Isolation and characterization of a nitrobenzene degrading yeast strain from activated sludge

Strain Z1 was isolated from nitrobenzene-contaminated sludge. Strain Z1 was able to utilize nitrobenzene as a sole source of carbon, nitrogen, and energy under aerobic condition. Based on the morphology, physiological biochemical characteristics, and 26S rDNA D1/D2 domain sequence, strain Z1 was identified as Rhodotorula mucilaginosa. Strain Z1 mineralized up to 450mg L(-1) nitrobenzene. Kinetics of nitrobenzene degradation was described using the Andrews equation. The kinetic parameters were as follows: q(max)=1.50h(-1), K(s)=31.31mg L(-1), and K(i)=101.34mg L(-1). Strain Z1 had a high-salinity tolerance. It degraded nitrobenzene effectively in 5% NaCl (quality concentration). Even in the presence of aniline or phenol, strain Z1 degraded nitrobenzene efficiently. Strain Z1 therefore could be an excellent candidate for the bio-treatment of nitrobenzene industrial wastewaters.     

Phenol degradation by Aureobasidium pullulans FE13 isolated from industrial effluents

The degradation of phenol (2-30 mM) by free cells and by alginate-immobilized cells of Aureobasidium pullulans FE13 isolated from stainless steel effluents was studied in batch cultures with saline solution not supplemented with nutrients or yeast extract. The rate at which the immobilized cells degrade phenol was similar to the rate at which the suspended cells could degrade phenol, for a concentration of up to 16 mM of phenol. The maximum phenol volumetric degradation rate for 16 mM phenol was found to be 18.35 mg l(-1)h(-1) in the assays with free cells and 20.45 mg l(-1)h(-1) in the assays with alginate-immobilized cells, 18 mM phenol and cellular concentration of 0.176 g/l. At concentrations higher than this, an inhibitory effect was observed, resulting in the lowering of the phenol degradation rates. The immobilization was detrimental to the catechol 1,2-dioxygenase activity. However, the immobilized cells remained viable for a longer period, increasing the efficiency of phenol degradation. The yeast showed catechol 1,2-dioxygenase activity only after growth in the phenol, which was induced at phenol concentrations as low as 0.05 mM and up to 25 mM at 45 h of incubation at 30 degrees C. Phenol concentrations higher than 6mM were inhibitory to the enzyme. Addition of glucose, lactate, succinate, and benzoate reduced the rate at which phenol is consumed by cells. Our results suggest that inoculants based on immobilized cells of A. pullulans FE13 has potential application in the biodegradation of phenol and possibly in the degradation of other related aromatic compounds.     

Degradation of phenol and TCE using suspended and chitosan-bead immobilized Pseudomonas putida

The degradability of phenol and trichloroethene (TCE) by Pseudomonas putida BCRC 14349 in both suspended culture and immobilized culture systems are investigated. Chitosan beads at a size of about 1-2mm were employed to encapsulate the P. putida cells, becoming an immobilized culture system. The phenol concentration was controlled at 100 mg/L, and that of TCE was studied from 0.2 to 20 mg/L. The pH, between 6.7 and 10, did not affect the degradation of either phenol or TCE in the suspended culture system. However, it was found to be an important factor in the immobilized culture system in which the only significant degradation was observed at pH >8. This may be linked to the surface properties of the chitosan beads and its influence on the activity of the bacteria. The transfer yield of TCE on a phenol basis was almost the same for the suspended and immobilized cultures (0.032 mg TCE/mg phenol), except that these yields occurred at different TCE concentrations. The transfer yield at a higher TCE concentration for the immobilized system suggested that the cells immobilized in carriers can be protected from harsh environmental conditions. For kinetic rate interpretation, the Monod equation was employed to describe the degradation rates of phenol, while the Haldane's equation was used for TCE degradation. Based on the kinetic parameters obtained from the two equations, the rate for the immobilized culture systems was only about 1/6 to that of the suspended culture system for phenol degradation, and was about 1/2 for TCE degradation. The slower kinetics observed for the immobilized culture systems was probably due to the slow diffusion of substrate molecules into the beads. However, compared with the suspended cultures, the immobilized cultures may tolerate a higher TCE concentration as much less inhibition was observed and the transfer yield occurred at a higher TCE concentration.     

