Interaction of tetrahydrofuran and methyl tert-butyl ether in waste gas treatment by a biotrickling filter bioaugmented with Piscinibacter caeni MQ-18 and Pseudomonas oleovorans DT4.

Bioaugmented biotrickling filter (BTF) seeded with Piscinibacter caeni MQ-18, Pseudomonas oleovorans DT4, and activated sludge was established to investigate the treatment performance and biodegradation kinetics of the gaseous mixtures of tetrahydrofuran (THF) and methyl tert-butyl ether (MTBE). Experimental results showed an enhanced startup performance with a startup period of 9 d in bioaugmented BTF (25 d in control BTF seeded with activated sludge). The interaction parameter I2,1 of control (7.462) and bioaugmented BTF (3.267) obtained by the elimination capacity-sum kinetics with interaction parameter (EC-SKIP) model indicated that THF has a stronger inhibition of MTBE biodegradation in the control BTF than in the bioaugmented BTF. Similarly, the self-inhibition EC-SKIP model quantified the positive effects of MTBE on THF biodegradation, as well as the negative effects of THF on MTBE biodegradation and the self-inhibition of MTBE and THF. Metabolic intermediate analysis, real-time quantitative polymerase chain reaction, biofilm-biomass determination, and high-throughput sequencing revealed the possible mechanism of the enhanced treatment performance and biodegradation interactions of MTBE and THF.

Enhancing Chlorobenzene Biodegradation by Delftia tsuruhatensis Using a Water-Silicone Oil Biphasic System.

In this study, a water-silicone oil biphasic system was developed to enhance the biodegradation of monochlorobenzene (CB) by Delftia tsuruhatensis LW26. Compared to the single phase, the biphasic system with a suitable silicone oil fraction (v/v) of 20% allowed a 2.5-fold increase in the maximum tolerated CB concentration. The CB inhibition on D. tsuruhatensis LW26 was reduced in the presence of silicone oil, and the electron transport system activity was maintained at high levels even under high CB stress. Adhesion of cells to the water-oil interface at the water side was observed using confocal laser scanning microscopy. Nearly 75% of cells accumulated on the interface, implying that another interfacial substrate uptake pathway prevailed besides that initiated by cells in the aqueous phase. The 8-fold increase in cell surface hydrophobicity upon the addition of 20% (v/v) silicone oil showed that silicone oil modified the surface characteristics of D. tsuruhatensis LW26. The protein/polysaccharide ratio of extracellular polymeric substances (EPS) from D. tsuruhatensis LW26 presented a 3-fold enhancement. These results suggested that silicone oil induced the increase in the protein content of EPS and rendered cells hydrophobic. The resulting hydrophobic cells could adhere on the water-oil interface, improving the mass transfer by direct CB uptake from silicone oil.

[Treatment of the Waste Gas Containing Methyl tert-Butyl Ether via a Biotrickling Filter].

The performance and microbial communities of methyl tert-butyl ether (MTBE) treatment using a biotrickling filter (BTF) that was inoculated with activated sewage sludge were investigated. The BTF successfully started up within 23 days when the inlet concentration of MTBE was 100 mg·m-3 and empty bed retention time was 60 s, with 70% removal efficiency (RE). Under steady-state conditions, an elimination capacity (EC) and a mineralization ratio of 13.47 g·(m3·h)-1 and 68% were achieved, respectively. The ECmax was 21.03 g·(m3·h)-1 according to the Haldane model, and a KS of 0.16 g·m-3 and KI of 0.99 g·m-3 were obtained. High-throughput sequencing was used to identify the community structure of the mixed microbial consortium in the BTF. The results indicated that Methylibium sp. (11.33%) and Blastocatella sp. (9.95%) were the dominant bacteria.

Piscinibacter caeni sp. nov., isolated from activated sludge.

