Catechol and phenol degradation by a methanogenic population of bacteria

An anaerobic population of bacteria became acclimated to catechol and phenol in 32 and 18 days, respectively. Evidence from carbon balance measurements indicates that the aromatic ring is cleaved and that the products are stoichiometrically fermentable to methane and carbon dioxide.     

Kinetics of methanogenic degradation of phenol by activated-carbon-supported and granular biomass

A continuous-feed recycle bioreactor was used to study the kinetics of methanogenic degradation of phenol at 35 degrees C by bacteria supported on a bed of granular activated carbon (GAC). At dilution rates well above the growth rate of the culture, the cells not only populated the GAC, but also formed a layer of granular biomass. This layer was stabilized by the presence of the GAC, and accounted for over half of the phenol-degrading activity in the bioreactor. The specific phenol degradation rates for GAC-attached biomass, suspended biomass, and granular biomass were all in the range 0.15 to 0.22 mg phenol/mg volatile solids per day as measured under pseudo-steady-state conditions. (c) 1992 John Wiley & Sons, Inc.     

Effects of some alkyl phenols on methanogenic degradation of phenol.

The effects of six phenolic compounds (o-, m-, and p-cresol and 2-, 3-, and 4-ethylphenol) on the anaerobic biodegradation of phenol was examined in batch methanogenic cultures. Results showed that ethylphenols were more inhibitory of phenol degradation than were cresols. The inhibitory effects of the three isomers of cresol and ethylphenol did not vary with the isomer but rather with the substituted functional group.     

Effects of some alkyl phenols on methanogenic degradation of phenol

The effects of six phenolic compounds (o-, m-, and p-cresol and 2-, 3-, and 4-ethylphenol) on the anaerobic biodegradation of phenol was examined in batch methanogenic cultures. Results showed that ethylphenols were more inhibitory of phenol degradation than were cresols. The inhibitory effects of the three isomers of cresol and ethylphenol did not vary with the isomer but rather with the substituted functional group.     

Separation of bacteria from a methanogenic wastewater population by utilizing a self-generating Percoll gradient

Bacteria from a methanogenic wastewater population could be separated with a self-generating density gradient of Percoll. The separation was performed by centrifugation for 30 min at 30000 g in a simple angle-head rotor. Three types of bacteria were concentrated to apparent homogeneity in different bands; these were attributed to the methanogens Methanosarcina and Methanothrix , and to the dissimilatory sulphate-reducing bacterium Desulfovibrio. The described technique will contribute to a rapid diagnosis of the bacterial types that are active in waste-water treatment.     

Anaerobic degradation of toluene and o-xylene by a methanogenic consortium.

Toluene and o-xylene were completely mineralized to stoichiometric amounts of carbon dioxide, methane, and biomass by aquifer-derived microorganisms under strictly anaerobic conditions. The source of the inoculum was creosote-contaminated sediment from Pensacola, Fla. The adaptation periods before the onset of degradation were long (100 to 120 days for toluene degradation and 200 to 255 days for o-xylene). Successive transfers of the toluene- and o-xylene-degrading cultures remained active. Cell density in the cultures progressively increased over 2 to 3 years to stabilize at approximately 10(9) cells per ml. Degradation of toluene and o-xylene in stable mixed methanogenic cultures followed Monod kinetics, with inhibition noted at substrate concentrations above about 700 microM for o-xylene and 1,800 microM for toluene. The cultures degraded toluene or o-xylene but did not degrade m-xylene, p-xylene, benzene, ethylbenzene, or naphthalene. The degradative activity was retained after pasteurization or after starvation for 1 year. Degradation of toluene and o-xylene was inhibited by the alternate electron acceptors oxygen, nitrate, and sulfate. Degradation was also inhibited by the addition of preferred substrates such as acetate, H2, propionate, methanol, acetone, glucose, amino acids, fatty acids, peptone, and yeast extract. These data suggest that the presence of natural organic substrates or contaminants may inhibit anaerobic degradation of pollutants such as toluene and o-xylene at contaminated sites.     

