Identification of a chlorosalicylic acid decarboxylase (CsaD) involved in decarboxylation of 3,6-DCSA from an anaerobic dicamba-degrading sludge.

3,6-Dichlorosalicylic acid (3,6-DCSA) is the demethylation metabolite of herbicide 3,6-dichloro-2-methoxy benzoic acid (dicamba). Previous studies have shown that anaerobic sludge further transformed 3,6-DCSA through decarboxylation and dechlorination. However, the anaerobe, enzyme, and gene involved in the anaerobic degradation of 3,6-DCSA are still unknown. In this study, an anaerobic sludge that efficiently degraded dicamba was enriched, and a 3,6-DCSA decarboxylase, designated chlorosalicylic acid decarboxylase (CsaD), was partially purified and identified from the anaerobic sludge. Metagenomic analysis showed that the csaD gene was located in a gene cluster of metagenome-assembled genome 8 (MAG8). MAG8 belonged to an uncultured order, OPB41, in the class Coriobacteriia of the phylum Actinobacteria, and its abundance increased approximately once during the enrichment process. CsaD was a non-oxidative decarboxylase in the amidohydrolase 2 family catalyzing the decarboxylation of 3,6-DCSA and 6-chlorosalicylic acid (6-CSA). Its affinity and catalytic efficiency for 3,6-DCSA were significantly higher than those for 6-CSA. This study provides new insights into the anaerobic catabolism of herbicide dicamba.IMPORTANCEDicamba, an important hormone herbicide, easily migrates to anoxic habitats such as sediment, ground water, and deep soil. Thus, the anaerobic catabolism of dicamba is of importance. Anaerobic bacteria or sludge demethylated dicamba to 3,6-DCSA, and in a previous study, based on metabolite identification, it was proposed that 3,6-DCSA be further degraded via two pathways: decarboxylation to 2,5-dichlorophenol, then dechlorination to 3-chlorophenol (3-CP); or dechlorination to 6-CSA, then decarboxylation to 3-CP. However, there was no physiological and genetic validation for the pathway. In this study, CsaD catalyzed the decarboxylation of both 3,6-DCSA and 6-CSA, providing enzyme-level evidence for the anaerobic catabolism of 3,6-DCSA through the two pathways. CsaD was located in MAG8, which belonged to an uncultured anaerobic actinomycetes order, OPB41, indicating that anaerobic actinomycetes in OPB41 was involved in the decarboxylation of 3,6-DCSA. This study provides a basis for understanding the anaerobic catabolism of dicamba and the demethylation product, 3,6-DCSA.

New insights on the degradation of polystyrene and polypropylene by larvae of the superworm Zophobas atratus and gut bacterial consortium enrichments obtained under different culture conditions.

This study aims to deepen knowledge of the biodegradation of plastics, focusing on polypropylene (PP) fabric from surgical masks and polystyrene (PS) by larvae of Zophobas atratus as well as of specialized bacterial consortia from their gut, which were obtained in different enrichment conditions (aerobic, anaerobic, presence or absence of combined nitrogen). Plastics ingested by larvae obtained in Spain did not show any signs of oxidation but only limited depolymerization, preferably from the lowest molecular weight chains. Gut microbiota composition changed as an effect of plastic feeding. Such differences were more evident in bacterial enrichment cultures, where the polymer type influenced the composition more than by culture conditions, with an increase in the presence of nitrogen-fixers in anaerobic conditions. PS and PP degradation by different enrichment cultures was confirmed under aerobic and anaerobic conditions by respirometry tests, with anaerobic conditions favouring a more active plastic degradation. In addition, exposure to selected bacterial consortia in aerobiosis induced limited surface oxidation of PS. This possibly indicates that different biochemical routes are being utilized in the anaerobic gut and in aerobic conditions to degrade the polymer.

Catabolism and interactions of syntrophic propionate- and acetate oxidizing microorganisms under mesophilic, high-ammonia conditions.

