Biodegradation Of Methyl Tertiary Butyl Ether Research Articles

Methyl tertiary butyl ether biodegradation by the bacterial consortium isolated from petrochemical wastewater and contaminated soils of Imam Khomeini Port Petrochemical Company (Iran)

Methyl tertiary butyl ether (MTBE) is a high soluble oxygenated supplement to fuel that may rapidly contaminate water and soil. Low biodegradability of this compound in the environment has led to more extensive efforts for finding powerful MTBE degrading microbial populations to be used in biological treatment of polluted waters and soils. The present study aimed to isolate MTBE degrading bacteria from petrochemical wastewater and soil of Mahsahar MTBE production site (Iran), and increasing their biodegradation potential in a bacterial consortium. Enrichment of degrading bacteria was done in basal salt media (BSM) containing 50–400 mg/l MTBE. The MTBE degradation pattern was assessed via gas chromatography (GC). The isolates were identified molecularly based on the sequence of 16S rRNA gene. The growth and degradation potential of bacterial consortium was optimized in different pH and temperatures. The best MTBE degrading bacteria were identified as Pseudomonas putida, Pseudomonas aeruginosa, and Enterobacter oryzendophyticus. The first order kinetics for MTBE degradation showed that the bacterial consortium was able to degrade 99.93% of MTBE during 15 days that was 6.08% higher than the most putative monoculture bacterium. The highest MTBE degradation by bacterial consortium was obtained at 30 °C and pH value of 7. The isolated bacterial consortium that involved novel MTBE degrading bacterial strains, decomposed high amounts of MTBE during 15 days of incubation in different temperatures and pH values without addition of co-catalytic substrate is proposed for cost effective removal of this pollutant from the environment.

Biodegradation of Methyl Tertiary Butyl Ether (MTBE) by a Microbial Consortium in a Continuous Up-Flow Packed-Bed Biofilm Reactor: Kinetic Study, Metabolite Identification and Toxicity Bioassays.

This study investigated the aerobic biodegradation of methyl tertiary-butyl ether (MTBE) by a microbial consortium in a continuous up-flow packed-bed biofilm reactor using tezontle stone particles as a supporting material for the biofilm. Although MTBE is toxic for microbial communities, the microbial consortium used here was able to resist MTBE loading rates up to 128.3 mg L-1 h-1, with removal efficiencies of MTBE and chemical oxygen demand (COD) higher than 90%. A linear relationship was observed between the MTBE loading rate and the MTBE removal rate, as well as between the COD loading rate and the COD removal rate, within the interval of MTBE loading rates from 11.98 to 183.71 mg L-1 h-1. The metabolic intermediate tertiary butyl alcohol (TBA) was not detected in the effluent during all reactor runs, and the intermediate 2-hydroxy butyric acid (2-HIBA) was only detected at MTBE loading rates higher than 128.3 mg L-1 h-1. The results of toxicity bioassays with organisms from two different trophic levels revealed that the toxicity of the influent was significantly reduced after treatment in the packed-bed reactor. The packed-bed reactor system used in this study was highly effective for the continuous biodegradation of MTBE and is therefore a promising alternative for detoxifying MTBE-laden wastewater and groundwater.

Biodegradation of MTBE by Bacteria Isolated From Oil Hydrocarbons- Contaminated Environments

Methyl tertiary butyl ether (MTBE) belongs to the group of gasoline oxygenates and persistent environment contaminants, and shows potential for biodegradation in aerobic and anaerobic conditions, through application of pure microbial cultures. Presented research shows that indigenous bacterial isolates 6sy and 24p, selected from oil hydrocarbons-contaminated environments, were capable of utilizing MTBE as sole carbon and energy source. Based on 16S rDNA sequence analysis, bacterial isolates 6sy and 24p were identified as Staphylococcus saprophyticus subsp. saprophyticus and Pseudomonas sp., respectively. The MTBE biodegradation rate was affected by longevity of incubation period and initial MTBE concentration. After 3 weeks of incubation at 25°C in a dark, the removal rates of initial 25 and 125 ppm MTBE concentrations by Staphylococcus saprophyticus 6sy were found to be 97, and 63%, respectively, while efficiency of Pseudomonas sp. in degradation of indicated concentrations was 96, and 40%, respectively. Both bacterial isolates were able to grow in MTBE-containing growth medium. Highest growth rate of bacterial isolates was observed at the end of incubation period. The presented results indicated the potential of these bacterial isolates in bioremediation of MTBE-contaminated environments.

