, dermal absorption is a route of exposure to be considered when assessing occupational risks. To investigate dermal penetration of chemicals, human skin samples are regarded as the gold standard. As human samples can be difficult to obtain, many experiments are performed with rat skins, and the results extrapolated to describe human percutaneous absorption. Here, we examined the characteristics of rat skin samples and compared absorption to that measured with in vitro human skin. The thickness of the stratum corneum layer in rat skin samples was found to be uniform when samples were excised from the animals' backs once they were at least 7-weeks-old. Overall, dorsal skin samples from mature rats could be reliably used to measure the flux of hydrophilic liquid molecules, such as N-methyl-2-pyrrolidone, and N,N-dimethylformamide. In contrast, with a solid lipophilic substance, bisphenol A, dissolved in acetone, the flux obtained with rat skin samples was 3-fold higher than that measured with human skin. Consequently, it does not appear relevant to use rat skin in place of human skin to measure absorption of solid lipophilic substances.

Molecular Mechanism of N,N-Dimethylformamide Degradation in Methylobacterium sp. Strain DM1.

N,N-Dimethylformamide (DMF) is one of the most common xenobiotic chemicals, and it can be easily emitted into the environment, where it causes harm to human beings. Herein, an efficient DMF-degrading strain, DM1, was isolated and identified as Methylobacterium sp. This strain can use DMF as the sole source of carbon and nitrogen. Whole-genome sequencing of strain DM1 revealed that it has a 5.66-Mbp chromosome and a 200-kbp megaplasmid. The plasmid pLVM1 specifically harbors the genes essential for the initial steps of DMF degradation, and the chromosome carries the genes facilitating subsequent methylotrophic metabolism. Through analysis of the transcriptome sequencing data, the complete mineralization pathway and redundant gene clusters of DMF degradation were elucidated. The dimethylformamidase (DMFase) gene was heterologously expressed, and DMFase was purified and characterized. Plasmid pLVM1 is catabolically crucial for DMF utilization, as evidenced by the phenotype identification of the plasmid-free strain. This study systematically elucidates the molecular mechanisms of DMF degradation by MethylobacteriumIMPORTANCE DMF is a hazardous pollutant that has been used in the chemical industry, pharmaceutical manufacturing, and agriculture. Biodegradation as a method for removing DMF has received increasing attention. Here, we identified an efficient DMF degrader, Methylobacterium sp. strain DM1, and characterized the complete DMF mineralization pathway and enzymatic properties of DMFase in this strain. This study provides insights into the molecular mechanisms and evolutionary advantage of DMF degradation facilitated by plasmid pLVM1 and redundant genes in strain DM1, suggesting the emergence of new ecotypes of Methylobacterium.

Dissection and characterization of the prokaryotic community during the long-term operation of a submerged anaerobic membrane bioreactor for the anaerobic treatment of N, N-dimethylformamide-containing wastewater with a co-cultured inoculum.

A submerged anaerobic membrane bioreactor (SAnMBR) was operated for the anaerobic treatment of wastewater containing approximately 2000 mg L-1N, N-dimethylformamide (DMF). Inoculated with a co-cultured inoculum, the SAnMBR obtained an excellent DMF removal under a low organic loading rate (OLR) of 3.14-4.16 g COD L-1 d-1. However, the elevation of OLR limited hydrolysis. While the co-cultured inoculum initially contains abundant DMF-hydrolyzing bacteria with potential to hydrolyze DMF into intermediates, such as Paracoccus, Hyphomicrobium, Burkholderia, Catellibacterium, Bacillus and Bradyrhizobium, since these bacteria are facultative anaerobes which survive anaerobically, they kept decaying rather than proliferating, resulting in the weakening of the DMF-hydrolyzing ability. Each re-inoculation of new sludge only temporarily revitalized hydrolysis activity for a short period. Due to the lack of nitrate, these bacteria were unable to proliferate. This suggests that a small dose of nitrate would help to enrich these bacteria and establish a stable DMF-degrading consortium.

Sensorineural hearing loss and volatile organic compound metabolites in urine.