The biodegradation pathway of triethylamine and its biodegradation by immobilized Arthrobacter protophormiae cells

A bacterial strain named R4 was isolated from a wastewater treatment pool containing triethylamine (TEA) as the sole source of carbon and nitrogen. Strain R4 was identified as Arthrobacter protophormiae based on 16S rRNA gene sequence analysis and morphological and physiological properties. The optimal pH, temperature and concentration of NaCl for TEA degradation by strain R4 were 7.0, 30°C and 0.5%, respectively. Strain R4 could completely degrade 100 mg l(-1) TEA to ammonia in 32 h, and could also effectively degrade diethylamine (DEA) and ethylamine (EA) to ammonia. The degradation of TEA was strongly inhibited by some metal ions (Cu(2+), Mn(2+), Zn(2+), Co(2+), Ni(2+) and Ag(+)) (1.0mM). Addition of either SO(4)(2-) or NH(4)(+) reduced the degradation efficiency of TEA by strain R4 to a certain extent. The inhibition became significant when the concentration of SO(4)(2-) and NH(4)(+) reached to 11 mM and 30 mM, respectively. Cell-free extracts prepared from cells grown in TEA exhibited TEA monooxygenase, DEA monooxygenase and EA monooxygenase activity. Here, we propose the metabolic pathway of TEA degradation in strain R4. The efficiency of TEA removal by immobilized cells of strain R4 was found to be equivalent to that of free cells. In addition, the immobilized cells could be reused without reduction in their ability to degrade TEA.     

Effect of nutrients on the biodegradation of tributyltin (TBT) by alginate immobilized microalga, Chlorella vulgaris, in natural river water

The removal and degradation of tributyltin (TBT) by alginate immobilized Chlorella vulgaris has been evidenced in our previously published work. The present study was further to investigate the effect of spiked nutrient concentrations on the TBT removal capacity and degradation in the same alginate immobilized C. vulgaris. During the 14-d experiment, compared to the control (natural river water), the spiked nutrient groups (50% or 100% nutrients of the commercial Bristol medium as the reference, marked as 1/2N or 1N) showed more rapid cell proliferation of microalgae and higher TBT removal rate. Moreover, significantly more TBT was adsorbed onto the alginate matrix, but less TBT was taken up by the algal cells of the nutrient groups than that of the control. Mass balance data showed that TBT was lost as inorganic tin in the highest degree in 1N group, followed by 1/2N group and the least was in the control, but the relative abundance of the intermediate products of debutylation (dibutyltin and monobutyltin) were comparable among three groups. In conclusion, the addition of nutrients in contaminated water stimulated the growth and physiological activity of C. vulgaris immobilized in alginate beads and improved its TBT degradation efficiency.     

Aerobic degradation of methyl tert-butyl ether in a closed symbiotic system containing a mixed culture of Chlorella ellipsoidea and Methylibium petroleiphilum PM1

The contamination of groundwater by methyl tert-butyl ether (MTBE) is one of the most serious environmental problems around the world. MTBE degradation in a closed algal-bacterial symbiotic system, containing a mixed culture of Methylibium petroleiphilum PM1 and Chlorella ellipsoidea, was investigated. The algal-bacterial symbiotic system showed increased MTBE degradation. The MTBE-degradation rate in the mixed culture (8.808 ± 0.007 mg l(-1) d(-1)) was higher than that in the pure bacterial culture (5.664 ± 0.017 mg l(-1) d(-1)). The level of dissolved oxygen was also higher in the mixed culture than that in the pure bacterial culture. However, the improved efficiency of MTBE degradation was not in proportional to the biomass of the alga. The optimal ratio of initial cell population of bacteria to algae was 100:1. An immobilized culture of mixed bacteria and algae also showed higher MTBE degradation rate than the immobilized pure bacterial culture. A mixed culture with algae and PM1 immobilized separately in different gel beads showed higher degradation rate (8.496 ± 0.636 mg l(-1) d(-1)) than that obtained with algae and PM1 immobilized in the same gel beads (5.424 ± 0.010 mg l(-1) d(-1)).     

