Recalcitrant Aromatic Compounds Research Articles

The Catabolic Network of Aromatoleum aromaticum EbN1T.

The denitrifying betaproteobacterium Aromatoleum aromaticum EbN1T is a facultative anaerobic degradation specialist and belongs to the environmental bacteria studied best on the proteogenomic level. This review summarizes the current state of knowledge about the anaerobic and aerobic degradation (to CO2) of 47 organic growth substrates (23 aromatic, 21 aliphatic, and 3 amino acids) as well as the modes of respiratory energy conservation (denitrification vs. O2-respiration). The constructed catabolic network is comprised of 256 genes, which occupy ∼7.5% of the coding regions of the genome. In total, 219 encoded proteins have been identified by differential proteomics, yielding a proteome coverage of ∼74% of the network. Its degradation section is composed of 31 peripheral and 4 central pathways, with several peripheral modules (e.g., for 4-ethylphenol, 2-phenylethylamine, indoleacetate, and phenylpropanoids) discovered only after the complete genome [Arch Microbiol. 2005 Jan;183(1):27-36] and a first proteomic survey [Proteomics. 2007 Jun;7(13):2222-39] of A. aromaticum EbN1T were reported. The activation of recalcitrant aromatic compounds involves a suite of biochemically intriguing reactions ranging from C-H-bond activation (e.g., ethylbenzene dehydrogenase) via carboxylation (e.g., acetophenone carboxylase) to oxidative deamination (e.g., benzylamine), reductive dearomatization (benzoyl-CoA), and epoxide-forming oxygenases (e.g., phenylacetyl-CoA). The peripheral reaction sequences are substrate-specifically induced, mediated by specific transcriptional regulators with in vivo response thresholds in the nanomolar range. While lipophilic substrates (e.g., phenolics) enter the cells via passive diffusion, polar ones require active uptake that is driven by specific transporters. Next to the protein repertoire for canonical complexes I-III, denitrification, and O2-respiration (low- and high-affinity oxidases), the genome encodes an Ndh-II, a tetrathionate reductase, two ETF:quinone oxidoreductases, and two Rnf-type complexes, broadening the electron transfer flexibility of the strain. Taken together, the detailed catabolic network presented here forms a solid basis for future systems biology-level studies with A. aromaticum EbN1T.

Heteroatoms-doped biochar derived from deciduous resource as persulfate catalysts for efficient degradation of phenol

Phenol, as the representative recalcitrant aromatic organic compounds (AOCs) in wastewater, has been listed as the priority pollutant due to its toxicity to organisms and ecosystem. Therefore, the exploration of efficient and practical technology for phenol elimination from wastewater has become an important research topic. Peroxymonosulfate-based advanced oxidation processes (PMS-based AOPs) have been developed to be an efficient phenol-degrading strategy. Thereinto, the development of high-performance and cost-effective catalyst is the crux of degrading pollutants in PMS-based AOPs. In this study, the ubiquitous ginkgo deciduous resource is expected to convert into low-cost heteroatoms self-doped biochar to activate PMS for the removal of phenol. The removal rate of phenol (10 mg/L) achieved 90% within 60 min in pH of 6 by using 1.0 mmol/L of PMS and 0.5 g/L of catalyst. The phenol degradation in the catalytic process was accomplished via •OH, SO4• −, 1O2, and electron-transfer. On the basis of the correlation analysis, oxygen functional groups and sulfur functional groups were considered as the active sites that may be the most relevant to removal efficiency. This study proposed a facile method for converting deciduous waste resource into catalyst for triggering efficient catalytic oxidation process to degrade organic pollutants.

Nitrifiers drive successions of particulate organic matter and microbial community composition in a starved macrocosm.