Modeling simultaneous hexavalent chromium reduction and phenol degradation by a defined coculture of bacteria

Based on the kinetics of Cr(VI) reduction by Escherichia coli ATCC 33456 and phenol degradation by Pseudomonas putida DMP-1, a mathematical model is developed to describe simultaneous Cr(VI) reduction and phenol degradation in the coculture of the two species. The developed model incorporates the toxicity effects of Cr(VI) and phenol on phenol degradation and Cr(VI) reduction in the coculture. The model illustrates the inhibitory effects of phenol on Cr(VI) reduction and Cr(VI) toxicity toward phenol degradation. The model also reveals the recoveries of the activities of the repressed bacterial cells with continuous Cr(VI) reduction and phenol degradation in the coculture. The model is capable of predicting simultaneous Cr(VI) reduction and phenol degradation within a broad range of Cr(VI) and phenol concentrations and under an appropriate composition of populations. However, the model simulates lower concentrations of phenol than experimental observations once Cr(VI) is reduced to a low level (<7 mg/L). The model simulation for Cr(VI) also deviates from experimental data when P. putida is outnumbered by E. coli by a ratio of 1:5. (c) 1995 John Wiley & Sons, Inc.     

Thermophilic anaerobic degradation of butyrate by a butyrate-utilizing bacterium in coculture and triculture with methanogenic bacteria

We studied syntrophic butyrate degradation in thermophilic mixed cultures containing a butyrate-degrading bacterium isolated in coculture with Methanobacterium thermoautotrophicum or in triculture with M. thermoautotrophicum and the TAM organism, a thermophilic acetate-utilizing methanogenic bacterium. Butyrate was beta-oxidized to acetate with protons as the electron acceptors. Acetate was used concurrently with its production in the triculture. We found a higher butyrate degradation rate in the triculture, in which both hydrogen and acetate were utilized, than in the coculture, in which acetate accumulated. Yeast extract, rumen fluid, and clarified digestor fluid stimulated butyrate degradation, while the effect of Trypticase was less pronounced. Penicillin G, d-cycloserine, and vancomycin caused complete inhibition of butyrate utilization by the cultures. No growth or degradation of butyrate occurred when 2-bromoethanesulfonic acid or chloroform, specific inhibitors of methanogenic bacteria, was added to the cultures and common electron acceptors such as sulfate, nitrate, and fumarate were not used with butyrate as the electron donor. Addition of hydrogen or oxygen to the gas phase immediately stopped growth and butyrate degradation by the cultures. Butyrate was, however, metabolized at approximately the same rate when hydrogen was removed from the cultures and was metabolized at a reduced rate in the cultures previously exposed to hydrogen.     

Inhibition of a methanogenic population by thioglycollate

Thioglycollate added at 3.2 mM inhibited methane (CH₄) production from propionate by a methanogenic bacterial population obtained from an anaerobic digester. Gas production from acetate was not inhibited at this concentration. Glucose added in a semi-synthetic medium was degraded in the presence of 3.2 mM thioglycollate but propionate accumulated in the medium and the methanol yield was reduced. The severity of inhibition of propionate degradation increased with further incubation.     

Assimilatory reduction of sulfate and sulfite by methanogenic bacteria.

A variety of sulfur-containing compounds were investigated for use as medium reductants and sulfur sources for growth of four methanogenic bacteria. Sulfide (1 to 2 mM) served all methanogens investigated well. Methanococcus thermolithotrophicus and Methanobacterium thermoautotrophicum Marburg and delta H grew well with S0, SO3(2-), or thiosulfate as the sole sulfur source. Only Methanococcus thermolithotrophicus was able to grow with SO4(2-) as the sole sulfur source. 2-Mercaptoethanol at 20 mM was greatly inhibitory to growth of Methanococcus thermolithotrophicus on SO4(2-) or SO2(2-) and Methanobacterium thermoautotrophicum Marburg on SO3(2-) but not to growth of strain delta H on SO3(2-). Sulfite was metabolized during growth by Methanococcus thermolithotrophicus. Sulfide was produced in cultures of Methanococcus thermolithotrophicus growing on SO4(2-), SO3(2-), thiosulfate, and S0. Methanobacterium thermoautotrophicum Marburg was successfully grown in a 10-liter fermentor with S0, SO3(2-), or thiosulfate as the sole sulfur source.     

Anaerobic degradation of coniferyl alcohol by methanogenic consortia.

