Batch phenol degradation by candida tropicalis and its fusant

Phenol degradation by Candida tropicalis and its fusant, which is produced using protoplast fusion as a selective technique, is evaluated under batch and high concentration conditions. The respirometric data show that oxygen uptake activities of both yeast strains peak at pH 7.0 and 32 degrees C, but the fusant is more active than the control strain. Although the data show that both yeast strains are capable of sustaining discernible degradation in the presence of phenol inhibition, however, the C. tropicalis fusant is capable of attaining better phenol degradation than the control strain and it is less susceptible to phenol inhibition. Under the conditions tested, C. tropicalis is completely inhibited at phenol concentrations >/=3,300 mg/L, whereas for the C. tropicalis fusant complete inhibition is absent until phenol concentrations are >/=4, 000 mg/L. The observed cell yields of both yeast strains are virtually identical and remain fairly constant at approximately 0.5 mg MLVSS/mg C6H5OH (MLVSS: mixed liquor volatile suspended solids). Copyright 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 60: 391-395, 1998.     

Single-culture aerobic granules with <Emphasis Type="Italic">Acinetobacter calcoaceticus</Emphasis>

Aerobic granules are cultivated by a single bacterial strain, Acinetobacter calcoaceticus, in a sequencing batch reactor (SBR). This strain presents as a good phenol reducer and an efficient auto coagulator in the presence of phenol, mediated by heat-sensitive adhesins proteins. Stable 2.3-mm granules were formed in the SBR following a 7-week cultivation. These granules exhibit excellent settling attributes and degrade phenol efficiently at concentrations of 250–2,000 mg l⁻¹. The corresponding phenol degradation rate reached 993.6 mg phenol g⁻¹ volatile suspended solids (VSS) day⁻¹ at 250 mg l⁻¹ phenol and 519.3 mg phenol g⁻¹ VSS day⁻¹ at 2,000 mg l⁻¹ phenol concentration. Meanwhile, free A. calcoaceticus cells were fully inhibited at phenol >1,500 mg l⁻¹. Denaturing gradient gel electrophoresis fingerprint profile demonstrated no genetic modification in the strain during aerobic granulation. The present single-strain granules showed long-term structural stability and performed high phenol degrading capacity and high phenol tolerance. The confocal laser scanning microscopic test revealed that live A. calcoaceticus cells principally distributed at 200–250 μm beneath the outer surface, with an extracellular polymeric substance layer covering them to defend phenol toxicity. Autoaggregation assay tests demonstrated the possibly significant role of secreted proteins on the formation of single-culture A. calcoaceticus granules.     

Optimal microbial adaptation routes for the rapid degradation of high concentration of phenol

Pseudomonas fluorescence KNU417 was able to degrade up to 700 mg/L of phenol in 65 h but could not degrade 1,000 mg/L of phenol. Phenol degradation rate was noticeably enhanced by pre-adaptation. In addition, the cell was able to degrade up to 1,300 mg/L of phenol by pre-adapting to 700 mg/L of phenol. Repeated adaptations to the same concentration of phenol showed negligible increase in degradation rate. Also, relatively low concentration of phenol (100–700 mg/L) required only one pre-adaptation while high concentration (1,000 mg/L) did two consecutive stepwise pre-adaptations for rapid degradation. Optimal adaptation routes were suggested for the fast phenol degradation. For example, 1,000 mg/L of phenol was degraded as fast as in 48 h when the cell was pre-adapted to 100 and 300 mg/L of phenol sequentially. The mechanism of adaptation was explained in terms of catechol 1,2-dioxygenase induction, related to aromatic ring cleavage.     

Aggregation of immobilized activated sludge cells into aerobically grown microbial granules for the aerobic biodegradation of phenol