Organic carbon produced by nitrifiers plays an important role in maintaining the microbial metabolism in the aphotic ocean layer with carbon and energy scarcity. However, the contribution of nitrifiers to organic carbon processing remains unclear. To explore how nitrification impacts the material cycle in the starved ecosystem, we set up an ultra-large volume, long-term incubation experiment. Seawater collected from the Halifax coastal ocean was pumped into the Aquatron Tower Tank located at Dalhousie University, Canada, and was incubated under dark conditions for 73days. The results indicated that the relative abundance of nitrifiers increased and nitrification was strengthened in the dark system where energy and organic carbon were scarce. The importance of nitrogenous compounds in particulate materials increased over the course of the incubation. Correlation analysis showed that the relative abundances of nitrifiers and particulate organic compounds containing nitrogen were significantly and positively correlated. Furthermore, network analysis suggested that metabolic processes related to nitrogenous and aromatic compounds are most important to particle associated bacteria. This study suggests that the nitrifiers could produce a series of organic compounds that result in the alteration of organic matter composition by promoting the degradation of recalcitrant aromatic compounds, which has important implications for organic matter processing in the starved dark ecosystem.

Characterization of 2,3,7,8-tetrachlorodibenzo-p-dioxin biodegradation by extracellular lignin-modifying enzymes from ligninolytic fungus

Ligninolytic fungi secrete extracellular lignin-modifying enzymes (LME) that degrade plant polymers for fungal nutrition but that are, because of their broad substrate specificity, also applicable for the degradation of many hazardous pollutants. Laccase is one of the most well characterized LME and is involved in the removal and degradation of recalcitrant aromatic compounds with or without the assistance of laccase-mediators. The Ligninolytic fungus Rigidoporus sp. FMD21 can degrade 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) with a half-life of 6.2 days. Using Rigidoporus sp. FMD21 crude extracellular enzyme extract (ExE) that mainly consisted of laccase, 77.4% of 2,3,7,8-TCDD was degraded within 36 days. The degradation rate did not depend on the 2,3,7,8-TCDD concentration in the tested range between 0.005 and 0.5 pgTEQ/μL. 2,3,7,8-TCDD was analysed by DR-CALUX® bioassay and the degradation was confirmed by GC-HRMS. In this study, we found evidence for cleavage of the diaryl ether bond in the 2,3,7,8-TCDD molecule and here we propose a new degradation mechanism in which 3,4-dichlorophenol is the main metabolite of 2,3,7,8-TCDD degradation by FMD21’s ExE. Six laccase-mediators were tested. Three of them 1-hydroxybenzotriazole (HBT), syringaldehyde (Syr) and violuric acid (Vio) showed an equipotent added effect on 2,3,7,8-TCDD degradation by ExE, however only in case of Vio a level of significance was reached. The others showed no effect or negatively impacted degradation. In conclusion, we have shown that Rigidoporus sp. FMD21 produces extracellular enzymes, mainly laccases that apparently are able to degrade the highly recalcitrant and most toxic 2,3,7,8-congener of TCDD via diaryl bond cleavage into 3,4-dichlorophenol.

Enzymatic C–H activation of aromatic compounds through CO2 fixation

The direct C-H carboxylation of aromatic compounds is an attractive route to the corresponding carboxylic acids, but remains challenging under mild conditions. It has been proposed that the first step in anaerobic microbial degradation of recalcitrant aromatic compounds is a UbiD-mediated carboxylation. In this study, we use the UbiD enzyme ferulic acid decarboxylase (Fdc) in combination with a carboxylic acid reductase to create aromatic degradation-inspired cascade reactions, leading to efficient functionalization of styrene through CO2 fixation. We reveal that rational structure-guided laboratory evolution can expand the substrate scope of Fdc, resulting in activity on a range of mono- and bicyclic aromatic compounds through a single mutation. Selected variants demonstrated 150-fold improvement in the conversion of coumarillic acid to benzofuran + CO2 and unlocked reactivity towards naphthoic acid. Our data demonstrate that UbiD-mediated C-H activation is a versatile tool for the transformation of aryl/alkene compounds and CO2 into commodity chemicals.

