Direct ammonium oxidation to nitrogen gas (Dirammox) in Alcaligenes strain HO-1: The electrode role.

It has been recently suggested that Alcaligenes use a previously unknown pathway to convert ammonium into dinitrogen gas (Dirammox) via hydroxylamine (NH2OH). This fact alone already implies a significant decrease in the aeration requirements for the process, but the process would still be dependent on external aeration. This work studied the potential use of a polarised electrode as an electron acceptor for ammonium oxidation using the recently described Alcaligenes strain HO-1 as a model heterotrophic nitrifier. Results indicated that Alcaligenes strain HO-1 requires aeration for metabolism, a requirement that cannot be replaced for a polarised electrode alone. However, concomitant elimination of succinate and ammonium was observed when operating a previously grown Alcaligenes strain HO-1 culture in the presence of a polarised electrode and without aeration. The usage of a polarised electrode together with aeration did not increase the succinate nor the nitrogen removal rates observed with aeration alone. However, current density generation was observed along a feeding batch test representing an electron share of 3% of the ammonium removed in the presence of aeration and 16% without aeration. Additional tests suggested that hydroxylamine oxidation to dinitrogen gas could have a relevant role in the electron discharge onto the anode. Therefore, the presence of a polarised electrode supported the metabolic functions of Alcaligenes strain HO-1 on the simultaneous oxidation of succinate and ammonium.

Partially shielded enzymes capable of processing large protein substrates.

We report the first method of enzyme protection enabling the production of partially shielded enzymes capable of processing substrates as large as proteins. We show that partially shielded sortase retains its transpeptidase activity and can perform bioconjugation reactions on antibodies. Moreover, a partially shielded trypsin is shown to outperform its soluble counterpart in terms of proteolytic kinetics. Remarkably, partial enzyme shielding results in a drastic increase in temporal stability of the enzyme.

Accumulation and Transformation of 2,2',4,4'-Tetrabrominated Diphenyl Ether (BDE47) by the Earthworm Metaphire vulgaris in Soil.

The accumulation and transformation of 2,2',4,4'-tetrabrominated diphenyl ether (BDE47), one congener of the flame retardants polybrominated diphenyl ethers (PBDEs), in soil-feeding fauna are still unknown. Using radioactivity tracer, we incubated 14C-labelled BDE47 in soil for 21 days in the presence and absence of the geophagous earthworm Metaphire vulgaris. BDE47 accumulated in the earthworm predominantly via oral ingestion of soil, giving a biota-soil accumulation factor (BSAF) value of 1.3 for radioactivity at the end of incubation, and was mostly located in intestine, followed by clitellum (organs region) and skin of earthworms. Accumulation was accompanied by significant decrease of BDE47 concentration in soil porewater and BDE47 mineralization in soil. BDE47 was transformed in the earthworm gut into two metabolites with higher polarities than BDE47. The results provide for the first time insights into accumulation and transformation of lower-brominated congeners of PBDEs in geophagous earthworms, being helpful for environmental risk assessment of PBDEs.

Fate of 4-bromodiphenyl ether (BDE3) in soil and the effects of co-existed copper.

The quantitative fate of polybrominated diphenyl ethers (PBDEs) in soil is unknown. Furthermore, the effects of co-contamination by toxic copper on the behavior of PBDEs have not been investigated. Using a 14C-tracer, we studied mineralization, metabolism, and formation of non-extractable residues (NERs) of one PBDE congener, i.e., the 4-bromodiphenyl ether (BDE3) in oxic soil for 50 days, without and with amendment of Cu (400 mg kg-1 soil dw). BDE3 rapidly dissipated with a half-life of 5.5 days and large amounts of CO2 (38.8 ± 0.3% of initial applied amount at the end of incubation) and NERs (42.5 ± 0.4%) were rapidly produced. One hydroxylated metabolite (4'-HO-BDE3) was formed (8.1 ± 0.6%) at the beginning of the incubation, but then decreased to 2.2 ± 0.4%. Only BDE3 occurred in physico-chemically entrapped NERs, amounting to 9.2 ± 0.7%, while only 4'-HO-BDE3 in ester-linked NERs (10.9 ± 0.7%). The addition of Cu strongly reduced the kinetics constants of the transformations (including dissipation, mineralization, and NER-formation), the predicted maximal amounts of mineralization, as well as covalent binding of 4'-HO-BDE3 to soil. The results provide first quantitative insights into the fate of low-brominated congeners of PBDEs in soil and indicate that co-contamination by Cu may increase the environmental risks of biodegradable PBDEs in soil by increasing their persistence.