Microbial inhibition by high ammonia concentrations is a recurring problem that significantly restricts methane formation from intermediate acids, i.e., propionate and acetate, during anaerobic digestion of protein-rich waste material. Studying the syntrophic communities that perform acid conversion is challenging, due to their relatively low abundance within the microbial communities typically found in biogas processes and disruption of their cooperative behavior in pure cultures. To overcome these limitations, this study examined growth parameters and microbial community dynamics of highly enriched mesophilic and ammonia-tolerant syntrophic propionate and acetate-oxidizing communities and analyzed their metabolic activity and cooperative behavior using metagenomic and metatranscriptomic approaches. Cultivation in batch set-up demonstrated biphasic utilization of propionate, wherein acetate accumulated and underwent oxidation before complete degradation of propionate. Three key species for syntrophic acid degradation were inferred from genomic sequence information and gene expression: a syntrophic propionate-oxidizing bacterium (SPOB) "Candidatus Syntrophopropionicum ammoniitolerans", a syntrophic acetate-oxidizing bacterium (SAOB) Syntrophaceticus schinkii and a novel hydrogenotrophic methanogen, for which we propose the provisional name "Candidatus Methanoculleus ammoniitolerans". The results revealed consistent transcriptional profiles of the SAOB and the methanogen both during propionate and acetate oxidation, regardless of the presence of an active propionate oxidizer. Gene expression indicated versatile capabilities of the two syntrophic bacteria, utilizing both molecular hydrogen and formate as an outlet for reducing equivalents formed during acid oxidation, while conserving energy through build-up of sodium/proton motive force. The methanogen used hydrogen and formate as electron sources. Furthermore, results of the present study provided a framework for future research into ammonia tolerance, mobility, aggregate formation and interspecies cooperation.

Insights on kraft lignin degradation in an anaerobic environment.

Lignin is an aromatic macromolecule and one of the main constituents of lignocellulosic materials. Kraft lignin is generated as a residual by-product of the lignocellulosic biomass industrial process, and it might be used as a feedstock to generate low molecular weight aromatic compounds. In this study, we seek to understand and explore the potential of ruminal bacteria in the degradation of kraft lignin. We established two consortia, KLY and KL, which demonstrated significant lignin-degrading capabilities. Both consortia reached maximum growth after two days, with KLY showing a higher growth and decolorization rate. Additionally, SEM analysis revealed morphological changes in the residual lignin from both consortia, indicating significant degradation. This was further supported by FTIR spectra, which showed new bands corresponding to the C-H vibrations of guaiacyl and syringyl units, suggesting structural transformations of the lignin. Taxonomic analysis showed enrichment of the microbial community with members of the Dickeya genus. Seven metabolic pathways related to lignin metabolism were predicted for the established consortia. Both consortia were capable of consuming aromatic compounds such as 4-hydroxybenzoic acid, syringaldehyde, acetovanillone, and syringic acid, highlighting their capacity to convert aromatic compounds into commercially valuable molecules presenting antifungal activity and used as food preservatives as 4-hydroxyphenylacetic, 3-phenylacetic, and phenylacetic acids. Therefore, the microbial consortia shown in the present work are models for understanding the process of lignin degradation and consumption in bacterial anaerobic communities and developing biological processes to add value to industrial processes based on lignocellulosic biomass as feedstock.

Mechanisms and high-value applications of phthalate isomers degradation pathways in bacteria.

Phthalate isomers are key intermediates in the biodegradation of pollutants including waste polyethylene terephthalate (PET) plastics and plasticizers. So far, an increasing number of phthalate isomer-degrading strains have been isolated, and their degradation pathways show significant diversity. In this paper, we comprehensively review the current status of research on the degrading bacteria, degradation characteristics, aerobic and anaerobic degradation pathways, and degradation genes (clusters) of phthalate isomers, and discuss the current shortcomings and challenges. Moreover, the degradation process of phthalate isomers produces many important aromatic precursor molecules, which can be used to produce higher-value derivative chemicals, and the modification of their degradation pathways holds good prospects. Therefore, this review also highlights the current progress made in modifying the phthalate isomer degradation pathway and explores its potential for high-value applications.

Neglected sludge solid phase in sludge pretreatment process: Physicochemical characterization and mechanism study of its role in anaerobic degradation.

