Pretreatment for potable reuse: Enhancing the biological removal of 1,4-dioxane from landfill leachate through cometabolism with tetrahydrofuran.

1,4-Dioxane is a probable human carcinogen and a persistent aquatic contaminant. Cometabolic biodegradation of 1,4-dioxane is a promising low-cost and effective treatment technology; however, further demonstration is needed for treating landfill leachate. This technology was tested in two full-scale moving bed biofilm reactors (MBBRs) treating raw landfill leachate with tetrahydrofuran selected as the cometabolite. The raw leachate contained on average 82 µg/L of 1,4-dioxane and before testing the MBBRs removed an average of 38% and 42% of 1,4-dioxane, respectively. First, tetrahydrofuran was added to MBBR 1, and 1,4-dioxane removal was improved to an average of 73%, with the control MBBR removing an average of 37% of 1,4-dioxane. During this period, an optimal dose of 2 mg/L of tetrahydrofuran was identified. Tetrahydrofuran was then fed to both MBBRs, where the 1,4-dioxane removal was on average 73% and 80%. Cometabolic treatment at the landfill significantly reduced the concentration of 1,4-dioxane received from the landfill at a downstream wastewater treatment and indirect potable reuse facility, reducing the load of 1,4-dioxane from 44% to 24% after the study. PRACTITIONER POINTS: Cometabolic degradation of leachate 1,4-dioxane with THF in MBBRs is a feasible treatment technology and a low-cost technique when retrofitting existing biological treatment facilities. The MBBRs can be operated at a range of temperatures, require no operational changes beyond THF addition, and operate best at a mass ratio of THF to 1,4-dioxane of 24. Source control of 1,4-dioxane significantly reduces the concentration of 1,4-dioxane in downstream wastewater treatment plants and potable reuse facilities.

Synergistic interactions in core microbiome Rhizobiales accelerate 1,4-dioxane biodegradation.

Next-generation sequencing (NGS) has revolutionized taxa identification within contaminant-degrading communities. However, uncovering a core degrading microbiome in diverse polluted environments and understanding its associated microbial interactions remains challenging. In this study, we isolated two distinct microbial consortia, namely MA-S and Cl-G, from separate environmental samples using 1,4-dioxane as a target pollutant. Both consortia exhibited a persistent prevalence of the phylum Proteobacteria, especially within the order Rhizobiales. Extensive analysis confirmed that Rhizobiales as the dominant microbial population (> 90 %) across successive degradation cycles, constituting the core degrading microbiome. Co-occurrence network analysis highlighted synergistic interactions within Rhizobiales, especially within the Shinella and Xanthobacter genera, facilitating efficient 1,4-dioxane degradation. The enrichment of Rhizobiales correlated with an increased abundance of essential genes such as PobA, HpaB, ADH, and ALDH. Shinella yambaruensis emerged as a key degrader in both consortia, identified through whole-genome sequencing and RNA-seq analysis, revealing genes implicated in 1,4-dioxane degradation pathways, such as PobA and HpaB. Direct and indirect co-cultivation experiments confirmed synergistic interaction between Shinella sp. and Xanthobacter sp., enhancing the degradation of 1,4-dioxane within the core microbiome Rhizobiales. Our findings advocate for integrating the core microbiome concept into engineered consortia to optimize 1,4-dioxane bioremediation strategies.

Isolation and characterization of pure cultures for metabolizing 1,4-dioxane in oligotrophic environments.