Enhanced thermal stability of laccase immobilized on monolayer-modified nanoporous Au

Nanoporous Au was fabricated by the dealloying of Au-Ag alloy in nitric acid, and was modified with a self-assembled monolayer (SAM) of 4-aminothiophenol, for the enhancement of high-temperature activity of immobilized laccase. Immobilized laccase exhibited much higher activity than that of free laccase at > 45 °C. SAM surface modification greatly improved the thermal stability and reusability of immobilized laccase. For example, little degradation in laccase activity when immobilized on SAM-modified nanoporous Au was observed after 2 h incubation at 50 °C. This suggested the nanoporous structure and SAM synergistically prevented the conformational change of laccase, and resulted in the enhancement of high-temperature activity.     

Preparation and characterization of monodispersed microfloccules of TiO₂ nanoparticles with immobilized multienzymes

Microfloccules of TiO(2) nanoparticles, on which glycerol-dehydrogenase (GDH), 1,3-propanediol-oxidoreductase (PDOR), and glycerol-dehydratase (GDHt) were coimmobilized, were prepared by adsorption-flocculation with polyacrylamide (PAM). The catalytic activity of immobilized enzyme in the glycerol redox reaction system, the enzyme leakage, stabilities of pH and temperature, as well as catalytic kinetics of immobilized enzymes relative to the free enzymes were evaluated. Enzyme loading on the microfloccules as much as 104.1 mg/g TiO(2) (>90% loading efficiency) was obtained under the optimal conditions. PAM played a key role for the formation of microfloccules with relatively homogeneous distribution of size and reducing the enzyme leakage from the microfloccules during the catalysis reaction. The stabilities of GDH against pH and temperature was significantly higher than that those of free GDH. Kinetic study demonstrated that simultaneous NAD(H) regeneration was feasible in glycerol redox system catalysted by these multienzyme microfloccules and the yield of 1, 3-popanediol (1, 3-PD) was up to 11.62 g/L. These results indicated that the porous and easy-separation microfloccules of TiO(2) nanoparticles with immobilized multienzymes were efficient in term of catalytic activity as much as the free enzymes. Moreover, compared with free enzyme, the immobilized multienzymes system exhibited the broader pH, higher temperature stability.     

Nitrogen-Doped Graphene Foam as a Metal-Free Catalyst for Reduction Reactions under a High Gravity Field

Herein, we report on the effect of a high gravity field on metal-free catalytic reduction, taking the nitrobenzene (NB) reduction and methylene blue (MB) degradation as model reactions in a high-gravity rotating tube reactor packed with three-dimensional (3D) nitrogen-doped graphene foam (NGF) as a metal-free catalyst. The apparent rate constant (k_app) of the metal-free catalytic reduction of NB in the rotating tube reactor under a high gravity level of 6484g (g = 9.81 m·s⁻²) was six times greater than that in a conventional stirred reactor (STR) under gravity. Computational fluid dynamics (CFD) simulations indicated that the improvement of the catalytic efficiency was attributed to the much higher turbulent kinetic energy and faster surface renewal rate in the high-gravity tube reactor in comparison with those in a conventional STR. The structure of the 3D metal-free catalysts was stable during the reaction process under a high gravity field, as confirmed by X-ray photoelectron spectroscopy (XPS) and Raman spectra. In the other model reaction, the rate of MB degradation also increased as the high gravity level increased gradually, which aligns with the result for the NB catalytic reduction system. These results demonstrate the potential to use a high-gravity rotating packed tube reactor for the process intensification of metal-free catalytic reduction reactions.     