AIMS: The aim of this study is to evaluate the utility of aerobically grown microbial granules for the biological treatment of phenol-containing wastewater. METHODS AND RESULTS: A column-type sequential aerobic sludge blanket reactor was inoculated with activated sludge and fed with phenol as the sole carbon source, at a rate of 1.5 g phenol l-1 d-1. Aerobically grown microbial granules first appeared on day 9 of reactor operation and quickly grew to displace the seed flocs as the dominant form of biomass in the reactor. These granules were compact and regular in appearance, and consisted of bacterial rods and cocci and fungi embedded in an extracellular polymeric matrix. The granules had a mean size of 0.52 mm, a sludge volume index of 40 ml g-1 and a specific oxygen utilization rate of 110 mg oxygen g VSS-1 h-1 (VSS stands for volatile suspended solids). Specific phenol degradation rates increased with phenol concentration from 0 to 500 mg phenol l-1, peaked at 1.4 g phenol g VSS-1 d-1, and declined with further increases in phenol concentration as substrate inhibition effects became important. CONCLUSIONS: Aerobically grown microbial granules were successfully cultivated in a reactor maintained at a loading rate of 1.5 g phenol l-1 d-1. The granules exhibited a high tolerance towards phenol. Significant rates of phenol degradation were attained at phenol concentrations as high as 2 g l-1. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to demonstrate the ability of aerobically grown microbial granules to degrade phenol. These granules appear to represent an excellent immobilization strategy for microorganisms to biologically remove phenol and other toxic chemicals in high-strength industrial wastewaters.     

Growth and phenol activity of Pseudomonas aeruginosa strain 101/1 in batch cultures

Strain 101/1, isolated from petroleum wastewater sediment was classified as Pseudomonas aeruginosa. In wild type condition the strain tolerated phenol in concentration 1,000 mg/L under aerobic conditions and 800 mg/L under denitrifying conditions. As a result of adaptation to phenol the resistance of the strain to the compound increased to 1,600 and 1,400 mg/L, respectively. Maximum phenol activity under aerobic and denitrifying conditions was 350 and 65 mg/L x day-1, respectively. Under denitrifying conditions a reduction in incubation temperature from 30 degrees C to 20 degrees C resulted in two-fold drop in phenol activity of the adapted strain and reduction in tolerance to phenol by 400 mg/L.     

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.     

Anaerobic biodegradation of phenol by <Emphasis Type="Italic">Candida albicans</Emphasis> PDY-07 in the presence of 4-chlorophenol

Biodegradation of phenol and 4-chlorophenol (4-cp) using pure culture of Candida albicans PDY-07 under anaerobic condition was studied. The results showed that the strain could completely degrade up to 1,800 mg/l phenol within 68 h. The capacity of the strain to degrade phenol was higher than that to degrade 4-cp. In the dual-substrate system, 4-cp intensely inhibited phenol biodegradation. Comparatively, low-concentration phenol from 25 to 150 mg/l supplied a carbon and energy source for Candida albicans PDY-07 in the early phase of biodegradation and accelerated the assimilation of 4-cp, which resulted in that 50 mg/l 4-cp was degraded within less time than that without phenol. While the biodegradation of 50 mg/l 4-cp was inhibited in the presence of 200 mg/l phenol. In addition, the intrinsic kinetics of cell growth and substrate degradation were investigated with phenol and 4-cp as single and dual substrates in batch cultures. The results demonstrated that the models adequately described the dynamic behaviors of biodegradation by Candida albicans PDY-07.     

Mutation of Candida tropicalis by irradiation with a He-Ne laser to increase its ability to degrade phenol

Candida tropicalis isolated from acclimated activated sludge was used in this study. Cell suspensions with 5 x 10(7) cells ml(-1) were irradiated by using a He-Ne laser. After mutagenesis, the irradiated cell suspension was diluted and plated on yeast extract-peptone-dextrose (YEPD) medium. Plates with approximately 20 individual colonies were selected, and all individual colonies were harvested for phenol biodegradation. The phenol biodegradation stabilities for 70 phenol biodegradation-positive mutants, mutant strains CTM 1 to 70, ranked according to their original phenol biodegradation potentials, were tested continuously during transfers. Finally, mutant strain CTM 2, which degraded 2,600 mg liter(-1) phenol within 70.5 h, was obtained on the basis of its capacity and hereditary stability for phenol biodegradation. The phenol hydroxylase gene sequences were cloned in wild and mutant strains. The results showed that four amino acids were mutated by irradiation with a laser. In order to compare the activity of phenol hydroxylase in wild and mutant strains, their genes were expressed in Escherichia coli BL21(DE3) and enzyme activities were spectrophotometrically determined. It was clear that the activity of phenol hydroxylase was promoted after irradiation with a He-Ne laser. In addition, the cell growth and intrinsic phenol biodegradation kinetics of mutant strain CTM 2 in batch cultures were also described by Haldane's kinetic equation with a wide range of initial phenol concentrations from 0 to 2,600 mg liter(-1). The specific growth and degradation rates further demonstrated that the CTM 2 mutant strain possessed a higher capacity to resist phenol toxicity than wild C. tropicalis did.     