1,4-Dioxane concentration in most contaminated water is much less than 1 mg/L, which cannot sustain the growth of most reported 1,4-dioxane-metabolizing pure cultures. These pure cultures were isolated following enrichment of mixed cultures at high concentrations (20 to 1,000 mg/L). This study is based on a different strategy: 1,4-dioxane-metabolizing mixed cultures were enriched by periodically spiking 1,4-dioxane at low concentrations (≤1 mg/L). Five 1,4-dioxane-metabolizing pure strains LCD6B, LCD6D, WC10G, WCD6H, and WD4H were isolated and characterized. The partial 16S rRNA gene sequencing showed that the five bacterial strains were related to Dokdonella sp. (98.3%), Acinetobacter sp. (99.0%), Afipia sp. (99.2%), Nitrobacter sp. (97.9%), and Pseudonocardia sp. (99.4%), respectively. Nitrobacter sp. WCD6H is the first reported 1,4-dioxane-metabolizing bacterium in the genus of Nitrobacter. The net specific growth rates of these five cultures are consistently higher than those reported in the literature at 1,4-dioxane concentrations <0.5 mg/L. Compared to the literature, our newly discovered strains have lower half-maximum-rate concentrations (1.8 to 8.2 mg-dioxane/L), lower maximum specific 1,4-dioxane utilization rates (0.24 to 0.47 mg-dioxane/(mg-protein ⋅ d)), higher biomass yields (0.29 to 0.38 mg-protein/mg-dioxane), and lower decay coefficients (0.01 to 0.02 d-1). These are characteristics of microorganisms living in oligotrophic environments.

Dual phytoremediation and biochar production by Eichhornia crassipes in hydroponic system receiving different 1,4-dioxane dosages.

This study focuses on applying phytoremediation as a low-effective and simple process to treat wastewater laden with 1,4 dioxane (DIOX). A floating macrophyte (Eichhornia crassipes) was cultivated under hydroponic conditions (relative humidity 50-67%, photoperiod cycle 18:6 h light/dark, and 28-33 °C) and subjected to different DIOX loads between 0.0 (control) and 11.5 mg/g fresh mass (FM). The aquatic plant achieved DIOX and chemical oxygen demand (COD) removal efficiencies of 76-96% and 67-94%, respectively, within 15 days. E. crassipes could tolerate elevated DIOX-associated stresses until a dose of 8.2 mg DIOX/g, which highly influenced the oxidative defense system. Malondialdehyde (MDA) content, hydrogen peroxide (H2O2), and total phenolic compounds (TPC) increased by 7.3, 8.4, and 4.5-times, respectively, in response to operating the phytoremediation unit at a DIOX load of 11.5 mg/g. The associated succulent value, proteins, chlorophyll-a, chlorophyll-b, and pigments dropped by 39.6%, 45.8%, 51.5%, 80.8%, and 55.5%, respectively. The suggested removal mechanism of DIOX by E. crassipes could be uptake followed by phytovolatilization, whereas direct photodegradation from sunlight contributed to about 19.36% of the total DIOX removal efficiencies. Recycling the exhausted E. crassipes for biochar production was a cost-efficient strategy, making the payback period of the phytoremediation project equals to 6.96 yr.

Eichhornia crassipes could be used in phytoremediation of 1,4 dioxane (DIOX)-laden water at DIOX load< 8.2 mg/g FM. E. crassipes removed 77­97% DIOX via uptake and phytovolatilization. Recycling exhausted-plant to produce biochar was cost-efficient with 7 yr-payback period.

Self-assembling a 1,4-dioxane-degrading consortium and identifying the key role of Shinella sp. through dilution-to-extinction and reculturing.

IMPORTANCE: Assembling a functional microbial consortium and identifying key degraders involved in the degradation of 1,4-dioxane are crucial for the design of synergistic consortia used in enhancing the bioremediation of 1,4-dioxane-contaminated sites. However, due to the vast diversity of microbes, assembling a functional consortium and identifying novel degraders through a simple method remain a challenge. In this study, we reassembled 1,4-dioxane-degrading microbial consortia using a simple and easy-to-operate method by combining dilution-to-extinction and reculture techniques. We combined differential analysis of community structure and metabolic function and confirmed that Shinella species have a stronger 1,4-dioxane degradation ability than Xanthobacter species in the enriched consortium. In addition, a new dioxane-degrading bacterium was isolated, Shinella yambaruensis, which verified our findings. These results demonstrate that DTE and reculture techniques can be used beyond diversity reduction to assemble functional microbial communities, particularly to identify key degraders in contaminant-degrading consortia.

1,4-Dioxane removal in nitrifying sand filters treating domestic wastewater: Influence of water matrix and microbial inhibitors.