TBA Production by Acid Hydrolysis of MTBE During Heated Headspace Analysis and Evaluation of a Base as a Preservative

AbstractAt room temperature (20° ± 3°C), purge and trap samplers provide poor sensitivity for analysis of the fuel oxygenates that are alcohols, such as tertiary butyl alcohol (TBA). Because alcohols are miscible or highly soluble in water, they are not efficiently transferred to a gas chromatograph for analysis. To improve the efficiency of transfer, the water in a purge and trap sampler can be heated. Alternatively, the sensitivity for TBA can be improved by preparing the sample in a heated static headspace sampler. In a heated water sample, the acid used as a preservative may cause chemical hydrolysis of methyl tertiary butyl ether (MTBE) to produce TBA. This effect is well illustrated in this paper using data collected by the U.S. Environmental Protection Agency Office of Research and Development on a plume of MTBE in California. Samples were analyzed using a static head‐space sampler heated to 80°C. The ground water samples were preserved in the field with HCl to a pH ≤ 2. The extent of MTBE hydrolysis to TBA during sample analysis varied from 19% to 87%; the average extent of hydrolysis was 59%. To confirm and document the importance of acid hydrolysis of MTBE at higher temperatures during sample preparation, the rate of hydrolysis of MTBE was measured at 80°C. At pH = 1, the rate of hydrolysis was 1.22/h, while the rate at pH = 2 was 0.15/h. Acid hydrolysis of MTBE during sample preparation in a heated headspace sampler can be avoided by preserving the sample with 1% (w/w) trisodium phosphate dodecahydrate (TSP) instead of using HCl, or by neutralizing the acid before analysis. In the presence of an acclimated microbial culture, TSP prevented biodegradation of MTBE, as well as benzene, toluene, ethylbenzene, and xylene compounds, in ground water at room temperature for 66 d. However, in a spike recovery experiment, TSP caused based catalyzed hydrolysis of bromomethane. It is not appropriate as a universal preservative.

Induction of Methyl Tertiary Butyl Ether (MTBE)-Oxidizing Activity in Mycobacterium vaccae JOB5 by MTBE

Alkane-grown cells of Mycobacterium vaccae JOB5 cometabolically degrade the gasoline oxygenate methyl tertiary butyl ether (MTBE) through the activities of an alkane-inducible monooxygenase and other enzymes in the alkane oxidation pathway. In this study we examined the effects of MTBE on the MTBE-oxidizing activity of M. vaccae JOB5 grown on diverse nonalkane substrates. Carbon-limited cultures were grown on glycerol, lactate, several sugars, and tricarboxylic acid cycle intermediates, both in the presence and absence of MTBE. In all MTBE-containing cultures, MTBE consumption occurred and tertiary butyl alcohol (TBA) and tertiary butyl formate accumulated in the culture medium. Acetylene, a specific inactivator of alkane- and MTBE-oxidizing activities, fully inhibited MTBE consumption and product accumulation but had no other apparent effects on culture growth. The MTBE-dependent stimulation of MTBE-oxidizing activity in fructose- and glycerol-grown cells was saturable with respect to MTBE concentration (50% saturation level = 2.4 to 2.75 mM), and the onset of MTBE oxidation in glycerol-grown cells was inhibited by both rifampin and chloramphenicol. Other oxygenates (TBA and tertiary amyl methyl ether) also induced the enzyme activity required for their own degradation in glycerol-grown cells. Presence of MTBE also promoted MTBE oxidation in cells grown on organic acids, compounds that are often found in anaerobic, gasoline-contaminated environments. Experiments with acid-grown cells suggested induction of MTBE-oxidizing activity by MTBE is subject to catabolite repression. The results of this study are discussed in terms of their potential implications towards our understanding of the role of cometabolism in MTBE and TBA biodegradation in gasoline-contaminated environments.