A yellowish-pigmented bacterial strain, designated as MQ-18T, was isolated from a sample of activated sludge collected from a pharmaceutical factory in Zhejiang, China. The strain was characterized through a polyphasic taxonomy approach. 16S rRNA gene sequence analysis demonstrated that strain MQ-18T showed high similarities to Piscinibacter defluvii SH-1T (99.7 %) and Piscinibacter aquaticus IMCC1728T (98.4 %), thereby suggesting that it belongs to the genus Piscinibacter. The DNA-DNA relatedness values of this strain to strains SH-1T and IMCC1728T were only 35.4 and 33.3 %, respectively. Cells of MQ-18T were Gram-negative, aerobic, motile, rod-shaped and non-spore forming. This strain exhibited growth at 25-37 °C (optimum: 30 °C) in the presence of 0-3.0 % (w/v) NaCl (optimum, 0 % NaCl) and at pH 5.0-8.0 (pH 7.0). The predominant fatty acids were C12 : 0 (5.5 %), C16 : 0 (33.7 %), summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c; 38.5 %), and summed feature 4 (anteiso-C17 : 1 B and/or iso C17 : 1 I; 11.6 %). The main quinone type was ubiquinone-8, and the major polyamines were cadaverine and putrescine. The major polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The DNA G+C content was 70.1 mol%. On the basis of its phylogenetic, phenotypic and physiological characteristics, strain MQ-18T is considered to represent a novel species of the genus Piscinibacter, for which the name Piscinibacter caeni sp. nov. is proposed. The type strain is MQ-18T (CCTCC AB 2017223T=JCM 32138T).

Comparative investigation on a hexane-degrading strain with different cell surface hydrophobicities mediated by starch and chitosan.

Bioremediation usually exhibits low removal efficiency toward hexane because of poor water solubility, which limits the mass transfer rate between the substrate and microorganism. This work aimed to enhance the hexane degradation rate by increasing cell surface hydrophobicity (CSH) of the degrader, Pseudomonas mendocina NX-1. The CSH of P. mendocina NX-1 was manipulated by treatment with starch and chitosan solution of varied concentrations, reaching a maximum hydrophobicity of 52%. The biodegradation of hexane conformed to the Haldane inhibition model, and the maximum degradation rate (ν max) of the cells with 52% CSH was 0.72 mg (mg cell)-1·h-1 in comparison with 0.47 mg (mg cell)-1·h-1 for cells with 15% CSH. The production of CO2 by high CSH cells was threefold higher than that by cells at 15% CSH within 30 h, and the cumulative rates of O2 consumption were 0.16 and 0.05 mL/h, respectively. High CSH was related to low negative charge carried by the cell surface and probably reduced the repulsive electrostatic interactions between hexane and microorganisms. The FT-IR spectra of cell envelopes demonstrated that the methyl chain was inversely proportional to increasing CSH values, but proteins exhibited a positive effect to CSH enhancement. The ratio of extracellular proteins and polysaccharides increased from 0.87 to 3.78 when the cells were treated with starch and chitosan, indicating their possible roles in increased CSH.

A newly isolated Pseudomonas putida S-1 strain for batch-mode-propanethiol degradation and continuous treatment of propanethiol-containing waste gas.

Pseudomonas putida S-1 was isolated from activated sludge. This novel strain was capable of degrading malodorous 1-propanethiol (PT). PT degradation commenced with no lag phase by cells pre-grown in nutrition-rich media, such as Luria-Bertani (LB), and PT-contained mineral medium at specific growth rates of 0.10-0.19 h(-1); this phenomenon indicated the operability of a large-scale cell culture. A possible PT degradation pathway was proposed on the basis of the detected metabolites, including dipropyl disulfide, 3-hexanone, 2-hexanone, 3-hexanol, 2-hexanol, S(0), SO4(2-), and CO2. P. putida S-1 could degrade mixed pollutants containing PT, diethyl disulfide, isopropyl alcohol, and acetaldehyde, and LB-pre-cultured cells underwent diauxic growth. Waste gas contaminated with 200-400 mg/m(3) PT was continuously treated by P. putida S-1 pre-cultured in LB medium in a completely stirred tank reactor. The removal efficiencies exceeded 88% when PT stream was mixed with 200 mg/m(3) isopropanol; by contrast, the removal efficiencies decreased to 60% as the empty bed residence time was shortened from 40 s to 20 s.