1,4-Dioxane is a recalcitrant pollutant in water and is ineffectively removed during conventional water and wastewater treatment processes. In this study, we demonstrate the application of nitrifying sand filters to remove 1,4-dioxane from domestic wastewater without the need for bioaugmentation or biostimulation. The sand columns were able to remove 61 ± 10% of 1,4-dioxane on average (initial concentration: 50 µg/L) from wastewater, outperforming conventional wastewater treatment approaches. Microbial analysis revealed the presence of 1,4-dioxane degrading functional genes (dxmB, phe, mmox, and prmA) to support biodegradation being the dominant degradation pathway. Adding antibiotics (sulfamethoxazole and ciprofloxacin), that temporarily inhibited the nitrification process during the dosing period, showed a minor effect in 1,4-dioxane removal (6-8% decline, p < 0.05), suggesting solid resilience of the 1,4-dioxane-degrading microbial community in the columns. Columns amended with sodium azide significantly (p < 0.05) depressed 1,4-dioxane removal in the early stage of dosing but followed by a gradual increase of the removal over time to >80%, presumably due to a shift in the microbial community toward azide-resistant 1,4-dioxane degrading microbes (e.g., fungi). This study demonstrated for the first time the resilience of the 1,4-dioxane-degrading microorganisms during antibiotic shocks, and the selective enrichment of efficient 1,4-dioxane-degrading microbes after azide poisoning. Our observation could provide insights into designing better 1,4-dioxane remediation strategies in the future.

Characterization of 1,4-dioxane degrading microbial community enriched from uncontaminated soil.

1,4-Dioxane is a contaminant of emerging concern that has been commonly detected in groundwater. In this study, a stable and robust 1,4-dioxane degrading enrichment culture was obtained from uncontaminated soil. The enrichment was capable to metabolically degrade 1,4-dioxane at both high (100 mg L-1) and environmentally relevant concentrations (300 µg L-1), with a maximum specific 1,4-dioxane degradation rate (qmax) of 0.044 ± 0.001 mg dioxane h-1 mg protein-1, and 1,4-dioxane half-velocity constant (Ks) of 25 ± 1.6 mg L-1. The microbial community structure analysis suggested Pseudonocardia species, which utilize the dioxane monooxygenase for metabolic 1,4-dioxane biodegradation, were the main functional species for 1,4-dioxane degradation. The enrichment culture can adapt to both acidic (pH 5.5) and alkaline (pH 8) conditions and can recover degradation from low temperature (10°C) and anoxic (DO < 0.5 mg L-1) conditions. 1,4-Dioxane degradation of the enrichment culture was reversibly inhibited by TCE with concentrations higher than 5 mg L-1 and was completely inhibited by the presence of 1,1-DCE as low as 1 mg L-1. Collectively, these results demonstrated indigenous stable and robust 1,4-dioxane degrading enrichment culture can be obtained from uncontaminated sources and can be a potential candidate for 1,4-dioxane bioaugmentation at environmentally relevant conditions. KEY POINTS: •1,4-Dioxane degrading enrichment was obtained from uncontaminated soil. • The enrichment culture could degrade 1,4-dioxane to below 10 µg L-1. •Low Ks and low cell yield of the enrichment benefit its application in bioremediation.

Remedial strategies for abating 1,4-dioxane pollution-special emphasis on diverse biotechnological interventions.

1,4-dioxane is a heterocyclic ether used as a polar industrial solvent and are released as waste discharges. 1,4-dioxane deteriorates health and quality, thereby attracts concern by the environment technologists. The need of attaining sustainable development goals have resulted in search of an eco-friendly and technically viable treatment strategy. This extensive review is aimed to emphasis on the (a) characteristics of 1,4-dioxane and their occurrence in the environment as well as their toxicity, (b) remedial strategies, such as physico-chemical treatment and advanced oxidation techniques. Special reference to bioremediation that involves diverse microbial strains and their mechanism are highlighted in this review. The role of macronutrients, stimulants and other abiotic cofactors in the biodegradation of 1,4-dioxane is discussed lucidly. We have critically discussed the inducible enzymes, enzyme-based remediation, distinct instrumental method of analyses to know the fate of intermediates produced from 1,4-dioxane biotransformation. This comprehensive survey also tries to put forth the different toxicity assessment tools used in evaluating the extent of detoxification of 1,4-dioxane achieved through biotransforming mechanism. Conclusively, the challenges, opportunities, techno-economic feasibility and future prospects of implementing 1,4-dioxane through biotechnological interventions are also discussed.

