Aerobic Degradation Of Phenol Research Articles

Microbial electricity-driven anaerobic phenol degradation in bioelectrochemical systems

Microbial electrochemical technologies have been extensively employed for phenol removal. Yet, previous research has yielded inconsistent results, leaving uncertainties regarding the feasibility of phenol degradation under strictly anaerobic conditions using anodes as sole terminal electron acceptors. In this study, we employed high-performance liquid chromatography and gas chromatography-mass spectrometry to investigate the anaerobic phenol degradation pathway. Our findings provide robust evidence for the purely anaerobic degradation of phenol, as we identified benzoic acid, 4-hydroxybenzoic acid, glutaric acid, and other metabolites of this pathway. Notably, no typical intermediates of the aerobic phenol degradation pathway were detected. One-chamber reactors (+0.4 V vs. SHE) exhibited a phenol removal rate of 3.5 ± 0.2 mg L−1 d−1, while two-chamber reactors showed 3.6 ± 0.1 and 2.6 ± 0.9 mg L−1 d−1 at anode potentials of +0.4 and + 0.2 V, respectively. Our results also suggest that the reactor configuration certainly influenced the microbial community, presumably leading to different ratios of phenol consumers and microorganisms feeding on degradation products.

Oxidative degradation of phenol by sulfidated zero valent iron under aerobic conditions: The effect of oxalate and tripolyphosphate ligands

After adding either organic or inorganic ligands, sulfidated nano-zero-valent iron (SnZVI) was used for aerobic degradation of phenol, and the effect of the ligand species on oxidation performance was investigated. We found that SnZVI hardly degraded phenol in the absence of ligand addition. Ligands initiated and promoted the degradation of pollutants by SnZVI. The data herein show that a characteristic inorganic ligand, tripolyphosphate (TPP), is more effective in enhancing oxidation than a characteristic organic ligand oxalate. In addition to the scavenging of reactive oxidants by the organic ligand, more ferrous ion (Fe(II)) dissolution from SnZVI in the TPP system is another cause for the superior enhancement by the inorganic ligand. In the oxalate system, as the sulfur content of SnZVI increased, the oxidation efficiency increased because FeS shell promoted the transfer of electrons to produce more reactive oxygen species (ROS). In TPP system, the effect of sulfur content on oxidation performance is more complex. The SnZVI with low sulfur content showed poor oxidation performance compared with that of nZVI. Further experiments proved that sulfidation might weaken the complexation of TPP with surface bound Fe, which would slow down the ionic Fe(II) dissolution rate. Therefore, sulfidation has the dual effects of enhancing electron transfer and inhibiting the complexation of inorganic ligands. In addition, the mechanisms of ROS generation in different ligand systems were investigated herein. Results showed that the critical ROS in both the oxalate and TPP systems are hydroxyl radicals, and that they are produced via one-electron activation of O2.

Kinetics of phenol degradation by selected bacterial strains with different genetic properties

The major environmental problem in the northeastern Estonia is the semi-coke mounds in oilshale industry areas, Alternatives to the chemical methods (sorption, ozonation) for removingxenobiotic compounds from leachate are biological methods, like bioaugmentation, where theproperly selected microorganisms are used. Determination of the kinetic constants (maximumspecific growth rates (µmax), lag times (A.), half saturation constants (Kso for oxygenatingactivity and Ksa for growth) and inhibition constants (Ki)) will give us information about therate of pollutant degradation and is the basis for the selection of the most effective bacteria forbioaugmentation. In phenol degradation the initial and rate-limiting enzyme is phenolhydroxylase, encoded by different genes, The aim of this work was to carry out a kineticstudy of the aerobic degradation of phenol using single strains (Pseudomonas mendocinaPC 1, P. fluorescens PC 18, PC20 and PC24) isolated from river water continuously pollutedwith phenolic compounds, The strains PC 1 and PC 18 contain genes for multicomponentphenol hydroxylase, whereas single-component phenol hydroxylase (pheBA operon)characterizes the strains PC20 and PC24, The phenol-oxygenating activity (Kso) was obtainedfrom substrate-dependent oxygen uptake data (oxygen concentrations were measured with aClark-type oxygen electrode) using Michaelis-Mentens model. Specific growth rates µ andlag times). were calculated from absorbance growth curves on phenol concentrations 0,2-10.6mM and the growth kinetic constants (µmax, Ksa, Ki) were estimated using Haldanes, Edwardsand Aiba-Edwards model. The Kso values for phenol were one order of magnitude lower instrains PC 1 and PC 18 than in strains PC20 and PC24.

