Acceleration of the bio-reduction of methyl orange by a magnetic and extracellular polymeric substance nanocomposite.

Extracellular electron transfer (EET) plays an important role in bio-reduction of environmental pollutants. Extracellular polymeric substances (EPS), a kind of biogenic macromolecule, contain functional groups responsible for acceleration of EET. In this study, azo dye-methyl orange (MO) was chosen as a model pollutant, and a Fe3O4 and EPS nanocomposite (Fe3O4@EPS) was prepared to evaluate its promotion on the bio-reduction of MO. The flower-like core-shell configuration of Fe3O4@EPS with a 12 nm of light layer of EPS was confirmed by TEM. The redox ability of EPS was well reserved on Fe3O4@EPS by FTIR and electrochemical test. The application of Fe3O4@EPS on sustained acceleration of MO decolorization were confirmed by batch experiments and anaerobic sequenced batch reactors. Due to biocompatibility of the biogenic shell, the as-prepared Fe3O4@EPS exhibited low toxic to microorganisms by the Live/dead cell test. Moreover, negligible leaching of EPS under high concentration of various anions and less than 10% of EPS was released under extreme acidic and basic pH condition. The results of study provided a new preparation method of biological intimate and environmentally friendly redox mediators and suggested a feasible way for its use on bio-reduction of pollutants.

Adsorption characteristics of methylene blue by a dye-degrading and extracellular polymeric substance -producing strain.

Biosorption of dye by microbes and the extracellular polymeric substances (EPS) were of great environmental significance, especially for the dye-degrading and EPS-producing strain. Previous studies were mainly focused on the adsorption capacities and regeneration properties of pure culture, few were on the biosorption of dyes by the dye-degraders and the contributions of EPS on adsorption. In this study, a dye-degrading and EPS-producing strain i.e., Klebsiella oxytoca was used to evaluate its removal capacity to methylene blue. The maximum adsorption capacity (qe) by the strain was calculated as 145 mg g-1, which is superior to many reported bio-adsorbents and some synthetic materials. Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy results suggested that CO, -NH2 and P-OH groups were involved in the adsorption. High pressure steam sterilization (HPSS) increased the hydrophilicity of cell wall but did not significantly change the cell structure. Compared with the dead resting cell (DRC), the relative higher qe obtained by live resting cell (LRC) possibly due to the loss of some cell structure during the HPSS process. Adsorption experiments by EPS-free LRC, confocal laser microscope and three-dimensional excitation-emission matrix fluorescence spectroscopy results confirmed that the EPS played a role in the adsorption of MB dye. The adsorption characteristics of the dye-degrader and the contributions of EPS on adsorption were investigated in detail in this study. The results were benefit for better understanding of the interaction mechanisms between the dye molecules and cells that before the biodegradation process, which were of great significance for the practical usage of residual sludge on removal of dyes.

Energy-saving preparation of a bioflocculant under high-salt condition by using strain Bacillus sp. and the interaction mechanism towards heavy metals.

A highly efficient bioflocculant, i.e., Na-Bsp was successfully prepared by using a tolerant strain-Bacillus sp. under high-salt condition without sterilization. Salt-containing medium was not infected by other strains throughout the whole incubation period in 168 h. The as-prepared Na-Bsp was found to be cation-dependent, exhibiting high flocculant efficiency (FE) i.e., 97.69 ± 0.61%, towards kaolin particles by aid of Fe3+. High FE values were well maintained under a wide pH range and/or boiled water treatment, likely because of the main constituent of polysaccharide. The presence of hydroxyl, carboxyl, and amine groups on the bioflocculant surface were possibly responsible for strong interactions with heavy metals. The adsorption capacities of Pb2+, Cu2+ and Cr6+ were 1000.0, 434.8 and 384.6 mg g-1, respectively. The changing of structure and configuration of bioflocculant during the metal adsorption were explored by the scanning electron microscope with electron energy loss spectroscopy and three-dimensional excitation-emission fluorescence spectrometry. This study provided a novel production method, whereby the conventional sterilization could be avoided, which is of great environmental significance for steam-saving. Furthermore, the as-prepared Na-Bsp exhibited high adsorption capacities toward heavy metals, which sheds lights on its potential usage as an alternative adsorbent.

Simultaneous nitriles degradation and bioflocculant production by immobilized K. oxytoca strain in a continuous flow reactor.

High cost is one of the limiting factors in the industrial production of bioflocculant. Simultaneous preparation of bioflocculant from the contaminants in wastewater was considered as a potential approach to reduce the production cost. In this study, butyronitrile and succinonitrile were verified as sole nitrogen sources for the growth of strain K. oxytoca GS-4-08 in batch experiments. Moreover, more than 90 % of the mixed nitriles could be degraded in a continuous flow reactor, and the bioflocculant could be prepared simultaneously in the effluent. All the as-prepared bioflocculants exhibited high flocculation efficiencies of over 90 % toward Kaolin solution. FTIR and XPS results further unveiled that, the bioflocculant samples with abundance of carboxyl, amine and hydroxyl groups may play an important role on adsorption of Pd2+. The adsorption process could be well simulated by Freundlich model, and the Kf values were as high as 452.8 mg1-1/n l1/n g-1. The results obtained in this study not only confirm the technical feasibility for preparation of bioflocculant from various single nitrile and/or mixed nitriles, but also promise its economic feasibility.

Cloning and characterization of a Flavin-free oxygen-insensitive azoreductase from Klebsiella oxytoca GS-4-08.

OBJECTIVES: This study reports the identification of a new bacterial azoreductase (AzoR) from Klebsiella oxytoca GS-4-08, its heterologous production in Escherichia coli and the decolorization of azo dyes. RESULTS: The AzoR, as a flavin-free, oxygen-insensitive enzyme, has a molecular mass of 22 kDa and a high substrate specificity for methyl red (MR). In the presence of 5 mM of NADH, the enzyme activity for decolorization of 50 mg l-1 of MR reached 5.61 µmol min-1 mg protein-1. Double-reciprocal plots indicated that the NADH and MR reductions proceed by ping-pong mechanism. The calculated Vm was 8.17 µM min-1 mg protein-1, and the AzoR retained over 75% of relative activity under temperature range from 20 to 50 °C. CONCLUSION: This study for the first time identified an AzoR from Klebsiella oxytoca strain, which enabled efficient degradation of azo dyes by strain GS-4-08.