Electron donor addition for stimulating the microbial degradation of 1,4 dioxane by sequential batch membrane bioreactor: A techno-economic approach.

The presence of 1,4 dioxane in wastewater is associated with severe health and environmental issues. The removal of this toxic contaminant from the industrial effluents prior to final disposal is necessary. The study comprehensively evaluates the performance of sequential batch membrane bioreactor (MBR) for treating wastewater laden with 1,4 dioxane. Acetate was supplemented to the wastewater feed as an electron donor for enhancing and stimulating the microbial growing activities towards the degradation of 1,4 dioxane. The removal efficiency of 1,4 dioxane was maximized to 87.5 ± 6.8% using an acetate to dioxane (A/D) ratio of 4.0, which was substantially dropped to 31.06 ± 3.7% without acetate addition. Ethylene glycol, glyoxylic acid, glycolic acid, and oxalic acid were the main metabolites of 1,4 dioxane biodegradation using mixed culture bacteria. The 1,4 dioxane degrading bacteria, particularly the genus of Acinetobacter, were promoted to 92% at the A/D ratio of 4.0. This condition encouraged as well the increase of the main 1,4 dioxane degraders, i.e., Xanthomonadales (12.5%) and Pseudomonadales (9.1%). However, 50% of the Sphingobacteriales and 82.5% of Planctomycetes were reduced due to the inhibition effect of the 1,4 dioxane contaminate. Similarly, the relative abundance of Firmicutes, Verrucomicrobia, Chlamydiae, Actinobacteria, Chloroflexi, and Nitrospirae was reduced in the MBR at the A/D ratio of 4.0. The results derived from the microbial analysis and metabolites detection at different A/D ratios indicated that acetate supplementation (as an electron donor) maintained an essential role in encouraging the microorganisms to produce the monooxygenase enzymes responsible for the biodegradation process. Economic feasibility of such a MBR system showed that for a designed flow rate of 30 m3∙d-1, the payback period from reusing the treated wastewater would reach 6.6 yr. The results strongly recommend the utilization of mixed culture bacteria growing on acetate for removing 1,4 dioxane from the wastewater industry, achieving dual environmental and economic benefits.

Network and meta-omics reveal the cooperation patterns and mechanisms in an efficient 1,4-dioxane-degrading microbial consortium.

1,4-Dioxane is an emerging wastewater contaminant with probable human carcinogenicity. Our current understanding of microbial interactions during 1,4-dioxane biodegradation process in mixed cultures is limited. Here, we applied metagenomic, metatranscriptomic and co-occurrence network analyses to unraveling the microbial cooperation between degrader and non-degraders in an efficient 1,4-dioxane-degrading microbial consortium CH1. A 1,4-dioxane-degrading bacterium, Ancylobacter polymorphus ZM13, was isolated from CH1 and had a potential of being one of the important degraders due to its high relative abundance, highly expressed monooxygenase genes tmoABCDEF and high betweenness centrality of networks. The strain ZM13 cooperated obviously with 6 bacterial genera in the network, among which Xanthobacter and Mesorhizobium could be involved in the intermediates metabolism with responsible genes encoding alcohol dehydrogenase (adh), aldehyde dehydrogenase (aldh), glycolate oxidase (glcDEF), glyoxylate carboligase (gcl), malate synthase (glcB) and 2-isopropylmalate synthase (leuA) differentially high-expressed. Also, 1,4-dioxane facilitated the shift of biodiversity and function of CH1, and those cooperators cooperated with ZM13 in the way of providing amino acids or fatty acids, as well as relieving environmental stresses to promote biodegradation. These results provide new insights into our understandings of the microbial interactions during 1,4-dioxane degradation, and have important implications for predicting microbial cooperation and constructing efficient and stable synthetic 1,4-dioxane-degrading consortia for practical remediation.