The low anaerobic digestion efficiency of the solid phase separated from pre-treated sludge indicates the need to explore other suitable resource utilization pathways for sludge solid phase. However, there is a lack of comprehensive and in-depth research on the physicochemical properties of sludge solid phase. This study comprehensively analyzes the characteristics of sludge solid phase and elucidates the mechanism of sludge solid phase in the anaerobic degradation of toxic wastewater. The results show that the surface free energy of sludge solid phase after different pre-treatments is mainly contributed by Lewis acid-base hydration free energy. The distribution of proteins on the surface of sludge solid phase plays a major role in the adhesion between sludge solid particles. Metal ions in the sludge solid phase are mainly present in the exchange state, followed by the carbonate state and the organics-bound state. The sludge solid phase obtained by sludge pH 12 + 150 °C treatment has the highest conductivity (1.36 mS/m) and capacitance (25.51 µF/g), mainly due to the presence of melanoidins in the sludge solid phase, which has similar semiquinone radicals to humic acids, thus increasing conductivity. The addition of sludge solid phase promotes an increase in cumulative methane production and rate of methane production. The sludge solid phase might play a role of an auxiliary carbon source acting as an adsorbent to buffer against toxicity inhibition and facilitate electron transfer. This study reveals the characteristics of sludge solid phase and its role in anaerobic digestion, providing theoretical guidance for finding suitable resource utilization pathways for sludge solid phase.

Ultrafiltration fractionation of potentially inhibitory substances of hydrothermal liquefaction aqueous phase derived from municipal sludge.

Hydrothermal liquefaction (HTL) is a promising thermo-chemical technology for municipal sludge treatment due to its potential for biocrude oil recovery and minimizing biosolids management costs. However, the process generates a high volume of an aqueous byproduct that needs to be treated due to its high chemical oxygen demand (COD) and various organic and inorganic compounds. Although the aqueous phase is known to contain recalcitrant and potentially inhibitory substances that may affect its biological treatment, their molecular weight distribution (MwD) and its impact on anaerobic biodegradability are poorly understood. Ultrafiltration (UF) was conducted to fractionate HTL aqueous into different molecular weight (Mw) fractions using 300, 100, 10, and 1 kDa membranes. Mesophilic biochemical methane potential (BMP) assays were conducted to assess the anaerobic biodegradability of each fraction, and the first-order model was used to calculate the degradation kinetics of potential inhibitory compounds. The highest percentage of organics (65 %) was found in the Mw<1 kDa range, whereas the 10>Mw>1 kDa had the lowest percentage (8 %). There was no significant difference in the cumulative specific methane produced from various Mw fractions (p>0.05). The Mw<1 kDa fraction had the highest first-order specific methane production rate (0.53 day-1), whereas the unfiltered HTL had the lowest (0.38 day-1). Although UF fractionation increased the rate of anaerobic degradation of HTL aqueous for the Mw<1 kDa fraction, the observed methane potential was only 55 % of the theoretical value. This implies that 45 % of COD remains undegraded even after permeation through the lowest Mw cut-off membrane. Therefore, further characterization of HTL aqueous is needed for compounds with molecular weights below 1 kDa to fully understand the nature of inhibitory organics and their impact on anaerobic digestion. Furthermore, pretreatments utilizing techniques such as adsorption and advanced oxidation may be necessary to enhance the specific methane yields from various HTL aqueous fractions, thereby bringing them closer to the theoretical yield.

New perspectives on the anaerobic degradation of BTEX: Mechanisms, pathways, and intermediates.

Aromatic hydrocarbons like benzene, toluene, xylene, and ethylbenzene (BTEX) can escape into the environment from oil and gas operations and manufacturing industries posing significant health risks to humans and wildlife. Unlike conventional clean-up methods used, biological approaches such as bioremediation can provide a more energy and labour-efficient and environmentally friendly option for sensitive areas such as nature reserves and cities, protecting biodiversity and public health. BTEX contamination is often concentrated in the subsurface of these locations where oxygen is rapidly depleted, and biodegradation relies on anaerobic processes. Thus, it is critical to understand the anaerobic biodegradation characteristics as it has not been explored to a major extent. This review presents novel insights into the degradation mechanisms under anaerobic conditions and presents a detailed description and interconnection between them. BTEX degradation can follow four activation mechanisms: hydroxylation, carboxylation, methylation, and fumarate addition. Hydroxylation is one of the mechanisms that explains the transformation of benzene into phenol, toluene into benzyl alcohol or p-cresol, and ethylbenzene into 1-phenylethanol. Carboxylation to benzoate is thought to be the primary mechanism of degradation for benzene. Despite being poorly understood, benzene methylation has been also reported. Moreover, fumarate addition is the most widely reported mechanism, present in toluene, ethylbenzene, and xylene degradation. Further research efforts are required to better elucidate new and current alternative catabolic pathways. Likewise, a comprehensive analysis of the enzymes involved as well as the development of advance tools such as omic tools can reveal bottlenecks degradation steps and create more effective on-site strategies to address BTEX pollution.