Bioremediation of methyl tertiary-butyl ether (MTBE) by an innovative biofilter

Methyl tertiary-butyl ether (MTBE) is a synthetic chemical used in unleaded gasoline as an additive to reduce levels of ozone and carbon monoxide from auto exhaust. Due to its chemical and recalcitrant properties, MTBE has caused groundwater contamination worldwide. A laboratory-scale biofilter made of a natural fiber (kenaf) mat and inoculated with MTBE-degrading microorganisms, was evaluated for MTBE removal efficiency. Operational parameters of oxygen flow rate, hydraulic retention time (HRT), yeast extract and initial MTBE concentration were varied and MTBE removal efficiencies determined. Four kinetic models were evaluated to describe the MTBE removal in the reactor. Formaldehyde and tertiary butyl alcohol (the most two reported MTBE biodegradation byproducts) were not found in the effluent; instead, carbon dioxide was monitored as the end product based on the results of a metabolic mass balance evaluation. Toxicity of treated effluent was evaluated by employing the Microtox acute toxicity test and comparing that to the influent.

Cometabolism ofMethyl tertiary Butyl Ether and Gaseous n -Alkanes by Pseudomonas mendocina KR-1 Grown on C 5 toC 8 n -Alkanes

Pseudomonas mendocina KR-1 grew well on toluene, n-alkanes (C5 to C8), and 1 degrees alcohols (C2 to C8) but not on other aromatics, gaseous n-alkanes (C1 to C4), isoalkanes (C4 to C6), 2 degrees alcohols (C3 to C8), methyl tertiary butyl ether (MTBE), or tertiary butyl alcohol (TBA). Cells grown under carbon-limited conditions on n-alkanes in the presence of MTBE (42 micromoles) oxidized up to 94% of the added MTBE to TBA. Less than 3% of the added MTBE was oxidized to TBA when cells were grown on either 1 degrees alcohols, toluene, or dextrose in the presence of MTBE. Concentrated n-pentane-grown cells oxidized MTBE to TBA without a lag phase and without generating tertiary butyl formate (TBF) as an intermediate. Neither TBF nor TBA was consumed by n-pentane-grown cells, while formaldehyde, the expected C1 product of MTBE dealkylation, was rapidly consumed. Similar Ks values for MTBE were observed for cells grown on C5 to C8 n-alkanes (12.95 +/- 2.04 mM), suggesting that the same enzyme oxidizes MTBE in cells grown on each n-alkane. All growth-supporting n-alkanes (C5 to C8) inhibited MTBE oxidation by resting n-pentane-grown cells. Propane (Ki = 53 micromoles) and n-butane (Ki = 16 micromoles) also inhibited MTBE oxidation, and both gases were also consumed by cells during growth on n-pentane. Cultures grown on C5 to C8 n-alkanes also exhibited up to twofold-higher levels of growth in the presence of propane or n-butane, whereas no growth stimulation was observed with methane, ethane, MTBE, TBA, or formaldehyde. The results are discussed in terms of their impacts on our understanding of MTBE biodegradation and cometabolism.

Use of compound-specific stable carbon isotope analyses to demonstrate anaerobic biodegradation of MTBE in groundwater at a gasoline release site.