An efficient start-up strategy of a continuous stirred tank bioreactor with biofilm

The paper concerns start-up of a continuous stirred tank bioreactor with biofilm for aerobic microbiological degradation of a toxic organic compound. Method for the bioreactor start-up was proposed, which is based on using a proportional controller for the control by flow rate. Numerical simulations were performed using a heterogeneous mathematical model of the bioreactor, which takes into account three phases: gas, liquid and biofilm. It was shown that using the proposed strategy results in obtaining even over 8-fold decrease of time, after which a stationary, low concentration of a toxic compound is obtained. Moreover, it decreases the risk of anoxia of microorganisms related to considerable decrease in dissolved oxygen concentration, which takes place during a standard start-up method. The efficiency of the proposed method was shown on the example of aerobic degradation of phenol for different values of the substrate concentration in the feed stream and for different values of mean residence time of the liquid in the bioreactor.

INHIBITION KINETICS DURING THE OXIDATION OF BINARY MIXTURES OF PHENOL WITH CATECHOL, RESORCINOL AND HYDROQUINONE BY PHENOL ACCLIMATED ACTIVATED SLUDGE

In this work the aerobic degradation of phenol (PH), catechol (CA), resorcinol (RE), hydroquinone (HY) and of the binary mixtures PH+CA, PH+RE, PH+HY by phenol-acclimated activated sludge was studied. Single substrate experiments show a Haldane-type dependence of the respiration rate on PH, RE and HY, while CA corresponded to the Monod model. Binary substrate experiments demonstrated that the presence of a second substrate only affected the kinetics, but not the stoichiometry of the oxidation of the compounds tested. While CA inhibited the oxidation of PH, PH inhibited the oxidation of RE and HY. A mathematical model was developed to represent the aerobic biodegradation of the phenolic compounds tested. The agreement between the proposed model and the experimental data indicates that the proposed model can be useful for predicting substrate and dissolved oxygen concentrations in bioreactors treating phenolic wastewaters.

Modelling the change in the oxidation coefficient during the aerobic degradation of phenol by acclimated activated sludge

In this work the aerobic degradation of phenol by acclimated activated sludge was studied. Results demonstrate that while the phenol removal rate by acclimated activated sludge follows the Monod model, the oxygen uptake rate obeys a Haldane-type equation. The phenol oxidation coefficient obtained at different intial phenol concentrations ranged from 1.9 to 2.6 mol O2 · mol-1 phenol. A mathematical model based on a simplified version of the catalytic mechanism of the enzyme phenol 2-monooxygenase was developed to predict transient phenol concentrations and oxygen requirements by phenol-acclimated activated sludge in batch reactors under different initial phenol concentrations. The proposed model not only adequately represents the experimental results of the present paper, but also those reported by other authors. Particular cases of the proposed model are discussed.Keywords: oxygen uptake rate; phenol; biodegradation; biokinetic model; respirometry

Stoichiometry and kinetic of the aerobic oxidation of phenolic compounds by activated sludge

The aerobic degradation of phenol (PH), catechol (CA), resorcinol (RE), pyrogallol (PY), and hydroquinone (HY) by phenol-acclimated activated sludge was investigated. A Haldane-type dependence of the respiration rate on PH, RE, and HY was observed; CA and PY exhibited a biphasic respiration pattern. According to the initial biodegradation rate, tested compounds were ordered as follows: CA>PH>>PYRE>HY. Also, they exhibited the following degree of toxicity to their own degradation: PY>>CARE>>PH>HY. Oxidation coefficients for PH, PY, RE, and HY were constant as a function of the consecutive additions of the compound. Conversely, an increase of YO/S from 1 to 1.5molO2molCA−1 was observed during repeated additions of CA. The role of some enzymes involved in the aerobic degradation pathways of the tested compounds is discussed and related to the obtained results.