Environmental Potential for Microbial 1,4-Dioxane Degradation Is Sparse despite Mobile Elements Playing a Role in Trait Distribution.

1,4-Dioxane (dioxane) is an emerging contaminant of concern for which bioremediation is seen as a promising solution. To date, eight distinct gene families have been implicated in dioxane degradation, though only dioxane monooxygenase (DXMO) from Pseudonocardia dioxanivorans is routinely used as a biomarker in environmental surveys. In order to assess the functional and taxonomic diversity of bacteria capable of dioxane degradation, we collated existing, poorly-organized information on known biodegraders to create a curated suite of biomarkers with confidence levels for assessing 1,4-dioxane degradation potential. The characterized enzyme systems for dioxane degradation are frequently found on mobile elements, and we identified that many of the curated biomarkers are associated with other hallmarks of genomic rearrangements, indicating lateral gene transfer plays a role in dissemination of this trait. This is contrasted by the extremely limited phylogenetic distribution of known dioxane degraders, where all representatives belong to four classes within three bacterial phyla. Based on the curated set of expanded biomarkers, a search of more than 11,000 publicly available metagenomes identified a sparse and taxonomically limited distribution of potential dioxane degradation proteins. Our work provides an important and necessary structure to the current knowledge base for dioxane degradation and clarifies the potential for natural attenuation of dioxane across different environments. It further highlights a disconnect between the apparent mobility of these gene families and their limited distributions, indicating dioxane degradation may be difficult to integrate into a microorganism's metabolism. IMPORTANCE New regulatory limits for 1,4-dioxane in groundwater have been proposed or adopted in many countries, including the United States and Canada, generating a direct need for remediation options as well as better tools for assessing the fate of dioxane in an environment. A comprehensive suite of biomarkers associated with dioxane degradation was identified and then leveraged to examine the global potential for dioxane degradation in natural and engineered environments. We identified consistent differences in the dioxane-degrading gene families associated with terrestrial, aquatic, and wetland environments, indicating reliance on a single biomarker for assessing natural attenuation of dioxane is likely to miss key players. Most environments do not currently host the capacity for dioxane degradation-the sparse distribution of dioxane degradation potential highlights the need for bioaugmentation approaches over biostimulation of naturally occurring microbial communities.

Effect of biostimulation and bioaugmentation on biodegradation of high concentrations of 1,4-dioxane.

1,4-Dioxane is a pervasive and persistent contaminant in numerous aquifers. Although the median concentration in most contaminant plumes is in the microgram per liter range, a subset of sites have contamination in the milligram per liter range. Most prior studies that have examined 1,4-dioxane concentrations in the hundreds of milligrams per liter range have been performed with industrial wastewater. The main objective of this study was to evaluate aerobic biodegradation of 1,4-dioxane in microcosms prepared with soil and groundwater from a site where concentrations range from ~ 1500 mg·L-1 in the source zone, to 450 mg·L-1 at a midpoint of the groundwater plume, and to 6 mg·L-1 at a down-gradient location. Treatments included biostimulation with propane, addition of propane and a propanotrophic enrichment culture (ENV487), and unamended. The highest rates of biodegradation for each location in the plume occurred in the bioaugmented treatments, although indigenous propanotrophs also biodegraded 1,4-dioxane to below 25 µg·L-1. Nutrient additions were required to sustain biodegradation of propane and cometabolism of 1,4-dioxane. Among the unamended treatments, biodegradation of 1,4-dioxane was detected in the mid-gradient microcosms. An isolate was obtained that grows on 1,4-dioxane as a sole source of carbon and energy and identified through whole-genome sequencing as Pseudonocardia dioxivorans BERK-1. In a prior study, the same strain was isolated from an aquifer in the southeastern United States. Monod kinetic parameters for BERK-1 are similar to those for strain CB1190.