Degradation of pretilachlor and fenclorim and effects of these compounds on bacterial communities under anaerobic condition.

Pretilachlor and safener fenclorim are the main components of herbicides widely applied to control weeds. Although some pure cultures of bacteria and fungi which degraded these compounds under aerobic conditions were isolated, no isolated pretilachlor- and fenclorim-degrading bacterial strains under anaerobic condition had been available. In this study, the degradation of these compounds and the effects of them on bacterial community structures were investigated under anaerobic conditions. The dissipation rates of pretilachlor and fenclorim in slurries were in the order: soil from paddy field ≈ sediment from river > sediment from mangrove. Moreover, three pretilachlor-degrading bacterial strains (Pseudomonas sp. Pr1, Proteiniclasticum sp. Pr2 and Paracoccus denitrificans Pr3) and two fenclorim-degrading strains (Dechloromonas sp. Fe1 and Ralstonia pickettii Fe2) isolated from a slurry of paddy soil utilized the substrates as sole carbon and energy sources under anaerobic conditions. The degradation of pure pretilachlor and fenclorim at various concentrations by corresponding mixed pure cultures followed the Michaelis-Menten model, with the maximum degradation was 3.10 ± 0.31 µM/day for pretilachlor, and 2.08 ± 0.18 µM/day for fenclorim. During the degradation, 2-chloro-N-(2,6-diethylphenyl) acetamide and 2,6-dimethylaniline were produced in pretilachlor degradation, and benzene was a product of fenclorim degradation. The synergistic degradation of both substrates by all isolated bacteria reduced the metabolites concentrations accumulated in media. This study provides valuable information on effects of pretilachlor and fenclorim on bacterial communities in soil and sediments, and degradation of these substrates by isolated bacteria under anaerobic condition.

Differential degradation of petroleum hydrocarbons by Shewanella putrefaciens under aerobic and anaerobic conditions.

The complexity of crude oil composition, combined with the fluctuating oxygen level in contaminated environments, poses challenges for the bioremediation of oil pollutants, because of compound-specific microbial degradation of petroleum hydrocarbons under certain conditions. As a result, facultative bacteria capable of breaking down petroleum hydrocarbons under both aerobic and anaerobic conditions are presumably effective, however, this hypothesis has not been directly tested. In the current investigation, Shewanella putrefaciens CN32, a facultative anaerobic bacterium, was used to degrade petroleum hydrocarbons aerobically (using O2 as an electron acceptor) and anaerobically (using Fe(III) as an electron acceptor). Under aerobic conditions, CN32 degraded more saturates (65.65 ± 0.01%) than aromatics (43.86 ± 0.03%), with the following order of degradation: dibenzofurans > n-alkanes > biphenyls > fluorenes > naphthalenes > alkylcyclohexanes > dibenzothiophenes > phenanthrenes. In contrast, under anaerobic conditions, CN32 exhibited a higher degradation of aromatics (53.94 ± 0.02%) than saturates (23.36 ± 0.01%), with the following order of degradation: dibenzofurans > fluorenes > biphenyls > naphthalenes > dibenzothiophenes > phenanthrenes > n-alkanes > alkylcyclohexanes. The upregulation of 4-hydroxy-3-polyprenylbenzoate decarboxylase (ubiD), which plays a crucial role in breaking down resistant aromatic compounds, was correlated with the anaerobic degradation of aromatics. At the molecular level, CN32 exhibited a higher efficiency in degrading n-alkanes with low and high carbon numbers relative to those with medium carbon chain lengths. In addition, the degradation of polycyclic aromatic hydrocarbons (PAHs) under both aerobic and anaerobic conditions became increasingly difficult with increased numbers of benzene rings and methyl groups. This study offers a potential solution for the development of targeted remediation of pollutants under oscillating redox conditions.

Modeled Pathways and Fluxes of PCB Dechlorination by Redox Potentials.