Biodegradation of high phenol containing synthetic wastewater by an aerobic fixed bed reactor

The continuous aerobic degradation of phenol, mixed with readily degradable synthetic wastewater was studied over a period of 400 days at 25 ± 5 °C temperature in a fixed bed biofilm reactor using ‘Liapor’ clay beads as packing material. The phenol concentration added to the reactor ranged from 0.19 to 5.17 g/l and was achieved by a gradual increase of phenol in wastewater, thus adapting the microbial flora to high contaminant concentrations. A maximal removal rate of 2.92 g phenol/(l d) at a hydraulic retention time (HRT) of 0.95 days and a total organic loading rate (OLR) of 15.3 g COD/(l d) with a phenol concentration of 4.9 g/l was observed. However, this was not a stable rate at such high phenol loading. At the end of reactor operation on day 405, the phenol removal rate was 2.3 g/(l d) at a influent phenol concentration of 4.9 g/l. There were no phenol intermediates present in the reactor, as evident from corresponding COD, phenol removal and the absence of fatty acids. Omission of organic nitrogen compounds or of urea in influent feed was not favourable for optimal phenol removal. The phenol degradation profile that was studied in shake flasks indicated that the presence of a acetate which represent as an intermediate of phenol degradation retarded the phenol degradation. The highest phenol degradation rate observed in batch assays was 3.54 g/(l d).

Effect of Co‐Substrates on Aerobic Phenol Degradation by Acclimatized and Non‐acclimatized Enrichment Cultures

AbstractAerobic degradation of 7 mmol/L phenol in the presence of alternative carbon sources (7 mmol/L glucose or acetate or 1–2 mmol/L 2‐chlorophenol) was investigated using non‐acclimatized and acclimatized sewage sludges and enrichment cultures. The substrates represented an intermediate of phenol degradation (acetate), an independent substrate (glucose) or a “precursor‐substrate” of phenol degradation (2‐chlorophenol). Bacteria from sewage sludge, not pre‐adapted to phenol (2 mmol/L), rapidly respired acetate and glucose in the presence of phenol, whereas phenol was only bioconverted to any unknown aromatic metabolite after 24 h. In the presence of phenol and 2‐chlorophenol, no removal of both substances was observed when using the unacclimatized sludge. Sludge that was acclimatized to the degradation of phenol showed an initial preference for easily degradable co‐substrates such as glucose or acetate with only a slow concomitant respiration of phenol. Respiration of phenol increased rapidly after the co‐substrates were depleted. The highest phenol degradation rates were 51.6 mmol/L d, when phenol was the sole carbon substrate. Vice versa, phenol was preferentially respired in the presence of a less easily degradable co‐substrate such as 2‐chlorophenol at a rate of around 7 mmol/L d. Further studies with an enrichment culture that was obtained after 7 successive transfers of phenol‐adapted sludge into mineral medium with phenol as the only carbon source indicated that the acetate and glucose‐degrading capabilities were diminished or almost completely lost. In these enrichment cultures, phenol degradation was not affected by the presence of glucose, but glucose was not degraded. In contrary, the presence of acetate slightly slowed down the phenol degradation rate of the enrichment culture. Growth of the microorganisms apparently occurred at the expense of phenol and acetate respiration. The result of this work may be of practical importance in determining the feeding strategy, which is the key factor for most biological wastewater treatment systems. When acetate was present together with phenol in a wastewater, the phenol degradation rates were influenced by acetate, since acetate was an intermediate of phenol degradation. Glucose as an “independent substrate” was apparently degraded by other bacteria via acetate, and in this way it also influenced the phenol degradation rates. Glucose‐degrading bacteria could be “washed out” from the acclimatized sludge during several transfers into mineral medium with phenol as the sole carbon source. If later on, glucose was added again, it remained undegraded and did not influence phenol degradation. 2‐Chlorophenol degradation also requires other bacteria than phenol degraders.