Currently it is unclear if natural attenuation is an appropriate remedial approach for groundwater impacted by methyl tertiary butyl ether (MTBE). Site-characterization data at most gasoline release sites are adequate to evaluate attenuation in MTBE concentrations over time or distance. But, demonstrating natural biodegradation of MTBE requires laboratory microcosm studies, which could be expensive and time-consuming. Recently, compound-specific carbon isotope ratio analyses (13C/12C expressed in delta13C notation) have been used to demonstrate aerobic biodegradation of MTBE in laboratory incubations. This study explored the potential of this approach to distinguish MTBE biodegradation from other abiotic processes in an anaerobic groundwater plume that showed extensive decrease in MTBE concentrations. To our knowledge, this is the first study to use delta13C of MTBE data in groundwater and laboratory microcosms to demonstrate anaerobic biodegradation of MTBE. The delta13C of MTBE in monitoring wells increased by up to 31 per thousand (-25.5 per thousand to +5.5 per thousand) along with a 40-fold decrease in MTBE concentrations. Anaerobic incubations in laboratory microcosms indicated up to 20-fold reduction in MTBE concentrations with a corresponding increase in delta13C of MTBE of up to 33.4 per thousand (-28.7 per thousand to +4.7 per thousand) in live microcosms. Little enrichment was observed in autoclaved controls. These results demonstrate that anaerobic biodegradation was the dominant natural attenuation mechanism for MTBE at this site. The estimated isotopic enrichment factors (epsilon(field) = -8.10 per thousand and epsilon(lab) = -9.16 per thousand) were considerably larger than the range (-1.4 per thousand to -2.4 per thousand) previously reported for aerobic biodegradation of MTBE in laboratory incubations. These observations strongly suggest that delta13C of MTBE could be potentially useful as an "indicator" of in-situ MTBE biodegradation.

A Field Application of Hydrogen-Releasing Compound (HRCTM) for the Enhanced Bioremediation of Methyl Tertiary Butyl Ether (MTBE)

Due to a greater understanding of the behavior of the fuel oxygenate Methyl Tertiary Butyl Ether (MTBE) in groundwater, the United States Environmental Protection Agency (EPA) and the American Petroleum Institute (API) recently have acknowledged the need for the development and application of additional remedial strategies to address the more extensive, longer lived, and faster moving dissolved MTBE plumes often associated with oxygenated fuel releases (API, 2000 and USEPA, 2000a).The need for alternative methods for managing dissolved MTBE plumes is particularly evident in the case of the Upper Glacial aquifer of Long Island, New York. Hydrogeologic conditions in the this water table aquifer (i. e., high hydraulic conductivity, high average pore velocities, low organic carbon, and high rates of recharge) have been found to contribute to the formation of extensive, long, narrow, and three-dimensional dissolved MTBE plumes that plunge into the aquifer in response to recharge (Weaver et. al. 1999). The characteristics of MTBE plumes in the Upper Glacial aquifer in combination with abundant sensitive receptors (mainly drinking water supply wells), often renders monitored natural attenuation (MNA) plume management strategies inappropriate, resulting in the need for plume control, frequently via pumping and treating (NYSDEC, 2000). In such cases, remedial costs can rise well beyond those associated with similar fuel releases that did not contain MTBE (USEPA, 1998a). Consequently, the application of remedial technologies for MTBE other than MNA, or pumping and treating, are of great interest to those responsible for the management of dissolved MTBE plumes on Long Island or in similar hydrogeologic settings.An alternative strategy for the remediation of dissolved MTBE plumes was recently field tested at an oxygenated fuel spill site on Long Island. The strategy was enhanced biodegradation via the application of Hydrogen Release Compound (HRCTM). HRCTM is a form of polylactate ester that slowly releases biodegradation stimulating constituents into the aquifer and has been shown in other studies to foster methanogenic conditions that advance the reductive dechlorina-tion of perchloroethene (PCE) and trichloroethene (TCE) (Koenigsberg, 1998). Numerous reports have been written that discuss the biodegradation of MTBE under aerobic conditions, as well as microcosm studies in which MTBE biodegradation was observed under anaerobic conditions. However, there are limited reports that document the natural anaerobic biodegradation of dissolved MTBE (McLoughlin, 2000). Despite the lack of documented natural anaerobic biodegradation of MTBE, it has been observed that MTBE transport often occurs under anoxic conditions at oxygenated fuel releases as the result of the biodegradation of other fuel constituents, such as benzene, toluene, ethylbenzene and xylene (BTEX), which deplete the available dissolved oxygen as well as other electron acceptors (nitrate, ferric iron, manganese, etc.) (USEPA, 2000c and API, 1996). Therefore, an anaerobic biodegradation strategy is attractive due to its synergy with the existing geochemical conditions. Consequently, the study was conceived and designed to test the ability of HRC(tm) to foster the anaerobic bio-degradation of MTBE under methano-genic conditions (McLoughlin, 2000).The application of HRC(tm) did result in the formation of a large area of enhanced reducing conditions in the vicinity and down gradient of the application zone. However, under these site conditions, the HRC(tm) application did not induce measurable methanogenic conditions with the associated elevated dissolved hydrogen concentrations required for significant MTBE anaerobic biodegradation. The high hydraulic conductivity and high average pore velocity at the site were likely responsible. Despite this, the study can be viewed as a success since much was learned that can be used in future studies of anaerobic biodegradation of MTBE and the application of HRC(tm).