Dechlorination is one of the main processes for the natural degradation of polychlorinated biphenyls (PCBs) in an anaerobic environment. However, PCB dechlorination pathways and products vary with PCB congeners, types of functional dechlorinating bacteria, and environmental conditions. The present study develops a novel model for determining dechlorination pathways and fluxes by tracking redox potential variability, transforming the complex dechlorination process into a stepwise sequence. The redox potential is calculated via the Gibbs free energy of formation, PCB concentrations in reactants and products, and environmental conditions. Thus, the continuous change in the PCB congener composition can be tracked during dechlorination processes. The new model is assessed against four measurements from several published studies on PCB dechlorination. The simulation errors in all four measurements are calculated between 2.67 and 35.1% under minimum (n = 0) and maximum (n = 34) numbers of co-eluters, respectively. The dechlorination fluxes for para-dechlorination pathways dominate PCB dechlorination in all measurements. Furthermore, the model also considers multiple-step dechlorination pathways containing intermediate PCB congeners absent in both the reactants and the products. The present study indicates that redox potential might be an appropriate indicator for predicting PCB dechlorination pathways and fluxes even without prior knowledge of the functional dechlorinating bacteria.

A novel co-metabolic mode with Spirulina powder in enhancing the anaerobic degradation of typical nitrogen heterocyclic compounds.

Spirulina powder emerged as a novel and suitable co-metabolism substance significantly enhancing the anaerobic degradation of specific nitrogen heterocyclic compounds. On the addition of 1.0 mg/L of Spirulina powder, the reactor demonstrated optimal degradation efficiency for quinoline and indole, achieving ratios of 99.77 ± 1.83% and 99.57 ± 1.98%, respectively. Moreover, the incorporation of Spirulina powder resulted in increased concentrations of mixed liquor suspended solids, mixed liquor volatile suspended solids, proteins, and polysaccharides in anaerobic sludge. In addition, Spirulina powder led to reduced levels of Acinetobacter and enriched Aminicenantes genera incertae sedis, Levilinea, and Longilinea. The analysis of the archaeal community structure confirmed that the addition of Spirulina powder increased archaeal sequences, fostering greater richness and diversity in the archaeal community.

Microplastic release and sulfate reduction response in the early stage of a simulated landfill.

Landfills are essential facilities for treating and disposing municipal solid waste. They emit sulfur-containing odors and serve as an important sink for a new type of pollutant called microplastics (MPs). This study focused on the initial stage of anaerobic degradation to establish the relationship between the release of MPs and odor generation. Our findings show the rapid release of MPs into the leachate in the early stage of landfill and their predominant accumulation in the leachate sediment. The circulating leachate contained 1.45 times higher concentrations of MPs than the noncirculating leachate, with a peak concentration of 39 items·L-1. In addition, fragmentation of MPs occurred. The percentage of MPs with particle sizes of 2.5-5 mm decreased from 66.70 % to 22.32 %, while those measuring 0.1-0.5 mm increased by 33.12 %. A positive correlation was observed between MP release and sulfate reduction. Although leachate circulation increased the release of MPs, it also reduced the overall release time and total amount of MPs exported from the landfill. Compared with the initial landfill waste, the leachate operation mode, regardless of circulation, resulted in a 6.15-8.93-fold increase in MP release. These findings provide a valuable foundation for the simultaneous regulation of traditional pollutant odor and new pollutants (MPs) in landfills.

Sensitive and selective phenol sensing in denitrifying Aromatoleum aromaticum EbN1T.

IMPORTANCE: Aromatic compounds are globally abundant organic molecules with a multitude of natural and anthropogenic sources, underpinning the relevance of their biodegradation. A. aromaticum EbN1T is a well-studied environmental betaproteobacterium specialized on the anaerobic degradation of aromatic compounds. The here studied responsiveness toward phenol in conjunction with the apparent high ligand selectivity (non-promiscuity) of its PheR sensor and those of the related p-cresol (PcrS) and p-ethylphenol (EtpR) sensors are in accord with the substrate-specificity and biochemical distinctiveness of the associated degradation pathways. Furthermore, the present findings advance our general understanding of the substrate-specific regulation of the strain's remarkable degradation network and of the concentration thresholds below which phenolic compounds become essentially undetectable and as a consequence should escape substantial biodegradation. Furthermore, the findings may inspire biomimetic sensor designs for detecting and quantifying phenolic contaminants in wastewater or environments.

Bisphenol A monitoring during anaerobic degradation of papers with thermochromic prints in soil.