疏水性光催化剂的制备及其催化动力学

采用十二烷基硫酸钠(SDS)为改性剂对TiO₂进行了疏水改性, FTIR表征表明催化剂表面具有CC和CH烷基疏水基团. 同时采用CuO对催化剂进行了可见光响应改性, UV-Vis表征表明催化剂具有良好的可见光响应性能, 吸收边红移至830nm以上. 三维荧光扫描发现随着CuO和SDS的加入,催化剂的空穴电子分离效果迅速提高. 以硝基苯为处理对象, 考察了体系pH值、污染物初始浓度、催化剂用量和光照强度对光催化反应过程的影响, 建立了动力学模型. 低浓度条件下, 通过模型计算的动力学常数相对误差范围为-16.5％～-4.5％; 高浓度条件下为-11.3％～4.6％.     

Photodegradation of nitrobenzene using 172 nm excimer UV lamp

Photodegradation of nitrobenzene (NB) using an excimer UV lamp at a wavelength of 172 nm is investigated. Experimental results show that high concentration nitrobenzene can be efficiently degraded with irradiation by excimer UV lamp, and confirm that degradation of nitrobenzene is more efficient by UV/H(2)O(2) combination than UV only. In the case of using UV only, 60 min of treatment was found to be sufficient to degrade the major part of NB solution with a concentration of less than 4mM. In the case of using the combination of UV/H(2)O(2) with a H(2)O(2) concentration of 7:1 molar ratio to NB, 4.07 mM NB solution drastically decreased to 0.41 mM after treatment for only 20 min. Degradation intermediate products are identified by analyzing the degradation products with GC/HRMS and possible degradation pathways of nitrobenzene are suggested.     

Performance of pulsed plate bioreactor for biodegradation of phenol

Biodegradation of phenol was carried out using Nocardia hydrocarbonoxydans immobilised on glass beads, in a pulsed plate bioreactor. The effect of operating parameters like frequency of pulsation and amplitude of pulsation on the performance of pulsed plate bioreactor for biodegradation of phenol in a synthetic wastewater containing 500ppm phenol was studied. Axial concentration profile measurements revealed that the pulsed plate bioreactor shows continuous stirred tank behaviour. As the amplitude was increased, percentage degradation increased, reaching 100% at amplitude of 4.7cm and higher. Introduction of pulsation is found to increase the percentage degradation. Percentage degradation has increased with increase in frequency and 100% degradation was achieved at 0.5s(-1) and above. Biofilms developed in a non-pulsed bioreactor were thicker than those in the pulsed plate bioreactor. But biofilm thickness remained almost constant with increasing frequency. Biofilm density was found to be influenced by pulsation. The time required to reach steady state was more for pulsed reactor than the non-pulsed reactor and this start-up time had increased with increase in frequency of pulsation. The performance studies reveal that the pulsed plate bioreactor with immobilized cells has the potential to be an efficient bioreactor for wastewater treatment.     

Study of the effect of some organic solvents on the activity and stability of glucose oxidase

The relative activity and thermal stability of native and covalently immobilized glucose oxidase (GOD) onto polyamide-6 (PA-6) membrane was studied at temperatures of 28, 45 and 60 °C for 10 h and in the presence of organic solvents (methylol, ethyleneglycol, glycerol) with concentrations 10%, 30% and 60%. It was proven that immobilized GOD had better stability than the native one in all the three organic solvents. At 28 °C, the strongest activating and stabilizing effect on the free GOD was observed with 10% methylol and on the immobilized GOD with 10% and 30% methylol. The addition of certain concentrations of ethyleneglycol and glycerol to the enzyme at higher temperatures was found to stabilize the enzyme molecule. At temperatures of 45 and 60 °C, the strongest stabilizing effect on both forms of the enzyme was exerted by glycerol (optimal stabilizing concentration of 10%). It was concluded that the most stable form of the enzyme was GOD covalently immobilized onto PA-6 membrane in the presence of 10% solution of glycerol.     