Iron acquisition system of Sphingobium sp. strain SYK-6, a degrader of lignin-derived aromatic compounds

Iron, an essential element for all organisms, acts as a cofactor of enzymes in bacterial degradation of recalcitrant aromatic compounds. The bacterial family, Sphingomonadaceae comprises various degraders of recalcitrant aromatic compounds; however, little is known about their iron acquisition system. Here, we investigated the iron acquisition system in a model bacterium capable of degrading lignin-derived aromatics, Sphingobium sp. strain SYK-6. Analyses of SYK-6 mutants revealed that FiuA (SLG_34550), a TonB-dependent receptor (TBDR), was the major outer membrane iron transporter. Three other TBDRs encoded by SLG_04340, SLG_04380, and SLG_10860 also participated in iron uptake, and tonB2 (SLG_34540), one of the six tonB comprising the Ton complex which enables TBDR-mediated transport was critical for iron uptake. The ferrous iron transporter FeoB (SLG_36840) played an important role in iron uptake across the inner membrane. The promoter activities of most of the iron uptake genes were induced under iron-limited conditions, and their regulation is controlled by SLG_29410 encoding the ferric uptake regulator, Fur. Although feoB, among all the iron uptake genes identified is highly conserved in Sphingomonad strains, the outer membrane transporters seem to be diversified. Elucidation of the iron acquisition system promises better understanding of the bacterial degradation mechanisms of aromatic compounds.

Comprehensive chemical analysis and characterization of heavy oil electric desalting wastewaters in petroleum refineries

Large quantities of highly polluted point-source wastewaters (EDWs) are generated from electric desalting process of heavy oils (HOs), resulting in severe impacts on the efficiency of wastewater treatment plants in petroleum refineries. In the present study, a comprehensive chemical analysis and characterization of EDWs of two typical Chinese heavy oils, Liaohe heavy oil (LHO) and Karamy heavy oil (KHO), were investigated using Daqing light oil (DLO) as a control. The HO-EDWs (LHO-EDW and KHO-EDW) show high pollutants contents with complicated compositions, more polar dissolved organic pollutants (DOPs), strong emulsion stability and high acute biotoxicity towards Vibrio fischeri, compared to DLO-EDW. LHO-EDW and KHO-EDW have nearly equal pollutants contents but different compositions and distributions, where more types of DOPs exist in KHO-EDW. Large amounts of biologically recalcitrant aromatic compounds, as well as heteroatomic compounds such as CHO, CHOS and CHON species, extensively distribute in HO-EDWs. The organic nitrogen compounds (e.g., anilines and N2–3Ox, N1OxS1) in KHO-EDW most probably contribute to and thus leading to elevated levels of acute biotoxicity. Additionally, highly dispersed colloidal, micron-sized particles and polar compounds promote the emulsification and stabilization of HO-EDWs. These results can guide the development of pretreatment technologies for HO-EDWs, thus improving the treatment and management of heavy oil refineries' wastewater streams.

Degradation strategies and associated regulatory mechanisms/features for aromatic compound metabolism in bacteria.

As a result of anthropogenic activity, large number of recalcitrant aromatic compounds have been released into the environment. Consequently, microbial communities have adapted and evolved to utilize these compounds as sole carbon source, under both aerobic and anaerobic conditions. The constitutive expression of enzymes necessary for metabolism imposes a heavy energy load on the microbe which is overcome by arrangement of degradative genes as operons which are induced by specific inducers. The segmentation of pathways into upper, middle and/or lower operons has allowed microbes to funnel multiple compounds into common key aromatic intermediates which are further metabolized through central carbon pathway. Various proteins belonging to diverse families have evolved to regulate the transcription of individual operons participating in aromatic catabolism. These proteins, complemented with global regulatory mechanisms, carry out the regulation of aromatic compound metabolic pathways in a concerted manner. Additionally, characteristics like chemotaxis, preferential utilization, pathway compartmentalization and biosurfactant production confer an advantage to the microbe, thus making bioremediation of the aromatic pollutants more efficient and effective.

