Evaluation of anionic surfactant removal in anaerobic reactor with Fe(III) supplementation.

The objective of this study was to evaluate the removal of linear alkylbenzene sulfonate (LAS) associated with Fe(III) supplementation using an expanded granular sludge bed (EGSB) reactor. The reactor was inoculated with a granular sludge and fed with synthetic wastewater containing a specific LAS load rate (SLLR) of 1.5 mg gVS-1 d-1 (∼16.4 mgLAS L-1 influent) and supplied with 7276 µMol L-1 of Fe(III). The biomasses from the inoculum and at the end of the EGSB-Fe operation (127 days) were characterized using 16S rRNA Ion Tag sequencing. An increase of 20% in the removal efficiency was observed compared to reactors without Fe(III) supplementation that was reported in the literature, and the LAS removal was approximately 84%. The Fe(III) reduction was dissimilatory (the total iron concentration in the influent and effluent were similar) and reached approximately 64%. The higher Fe(III) reduction and LAS removal were corroborated by the enrichment of genera, such as Shewanella (only EGSB-Fe - 0.5%) and Geobacter (1% - inoculum; 18% - EGSB-Fe). Furthermore, the enrichment of genera that degrade LAS and/or aromatic compounds (3.8% - inoculum; 29.6% - EGSB-Fe of relative abundance) was observed for a total of 20 different genera.

Editor's Highlight: Perfluorooctane Sulfonate-Choline Ion Pair Formation: A Potential Mechanism Modulating Hepatic Steatosis and Oxidative Stress in Mice.

The mechanisms underlying perfluorooctane sulfonate (PFOS)-induced steatosis remain unclear. The hypothesis that PFOS causes steatosis and other hepatic effects by forming an ion pair with choline was examined. C57BL/6 mice were fed either a control diet or a marginal methionine/choline-deficient (mMCD) diet, with and without 0.003, 0.006, or 0.012% potassium PFOS. Dietary PFOS caused a dose-dependent decrease in body weight, and increases in the relative liver weight, hepatic triglyceride concentration and serum markers of liver toxicity and oxidative stress. Some of these effects were exacerbated in mice fed the mMCD diet supplemented with 0.012% PFOS compared with those fed the control diet supplemented with 0.012% PFOS. Surprisingly, serum PFOS concentrations were higher while liver PFOS concentrations were lower in mMCD-fed mice compared with corresponding control-fed mice. To determine if supplemental dietary choline could prevent PFOS-induced hepatic effects, C57BL/6 mice were fed a control diet, or a choline supplemental diet (1.2%) with or without 0.003% PFOS. Lipidomic analysis demonstrated that PFOS caused alterations in hepatic lipid metabolism in the PFOS-fed mice compared with controls, and supplemental dietary choline prevented these PFOS-induced changes. Interestingly, dietary choline supplementation also prevented PFOS-induced oxidative damage. These studies are the first to suggest that PFOS may cause hepatic steatosis and oxidative stress by effectively reducing the choline required for hepatic VLDL production and export by forming an ion pair with choline, and suggest that choline supplementation may prevent and/or treat PFOS-induced hepatic steatosis and oxidative stress.

Interaction of microbial sulphate reduction and methanogenesis in oil sands tailings ponds.

Anaerobic turnover of organic compounds in oil sands tailings ponds is accomplished by a complex microbial consortium. We examined major electron accepting processes in mature fine tailings (MFT). Beside methanogenesis and sulphate reduction, microbial iron reduction was an important process of anaerobic respiration. Microbial numbers and activity were comparable to those reported for natural lakes. To understand metabolic interactions of indigenous methanogenic and sulphate-reducing communities, we conducted a 6 month microcosm experiment with MFT supplemented with easily available carbon sources and molybdate and/or 2-bromoethane sulphonate (BES) as specific inhibitors for sulphate reduction and methanogenesis. Methanogenesis increased when microcosms were supplemented with extra carbon, but was completely inhibited by the addition of BES. Molybdate not only inhibited sulphate reduction, but also methanogenesis, indicating a positive relation between the two processes. The turnover of extra carbon sources differed between microcosms treated with molybdate and BES. Acetate and propionate were not consumed in microcosms amended with molybdate, indicating that sulphate-reducing bacteria (SRB) were responsible for their metabolisation, and that methane was rather produced by hydrogenotrophic methanogens. In microcosms without molybdate, acetate transiently accumulated, indicating the activity of both incomplete and complete oxidizing SRB. Ethanol and lactate were also consumed in the simultaneous presence of BES and molybdate, demonstrating the occurrence of other anaerobic processes. Biomass increased by the addition of extra carbon, mainly due to a relative increase in the proportion of SRB. The addition of extra carbon lowered the degradation of BTEX compounds.

Carryover of perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) from soil to plant and distribution to the different plant compartments studied in cultures of carrots (Daucus carota ssp. Sativus), potatoes (Solanum tuberosum), and cucumbers (Cucumis Sativus).