Isolation and characterization of phenol-degrading yeasts from an oil refinery wastewater in Brazil

This study investigated the aerobic degradation of phenol by yeast strains isolated from an oil refinery wastewater from the Northeast of Brazil. The samples displayed low fungal diversity, as only yeast colonies were detected on Sabouraud dextrose agar containing chloramphenicol 0.05% (w/v). Among the isolates, three yeast strains were selected to be evaluated for their potential for degrading high phenol concentrations. These species were identified through morphological and biochemical characteristics as Candida tropicalis, C. rugosa, and Pichia membranaefaciens. Although the strains were able to degrade the phenol concentration present in the wastewater, which was 7 mg l(-1), only C. tropicalis was capable of growing at high concentrations of phenol such as 500 mg l(-1 )and 1,000 mg l(-1) in a mineral medium containing this pollutant as the only carbon source. C. rugosa and P. membranaefaciens were inhibited in the presence of 500 mg l(-1) of phenol. However, a longer incubation time was needed for C. tropicalis strain to degrade 1,000 mg l(-1) of phenol compared to the time required to degrade 500 mg l(-1). Moreover, the strain released a significant amount of polysaccharide biosurfactant in the medium probably to minimize the toxic effect of the high phenol concentration. When challenged with 1,500 and 2,000 mg l(-1 )of phenol, C. tropicalis was unable to grow at the tested conditions. The results indicate that this strain of C. tropicalis can be considered both a good phenol-degrader and biosurfactant-producer. Application of this strain might be useful in bioremediation activities or treatment of phenol-polluted wastewater.

Continuous aerobic phenol degradation by defined mixed immobilized culture in packed bed reactors

A defined mixed culture of Pseudomonas putida, Commamonas testosteroni and Candida tropicalis was immobilized by adsorption on polyurethane foam, cocoa-fibers, expanded slate and sintered glass. Packed bed reactors were used for long-term continuous phenol biodegradations. Loading experiments were done to study the impact of the following parameters: (1) hydraulic retention time, (2) dissolved oxygen concentration, and (3) elimination of the oxygen limitation. After the acclimation period (approximately 10 d), the loading test with the individual packings showed the following maximum degradation rates: sintered glass 34, polyurethane foam 12, expanded slate 11.5, and cocoa-fibers 7.7 kg m(-3) d(-1). All these values were reached at a removal efficiency >99 % and with oxygen in excess. Under these conditions, the pH of the diluted unbuffered medium in the reactor effluent was 3.2-4.0 and no incompletely oxidized metabolic intermediates were found. The free cell concentration in the effluent increased after the phenol overloading time period.

Rhodococcus phenolicus sp. nov., a novel bioprocessor isolated actinomycete with the ability to degrade chlorobenzene, dichlorobenzene and phenol as sole carbon sources

The aerobic degradation of phenol, chlorobenzene and dichlorobenzene as a sole carbon source has been observed in bacterial Gram-positive strain G2P T isolated from a wastewater bioprocessor. Cells display branching mycelia fragmenting into rod and coccoid elements when grown on TSA. Aerial hyphae formation occurs when grown on phenol and chlorinated aromatics as the sole carbon source. Growth was observed at up to 0.75% phenol as a sole carbon source, indicating a strong tolerance for the compound. The 16S rRNA gene sequence shares the greatest similarity with members of the Rhodococcus genus, with the closest shared nucleotide identity of 98% with the aromatic toxin degrading bacteria Rhodococcus zopfii DSM 44108 T. Neighbor-joining and parsimony analysis of Corynebacterineae 16S rRNA gene sequences consistently places strain G2P T in a clade shared with R. zopfii within the Rhodococcus rhodochrous subclade. Based on a unique polyphasic profile involving phenotypic, ribosomal DNA sequence analysis, DNA–DNA hybridization, mol% DNA G+C content and fatty acid composition, G2P T is proposed to represent a previously uncharacterized, novel species in the genus Rhodococcus. The name Rhodococcus phenolicus is proposed for the isolate with the type strain G2P T (=DSM 44812) (=NRL B-24343).