Pentachlorophenol contaminated groundwater bioremediation using immobilized Sphingomonas cells inoculation in the bioreactor system

Pentachlorophenol (PCP) has been used as a wood preservative for more than 100 years. The extensive use of PCP has widely contaminated soil and groundwater. PCP is toxic to living organisms. The main objective of this research was to inoculate the pure PCP-degrading bacterium strain Sphingomonas chlorophenolica PCP-1, isolated from PCP-contaminated soils, into PCP-contaminated groundwater for remediation purposes. The factors that influenced the bioremediation were explored with batch experiments using the inoculated immobilized and suspended cells as inoculation. A biological treatment system inoculated with immobilized cells was set up to estimate the microbial capability to degrade PCP. The results indicated that the suspended and immobilized cells could be inoculated into PCP-contaminated groundwater without adding other supplementary nitrogen and phosphate sources in batch conditions. Moreover, PCP decomposition was accompanied with released Cl- and decreasing pH value. The optimum HRT in the bioreactor system was 12.6h. PCP removal in the bioreactor remained stable and PCP removal efficiency was higher than 92% at this phase. Furthermore, PCP concentration in the biotreatment system effluent remained undetectable. It is possible to bioremediate PCP-contaminated groundwater using immobilized S. chlorophenolica PCP-1 cells in a bioreactor system. The proposed biological treatment system could be maintained for at least for 2 months.     

Co3+掺杂LaFeO3的制备及光催化降解硝基苯的性能研究

以La(NO3)3、Fe(NO3)3.9H2O和Co(NO3)2.6H2O为原料,采用柠檬酸络合法制备了纯LaFeO3和掺杂Co3+的LaFeO3纳米粉体,通过XRD和SEM等手段对样品进行表征,以硝基苯为目标降解物,考察催化剂的光催化降解活性。研究表明:煅烧温度为800℃时制备的LaFeO3样品对硝基苯溶液的脱色率达47.02%;而采用Co3+的掺杂增强了LaFeO3的光吸收特性,对LaFeO3的光催化性能有较大促进作用,且掺杂量为5%时效果最好,光降解时间1h时,对硝基苯的降解率可达76.26%。     

Characterizations of immobilized neutral lipase on chitosan nano-particles

Characterizations of chitosan nano-particles prepared by the ionization gelation method were discussed in this paper. The optimal pH of an immobilized enzyme was 7.5. The optimal temperature of the immobilized enzyme was 45 °C. The thermal, operational and storage stabilities of an enzyme were improved after they were immobilized on chitosan nano-particles. The chitosan nano-particles were applied to protect the activity of free neutral lipase. They could improve the enzyme activity by 13.17% than that of the free neutral lipase. The Michaelis constant K_m of the immobilized enzyme was disclosed as 0.37 × 10² g/l by the Line-weaver Burk plot at 40 °C.     

Immobilization of Trichoderma viride for enhanced methylene blue biosorption: Batch and column studies

An efficient dye biosorbent was developed by entrapping a fungus mold, Trichoderma viride, within loofa sponge (LS) matirx. Immobilization enhanced the sorption of dye by 30% at equilibrium as compared with T. viride free biomass (TVFB). The maximum dye biosorption capacity of T. viride immobilized onto loofa sponge (TVILS) and TVFB was found to be 201.52 and 155.06 mg g⁻¹ biomass, respectively. The kinetics of dye removal by TVILS was rapid, with 84.3% sorption within the first 30 min and equilibrium after 90 min, whereas sorption by TVFB was slower as 61.4% dye was removed in first 30 min and equilibrium was achieved in 120 min. Biosorption kinetics and equilibria followed the pseudo-second-order and Langmuir adsorption models. FTIR spectroscopy of T. viride biomass showed that amine, hydroxyl, carbonyl and amide bonds were involved in the sorption of dye. Dye desorption from dye-laden TVILS with 0.1 M HCl was 99%. Regenerated TVILS was reusable without any appreciable decrease in its biosorption capacity during five repeated cycles. The dye removing capacity of TVILS in a continuous-flow column bioreactor was better than in batch-scale procedures. The study shows that TVILS has the potential of application as an efficient biosorbent for the removal of methylene blue from aqueous solutions.     

