Methanol suppression of trichloroethylene degradation by Methylosinus trichosporium (OB3b) and methane-oxidizing mixed cultures.
ABSTRACT The effect of methanol on trichloroethylene (TCE) degradation by mixed and pure methylotrophic cultures was examined in batch culture experiments. Methanol was found to relieve growth inhibition of Methylosinus trichosporium (OB3b) at high (14 mg/L) TCE concentrations. Degradation of TCE was determined by both radiolabeling and gas chromatography techniques. When cultures were grown on methanol over 10 to 14 d with 0.3 mg/L TCE, OB3b degraded 16.89 +/- 0.82% (mean +/- SD) of the TCE, and a mixed culture (DT type II) degraded 4.55 +/- 0.11%. Mixed culture (JS type I) degraded 4.34 +/- 0.06% of the TCE. When grown on methane with 0.3 mg/L TCE, 32.93 +/- 2.01% of the TCE was degraded by OB3b, whereas the JS culture degraded 24.3 +/- 1.38% of the TCE, and the DT culture degraded 34.3 +/- 2.97% of the TCE. The addition of methanol to cultures grown on methane reduced TCE degradation to 16.21 +/- 1.17% for OB3b and to 5.08 +/- 0.56% for JS. Although methanol reduces the toxicity of TCE to the cultures, biodegradation of TCE cannot be sustained in methanol-grown cultures. Since high TCE concentrations appear to inhibit methane uptake and growth, we suggest the primary toxicity of TCE is directed towards the methane monooxygenase.
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ABSTRACT: Trichloroethylene (TCE), a common groundwater contaminant, is a suspected carcinogen that is highly resistant to aerobic biodegradation. An aerobic, methane-oxidizing bacterium was isolated that degrades TCE in pure culture at concentrations commonly observed in contaminated groundwater. Strain 46-1, a type I methanotrophic bacterium, degraded TCE if grown on methane or methanol, producing CO(2) and water-soluble products. Gas chromatography and C radiotracer techniques were used to determine the rate, methane dependence, and mechanism of TCE biodegradation. TCE biodegradation by strain 46-1 appears to be a cometabolic process that occurs when the organism is actively metabolizing a suitable growth substrate such as methane or methanol. It is proposed that TCE biodegradation by methanotrophs occurs by formation of TCE epoxide, which breaks down spontaneously in water to form dichloroacetic and glyoxylic acids and one-carbon products.Applied and Environmental Microbiology 05/1988; 54(4):951-6. · 3.68 Impact Factor
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ABSTRACT: Over 20 new cultures of methane-utilizing microbes, including obligate (types I and III) and facultative methylotrophic bacteria were isolated. In addition to their ability to oxidize methane to methanol, resting cell-suspensions of three distinct types of methane-grown bacteria (Methylosinus trichosporium OB3b [type II, obligate]; Methylococcus capsulatus CRL M1 NRRL B-11219 [type I, obligate]; and Methylobacterium organophilum CRL-26 NRRL B-11222 [facultative]) oxidize C2 to C4 n-alkenes to their corresponding 1,2-epoxides. The product 1,2-epoxides are not further metabolized and accumulate extracellularly. Methanol-grown cells do not have either the epoxidation or the hydroxylation activities. Among the substrate gaseous alkenes, propylene is oxidized at the highest rate. Methane inhibits the epoxidation of propylene. The stoichiometry of the consumption of propylene and oxygen and the production of propylene oxide is 1:1:1. The optimal conditions for in vivo epoxidation are described. Results from inhibition studies indicate that the same monooxygenase system catalyzes both the hydroxylation and the epoxidation reactions. Both the hydroxylation and epoxidation activities are located in the cell-free particulate fraction precipitated between 10,000 and 40,000 x g centrifugation.Applied and Environmental Microbiology 08/1979; 38(1):127-34. · 3.68 Impact Factor
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ABSTRACT: Trichloroethylene (TCE) was metabolized by cytochrome P-450 containing mixed-function oxidase systems to chloral (2,2,2,-trichloroacetaldehyde), glyoxylic acid, formic acid, CO, and TCE oxide. TCE oxide was synthesized, and its breakdown products were analyzed. Under acidic aqueous conditions the primary products were glyoxylic acid and dichloracetic acid. The primary compounds formed under neutral or basic aqueous conditions were formic acid and CO. TCE oxide did not form chloral in any of these or other aqueous systems, even when iron salts, ferriprotoporphyrin IX, or purified cytochrome P-450 was present. Ferric iron salts catalyzed the rearrangement of TCE oxide to chloral only in CH2Cl2 or CH3CN. A 500-fold excess of iron was required for complete conversion. A kinetic model involving the zero-order oxidation of TCE to TCE oxide by cytochrome P-450 and the first-order degradation of the epoxide was used to test the hypothesis that TCE oxide is an obligate intermediate in the conversion of TCE to other metabolites. Kinetic constants fo the breakdown of TCE oxide and for the oxidative metabolism of TCE to stable metabolites were used to predict epoxide concentrations required to support the obligate intermediacy of TCE oxide. The maximum levels of TCE oxide detected in systems using microsomal fractions and purified cytochrome P-450 were 5-28-fold lower than those predicted from the model. The kinetic data and the discrepancies between the observed metabolites and TCE oxide breakdown products support the view that the epoxide is not an obligate intermediate in the formation of chloral, and an alternative model is presented in which chlorine migration occurs in an oxygenated TCE-cytochrome P-450 transition state.Biochemistry 04/1982; 21(5):1090-7. · 3.38 Impact Factor