Mesophilic and thermophilic anaerobic digestion of aqueous phase generated from hydrothermal liquefaction of cornstalk: Molecular and metabolic insights

The critical challenge of hydrothermal liquefaction (HTL) for bio-oil production from biomass is the production of large amounts of aqueous products (HTL-AP) with high organic contents. The present study investigated the anaerobic digestion (AD) performances of HTL-AP under both thermophilic and mesophilic conditions, and molecular and metabolic analysis were conducted to provide insights into the different performances. The results showed that thermophilic AD had lower COD removal efficiency compared to mesophilic AD (45.0% vs. 61.6%). Liquid chromatography coupled with organic carbon detection and organic nitrogen (LC-OCD-OND) analysis showed that both high molecular weight (HMW) and low molecular weight (LMW) compounds were degraded to some extent and more LMW acids (LMWA) and recalcitrant aromatic compounds were degraded in the mesophilic reactor, which was the main reason of higher COD removal efficiency. Phenyl compounds (e.g. phenol and 2 methoxyphenol), furans and pyrazines were the recalcitrant chemicals detected through GC-MS analysis. Fourier transform ion cyclone resonance mass spectrometry (FT-ICR-MS) analysis demonstrated the complexity of HTL-AP and the proportions of phenolic or condensed aromatic compounds increased especially in the thermophilic effluents. Metabolites analysis showed that the reasons contributing to the differences of mesophilic and thermophilic AD were not only related to the degradation of organic compounds (e.g. benzoate degradation via CoA ligation) in HTL-AP but also related to the microbial autogenesis (e.g. fatty acid biosynthesis) as well as the environmental information processing. In addition, the enrichment of Mesotoga, responsible for the high degradation efficiency of LMWA, and Pelolinea, involved in the degradation of phenyl compounds, were found in mesophilic reactor, which was consistent with higher removal of corresponding organics.

Spectroscopic Characterisation of the Naphthalene Dioxygenase from Rhodococcus sp. Strain NCIMB12038.

Polycyclic aromatic hydrocarbons (PAHs), such as naphthalene, are potential health risks due to their carcinogenic and mutagenic effects. Bacteria from the genus Rhodococcus are able to metabolise a wide variety of pollutants such as alkanes, aromatic compounds and halogenated hydrocarbons. A naphthalene dioxygenase from Rhodococcus sp. strain NCIMB12038 has been characterised for the first time, using electron paramagnetic resonance (EPR) spectroscopy and UV-Vis spectrophotometry. In the native state, the EPR spectrum of naphthalene 1,2-dioxygenase (NDO) is formed of the mononuclear high spin Fe(III) state contribution and the oxidised Rieske cluster is not visible as EPR-silent. In the presence of the reducing agent dithionite a signal derived from the reduction of the [2Fe-2S] unit is visible. The oxidation of the reduced NDO in the presence of O2-saturated naphthalene increased the intensity of the mononuclear contribution. A study of the “peroxide shunt”, an alternative mechanism for the oxidation of substrate in the presence of H2O2, showed catalysis via the oxidation of mononuclear centre while the Rieske-type cluster is not involved in the process. Therefore, the ability of these enzymes to degrade recalcitrant aromatic compounds makes them suitable for bioremediative applications and synthetic purposes.

Biodegradation of phenol, anthracene and acenaphthene singly and consortium culture of indigenous microorganism isolates from underground coal gasification area

The gasification of Coal has a potential for long term energy supply in the future by conversion of non-mineable coal seams to syngas for various industrial applications. However, recalcitrant aromatic compounds such as phenol, anthracene, and acenaphthene from UGC are potential to contaminate groundwater, soil and the broader environment. Therefore, the polluted environment required bioremediation. The purpose of this study was to determine the capability of Bacillus velezensis and Stenotropromonas maltophilia isolated from the area of Underground Coal Gasification (UCG) on biodegradation of phenol, anthracene, and acenaphthene in the single and mixed cultures. The degradation of pollutants in the concentration range of 250-500 ppm evaluated by measuring of the ability of bacteria to grow in single component supplemented mineral salt media, screening tests for hydrocarbon-degrading bacteria and biodegradation ability by gravimetric analysis method. The results showed biodegradability of phenol, anthracene, and acenaphthene at concentration 500 ppm after 14 days by S. maltophilia were 60,2%, 80,4%, and 82,5%. B. velezensis showed 79% of biodegradability of phenol, anthracene 79,8%, and acenaphthene 81,1%. The consortium of both isolates has the highest percentage of phenol, anthracene, and acenapthene biodegradation 83,4 %, 83, 53 % and 83,47 % respectively.