一株耐高盐热带假丝酵母菌降解苯酚的研究

从含酚废水处理池污泥中驯化分离得到一株能以苯酚为唯一碳源的菌株FD-1。经18SrDNA和ITS序列的BLAST比对及系统发育分析,鉴定FD-1为热带假丝酵母(Candida tropicalis)的近缘种。FD-1对苯酚的降解能力较强,能够完全降解浓度为1 000mg·L-1的苯酚溶液。初步确定了FD-1在降解苯酚溶液时的最适温度为30~35℃,pH为6.0~7.0,并且通过探讨加入无机盐、培养基原料以及改变接种量三个因素对苯酚降解的影响,其耐受盐的浓度可达5%,对实践中应用微生物降解含酚废水具有积极的意义。     

Mass transfer limitation of sulfate in methanogenic aggregates

The role of mass transfer limitation of sulfate as a factor governing the competition between sulfate reducing and methane producing bacteria in methanogenic aggregates was theoretically evaluated by the calculation of steady-state sulfate microprofiles using a reference set of parameters obtained from the literature. The shooting method was used as a numerical technique for solving the mathematical model. The effect of the parameters on mass transport limitation was tested by varying each reference value of the parameters with a factor of 3. Sulfate limitation within granules prevailed at moderate (0.1 kg m(-3)) and low sulfate concentrations in the bulk liquid, at high maximum sulfate utilization rates (3.73 x 10(-5) kg SO(4) (2-) kg(-1) VSS S(-1) or biomass concentrations (40 KG VSS m(-3)), and in large aggregates (radius of 7.5 10(-4) m). The effective diffusion coefficient of sulfate and the affinity constant were less determinative for the penetration depth of sulfate within a granule. (c) 1994 John Wiley & Sons, Inc.     

An isolated Candida albicans TL3 capable of degrading phenol at large concentration

An isolated yeast strain was grown aerobically on phenol as a sole carbon source up to 24 mM; the rate of degradation of phenol at 30 degrees C was greater than other microorganisms at the comparable phenol concentrations. This microorganism was further identified and is designated Candida albicans TL3. The catabolic activity of C. albicans TL3 for degradation of phenol was evaluated with the K(s) and V(max) values of 1.7 +/- 0.1 mM and 0.66 +/- 0.02 micromol/min/mg of protein, respectively. With application of enzymatic, chromatographic and mass-spectrometric analyses, we confirmed that catechol and cis,cis-muconic acid were produced during the biodegradation of phenol performed by C. albicans TL3, indicating the occurrence of an ortho-fission pathway. The maximum activity of phenol hydroxylase and catechol-1,2-dioxygenase were induced when this strain grew in phenol culture media at 22 mM and 10 mM, respectively. In addition to phenol, C. albicans TL3 was effective in degrading formaldehyde, which is another major pollutant in waste water from a factory producing phenolic resin. The promising result from the bio-treatment of such factory effluent makes Candida albicans TL3 be a potentially useful strain for industrial application.     

Modeling of local dynamic behavior of phenol degradation in an internal loop airlift bioreactor by yeast Candida tropicalis

A coupled computational fluid dynamic (CFD) model, combining hydrodynamics with biochemical reactions, was developed to simulate the local transient flow patterns and the dynamic behaviors of cell growth and phenol biodegradation by yeast Candida tropicalis in an internal loop airlift reactor (ILALR). To validate this proposed model effectively, the simulated local hydrodynamic characteristics of the gas-mineral salt medium solution (gas-liquid) two-phase system, at a phenol concentration of 1,200 mg L(-1) and no presence of cells, was experimentally investigated in the ILALR using laser Doppler anemometer (LDA) measurements and conductivity probe. Furthermore, the validation of the simulated phenol biodegradation behavior by C. tropicalis at different initial concentrations of phenol and cell was also carried out in the ILALR. The time-averaged and transient results of the model simulations illustrated a satisfactory agreement with the experimental data. Finally, the local instantaneous flow and phenol biodegradation features, including gas holdup, gas velocity, liquid velocity, cell concentration, and phenol concentration inside the ILALR were successfully predicted by the proposed model.     