Response surface algorithm for improved biotransformation of 1,4-dioxane using Staphylococcus capitis strain AG.

The present investigation reports the biotransformation of an endrocrine disrupting agent; 1,4-dioxane through bacterial metabolism. Initially, potential bacterial isolates capable of surviving with minimum 1,4-dioxane were screened from industrial wastewater. Thereafter, screening was done to isolate a bacteria which can biotransform higher concentration (1000 mg/L) of 1,4-dioxane. Morphological and biochemical features were examined prior establishing their phylogenetic relationships and the bacterium was identified as Staphylococcus capitis strain AG. Biotransformation experiments were tailored using response surface tool and predictions were made to elucidate the opimal conditions. Critical factors influencing bio-transformation efficiency such as tetrahydrofuran, availability of 1,4-dioxane and inoculum size were varied at three different levels as per the central composite design for ameliorating 1,4-dioxane removal. Functional attenuation of 1,4-dioxane by S. capitis strain AG were understood using spectroscopic techniques were significant changes in the peak positions and chemical shifts were visualized. Mass spectral profile revealed that 1.5 (% v/v) S. capitis strain AG could completely (∼99%) remove 1000 mg/L 1,4-dioxane, when incubated with 2 µg/L tetrahydrofuran for 96 h. The toxicity of 1,4-dioxane and biotransformed products by S. capitis strain AG were tested on Artemia salina. The results of toxicity tests revealed that the metabolic products were less toxic as they exerted minimal mortality rate after 48 h exposure. Thus, this research would be the first to report the response prediction and precise tailoring of 1,4-dioxane biotransformation using S. captis strain AG.

Xanthobacter dioxanivorans sp. nov., a 1,4-dioxane-degrading bacterium.

A Gram-stain-negative bacterium, designated as YN2T, that is capable of degrading 1,4-dioxane, was isolated from active sludge collected from a wastewater treatment plant in Harbin, PR China. Cells of strain YN2T were aerobic, motile, pleomorphic rods, mostly twisted, and contained the water-insoluble yellow zeaxanthin dirhamnoside. Strain YN2T grew at 10-40 °C (optimum, 30 °C), pH 5.0-8.0 (pH 7.0) and with 0-1 % (w/v) NaCl (0.1 %). It also could grow chemolithoautotrophically and fix N2 when no ammonium or nitrate was supplied. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain YN2T belongs to the genus Xanthobacter and shares the highest pairwise identity with Xanthobacter autotrophicus 7cT (98.6 %) and Xanthobacter flavus 301T (98.4 %). The major respiratory quinone was ubiquinone-10. Chemotaxonomic analysis revealed that the strain possesses C16 : 0, C19 : 0 cyclo ω8c and C18 : 1 ω7c as the major fatty acids. The DNA G+C content was 67.95 mol%. Based on genome sequences, the DNA-DNA hybridization estimate values between strain YN2T and X. autotrophicus 7cT, X. flavus 301T and X. tagetidis TagT2CT (the only three species of Xanthobacter with currently available genomes) were 31.70, 31.30 and 28.50 %; average nucleotide identity values were 85.23, 84.84 and 83.59 %; average amino acid identity values were 81.24, 80.23 and 73.57 %. Based on its phylogenetic, phenotypic, and physiological characteristics, strain YN2T is considered to represent a novel species of the genus Xanthobacter, for which the name Xanthobacter dioxanivorans sp. nov. is proposed. The type strain is YN2T (=CGMCC 1.19031T=JCM 34666T).

Adaption and Degradation Strategies of Methylotrophic 1,4-Dioxane Degrading Strain Xanthobacter sp. YN2 Revealed by Transcriptome-Scale Analysis.