[Removal of Waste Gas Containing Mixed Chlorinated Hydrocarbons by the Biotrickling Filter].

An experimental investigation on purification of waste gas contaminated with a mixture of dichloromethane (DCM) and dichloroethane(1,2-DCA) was conducted in a biotrickling filter (BTF) inoculated with activated sludge of pharmaceuticals industry. Stable removal efficiency(RE) above 80% for DCM and above 75% for 1,2-DCA were achieved after 35 days, indicating that biofilm was developed. The best elimination capacity (EC) of DCM and 1,2-DCA were 13 g.(m3.h)-1 and 10 g.(m3.h)-1 respectively. And there was a linear relationship between the production of CO2 and mixed gas EC, the maximum mineralization rate of mixed gas stabled at 61. 2%. The interaction test indicated that DCM and 1,2-DCA would inhibit with each other. The changing of biomass of BTF during the operation process was also been studied.

Biodegradation kinetics of tetrahydrofuran, benzene, toluene, and ethylbenzene as multi-substrate by Pseudomonas oleovorans DT4.

The biodegradation kinetics of tetrahydrofuran, benzene (B), toluene (T), and ethylbenzene (E) were systematically investigated individually and as mixtures by a series of aerobic batch degradation experiments initiated by Pseudomonas oleovorans DT4. The Andrews model parameters, e.g., maximum specific growth rates (µmax), half saturation, and substrate inhibition constant, were obtained from single-substrate experiments. The interaction parameters in the sum kinetics model (SKIP) were obtained from the dual substrates. The µmax value of 1.01 for tetrahydrofuran indicated that cell growth using tetrahydrofuran as carbon source was faster than the growth on B (µmax, B = 0.39) or T (µmax, T = 0.39). The interactions in the dual-substrate experiments, including genhancement, inhibition, and co-metabolism, in the mixtures of tetrahydrofuran with B or T or E were identified. The degradation of the four compounds existing simultaneously could be predicted by the combination of SKIP and co-metabolism models. This study is the first to quantify the interactions between tetrahydrofuran and BTE.

[Biodegradation of tetrahydrofuran by combined immobilized of Pseudomonas oleovorans DT4].

A new composite matrix, calcium alginate (CA) coupled with activated carbon fiber (ACF) was designed to immobilize the cells of Pseudomonas oleovorans DT4 for tetrahydrofuran (THF) degradation. The average removal rate of the CA-ACF immobilized cells reached 24.0 mg x (L x h)(-1) with an initial THF concentration of 360 mg x L(-1) when the concentration of CA and ACF was 3% and 1.5% respectively. The mechanical strength of the mobilized cells was also significantly improved with the addition of ACF. Compared to the free suspended cells, higher stable removal efficiency (more than 80%) of CA-ACF cells was detected under different conditions of temperature and pH. The feasibility of the newly designed matrix was also reflected by the repeated batch degradation which showed that the removal activity decreased insignificantly after 80 cycles with the modified reaction system (PNS).

Biodegradation of tetrahydrofuran by Pseudomonas oleovorans DT4 immobilized in calcium alginate beads impregnated with activated carbon fiber: mass transfer effect and continuous treatment.

A novel entrapment matrix, calcium alginate (CA) coupled with activated carbon fiber (ACF), was prepared to immobilize Pseudomonas oleovorans DT4 for degrading tetrahydrofuran (THF). The addition of 1.5% ACF increased the adsorption capacity of the immobilized bead, thus resulting in an enhanced average removal rate of 30.3mg/(Lh). The synergism between adsorption and biodegradation was observed in the hybrid CA-ACF beads instead of in the system comprising CA beads and freely suspended ACF. The effective diffusion coefficient of the CA-ACF bead was not significantly affected by bead size, but the bead's value of 1.14×10(-6)cm(2)/s (for the bead diameter of 0.4 cm) was larger than that of the CA bead by almost one order of magnitude based on the intraparticle diffusion-reaction kinetics analysis. Continuous treatment of the THF-containing wastewater was succeeded by CA-ACF immobilized cells in a packed-bed reactor for 54 d with a >90% removal efficiency.