INVESTIGACIÓN DE UNA lacasa NOVEDOSA PRODUCTORA DE UNA ESPECIE FÚNGICA Y SU ESPECIFICIDAD SOBRE EL SUBSTRATO POR MÉTODOS HÚMEDOS Y SECOS

Fungal laccase has potential to degrade organic pollutants of waste water in the presence of redox mediators. Choice of better enzyme-redox mediator system can enhance the degradation of recalcitrant aromatic compounds. Present study aimed at the screening and identification of laccase producing fungal strain. Evaluate its substrate specificity to understand the interaction between the enzyme and different substrates. A total of 30 fungal strains were isolated from air, soil and decaying wood of mangifera indica and screened for their ability to produce laccase. The enzyme was produced on sawdust, a cheap substrate under static condition. Specie identification of the most potent strain was carried out by DNA barcoding. Tree building tool was used to get the phylogenetic relationship. Best enzyme substrate complex was determined by combining wet and dry lab investigations using different substrates. Predicted protein model was generated and used for docking analysis with the mediators. Only 15 strains exhibited laccase producing capability and the strain MFw3 showed highest enzyme activity 297±1 U/ml/min. Phylogenetic tree analysis showed the novelity of Aspergillus flavus MFw3 (GenBank accession number: MF167358). Bioinformatics tools were used to get the best enzyme/substrate complex and ABTS was found best substrate with minimum dock score (30.2). Our results showed that MFw3 is a novel strain and has great potential for laccase production by using ABTS as a substrate.

Community proteogenomics reveals the systemic impact of phosphorus availability on microbial functions in tropical soil.

Phosphorus is a scarce nutrient in many tropical ecosystems, yet how soil microbial communities cope with growth-limiting phosphorus deficiency at the gene and protein levels remains unknown. Here, we report a metagenomic and metaproteomic comparison of microbial communities in phosphorus-deficient and phosphorus-rich soils in a 17-year fertilization experiment in a tropical forest. The large-scale proteogenomics analyses provided extensive coverage of many microbial functions and taxa in the complex soil communities. A greater than fourfold increase in the gene abundance of 3-phytase was the strongest response of soil communities to phosphorus deficiency. Phytase catalyses the release of phosphate from phytate, the most recalcitrant phosphorus-containing compound in soil organic matter. Genes and proteins for the degradation of phosphorus-containing nucleic acids and phospholipids, as well as the decomposition of labile carbon and nitrogen, were also enhanced in the phosphorus-deficient soils. In contrast, microbial communities in the phosphorus-rich soils showed increased gene abundances for the degradation of recalcitrant aromatic compounds, transformation of nitrogenous compounds and assimilation of sulfur. Overall, these results demonstrate the adaptive allocation of genes and proteins in soil microbial communities in response to shifting nutrient constraints.

Mechanism of aerobic denitrifiers and calcium nitrate on urban river sediment remediation

Calcium nitrate, coupled with aerobic denitrifiers, were applied to reduce nitrogen and organic pollutants of urban river sediment. 115-day remediation tests showed that total nitrogen in the sediment decreased by 16.5%, while total transferable nitrogen increased from 0.097 to 0.109 mg/g. Increased urease activity and decreased protease showed a potential for further nitrogen degradation. Although the total organic carbon (TOC) content decreased slightly, the predominant organic pollutants were removed or converted to low molecular weight compounds through microbial biodegradation. Decreased phenol oxidase activity exhibited that low concentrations of recalcitrant aromatic compounds remained in the sediment. The high-throughput sequencing results reveals that the community diversity of bacteria markedly increased. Compared with the addition of calcium nitrate or aerobic denitrifiers alone, genera mainly removed nitrogen and organic compounds, especially recalcitrant compounds, following the treatment with the combination technology. Moreover, the concentrations of nitrogen and organic matter did not increase significantly in the overlying water, which was free of undesirable secondary pollution. During the process of remediation, dosing with nitrate may provide a more favorable environment for aerobic denitrifiers and indigenous microbes in the sediment. Furthermore, the introduced aerobic denitrifiers may improve nitrogen removal and have a collaborative effect with indigenous microbes.