Biodegradation of 1,4-dioxane (dioxane) contamination has gained much attention for decades. In our previous work, we isolated a highly efficient dioxane degrader, Xanthobacter sp. YN2, but the underlying mechanisms of its extraordinary degradation performance remained unresolved. In this study, we performed a comparative transcriptome analysis of YN2 grown on dioxane and citrate to elucidate its genetic degradation mechanism and investigated the transcriptomes of different dioxane degradation stages (T0, T24, T48). We also analyzed the transcriptional response of YN2 over time during which the carbon source switched from citrate to dioxane. The results indicate that strain YN2 was a methylotroph, which provides YN2 a major advantage as a pollutant degrader. A large number of genes involved in dioxane metabolism were constitutively expressed prior to dioxane exposure. Multiple genes related to the catabolism of each intermediate were upregulated by treatment in response to dioxane. Glyoxylate metabolism was essential during dioxane degradation by YN2, and the key intermediate glyoxylate was metabolized through three routes: glyoxylate carboligase pathway, malate synthase pathway, and anaplerotic ethylmalonyl-CoA pathway. Genes related to quorum sensing and transporters were significantly upregulated during the early stages of degradation (T0, T24) prior to dioxane depletion, while the expression of genes encoding two-component systems was significantly increased at late degradation stages (T48) when total organic carbon in the culture was exhausted. This study is the first to report the participation of genes encoding glyoxalase, as well as methylotrophic genes xoxF and mox, in dioxane metabolism. The present study reveals multiple genetic and transcriptional strategies used by YN2 to rapidly increase biomass during growth on dioxane, achieve high degradation efficiency and tolerance, and adapt to dioxane exposure quickly, which provides useful information regarding the molecular basis for efficient dioxane biodegradation.

Cometabolic degradation of 1,4-dioxane by a tetrahydrofuran-growing Arthrobacter sp. WN18.

1,4-Dioxane (dioxane), an emerging groundwater contaminant, is frequently detected in landfill leachates with its structural analog, tetrahydrofuran (THF). Along with undesirable leakage of landfill leachates, dioxane and THF inevitably percolate into groundwater leading to a broader region of contamination. Cometabolic bioremediation is an effective approach to manage commingled THF and dioxane pollution. In this study, a newly isolated bacterium Arthrobacter sp. WN18 is able to co-oxidize dioxane with THF as the primary substrate. Meanwhile, the THF-induced thmADBC gene cluster was responsible for the dioxane degradation rate indicating THF monooxygenase is the essential enzyme that initializing α-hydroxylation of THF and dioxane. Further, γ-butyrolactone and HEAA were characterized as the key metabolites of THF and dioxane, respectively. In addition, WN18 can tolerate the inhibition of trichloroethylene (5.0 mg/L) as a representative of co-existing leachate constituent, and sustain its activity at various pH (5-11), temperatures (15-42 °C), and salinities (up to 4%, as NaCl wt). Like other Arthrobacter species, WN18 also exhibited the capability of fixing nitrogen. All this evidence indicates the feasibility and advantage of WN18 as a thmADBC-catalyzed inoculator to bioremediate co-contamination of THF and dioxane.

Kinetic characterization of purified laccase from Trametes hirsuta: a study on laccase catalyzed biotransformation of 1,4-dioxane.

OBJECTIVE: Laccase is one of the best known biocatalysts which degrade wide varieties of complex molecules that are both non-cyclic and cyclic in structure. The study focused on enzyme kinetics of a purified laccase from Trametes hirsuta L. fungus and its application on biotransformation of a carcinogenic molecule 1,4-dioxane. RESULTS: Laccase was purified from white-rot fungus T. hirsuta L. which showed specific activity of 978.34 U/mg after the purification fold of 54.08. The stable laccase activity (up to 16 h) is shown at 4-6 pH and 20-40 °C temperature range. The purified enzyme exhibited significant stability for 10 metal ions up to 10 mM concentration, except for Fe2+ and Hg2+. The Cu2+ ion induced laccase activity up to 142% higher than the control at 10 mM concentration. The laccase enzyme kinetic parameters Km was 20 ± 5 µM and 400 ± 60 µM, whereas Kcat was 198.29 ± 0.18/s and 80.20 ± 1.59/s for 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and guaiacol respectively. The cyclic ether 1,4-dioxane (100 ppm) was completely degraded in presence of purified laccase within 2 h of incubation and it was confirmed by HPLC and GC analysis. The oxidation reaction was accelerated by 25, 22, 6 and 19% in presence of 1 mM syringaldehyde, vanillin, ABTS and guaiacol mediators respectively. CONCLUSIONS: In this study, fungal laccase (a natural biocatalyst) based degradation of synthetic chemical 1,4-dioxane was reported for the first time. This method has added advantages over the multiple methods reported earlier being a natural remedy.