Biodegradation of Phenol: Mechanisms and Applications

Phenol, or hydroxybenzene, is both a synthetically and naturally produced aromatic compound. Microorganisms capable of degrading phenol are common and include both aerobes and anaerobes. Many aerobic phenol-degrading microorganisms have been isolated and the pathways for the aerobic degradation of phenol are now firmly established. The first steps include oxygenation of phenol by phenol hydroxylase enzymes to form catechol, followed by ring cleavage adjacent to or in between the two hydroxyl groups of catechol. Phenol hydroxylases ranging from simple flavoprotein monooxygenases to multicomponent hydroxylases, as well as the genes coding for these enzymes, have been described for a number of aerobic phenol-degrading microorganisms. Phenol can also be degraded in the absence of oxygen. Our knowledge of this process is less advanced than that of the aerobic process, and only a few anaerobic phenol-degrading bacteria have been isolated to date. Convincing evidence from both pure culture studies with the denitrifying organism Thauera aromatica K172 and with two Clostridium species, as well as from mixed culture studies, indicates that the first step in anaerobic phenol degradation is carboxylation in the para-position to form 4-hydroxybenzoate. Following para-carboxylation, thioesterification of 4-hydroxybenzoate to co-enzyme A allows subsequent ring reduction, hydration, and fission. Para-carboxylation appears to be involved in the anaerobic degradation of a number of aromatic compounds. Numerous practical applications exist for microbial phenol degradation. These include the exploitation of indigenous anaerobic phenol-degrading bacteria in the in situ bioremediation of creosote-contaminated subsurface environments, and the use of phenol as a co-substrate for indigenous aerobic phenol-degrading bacteria to enhance in situ biodegradation of chlorinated solvents.

Variation in stable carbon isotope fractionation during aerobic degradation of phenol and benzoate by contaminant degrading bacteria

Variation in the natural abundance stable carbon isotope composition of respired CO 2 and biomass has been measured for two types of aerobic bacteria found in contaminated land sites. Pseudomonas putida strain NCIMB 10015 was cultured on phenol and benzoate and Rhodococcus sp. I 1 was cultured on phenol. Results indicate that aerobic isotope fractionations of differing magnitudes occur during aerobic biodegradation of these substrates with an isotopic depletion in the CO 2 (Δ 13C phenol-CO2) as much as 3.7‰ and 5.6‰ for Pseudomonas putida and Rhodococcus sp. I 1 respectively. This observation has significant implications for the use of a stable isotope mass balance approach in monitoring degradation processes that rely on indigenous bacterial populations. The effects of the metabolic pathway utilised in degradation and inter-species variation on the magnitude of isotope fractionation are discussed. Possible explanations for the observed isotope fractionation include differences in the metabolic pathways utilised by the organisms and differences in specific growth rates and physiology.

Isolation and characterization of phenol-degrading denitrifying bacteria.

Phenol is a man-made as well as a naturally occurring aromatic compound and an important intermediate in the biodegradation of natural and industrial aromatic compounds. Whereas many microorganisms that are capable of aerobic phenol degradation have been isolated, only a few phenol-degrading anaerobic organisms have been described to date. In this study, three novel nitrate-reducing microorganisms that are capable of using phenol as a sole source of carbon were isolated and characterized. Phenol-degrading denitrifying pure cultures were obtained by enrichment culture from anaerobic sediments obtained from three different geographic locations, the East River in New York, N.Y., a Florida orange grove, and a rain forest in Costa Rica. The three strains were shown to be different from each other based on physiologic and metabolic properties. Even though analysis of membrane fatty acids did not result in identification of the organisms, the fatty acid profiles were found to be similar to those of Azoarcus species. Sequence analysis of 16S ribosomal DNA also indicated that the phenol-degrading isolates were closely related to members of the genus Azoarcus. The results of this study add three new members to the genus Azoarcus, which previously comprised only nitrogen-fixing species associated with plant roots and denitrifying toluene degraders.