A vegetation study was carried out to investigate the carryover of Perfluorooctanoic Acid (PFOA) and Perfluorooctane Sulfonate (PFOS) from soil mixed with contaminated sewage sludge to potato, carrot, and cucumber plants. Analysis was done by liquid-extraction using acetonitrile with dispersive SPE cleanup and subsequent HPLC-MS/MS. In order to assess the transfer potential from soil, transfer factors (TF) were calculated for the different plant compartments: TF = [PFC](plant (wet substance))/[PFC](soil (dry weight)). The highest TF were found for the vegetative plant compartments with average values for PFOS below those for PFOA: cucumber, 0.17 (PFOS), 0.88 (PFOA); potato, 0.36 (PFOS), 0.40 (PFOA); carrot, 0.38 (PFOS), 0.53 (PFOA). Transfer of PFOA and PFOS into potato peelings (average values of TF: PFOA 0.03, PFOS 0.04) exceeded the carryover to the peeled tubers (PFOA 0.01, PFOS < 0.01). In carrots, this difference did not occur (average values of TF: PFOA 0.04, PFOS 0.04). Transfer of PFOS into the unpeeled cucumbers was low and comparable to that of peeled potatoes (TF < 0.01). For PFOA, it was higher (TF: 0.03).

Production and characterization of methyl ester sophorolipids with 22-carbon-fatty acids.

In this study, various fatty acids of 18 and 22 carbons were used as substrates to produce sophorolipids in Candida bombicola ATCC 22214. Methyl esterification of erucic acid and C22-enriched rapeseed oil generated mainly acidic sophorolipids with different degrees of saturation in the fatty acid chain, which was demonstrated by structural analysis using HPLC, LC-MS and NMR spectroscopy. Regardless of methyl esterification, the rapeseed oil served as the best substrate for high production of sophorolipids among the tested. Methyl esterification also had no noticeable effect on the interfacial properties of sophorolipids. However, 22 carbons introduced in the fatty acid chain increased the hydrophobicity of sophorolipids, and therefore improved surface-active properties and biodegradability.

Effect of temperature and organic nutrients on the biodegradation of linear alkylbenzene sulfonate (LAS) during the composting of anaerobically digested sludge from a wastewater treatment plant.

Limits on the application of biosolids (anaerobically processed sludges from wastewater treatment plants) as fertilizers for the amendment of soil are becoming greater because of the accumulation of recalcitrant substances, making necessary the use of techniques that bring the concentration of xenobiotics to lower concentrations than those permitted. In general, the biosolids composting process is sufficient to reduce the usual concentration of linear alkylbenzene sulfonates (LAS) to low levels. In this work, an assessment is made on the effect of temperature in the capacity of enriched bacterial populations to biodegrade LAS, together with the influence that the available nutrients may have in the biodegradation of these compounds. The results show that the microbial metabolism of LAS was not observed in the thermophilic range. The optimum temperature for the biodegradation of LAS appears to be around 40 degrees C, this is, the lowest assayed here, and at this temperature the differences in the biodegradation of LAS among the nutritionally supplemented cultures are small.

Display of active subtilisin 309 on phage: analysis of parameters influencing the selection of subtilisin variants with changed substrate specificity from libraries using phosphonylating inhibitors.

Many attempts have been made to endow enzymes with new catalytic activities. One general strategy involves the creation of random combinatorial libraries of mutants associated with an efficient screening or selection scheme. Phage display has been shown to greatly facilitate the selection of polypeptides with desired properties by establishing a close link between the polypeptide and the gene that encodes it. Selection of phage displayed enzymes for new catalytic activities remains a challenge. The aim of this study was to display the serine protease subtilisin 309 (savinase) from Bacillus lentus on the surface of filamentous fd phage and to develop selection schemes that allow the extraction of subtilisin variants with a changed substrate specificity from libraries. Subtilisins are produced as secreted preproenzyme that mature in active enzyme autocatalytically. They have a broad substrate specificity but exhibit a significant preference for hydrophobic residues and very limited reactivity toward charged residues at the P4 site in the substrate. Here, we show that savinase can be functionally displayed on phage in the presence of the proteic inhibitor CI2. The free enzyme is released from its complex with CI2 upon addition of the anionic detergent LAS. The phage-enzyme can be panned on streptavidin beads after labelling by reaction with (biotin-N-epsilon-aminocaproyl-cystamine-N'-glutaryl)-l-Ala-l-Ala-l-P ro-Phe(P)-diphenyl ester. Reactions of libraries, in which residues 104 and 107 forming part of the S4 pocket have been randomised, with (biotin-N-epsilon-aminocaproyl-cystamine-N'-glutaryl)-alpha-l-Lys-l-A la-l-Pro-Phe(P)-diphenylester allowed us to select enzymes with increased specific activity for a substrate containing a lysine in P4. Parameters influencing the selection as for instance the efficiency of maturation of mutant enzymes in libraries have been investigated.