Enhanced chlorobenzene removal by internal magnetic field through initial cell adhesion and biofilm formation.

Magnetic fields (MF) have been proven efficient in bioaugmentation, and the internal MFs have become competitive because they require no configuration, despite their application in waste gas treatment remaining largely unexplored. In this study, we firstly developed an intensity-regulable bioaugmentation with internal MF for gaseous chlorobenzene (CB) treatment with modified packing in batch bioreactors, and the elimination capacity increased by up to 26%, surpassing that of the external MF. Additionally, the microbial affinity to CB and the packing surface was enhanced, which was correlated with the ninefold increased secreted ratio of proteins/polysaccharides, 43% promoted cell surface hydrophobicity, and half reduced zeta potential. Furthermore, the dehydrogenase content was promoted over 3 times, and CB removal steadily increased with the rising intensity indicating enhanced biofilm activity and reduced CB bioimpedance; this was further supported by kinetic analysis, which resulted in improved cell adhesive ability and biological utilisation of CB. The results introduced a novel concept of adjustable magnetic bioaugmentation and provided technical support for industrial waste gas treatments. KEY POINTS: • Regulable magnetic bioaugmentation was developed to promote 26% chlorobenzene removal • Chlorobenzene mineralisation was enhanced under the magnetic field • Microbial adhesion was promoted through weakening repulsive forces.

Airlift two-phase partitioning bioreactor for dichloromethane removal: Silicone rubber stimulated biodegradation and its auto-circulation.

Solid non-aqueous phases (NAPs), such as silicone rubber, have been used extensively to improve the removal of volatile organic compounds (VOCs). However, the removal of VOCs is difficult to be further improved because the poor understanding of the mass transfer and reaction processes. Further, the conventional reactors were either complicated or uneconomical. In view of this, herein, an airlift bioreactor with silicone rubber was designed and investigated for dichloromethane (DCM) treatment. The removal efficiency of Reactor 1 (with silicone rubber) was significantly higher than that of Reactor 2 (without silicone rubber), with corresponding higher chloride ion and CO2 production. It was found that Reactor 1 achieved a much better DCM shock tolerance capability and biomass stability than Reactor 2. Silicone rubber not only enhanced the mass transfer in terms of both gas/liquid and gas/microbial phases, but also decreased the toxicity of DCM to microorganisms. Noteworthily, despite the identical inoculum used, the relative abundance of potential DCM-degrading bacteria in Reactor 1 (91.2%) was much higher than that in Reactor 2 (24.3%) at 216 h. Additionally, the silicone rubber could be automatically circulated in the airlift bioreactor due to the driven effect of the airflow, resulting in a significant reduction of energy consumption.

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.

Characteristics of a novel thermophilic heterotrophic bacterium, Anoxybacillus contaminans HA, for nitrification-aerobic denitrification.

A strain bacterium that is thermophilic, heterotrophic nitrifying, and aerobic denitrifying was isolated and identified as Anoxybacillus contaminans HA for the first time. The identification was based on morphological and physiological characterizations, together with phylogenetic analysis of 16S rDNA sequence. The strain possessed excellent tolerance to high temperatures, with 55 °C as its optimum and 60 °C as viable. Moreover, NH4 (+)-N and NO3 (-)-N could be efficiently removed under thermophilic and solely aerobic conditions, with little intermediate accumulation. Average removal efficiencies of NH4 (+)-N and NO3 (-)-N at 55 °C reached 71.0 and 74.7 %, respectively, with removal rates of 5.83 and 32.08 mg l(-1) h(-1), respectively. Single-factor experiments suggested that the optimal conditions for both heterotrophic nitrification and aerobic denitrification were glucose as carbon source, NH4 (+)-N range of 50-200 mg l(-1), and wide NO3 (-)-N range of 200-1000 mg l(-1). These results indicated that strain HA had heterotrophic nitrification and aerobic denitrification abilities, as well as the notable ability to remove ammonium under thermophilic condition. Thus, this strain has potential application in waste-gas treatment.

Effects of 1,2,4-trichlorobenzene on the enzyme activities and ultrastructure of earthworm Eisenia fetida.