PURPOSE: Oxidative stress in the auditory system contributes to acquired sensorineural hearing loss. Systemic oxidative stress, which may predict auditory oxidative stress, can be assessed by measuring volatile organic compound metabolite concentrations in urine. The purpose of this retrospective study was to determine if hearing decreased in those with higher concentrations of urinary volatile organic compound metabolites. MATERIALS AND METHODS: Audiometric, demographic, and metabolite concentration data were downloaded from the 2011-2012 cycle of the U.S. National Health and Nutritional Examination Survey. Participants were first grouped by reported noise exposure. For each metabolite, an analysis of covariance was used to look for differences in age-adjusted hearing loss among urinary volatile organic compound metabolite concentration groups. Participants were grouped into quartiles based on concentration for each metabolite separately because many individuals were at the lower limit of concentration detection for several metabolites, leading to a non-normal distribution. RESULTS: Age-adjusted high-frequency pure-tone thresholds were significantly (FDR < 0.05) increased by about 3 to 4 dB in high concentration quartile groups for five metabolites. All five metabolites were glutathione-dependent mercapturic acids. The parent compounds of these metabolites included acrylonitrile, 1,3 butadiene, styrene, acrylamide, and N,N-dimethylformamide. Significant associations were only found in those with no reported noise exposure. CONCLUSIONS: Urinary metabolites may help to explain susceptibility to oxidative stress-induced hearing loss.

Anaerobic treatment of N,N-dimethylformamide-containing high-strength wastewater by submerged anaerobic membrane bioreactor with a co-cultured inoculum.

The anaerobic treatment of wastewater containing approximately 2000 mg L-1N,N-dimethylformamide (DMF) was conducted by a lab-scale submerged anaerobic membrane bioreactor (SAnMBR). The inoculum consisted of aerobic DMF-hydrolyzing activated sludge (DAS) and anaerobic digested sludge (ADS). A rapid start-up was achieved with thorough DMF methanogenic degradation on the first day. The results of a 250-day long-term experiment demonstrated that under a low organic loading rate (OLR) of 3.14-4.16 g COD L-1 d-1, SAnMBR maintained excellent DMF removal efficiency along with high methane conversion. However, the elevation of OLR significantly limited DMF hydrolysis. When OLR exceeded 6.54 g COD L-1 d-1, both removal efficiency and methane production dramatically dropped. The DMF-hydrolyzing bacteria originating from the DAS gradually decayed under the anaerobic condition, resulting in the weak hydrolysis of DMF. The shortening of hydraulic retention time (HRT) is not recommended for the SAnMBR because severe membrane fouling occurred when HRT was shortened to 8 h. To handle high OLRs, an appropriate solution is to maintain a low F/M ratio by increasing both the influent DMF concentration and sludge concentration. The high CH4 content in the biogas, exceeding 85%, was shown to be the reason for the suitability of anaerobic treatment to DMF. Some improvements which would help to maintain the effective hydrolysis are proposed: a side-stream system to replenish DAS to the SAnMBR is helpful; slight dosage of nitrate could also help to enrich the DMF-hydrolyzing bacteria; and the co-digestion of DMF and other organics might be convenient to establish a stable DMF-degrading consortium.

Insights into the methanogenic degradation of N, N-dimethylformamide: The functional microorganisms and their ecological relationships.

The methanogenic degradation of N, N-dimethylformamide (DMF) was investigated using anaerobic digested sludge (ADS), aerobic activated sludge (AAS) and co-cultured sludge (CCS), respectively. Both the metabolic pathway and the corresponding microorganisms which function in the methanogenic degradation of DMF were elucidated. DMF was unable to be degraded anaerobically by ADS due to the lack of DMF-hydrolyzing bacteria. DMF can be effectively degraded by AAS, however, no methane was recovered under the aerobic condition. The co-culture of DMF-hydrolyzing bacteria and methanogens in the CCS allowed for both hydrolysis of DMF and methane production to proceed successfully under the anaerobic condition, realizing the complete conversion from DMF to methane. However, a niche overlap due to the competition for the intermediates lowered the abundance of DMF-hydrolyzing bacteria. The introduction of nitrate, timely replenishment of AAS, micro-aeration and co-digestion were likely to maintain a high abundance of DMF-hydrolyzing bacteria to ensure an effective hydrolysis.

Study on the potential way of hepatic cytotoxicity of N,N-dimethylformamide.

The intermediate metabolites and redox status imbalance were supported as the two major points for N,N-dimethylformamide (DMF)-induced hepatotoxicity. However, the potential mechanism has not yet been concerned. By applying two inhibitors, this study tried to seek the major role in DMF-induced toxicity on HL7702 cell. We observed that DMF induced cell apoptosis through mitochondrial-dependent and p53 pathway. Inhibition reactive oxygen species by catalase remarkably attenuated the mitochondrial transmembrane potential (MMP), apoptotic proteins, and apoptosis. On the contrary, it reduced the biodegradation rate of DMF by coincubation with CYP2E1 antagonist (DDC) partially reduced late apoptosis. However, the change in MMP, the ratio of Bax to Bcl-xl, and cleaved-caspase 9 was not attenuated by DDC. The pathway in DDC coincubation groups was related to the p53 rather than the mitochondrial pathway. Restoring the redox balance during biodegradation is much more effective than attenuating the metabolite rate of DMF. This study may provide a suitable prevention method to occupational workers.