Pseudoestrogene bisphenol A (BPA) can be important ingredient of thermochromic inks, increasingly used materials in thermal printing paper, security printing, advertising, design and as temperature indicators in medicine and food industry. BPA mass fraction in thermochromic inks can be up to several percent. Hence, disposal of items with thermochromic prints pose a risk of environmental pollution. In this work BPA mass fraction was monitored during anaerobic degradation of papers with thermochromic prints in soil in both matrices: papers and soil. The degradation conditions simulated deeper layers of waste at a landfill site. Six types of papers with prints of thermochromic ink containing 2% of BPA were subjected to anaerobic degradation over up to 150 days. Initial mass fractions of BPA in papers decreased form (126-460) µg/g to (

Anaerobic degradation of organic carbon supports uncultured microbial populations in estuarine sediments.

BACKGROUND: A large proportion of prokaryotic microbes in marine sediments remains uncultured, hindering our understanding of their ecological functions and metabolic features. Recent environmental metagenomic studies suggested that many of these uncultured microbes contribute to the degradation of organic matter, accompanied by acetogenesis, but the supporting experimental evidence is limited. RESULTS: Estuarine sediments were incubated with different types of organic matters under anaerobic conditions, and the increase of uncultured bacterial populations was monitored. We found that (1) lignin stimulated the increase of uncultured bacteria within the class Dehalococcoidia. Their ability to metabolize lignin was further supported by the presence of genes associated with a nearly complete degradation pathway of phenolic monomers in the Dehalococcoidia metagenome-assembled genomes (MAGs). (2) The addition of cellulose stimulated the increase of bacteria in the phylum Ca. Fermentibacterota and family Fibrobacterales, a high copy number of genes encoding extracellular endoglucanase or/and 1,4-beta-cellobiosidase for cellulose decomposition and multiple sugar transporters were present in their MAGs. (3) Uncultured lineages in the order Bacteroidales and the family Leptospiraceae were enriched by the addition of casein and oleic acid, respectively, a high copy number of genes encoding extracellular peptidases, and the complete ß-oxidation pathway were found in those MAGs of Bacteroidales and Leptospiraceae, respectively. (4) The growth of unclassified bacteria of the order Clostridiales was found after the addition of both casein and cellulose. Their MAGs contained multiple copies of genes for extracellular peptidases and endoglucanase. Additionally, 13C-labeled acetate was produced in the incubations when 13C-labeled dissolved inorganic carbon was provided. CONCLUSIONS: Our results provide new insights into the roles of microorganisms during organic carbon degradation in anaerobic estuarine sediments and suggest that these macro and single molecular organic carbons support the persistence and increase of uncultivated bacteria. Acetogenesis is an additional important microbial process alongside organic carbon degradation. Video Abstract.

Naphthalene Carboxylation in the Sulfate-Reducing Enrichment Culture N47 Is Proposed to Proceed via 1,3-Dipolar Cycloaddition to the Cofactor Prenylated Flavin Mononucleotide.

Polycyclic aromatic hydrocarbons are persistent pollutants of anthropogenic or natural origin in the environment and accumulate in anoxic habitats. In this study, we investigated the mechanism of the enzyme naphthalene carboxylase as a model reaction for polycyclic aromatic hydrocarbon activation by carboxylation. An enzyme assay was established with cell extracts of the highly enriched culture N47. In assays without addition of ATP, naphthalene carboxylase catalyzed a stable isotope exchange of the carboxyl group of naphthoate with 13C-labeled bicarbonate buffer, which can only occur via a partial backwards reaction of the naphthalene carboxylase reaction to an intermediate that does not include the carboxyl group. Hence, a new carboxyl group from the labeled bicarbonate is added upon forward reaction to the naphthoate. This indicates that the reaction mechanism consists of two or more steps and that at least the latter steps are reversible and ATP independent. Naphthalene carboxylation assays were carried out in deuterated buffer and revealed the incorporation of 0, 1, 2, or 3 deuterium atoms in the final product naphthoyl-coenzyme A, indicating that the reaction is fully reversible. Putative reaction mechanisms were tested by quantum mechanical calculations. The proposed mechanism of the reaction consists of three steps: the activation of the naphthalene by 1,3-dipolar cycloaddition of the cofactor prFMN to naphthalene, release of a proton and rearomatization producing a stable intermediate, and a carboxylation with a reverse 1,3-dipolar cycloaddition and cleavage of the bond to the cofactor producing 2-naphthoate. IMPORTANCE Pollution with polycyclic aromatic hydrocarbons poses a great hazard to humans and animals, with considerable long-term effects. The anaerobic degradation of polycyclic aromatic hydrocarbons in anoxic zones and anaerobic growth of such organisms is very slow, leading to only poor investigation of the degradation pathways, so far. In this work, we elucidated the mechanism of naphthalene carboxylase, a key enzyme in anaerobic naphthalene degradation. This is the first mechanism proposed for a carboxylase targeting nonsubstituted (polycyclic) aromatic compounds and can serve as a model for the initial activation reaction in the anaerobic degradation of benzene or nonsubstituted polycyclic aromatic hydrocarbons, as well as similar enzymatic reactions from the expanding class of UbiD-like (de)carboxylases.