Adult Eisenia fetida earthworms were exposed to sub-lethal concentrations of 1,2,4-trichlorobenzene to assess the toxicity of contaminated soils. The LC50 of 1,2,4-TCB at 7 and 14 d were 945±175 and 890±169 mg kg⁻¹. A lower dose of 1,2,4-TCB (25 mg g⁻¹) had stimulatory effects on SOD and AChE activities, but AChE activities were significantly inhibited at higher treatment levels (100 and 400 mg g⁻¹). SOD activities increased after 2, 7, and 14 d of exposure, but AChE activities were significantly inhibited at 400 mg/kg 1,2,4-TCB after 2, 7, and 14 d of exposure, and the inhibition rates were 25.41%, 28.65%, and 25.05%. Ultrastructural observation of the intestinal epithelium at three concentrations (control, 50, and 400 mg g⁻¹) revealed that the epicuticle, cuticle layer, and microvilli were damaged with increasing 1,2,4-TCB concentration. At 400 mg kg⁻¹, mitochondria were seriously injured and the smooth endoplasmic reticulum was seriously dilated.

Promoting effect of plant hormone gibberellin on co-metabolism of sulfamethoxazole by microalgae Chlorella pyrenoidosa.

In this study, sodium acetate (NaAC) as a co-substrate effectively promoted the metabolism of sulfamethoxazole (SMX) by microalgae Chlorella pyrenoidosa. In the cultivation supplied with 5.0 and 10.0 g L-1 NaAC, 51.1% and 61.2% SMX was removed, respectively. On this basis, the improvement effect of plant hormone gibberellin (GA3) on SMX removal by 5 g L-1 NaAC supplied as co-substrate was further investigated. The results showed that biodegradation played decisive role in the removal of SMX. As a plant hormone, GA3 effectively improved the co-metabolic removal efficiency of SMX by C. pyrenoidosa. Especially when GA3 dosage reached 10.0 and 50.0 mg L-1, C. pyrenoidosa showed a very high SMX removal rate of 83.5% and 95.3%, respectively. Transcriptome analysis showed that GA3 promoted the removal of SMX by C. pyrenoidosa was the result of the combined action of exogenous and endogenous plant hormones.

Improving sedimentation and lipid production of microalgae in the photobioreactor using saline wastewater.

Saline wastewater was used in this study to culture freshwater microalgae Chlorella pyrenoidosa in sequencing batch photobioreactor to improve the sedimentation and lipid production of algal cells. Influent salinity of 0.5% or above effectively promoted the sedimentation of microalgae in the settling stage of photobioreactor, and greatly reduced the algal biomass in effluent. The mechanism of the saline wastewater in improving the sedimentation of microalgae included decreasing zeta potential, increasing cell particle size and promoting extracellular polymeric substances synthesis, which varied with influent salinity. Saline wastewater also promoted the lipid accumulation in microalgae. Lipid content of microalgae increased with increasing influent salinity. However, the growth of microalgae was greatly inhibited at the influent salinity of 2.0% and 3.0%. Therefore, the PBR with influent salinity of 1.0% achieved the highest productivity of microalgae lipid. The saturation of fatty acids of microalgae gradually increased with increasing influent salinity.

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.

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.

Mixotrophic cultivation of microalgae coupled with anaerobic hydrolysis for sustainable treatment of municipal wastewater in a hybrid system of anaerobic membrane bioreactor and membrane photobioreactor.

This study investigated the possibility of coupling anaerobic hydrolysis in an anaerobic membrane bioreactor (AnMBR) with mixotrophic microalgae cultivation in a membrane photobioreactor (MPBR) for the sustainable treatment of municipal wastewater. Using the hydrolyzed wastewater discharged from AnMBR, Chlorella pyrenoidosa in MPBR grew in a mixotrophic mode and realized rapid growth. During the stable operation, MPBR achieved average carbon capture rate of 42.82 mg L-1 d-1 and algal lipid production rate of 19.66 mg L-1 d-1. The average reduction in TN, TP, and TOC during stable operation was 96.7%, 98.0%, and 95.9%, respectively. Mass balance analysis showed that the overall system captured 14.76 mg of carbon from the atmosphere per liter of wastewater treated. Therefore, this AnMBR-MPBR hybrid system simultaneously realized advanced treatment of municipal wastewater, efficient production of algal lipid, and carbon capture from atmosphere, and thus has a good potential in the sustainable treatment of municipal wastewater.