Vertically integrated human P450 and oxygen sensing film for the assays of P450 metabolic activities

An assaying method of cytochrome P450 (P450 or CYP) monooxygenase activities for toxicological evaluation of drugs and environmental pollutants was developed by immobilizing P450 on an oxygen sensoring layer. Membrane fractions from E. coli expressing human P450 were entrapped in agarose or silica-based gels and immobilized on 96-well microarrays having an oxygen sensing film at the bottom. The oxygen sensing film was made of an organically modified silica film (ORMOSIL) doped with Tris(4,7-diphenyl-1,10-phenanthroline) ruthenium dichloride (Ru(dpp)(3)Cl(2)). P450 activity toward the substrates was monitored through the fluorescence intensity enhancement due to the oxygen consumption by the metabolic reactions. For the metabolism of chlortoluron, a selective herbicide used to control grass weeds, CYP1A1 immobilized in agarose gel showed a higher activity and stability compared with those in silica gels and free suspensions. The luminescence changing rate evaluated by the dynamic transient method (DTM) could be correlated with the substrate concentration. We also compared the metabolic responses of human P450s (CYP1A1,CYP2C8, CYP2E1, CYP3A4) toward various substances. The use of immobilized P450 on an oxygen sensing layer provides a versatile assaying platform owing to the following features. First, the oxygen sensor can detect metabolic reactions of any P450 species, in contrast with assays using fluorogenic substrates. Second, vertical integration of the oxygen sensor and immobilized P450 enhanced the sensitivity because of the effective depletion of oxygen in the vicinity of the oxygen sensing layer. Third, immobilization enables repeated use of P450 by replacing the substrate solutions using a flow cell. Furthermore, the activity of immobilized P450 was retained at least for 3 weeks at 4 °C, suggesting its long-term stability, which is an additional attractive feature.     

Facile synthesis of α-Fe2O3 nanodisk with superior photocatalytic performance and mechanism insight

Intrinsic short hole diffusion length is a well-known problem for α-Fe₂O₃ as a visible-light photocatalytic material. In this paper, a nanodisk morphology was designed to remarkably enhance separation of electron-hole pairs of α-Fe₂O₃. As expected, α-Fe₂O₃ nanodisks presented superior photocatalytic activity toward methylene blue degradation: more than 90% of the dye could be photodegraded within 30 min in comparison with a degradation efficiency of 50% for conventional Fe₂O₃ powder. The unique multilayer structure is thought to play a key role in the remarkably improved photocatalytic performance. Further experiments involving mechanism investigations revealed that instead of high surface area, ·OH plays a crucial role in methylene blue degradation and that O^·2− may also contribute effectively to the degradation process. This paper demonstrates a facile and energy-saving route to fabricating homogenous α-Fe₂O₃ nanodisks with superior photocatalytic activity that is suitable for the treatment of contaminated water and that meets the requirement of mass production.     

Immobilized-OPG-Fc on a titanium surface inhibits RANKL-dependent osteoclast differentiation in vitro

The purpose of the present study was to examine the effect of osteoprotegerin (OPG)-Fc fusion protein immobilized on a titanium surface on the initial differentiation of osteoclast precursor RAW264.7 cells. These cells were cultured on titanium specimens over which OPG-Fc was immobilized. The enhancement of tartrate-resistant acid phosphatase (TRAP) and cathepsin K mRNA expression in RAW264.7 cells exposed to receptor activator of NF-κB ligand (RANKL) stimulation on OPG-Fc-coated titanium was significantly lower than that in RAW264.7 cells exposed to RANKL on titanium specimens without immobilized OPG-Fc (ANOVA, P < 0.01). Preincubation of OPG-Fc-coated titanium, in a medium supplemented with 10% fetal bovine serum at 37°C for two days before the cells were seeded, had no significant effect on the decrease in mRNA expression (ANOVA, P < 0.01). Taken together, these results indicate that OPG-Fc immobilized on a titanium surface blocks the differentiation of RAW264.7 cells induced by RANKL stimulation.