Hydrophobicity-responsive engineered mesoporous silica nanoparticles: application in the delivery of essential nutrients to bacteria combating oil spills.

Facile chemical modification of mesoporous silica particles allows the production of gated reservoir systems capable of hydrophobicity-triggered release. Applied to the delivery of nutrients specifically to an oil phase, the systems developed have been shown to reliably assist the bacterial degradation of hydrocarbons. The gated system developed, made of C18 hydrocarbon chains, is demonstrated to be in a closed collapsed state in an aqueous environment, yet opens up through solvation by lipophilic alkanes and releases its content on contact with the oil phase.

Reversibility of enzymatic reactions might limit biotransformation of organic micropollutants.

Biotransformation of many organic micropollutants (OMPs) in sewage treatment plants is incomplete leading to their release into the environment. Recent findings suggest that thermodynamic aspects of the reaction as chemical equilibrium limit biotransformation, while kinetic parameters have a lower influence. Reversibility of enzymatic reactions might result in a chemical equilibrium between the OMP and the transformation product, thus impeding a total removal of the compound. To the best of our knowledge, no study has focused on proving the reversible action of enzymes towards OMPs so far. Therefore, we aimed at demonstrating this hypothesis through in vitro assays with bisphenol A (BPA) in the presence of kinase enzymes, namely acetate kinase and hexokinase, which are key enzymes in anaerobic processes. Results suggest that BPA is phosphorylated by acetate kinase and hexokinase in the presence of ATP (adenosine 5-triphosphate), but when the concentration of this co-substrate decreases and the enzymes loss their activity, the backward reaction occurs, revealing a reversible biotransformation mechanism. This information is particularly relevant to address new removal strategies, which up to now were mainly focused on modifying the kinetic parameters of the reaction.

Combined speciation analysis by X-ray absorption near-edge structure spectroscopy, ion chromatography, and solid-phase microextraction gas chromatography-mass spectrometry to evaluate biotreatment of concentrated selenium wastewaters.

In this study we evaluate the potential of anaerobic granular sludge as an inoculum for the bioremediation of selenium-contaminated waters using species-specific analytical methods. Solid species formed by microbial reduction were investigated using X-ray absorption near-edge structure (XANES) spectroscopy at the selenium K-edge. Furthermore, dissolved selenium species were specifically determined by ion chromatography (IC) and solid-phase microextraction gas chromatography-mass spectrometry (SPME-GC-MS). Least-squares linear combination of the XANES spectra for samples incubated with the highest selenate/selenite concentrations (10(-3) M) show the predominance of elemental selenium and a Se(-I) selenide, such as ferroselite, the thermodynamically most stable iron selenide. In contrast, elemental selenium and Se(-II) selenides are the main species detected at the lower selenate/selenite concentrations. In each repeated fed batch incubation, most aqueous selenite anions were converted into solid selenium species, regardless of the type of electron donor used (acetate or H(2)/CO(2)) and the selenium concentration applied. On the other hand, at higher concentrations of selenate (10(-4) and 10(-3) M), significant amounts of the oxyanion remained unconverted after consecutive incubations. SPME-GC-MS demonstrated selenium alkylation with both electron donors investigated, as dimethyl selenide (DMSe) and dimethyl diselenide (DMDSe). Selenite was even more alkylated in the presence of H(2)/CO(2) (maximum 2156 µg of Se/L of DMSe + DMDSe) as compared to acetate (maximum 50 µg of Se/L). In contrast, selenate was less alkylated using both electron donors (maximum 166 and 3 µg of Se/L, respectively). The high alkylation potential for selenite limits its bioremediation in selenium laden waters involving H(2)/CO(2) as the electron donor despite the fact that nontoxic elemental selenium and thermodynamically stable metal selenide species are formed.