Simulation of oxygen concentration profile in calcium alginate gel beads entrapping microbes during biological phenol degradation

Numerical simulation was carried out to elucidate the effects of gel-bead diameter, initial cell density, initial phenol concentration, and oxygen concentration in the bulk liquid on the change in the oxygen concentration profile in a gel bead during biological aerobic phenol degradation with activated sludge entrapped in the calcium alginate gel bead. The results of simulation show that oxygen-limiting occurs in the central parts of the gel beads under most conditions examined in this study. After phenol degradation starts, the oxygen concentration in the beads is found to be immediately reduced and slightly increased instantly, and then slowly decreased linearly to a minimum value as the degradation proceeds. Finally, the oxygen concentration rapidly recovers to the same level of the bulk liquid. For small-sized gel beads, at low initial cell densities no oxygen-limiting is found to occur even in the core of the gel beads due to the small mass transfer resistance for oxygen diffusion, low oxygen consumption rate, or both. The critical gel-bead diameter for freedom from oxygen depletion is about 1 mm. At high phenol concentrations, oxygen may penetrate to the central parts of the gel beads as a result of reduction of the oxygen consumption rate by substrate inhibition. The effectiveness factors for small-sized cell-loaded gel beads were found to be unity at relatively high phenol concentrations in the bulk liquid and to decrease with increasing cell density. Further, it is found that at higher initial cell densities or for large-sized gel beads a high cell density layer will be formed around the surface of the gel beads during phenol degradation periods.

A new bioreactor with a semi-fixed packing: investigation of degradation of phenol

A new bioreactor using a semi-fixed packing of frames with “sacks” made of a fabric of “Raschell” type stretched on them is proposed. The construction provides not only a large surface area of the biofilm carrier per unit volume of the apparatus, but also the possibility for an easy removal of the biomass after reaching a certain thickness of the biofilm increasing the gas velocity. Aerobic degradation of phenol in the new bioreactor, using microorganisms of the strain Pseudomonas putida, was studied. The experiments are carried out using water containing 0.7 g/l of phenol at a temperature of 27–30 °C. Different specific surface area of the packing (within 176 and 387 m2/m3) are studied. Degradation rates from 60 to 140 mg/(1 h) are attained. A retardation of the process at the end is observed, probably due to inhibition effect. This rate is 5–6 times higher than the rate observed when using free cells. At air velocity of 0.03–0.035 m/s (related to the total cross section of the bioreactor) the vibrations of the packing material lead to destruction and removal of the old biomass.

Biodegradation of Xenobiotics in a fixed bed reactor

AbstractThe specific example of an at‐source biological wastewater treatment has been elaborated and developed. A two‐phase fixed‐bed bioreactor system was tested for its efficiency using phenol as a test substance and then, the degradation of sodium anthraquinone‐2‐sulfonate (SAS) was studied and performed.The continuous aerobic degradation of phenol or SAS as the sole source of carbon was investigated. For SAS degradation with an immobilized mixed bacterial culture, industrial activated sludge was used as inoculum, while for phenol a domestic activated sludge from a municipal wastewater treatment plant and activated Actizym‐sludge prepared with Actizym® powder were applied. In our laboratory fixed bed reactor system the adapted mixed culture was able to degrade up to 0.17 g L−1 h−1 of SAS and 0.28 g L−1 h−1 of phenol at dilution rates of 0.149 h−1 and 0.059 h−1, respectively. The removal efficiency of our two‐phase continuous bioreactor system was about 99% of primary and 90% of ultimate SAS degradation.In the adapted bacterial culture degrading SAS, pure bacterial strains were isolated and identified; three were gram negative and six were gram positive.