Degradation of sec-hexylbenzene and its metabolites by a biofilm-forming yeast Trichosporon asahii B1 isolated from oil-contaminated sediments in Quangninh coastal zone, Vietnam

ABSTRACTThis article reports on the ability of yeast Trichosporon asahii B1 biofilm-associated cells, compared with that of planktonic cells, to transform sec-hexylbenzene and its metabolites. This B1 strain was isolated from a petroleum-polluted sediment collected in the QuangNinh coastal zones in Vietnam, and it can transform the branched aromatic hydrocarbons into a type of forming biofilm (pellicle) more efficiency than that the planktonic forms can. In the biofilm cultivation, seven metabolites, including acetophenone, benzoic acid, 2,3-dihydroxybenzoic acid, β-methylcinnamic acid, 2-phenylpropionic acid, 3-phenylbutyric acid, and 5-phenylhexanoic acid were extracted by ethyl acetate and analyzed by HPLC and GC-MS. In contrast, in the planktonic cultivation, only three of these intermediates were found. An individual metabolite was independently used as an initial substrate to prove its degradation by biofilm and planktonic types. The degradation of these products indicated that their inoculation with B1 biofilms was indeed higher than that observed in their inoculation with B1 planktonic cells. This is the first report on the degradation of sec-hexylbenzene and its metabolites by a biofilm-forming Trichosporon asahii strain. These results enhance our understanding of the degradation of branched-side-chain alkylbenzenes by T. asahii B1 biofilms and give a new insight into the potential role of biofilms formed by such species in the bioremediation of other recalcitrant aromatic compounds.

Time-series metagenomic analysis reveals robustness of soil microbiome against chemical disturbance.

Soil microbial communities have great potential for bioremediation of recalcitrant aromatic compounds. However, it is unclear which taxa and genes in the communities, and how they contribute to the bioremediation in the polluted soils. To get clues about this fundamental question here, time-course (up to 24 weeks) metagenomic analysis of microbial community in a closed soil microcosm artificially polluted with four aromatic compounds, including phenanthrene, was conducted to investigate the changes in the community structures and gene pools. The pollution led to drastic changes in the community structures and the gene sets for pollutant degradation. Complete degradation of phenanthrene was strongly suggested to occur by the syntrophic metabolism by Mycobacterium and the most proliferating genus, Burkholderia. The community structure at Week 24 (∼12 weeks after disappearance of the pollutants) returned to the structure similar to that before pollution. Our time-course metagenomic analysis of phage genes strongly suggested the involvement of the ‘kill-the-winner’ phenomenon (i.e. phage predation of Burkholderia cells) for the returning of the microbial community structure. The pollution resulted in a decrease in taxonomic diversity and a drastic increase in diversity of gene pools in the communities, showing the functional redundancy and robustness of the communities against chemical disturbance.

Draft Genome Sequence of the Lignin-Degrading Burkholderia sp. Strain LIG30, Isolated from Wet Tropical Forest Soil.

Burkholderia species are common soil Betaproteobacteria capable of degrading recalcitrant aromatic compounds and xenobiotics. Burkholderia sp. strain LIG30 was isolated from wet tropical forest soil and is capable of utilizing lignin as a sole carbon source. Here we report the draft genome sequence of Burkholderia sp. strain LIG30.

Transformation of iso-pentylbenzene by a biofilm-forming strain of Candida viswanathii TH1 isolated from oil-polluted sediments collected in coastal zones in Vietnam