Methyl tertiary-butyl ether (MTBE) biodegradation in batch and continuous upflow fixed-biofilm reactors

MTBE was effectively biodegraded under oxidizing environmental conditions in the presence of an acclimated mixed culture isolated from a petrochemical biotreater. MC-1 (Gram-positive coccus), MC-2 (Acinetobacter lwoffii) and MC-3 (Bacillus sp.) were present in the culture medium, with MC-1 being the predominant organism. The presence of other easily assimilated carbon sources in the culture medium influenced MTBE biodegradation. In batch studies, 62–73% MTBE was biodegraded in 144–192 hours at an initial concentration of 100 ppm as the sole carbon source, with the ORP ranging from 191–274 mv, and at a temperature of 29°C. The overall K rate constants and the specific k rate constants were evaluated using a first order rate equation. Mean values determined were 1.79×10-1 day-1 and 1.66×10-2 day-1/(mg/L cell mass), respectively. Continuous upflow fixed biofilm reactor studies were performed at retention times of 0.25, 0.5, and 1 day at an initial MTBE concentration of 150 ppm. Results indicated that approximately 53% MTBE was biodegraded for the 0.25 day retention time and 70% for both the 0.5 and 1.0 day retention times. Three kinetic models were evaluated for all experimental retention times. These included: Model I (Eckenfelder); Model II (Arvin) and Model III (first-order biphasic). Results indicated that model III yielded the highest and most consistent correlation coefficients for all retention times evaluated.

Biodegradation of Methyl tertiary-Butyl Ether (MTBE) and Other Fuel Oxygenates

Methyl tertiary-butyl ether (MTBE) has been added to gasoline in the U.S. for the last decade in order to meet the mandates of the 1990 Clean Air Act. This law decreed that gasoline sold in many locations must contain oxygenates to improve combustion and minimize air pollution. Unfortunately, this widespread use has led to the contamination of some soils and aquifers, and remediation is now required. Bioremediation has proven to be an environmentally responsible and cost-effective approach to remediating petroleum spills; this article reviews the potential that bioremediation may also be appropriate for remediating MTBE contamination. There is now good evidence that MTBE can be degraded by bacteria and fungi under aerobic conditions, and promising indications that the process also occurs under methanogenic and ferric iron-reducing conditions. Yet, apparently it is not a widespread phenomenon. The challenge is to find effective bioremediation strategies that maximize this biodegradation so that it can be used reliably in cleaning contaminated sites. Both simple biostimulation and more complex bioaugmentation protocols are being developed to meet this pressing need.