Anaerobic Degradation of the Invasive Weed Solidago canadensis L. (goldenrod) and Copper Immobilization by a Community of Sulfate-Reducing and Methane-Producing Bacteria.

The weed Solidago canadensis L. poses a global threat to the environment as it spreads uncontrollably on roadsides, in forests, fields, meadows, and farmland. Goldenrod emits toxic substances that suppress other plants on the site, displacing wild ones. Thus, goldenrod conquers huge areas very quickly. The use of herbicides and mechanical methods does not solve the problem of the spontaneous spread of goldenrod. On the other hand, many scientists consider goldenrod as a valuable source of biologically active substances: flavonoids, phenolic compounds, vitamins, etc. In this study, we consider Solidago plants as a promising, free (cheap), and renewable substrate for the production of methane gas. The goal of the study was to identify the main patterns of degradation of the Solidago canadensis L. plant by methane-producing and sulfate-reducing bacteria with methane gas production and simultaneous detoxification of toxic copper. The composition of the gas phase was monitored by gas chromatography. The pH and redox potential parameters were determined potentiometrically; metal concentrations were measured by photometry. The concentration of flavonoids, sugars and phenolic compounds in plant biomass was determined according to well-known protocols. As a result of the study, high efficiencies of methane degradation in the Solidago plant and copper detoxification were obtained. Methane yield has reached the value of 68.2 L kg-1 TS of Solidago canadensis L. biomass. The degradation coefficient (Kd) was also high at 21.4. The Cu(II) was effectively immobilized by methanogens and sulfate reducers during the goldenrod degradation at the initial concentrations of 500 mg L-1. Thus, a new method of beneficial application of invasive plants was presented. The result confirms the possibility of using methanogenic microorganisms to produce methane gas from invasive weeds and detoxification of toxic metals.

Degradation of high-strength acrylic acid wastewater with anaerobic granulation technology: A mini-review.

The anaerobic granulation technology has been successfully applied full-scale for treating high-strength recalcitrant acrylic acid wastewater. This mini-review highlighted the recalcitrance of acrylic acid and its biological degradation pathways. And then, the full-scale practices using anaerobic granulation technology for acrylic wastewater treatment were outlined. The granules are proposed to provide barriers for high-concentration acrylic acid to the embedded anaerobic microbes, maintaining its high degradation rate without apparent substrate inhibition. Based on this proposal, the prospects of applying anaerobic granulation technology to handle a wide range of high-strength recalcitrant wastewaters, to improve the current process performances, and to recover renewable resources were delineated. The anaerobic granulation for high-strength recalcitrant wastewater treatment is an emergent technology that can assist in fulfilling the appeals of the circular bioeconomy of modern society.

Stimulating Anaerobic Degradation of Butyrate via Syntrophomonas wolfei and Geobacter sulfurreducens: Characteristics and Mechanism.

Anaerobic digestion (AD) has been widely applied for the degradation of organic wastewater due to its advantages of high-load operation and energy recovery. However, some challenges, such as low treatment capacity and instability caused by the accumulation of volatile fatty acids, limit its further application. Here, S. wolfei and G. sulfurreducens were initially co-cultured in the anaerobic anode of bio-electrochemical system for degrading butyric acid. Butyrate degradation characteristics in different conditions were quantitatively described. Moreover, G. sulfurreducens simultaneously strengthened the consumption of H2 and acetic acid via direct interspecies electron transfer, thereby strengthening the degradation of butyric acid via a co-metabolic process. During butyrate degradation, the co-culture of S. wolfei and G. sulfurreducens showed more advantages than that of S. wolfei and methanogens. This present study provides a new perspective of butyrate metabolism, which was independent of methanogens in an AD process.