This work is aimed to assess the aerobic biotransformation of a branched side chain alkylbenzene, iso-pentylbenzene, by Candida viswanathii TH1. The yeast Candida viswanathii TH1 isolated from oil-polluted sediments collected in coastal zones in Vietnam exhibited as a strain that could better transform branched aromatic hydrocarbons in biofilm (pellicle) than in planktonic form. During incubation of TH1 as biofilm with iso-pentylbenzene, the seven intermediates produced were benzoic acid, phenylacetic acid, 2-methyl-4-phenyl-butan-1-ol, 2-hydroxy-phenylacetic acid, 2-methyl-4-phenylbutyric acid, succinic acid and iso-valerophenone as revealed by gas chromatography/mass spectra and high-performance liquid chromatography analyses. The occurrence of these intermediates showed that iso-pentylbenzene could be oxidized not only via mono- but also by a sub-terminal oxidation pathway. This is the first study on iso-pentylbenzene transformation by a biofilm-forming Candida viswanathii strain. The catabolic versatility of the biofilm-forming strain TH1 and its use for mono and sub-terminal oxidation during the transformation of iso-pentylbenzene enhance our understanding of the degradation of branched side chain phenylalkanes and give new insight into the potential role of such species in the transformation of other recalcitrant aromatic compounds.

Protective effect of vanilloids against chemical stress on the white-rot fungus Ganoderma lucidum

Bioremediation of contaminated sites by biosorption of pollutants onto a wide range of materials has emerged as a promising treatment for recalcitrant aromatic compounds or heavy metals. When adsorption occurs on living white-rot fungi mycelia, the pollutants may be degraded by ligninolytic enzymes. However, the survival of mycelia in harsh conditions is one of the drawbacks of those methodologies. In this study, it was demonstrated that culture media supplemented with several guaiacol derivatives (vanilloids) increased the resistance of Ganoderma lucidum E47 cultures to chemical stress by enhancing the adsorptive capacity of the extracellular mucilaginous material (ECMM). The toxicity of the fungicides gentian violet (GV), malachite green (MG) and clotrimazole, and the heavy metal Cadmium was noticeably diminished in fungal cultures supplemented with the guaiacol derivative vanillic acid (VA). No degradation of the tested compounds was detected. The activity of the oxidative enzymatic systems like laccase, a well-known oxidase associated to dye degradation, was only detectable after complete growth on plates. Extremely low concentrations of VA caused a significant protective effect, radial extension of the growth halo in plates supplemented with 0.0001 mM of VA plus GV was up to 20% to that obtained in control plates (without addition of GV and VA). Therefore, the protective effect could not be attributable to VA per se. ECMM separated from the mycelium exhibited a much higher increase in the adsorptive capacity when isolated from liquid cultures containing VA, while that obtained from unsupplemented cultures showed an almost null adsorptive capacity.

Comparison of 26 Sphingomonad Genomes Reveals Diverse Environmental Adaptations and Biodegradative Capabilities

Sphingomonads comprise a physiologically versatile group within the Alphaproteobacteria that includes strains of interest for biotechnology, human health, and environmental nutrient cycling. In this study, we compared 26 sphingomonad genome sequences to gain insight into their ecology, metabolic versatility, and environmental adaptations. Our multilocus phylogenetic and average amino acid identity (AAI) analyses confirm that Sphingomonas, Sphingobium, Sphingopyxis, and Novosphingobium are well-resolved monophyletic groups with the exception of Sphingomonas sp. strain SKA58, which we propose belongs to the genus Sphingobium. Our pan-genomic analysis of sphingomonads reveals numerous species-specific open reading frames (ORFs) but few signatures of genus-specific cores. The organization and coding potential of the sphingomonad genomes appear to be highly variable, and plasmid-mediated gene transfer and chromosome-plasmid recombination, together with prophage- and transposon-mediated rearrangements, appear to play prominent roles in the genome evolution of this group. We find that many of the sphingomonad genomes encode numerous oxygenases and glycoside hydrolases, which are likely responsible for their ability to degrade various recalcitrant aromatic compounds and polysaccharides, respectively. Many of these enzymes are encoded on megaplasmids, suggesting that they may be readily transferred between species. We also identified enzymes putatively used for the catabolism of sulfonate and nitroaromatic compounds in many of the genomes, suggesting that plant-based compounds or chemical contaminants may be sources of nitrogen and sulfur. Many of these sphingomonads appear to be adapted to oligotrophic environments, but several contain genomic features indicative of host associations. Our work provides a basis for understanding the ecological strategies employed by sphingomonads and their role in environmental nutrient cycling.