[Characteristics of biofilm phase during the long-term degradation of a toluene-contaminated gas stream using BTF].

This study analyzes the accumulation and distribution of biomass and changes in properties of biofilm in a long-term biotrickling filter (BTF) system in order to investigate the correlation between the biofilm phase properties and the performance. After a long-term operation of 130 days, the BTF showed a deterioration in degradation performance and an increase in pressure drop with a gradual increase of biofilm thickness and uneven distribution of biomass. Meanwhile, the porosity of the upper and lower layers decreased from 85% and 82% in the start-up period to 65% and 40%, respectively, as a result of the excessive accumulation of biomass and its non-uniform distribution. The AWCD values showed a decreasing trend indicating that the biological activity decreased with the aging of biofilm in the long-term BTF. The contents of total extracellular polymeric substances (EPS) and protein in the later period were twice as much as those in the start-up period. The value of protein to polysaccharide (PN/PS) ratio increased from 0.3 to 0.95, and showed positive correlation with the surface hydrophobicity of the biofilm, which increased from 33% to 73% accordingly. The mean molecular weight of EPS decreased during the operation of BTF. The result of FTIR further showed that the main chemical composition of EPS changed in the long-term BTF accordingly, possibly resulting in the deterioration of performance in the long-term BTF. The above experimental results could be very helpful for reducing the clogging and performance deterioration in long-term BTF.

[Biodegradation of methyl tert-butyl ether by stabilized immobilized Methylibium petroleiphilum PM1 cells and its biodegradation kinetics analysis].

Methylibium petroleiphilum PM1, which is capable of degrading methyl tert-butyl ether (MTBE) , was immobilized in calcium alginate gel beads. Several methods were explored to increase the strength of these gel beads. The central composite design analysis indicated that the introduction of 0.2 mol x L(-1) Ca2+ into the crosslinking solution, 1.38 mmol x L(-1) Ca2+ into the growth medium and 0.1% polyethyleneimine (PEI) as the chemical crosslinking agent could increase the stability of the Ca-alginate gel beads with no loss of biodegradation activity. The stabilized immobilized cells could be used 400 h continuously with no breakage and no bioactivity loss. Examination of scanning electron microscope demonstrated that a membrane surrounding the gel beads was formed and the cells could grow and breed well in the stabilized calcium alginate gel beads. Kinetic analysis of the gel bead-degradation indicated that the rate-limiting step was biochemical process instead of intraparticle diffusion process. The diameter of 3 mm affected the biodegradability less while high concentration of PEI induced much more serious mass transfer restraint.

Substrate interactions during the biodegradation of BTEX and THF mixtures by Pseudomonas oleovorans DT4.

The efficient tetrahydrofuran (THF)-degrading bacterium, Pseudomonas oleovorans DT4 was used to investigate the substrate interactions during the aerobic biotransformation of THF and BTEX mixtures. Benzene and toluene could be utilized as growth substrates by DT4, whereas cometabolism of m-xylene, p-xylene and ethylbenzene occurred with THF. In binary mixtures, THF degradation was delayed by xylene, ethylbenzene, toluene and benzene in descending order of inhibitory effects. Conversely, benzene (or toluene) degradation was greatly enhanced by THF leading to a higher degradation rate of 39.68 mg/(h g dry weight) and a shorter complete degradation time about 21 h, possibly because THF acted as an "energy generator". Additionally, the induction experiments suggested that BTEX and THF degradation was initiated by independent and inducible enzymes. The transient intermediate hydroquinone was detected in benzene biodegradation with THF while catechol in the process without THF, suggesting that P. oleovorans DT4 possessed two distinguished benzene pathways.