Enhanced biodegradation of n-hexane in a two-phase partitioning bioreactor inoculated with Pseudomonas mendocina NX-1 under chitosan stimulation.

Two-phase partitioning bioreactors (TPPBs) have been extensively used for volatile organic compounds (VOCs) removal. To date, most studies have focused on improving the mass transfer of gas phases/non-aqueous phases (NAPs)/aqueous phases, whereas the NAP/biological phases and gas/biological phases transfer has been neglected. Herein, chitosan was introduced into a TPPB to increase cell surface hydrophobicity (CSH) and improve the n-hexane mass transfer. The performance and stability of the TPPB with chitosan for n-hexane biodegradation were investigated, and it was found out that the TPPB with chitosan achieved maximum removal efficiency and elimination capacity of 80.6% and 26.5 g m-3 h-1, thereby reaching much higher values than those obtained without chitosan (61.3% and 15.2 g m-3 h-1). Chitosan not only obvio usly increased cell surface hydrophobicity and cell dry biomass on the surface of silicone oil, but might also allow hydrophobic cells in aqueous phases to directly capture and biodegrade n-hexane, resulting in an obvious improvement of mass transfer from the gas phase to biomass. Stability enhancement was another attractive advantage from chitosan addition. This study might provide a new strategy for the development of TPPB in the hydrophobic VOCs treatment.

[Removal of Nitrate Nitrogen by Microbial Photoelectrochemical Cell: PANI/TiO2-NTs as a Photoanode].

The use of microbial photoelectrochemical cells (MPECs) for the removal of contaminants is a cost-effective and environment-friendly method. Based on the preparation of polyaniline/titanium dioxide nanotube array (PANI/TiO2-NTs) composite photoelectrodes, an MPEC system comprising PANI/TiO2-NTs photoanode and biocathode was constructed and the removal performance of nitrate nitrogen (NO3--N) was studied. The experimental results showed that the PANI/TiO2-NT electrode exhibited the best photoelectric performance when the PANI loading time was 80 s. Compared with the TiO2-NTs electrode, the photocurrent density doubled. The light-driven MPEC system could realize autotrophic denitrification without an external voltage. The biodegradation of NO3--N conformed to the pseudo first-order kinetics. The higher the photoresponse current density, the better the denitrification performance of the system. When the initial concentration of NO3--N was 25 mg·L-1 and the photoresponse current density increased from 0.17 mA·cm-2 to 0.67 mA·cm-2, the average denitrification rate increased from 0.83 mg·(L·h)-1 to 2.83 mg·(L·h)-1. High-throughput sequencing of the biocathode microbial membranes revealed that Pseudomonas (27.37%) was the dominant bacteria. It was considered that the photogenerated electrons generated by the PANI/TiO2-NTs photoanode were transmitted to the cathode through an external circuit. Pseudomonas and other microorganisms with autotrophic denitrification and electrochemical activity directly used the electrons on the electrode as the sole electron donors for autotrophic denitrification reaction.

[Preparation of a Nutritional Slow-release Packing Material with Function Microorganisms and Its Characteristics Evaluation].

A nutritional slow-release packing material with function microorganisms (SC) was prepared using emulsification and the cross-linked method. Its potential as packing material in biotrickling filters (BTF) for butyl acetate removal was evaluated. The physicochemical properties show that the packing has a porosity of 92.6%, bulk density of 40.75 kg·m-3, surface area of 2.45 m2·g-1, and real density of 551.52 kg·m-3. The packing material contains hydrophilic groups (O-H, C O) on its surface and nutrient elements (N, P), which are distributed uniformly, with release rates of 22.35 and 8.36 mg·(L·d)-1, respectively. The biomass concentration of the packing (protein/packing) is 14.61 mg·g-1. After storage for 7 and 30 d, the microorganisms fixed on the packing material could still remove more than 96% of butyl acetate. The BTF using SC as packings reach stable performance within a short time (8 d) and the removal efficiency is maintained at 94% unless there nutrition is supplied or the pH is adjusted. The BTF with polyurethane as packing material need a longer time to start up and the removal efficiency decreases to 80% under the same operating conditions. High-throughput sequencing analysis shows that the fixed degrading stains are dominant during the whole operation and the microbial structure is more stable, which could sustain the stable removal of butyl acetate in BTF using SC.