Modifications at C(5) of 2-(2-Pyrrolidinyl)-Substituted 1,4-Benzodioxane Elicit Potent α4ß2 Nicotinic Acetylcholine Receptor Partial Agonism with High Selectivity over the α3ß4 Subtype.

A series of diastereomeric 2-(2-pyrrolidinyl)-1,4-benzodioxanes bearing a small, hydrogen-bonding substituent at the 7-, 6-, or 5-position of benzodioxane have been studied for α4ß2 and α3ß4 nicotinic acetylcholine receptor affinity and activity. Analogous to C(5)H replacement with N and to a much greater extent than decoration at C(7), substitution at benzodioxane C(5) confers very high α4ß2/α3ß4 selectivity to the α4ß2 partial agonism. Docking into the two receptor structures recently determined by cryo-electron microscopy and site-directed mutagenesis at the minus ß2 side converge in indicating that the limited accommodation capacity of the ß2 pocket, compared to that of the ß4 pocket, makes substitution at C(5) rather than at more projecting C(7) position determinant for this pursued subtype selectivity.

Features of In Vitro Degradation and Physical Properties of a Biopolymer and In Vivo Tissue Reactions in Comparison with Polypropylene.

We compared in vitro degradation and physical properties of polypropylene and a biodegradable polymer synthesized by electrospinning and consisting of 65% polycaprolactone and 35% polytrimethylene carbonate as a possible alternative material for use in surgery for pelvic floor muscle failure. Samples of the studied polymers were implanted to 10 male Wistar rats into the interfascial space on the back (polypropylene on the right side and biodegradable polymer on the left side). The synthesized biopolymer was characterized by elongation and tear resistance, similar to those of polypropylene. During the period from the third to the sixth month after implantation, the area of fibrosis around individual polypropylene and biopolymer fibers increased by 16.7 and 107.9%, respectively, while remaining reduced compared to polypropylene. The total fibrosis area in 6 months after implantation of polypropylene and biopolymer samples significantly increased by 18% (p=0.0097) and 48% (p=0.05), respectively, i.e. fibrosing processes were more intense in case of biopolymer. Induction of more pronounced fibrosis can be an advantage of the synthesized biopolymer when choosing the material for fabrication of implants and their use for correction of incompetence of the ligamentous and muscular apparatus.

Impact of physiologically relevant temperatures on dermal absorption of active substances - an ex-vivo study in human skin.

Skin temperature plays a certain role in the dermal absorption of substances, but the extent and mechanisms of skin temperatures-induced modulation in ranges caused by physiological thermoregulation or environmental conditions are largely unknown. The influence of dermal temperature on the absorption of the model lipophilic compound (anisole) and the model hydrophilic compounds (1,4-dioxane, ethanol) through human skin was investigated at three dermal temperatures (25, 32 and 39 °C) in an ex-vivo diffusion cell model. The substances were applied to the skin and transdermal penetration was monitored. All substances showed temperature dependent variations in their penetration behavior (3 h: 25-39 °C: 202-275% increase in cumulative, transdermally penetrated amounts). The relative differences in absorption in relation to temperature were greatest within 45 min after exposure (25-39 °C: 347-653% rise in cumulated penetration), although absolute amounts absorbed were small (45 min vs. 3 h: 4.5-14.5%). Regardless of blood circulation, skin temperature significantly influences the amount and kinetics of dermal absorption. Substance-dependent, temperature-related changes of the lipid layer order or the porous pathway may facilitate penetration. The early-stage modulation of transdermal penetration indicates transappendageal absorption, which may be relevant for short-term exposures. For both, toxicological evaluation and perfusion cell studies, it is important to consider the thermal influence on absorption or to perform the latter at a standardized temperature (32±1 °C).