Coniferyl alcohol was shown to be completely biodegradable to carbon dioxide and methane under strictly anaerobic culture conditions. The mineralization of 300 mg of the substrate per liter was observed in acclimated ferulic acid-degrading methanogenic consortia, as well as in anaerobic enrichments on coniferyl alcohol seeded with sewage sludge. Ferulic and phenylpropionic acids were detected in the cultures degrading coniferyl alcohol as the sole carbon and energy source, suggesting that this compound is oxidized to ferulic acid, which is then degraded as previously described.     

Anaerobic degradation of isovalerate by a defined methanogenic coculture

Isovalerate-oxidizing strictly aneerobic bacteria were isolated from marine sediment and sewage sludge in coculture with Desulfovibrio sp. Cells stained Gram positive and behaved Gram positive also in Gram classification with KOH. Isovalerate degradation depended on interspecies hydrogen transfer to syntrophic hydrogen-oxidizing sulfate reducers or methanogens. Isovalerate was the only substrate utilized and was fermented to 3 mol acetate and 1 mol hydrogen per mol substrate. The degradation pathway was studied by enzyme assays in crude cell extracts, and included acetyl-CoA dependent activation of isovalerate, oxidation to methylcrotonyl-CoA and carboxylation to methylgluta-conyl-CoA which is hydrated and cleaved to acetoacetate and acetyl-CoA. Studies with inhibitors and ionophores suggest that energy conservation with this organism depends on either acetate efflux-driven proton symport or on an ion-gradient driven carboxylation mechanism.     

Acidophilic degradation of methanol by a methanogenic enrichment culture

Abstract An acidophilic methanogenic enrichment culture was obtained in a continuous up-flow anaerobic sludge blanket reactor operated at pH 4.2 with methanol as the sole carbon source. The specific methylotrophic methanogenic activity of the enriched reactor sludge at pH 5 was 3.57 g COD g⁻¹ volatile suspended solids day⁻¹ and the apparent doubling time of the biomass was 15.8 h. Acidic conditions were obligatory, since the enrichment culture was not able to produce methane or to grow at pH 7. Based on morphological characteristics, the dominant methanogenic species in the enrichment culture was a Methanosarcina .     

Improved identification of methanogenic bacteria by fluorescence microscopy.

Methanogenic bacteria can be tentatively identified by fluorescence microscopy. This technique was improved by carefully selecting a series of excitation and barrier filters that matched the excitation and emission spectra of some unique coenzymes viz., F420 and F350, in methanogenic bacteria.     

Improved identification of methanogenic bacteria by fluorescence microscopy.

Methanogenic bacteria can be tentatively identified by fluorescence microscopy. This technique was improved by carefully selecting a series of excitation and barrier filters that matched the excitation and emission spectra of some unique coenzymes viz., F420 and F350, in methanogenic bacteria.     

Genome complexity of methanogenic bacteria.

The genome complexities of different methanogenic bacteria were investigated by using an optical method to study renaturation kinetics of single-stranded DNA. The observed genome sizes ranged from 1.0 X 10(9) to 1.8 X 10(9) daltons, which is a typical range for procaryotic cells. Melting profiles of the DNA of three methanogenic species from different families show fractions which have a higher A . T content than the average DNA of that species.     

Anaerobic degradation of phenol and phenol derivatives by Desulfobacterium phenolicum sp. nov.

A new sulfate-reducing bacterium was enriched and isolated from marine sediment with phenol as sole electron donor and carbon source. Strain Ph01 grew well in defined media without growth factors. Further aromatic compounds oxidized by strain Ph01 were benzoate, phenylacetate, 2-hydroxybenzoate, 4-hydroxybenzoate, 4-hydroxyphenylacetate, p-cresol, indole, anthranilic acid, and phenylalanine. Various fatty acids, alcohols and dicarboxylic acids were also utilized by strain Ph01. Sulfate and thiosulfate served as electron acceptors and were reduced to H₂S. Stoichiometric measurements with strain Ph01 showed complete oxidation of phenol to CO₂. Cytochromes and menaquinone MK-7(H₂) were present; desulfoviridin could not be detected. Strain Ph01 is described as type strain of the new species Desulfobacterium phenolicum.In further marine enrichments with 4-hydroxybenzoate, 4-hydroxyphenylacetate, p-cresol or o-cresol as substrates and sulfate as electron acceptor a variety of morphologically different sulfate-reducing bacteria developed. However, since the new isolate strain Ph01 was able to degrade all these aromatic compounds (except o-cresol) no further studies with the enrichment cultures were carried out.