Degradation of chlorobenzene by strain Ralstonia pickettii L2 isolated from a biotrickling filter treating a chlorobenzene-contaminated gas stream.

A Ralstonia pickettii species able to degrade chlorobenzene (CB) as the sole source of carbon and energy was isolated from a biotrickling filter used for the removal of CB from waste gases. This organism, strain L2, could degrade CB as high as 220 mg/L completely. Following CB consumption, stoichiometric amounts of chloride were released, and CO2 production rate up to 80.2% proved that the loss of CB was mainly via mineralization and incorporation into cell material. The Haldane modification of the Monod equation adequately described the relationship between the specific growth rate and substrate concentration. The maximum specific growth rate and yield coefficient were 0.26 h⁻¹ and 0.26 mg of biomass produced/mg of CB consumed, respectively. The pathways for CB degradation were proposed by the identification of metabolites and assay of ring cleavage enzymes in cell extracts. CB was degraded predominantly via 2-chlorophenol to 3-chlorocatechol and also partially via phenol to catechol with subsequent ortho ring cleavage, suggesting partially new pathways for CB-utilizing bacteria.

[Characteristics of tetrahydrofuran degradation by Pseudomonas oleovorans DT4].

A tetrahydrofuran (THF)-degrading strain Pseudomonas oleovorans DT4 was isolated from the activated sludge of a pharmaceutical plant. P. oleovorans DT4 was able to utilize THF as the sole carbon and energy source under aerobic condition. 5 mmol/L of THF could be completely degraded by 3.2 mg/L inoculums of P. oleovorans DT4 in 14 h at pH 7.2 and 30 degrees C, with the cells concentration increasing to 188.6 mg/L. After the complete consumption of THF, no TOC could be detected but IC reached the stable value of about 46 mg/L, with pH decreasing to 6.54, which indicated that the substance was totally mineralized by P. oleovorans DT4. The optimum conditions for THF biodegradation in shaking flasks were pH 7.5 and temperature 37 degrees C, respectively. Results from the oxygen control experiments revealed that the oxygen supply by shaking was the satisfactory growth condition. Additionally, as the important elements for DT4, Mg2+ and Ca2+ at concentrations of 0.80 mmol/L and 0.20 mmol/L, respectively, were suitable for THF degradation. All the results contribute to the efficient bioremediation for the THF contaminated.

[BTF performance treating a chlorobenzene-contaminated gas stream].

In this study, biotrickling filter (BTF) inoculated with acclimated sludge was established to treat waste gas containing chlorobenzene. The BTF performance, average well color development (AWCD) values and microbial community were examined in steady state. Results revealed BTF achieved removal efficiency more than 80% of chlorobenzene under the conditions of < 0.6 g x m(-3) inlet concentration and > 45 s EBRT. Therefore, BTF have an advantage in treating low-concentration waste gas containing chlorobenzene (< or = 0.6 g x m(-3)). The overall chlorobenzene elimination capacity reached a maximum of 70 g x (m3 x h)(-1) at an inlet load of 80 g x (m3 x h)(-1). The mass ratio of carbon dioxide produced to the BTo-X removed was approximately 1.92, which confirms complete degradation of chlorobenzene, given that some of the organic carbon consumed is also used for the microbial growth. The degradation of chlorobenzene in the BTF followed Michaelis-Menten kinetic model, the maximum specific degradation rate (r(max)) was 35.6 g x (m3 x h)(-1). The AWCD values indicated that the microorganisms in the BTF showed high the microbial metabolic activity. The PCR-DGGE fingerprinting analysis on biofilm samples in the BTF indicated that the microbial community had a relative stability and complexity during the steady-state phase. The stability and complexity of microbial community could contribute to the degradation and mineralization of chlorobenzene in BTF.

[Isolation, identification of 17beta-estradiol (E2)-degrading strain and its degradation characteristics].