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.

Desulfurization of various organic sulfur compounds and the mixture of DBT + 4,6-DMDBT by Mycobacterium sp. ZD-19.

A new isolated dibenzothiophene (DBT) desulfurizing bacterium, identified as Mycobacterium sp. ZD-19 can utilize a wide range of organic sulfur compounds as a sole sulfur source. Thiophene (TH) or benzothiophene (BTH) was completely degraded by strain ZD-19 within 10h or 42 h, and 100% DBT or 4,6-dimethyldibenzothiophene (4,6-DMDBT) was removed within 50h or 56 h, respectively. Diphenylsulfide (DPS) possessed the lowest desulfurization efficiencies with 60% being transformed within 50h and 80% at 90 h. The desulfurization activities of five substrates by resting cells are in order of TH>BTH>DPS>DBT>4,6-DMDBT. In addition, when DBT and 4,6-DMDBT were mixed, they could be simultaneously desulfurized by strain ZD-19. However, DBT appeared to be attacked prior to 4,6-DMDBT. The desulfurization rate of DBT or 4,6-DMDBT in mixture is lower than they are desulfurized separately, indicating that the substrate competitive inhibition is existent when DBT and 4,6-DMDBT are mixed.

Composition and functional group characterization of extracellular polymeric substances (EPS) in activated sludge: the impacts of polymerization degree of proteinaceous substrates.

Characteristics of extracellular polymeric substances (EPS) in activated sludge strongly depend on wastewater substrates. Proteinaceous substrates (ProS) present in heterogeneous polymeric form are intrinsic and important parts of wastewater substrates for microorganisms in activated sludge systems. However, correlations between ProS and characteristics of EPS are scarce. This study systematically explored the impacts of monomeric (Mono-), low polymeric (LoP-) and high polymeric (HiP-) ProS on compositions and functional groups of EPS in activated sludge. The results showed that the change of polymerization degree of ProS significantly altered the composition of EPS. Compared to EPSMono-ProS, the proportion of proteins in EPSLoP-ProS and EPSHiP-ProS increased by 12.8% and 27.7%, respectively, while that of polysaccharides decreased by 22.9% and 63.6%, respectively. Moreover, the proportion of humic compounds in EPSLoP-ProS and EPSHiP-ProS were ∼6 and ∼16-fold higher than that in EPSMono-ProS, respectively. The accumulation of humic compounds in EPS increased the unsaturation degree of EPS molecules, and thereby reduced the energy requirement for electrons transition of amide bonds and aromatic groups. Size exclusion chromatography (SEC) analyses detected more molecular clusters in EPSHiP-ProS, indicating more complex composition of EPS in HiP-ProS fed activated sludge. Spectroscopic characterization revealed the dominance of hydrocarbon, protein, polysaccharide and aromatic associated bonds in all three EPS. Nevertheless, with the increase of polymerization degree of ProS, the protein associated bonds (such as CONH, CO, NC, NH) increased, while the polysaccharide associated bonds (such as COC, COH, OCOH) decreased. This paper paves a path to understand the role of ProS in affecting the production and characteristics of EPS in biological wastewater treatment systems.

A solid composite microbial inoculant for the simultaneous removal of volatile organic sulfide compounds: Preparation, characterization, and its bioaugmentation of a biotrickling filter.

Volatile organic sulfide compounds (VOSCs) are usually resistant to biodegradation, thereby limiting the performance of traditional biotechnology dealing with waste gas containing such pollutants especially in mixture. In this study, a solid composite microbial inoculant (SCMI) was prepared to remove dimethyl sulfide (DMS) and propanethiol (PT). Given that the DMS degradation activity of Alcaligenes sp. SY1 is inducible and the PT-degradation activity of Pseudomonas putida S-1 is constitutive, different strategies are designed for cell cultivation to obtain high VOSC removal rates of SCMI. Compared with the microbial suspension, the prepared SCMI exhibited better storage stability at 4 and 25°C. Inoculation of the SCMI in biotrickling filters (BTFs) could effectively shorten the start-up period and enhance the removal performance. Microbial analysis by Illumina MiSeq indicated that Alcaligenes sp. SY1 and P. putida S-1 might be dominant and persistent among the microbial communities of the BTF during the operation.

[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.

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.