A novel bacterium capable of degrading 17beta-estradiol (E2) was isolated from the activated sludge collected from wastewater treatment plant of an oral contraceptives producing factory in Xianju, Zhejiang. According to its morphology, physiochemical characteristics and 16S rDNA sequence analysis, this strain was identified as Bacillus sp. The optimal pH and temperature for E2 biodegradation in shaking flasks were 7.5 and 30 degrees C, respectively. The enhanced action of peptone and Lab-Lemco were quite obvious; metal ions such as Ba2+, Zn2+, Sn2+, Cd2+, Cr2+, Pb2+ inhibited degradation, and the actions were ordinal increasing; the addition such as Ca2+, Fe2+, KNO and amylum had no obvious effect on the degradation of E2. Furthermore, this strain could degrade E2 completely within 7 days with initial concentration ranged from 0.5 to 50 mg/L and was capable of converting E2 to estrone (E1), then to nonestrogenic compounds, thus 95% of initial estrogenic activities was decreased within 15 days during the biodegradation process.

[Isolation, identification and conditions of bacterial strain capable to metabolize high concentrations of formaldehyde].

One bacterial strain capable to degrade and metabolize formaldehyde as a sole carbon source was isolated from soil. Based on the results of standard morphological identification, physiological and biochemical characters, and 16S rDNA sequence analysis, the strain was identified as Pseudomonas putida. After single factor test and orthogonal test, the optimal condition for formaldehyde degradation was determined as the follows: peptone 1.2 g/L, KH2PO4 4 g/L, K2HPO4 3 g/L, MgSO4 x 7H2O 0.2 g/L, trace elements solution 0.1 mL/L, temperature 30 degrees C, pH 8. Under the optimal conditions, the strain tolerance of original formaldehyde concentration was up to 6 g/L and 86% of formaldehyde was consumed after 54 h. It completely consumed 5 g/L formaldehyde after 46 h and degraded 100% of 4 g/L formaldehyde after 35 h.

[Oxidative transformation of 17beta-estradiol at the interface of delta-MnO2 and water].

The oxidative transformation of 17beta-estradiol (E2) by the delta-MnO2 was investigated in batch experiments, and the influence of pH and delta-MnO2 dosage on the reaction process was studied. The results suggested that E2 could be degraded by delta-MnO2, and the oxidation reaction deviated from the pseudo-first-order kinetics at a constant pH and with an enough excess of delta-MnO2. Delta-MnO2 dosage had a positive effect on oxidative transformation of E2. When c (E2) : c (MnO2) = 1:100, the delta-MnO2 dosage was enough for E2 degradation, and the removalrate of E2 was above 90% after 8h reaction. The pH value had a negative effect on E2 degradation. When at pH 4.0, E2 concentration was decreased sharply and removal rate reached 95% after 0.25 h reaction, while E2 was disappeared by 80% at pH 6.8 and by 75% at pH 9.0 even after 1 h reaction. Two products, estrone (E1) and 2-hydroxyestradiol, were detected by gas chromatography coupled with mass spectrometry (GC-MS), and the probable transformation pathway was proposed.

Aerobic degradation of methyl tert-butyl ether by a Proteobacteria strain in a closed culture system.

The contamination of methyl tert-butyl ether (MTBE) in underground waters has become a widely concerned problem all over the world. In this study, a novel closed culture system with oxygen supplied by H2O2 was introduced for MTBE aerobic biodegradation. After 7 d, almost all MTBE was degraded by a pure culture, a member of beta-Proteobacteria named as PM1, in a closed system with oxygen supply, while only 40% MTBE was degraded in one without oxygen supply. Dissolved oxygen (DO) levels of the broth in closed systems respectively with and without H2O2 were about 5-6 and 4 mg/L. Higher DO may improve the activity of monooxygemase, which is the key enzyme of metabolic pathway from MTBE to tert-butyl alcohol and finally to CO2, and may result in the increase of the degrading activity of PM1 cell. The purge and trap GC-MS result of the broth in closed systems showed that tert-butyl alcohol, isopronol and acetone were the main intermediate products.