高效降解棉酚菌株的选育及脱毒条件的研究

From mildewed cottonseed cake and stock cultUres of mold and yeast We select more than ten strains of yeastS and molds which can degrade cotton Phenol. At last we got four strains which can degrade cotton phenol highly effeted after mutagenized by physical and chemical factors and induced by cotton Phenol. They belong to Candida tropicalis, Torulopsis candida, Aspergillus flavus and ASPergillus niger. By small and medium size fermenfations, the content of dissociated cottom Phenol all reach safe criterion (…     

Biodegradation of phenol and <Emphasis Type="Italic">m</Emphasis>-cresol by <Emphasis Type="Italic">Candida albicans</Emphasis> PDY-07 under anaerobic condition

Strain Candida albicans PDY-07 was used to study the anaerobic biodegradation of phenol and m-cresol as single and dual substrates in batch cultures. The strain had a higher potential to degrade phenol than m-cresol. The cell growth kinetics of batch cultures with various initial m-cresol concentrations was investigated, and the Haldane kinetic model adequately described the dynamic behavior of cell growth on m-cresol. When cells grew on the mixture of phenol and m-cresol, substrate interactions were observed. Phenol inhibited the utilization of m-cresol; on the other hand, m-cresol also inhibited the degradation of phenol. However, the presence of low-concentration phenol enhanced m-cresol biodegradation; 100 mg/l m-cresol could be completely degraded within a shorter period of time than m-cresol alone in the presence of 150–300 mg/l phenol. The maximum m-cresol biodegradation rate was obtained at the existence of 200 mg/l phenol. Phenol was preferably utilized by the strain as a carbon and energy source. In addition, a sum kinetics model was used to describe the cell growth behavior in binary mixture of phenol and m-cresol, and the interaction parameters were determined. The model adequately predicted the growth kinetics and the interaction between the substrates.     

Degrading high-strength phenol using aerobic granular sludge

Aerobic granules were adopted to degrade high-strength phenol wastewater in batch experiments. The acclimated granules effectively degraded phenol at a concentration of up to 5,000 mg l⁻¹ without severe inhibitory effects. The biodegradation of phenol by activated sludge was inhibited at phenol concentrations >3,000 mg l⁻¹. The granules were composed of cells embedded in a compact extracellular matrix. After acid or alkaline pretreatment, the granules continued to degrade phenol at an acceptable rate. The polymerase chain reaction-denaturing gradient gel electrophoresis technique was employed to monitor the microbial communities of the activated sludge and the aerobic granules following their being used to treat high concentrations of phenol in batch tests.     

Characteristics of rapidly formed hydrogen-producing granules and biofilms

The physicochemical and microbiological characteristics of rapidly formed hydrogen-producing granules and biofilms were evaluated in the present study. Microbial species composition was examined using the 16S rDNA-based separation and sequencing techniques, and spatial distribution and internal structure of microbial components were evaluated by examining the confocal laser scanning microscope (CLSM) images. Phylogenetic analysis indicated that a pure culture of Clostridium pasteurianum-like bacterium (98% similarity) was found in microbial community of granules and biofilms. It is postulated that containing such a species favored the rapid immobilization of hydrogen-producing culture. Manure granules and biofilms secreted 24-35 mg extracellulous proteins and 142-175 mg extracellulous polysaccharides in each gram of culture (in VSS). Such a high productivity of extracellulous polymers (ECP), a bio-glue to facilitate cell-to-cell and/or cell-to-substratum interaction, may work as the driving forces for the immobilization of C. pasteurianum. As abundant proteins were noted in the granule cores, it can be derived that rapid formation of the hydrogen-producing granules could be due to the establishment of precursor protein-rich microbial nuclei.     

Simultaneous degradation of atrazine and phenol by Pseudomonas sp. strain ADP: effects of toxicity and adaptation

The strain Pseudomonas sp. strain ADP is able to degrade atrazine as a sole nitrogen source and therefore needs a single source for both carbon and energy for growth. In addition to the typical C source for Pseudomonas, Na(2)-succinate, the strain can also grow with phenol as a carbon source. Phenol is oxidized to catechol by a multicomponent phenol hydroxylase. Catechol is degraded via the ortho pathway using catechol 1,2-dioxygenase. It was possible to stimulate the strain in order to degrade very high concentrations of phenol (1,000 mg/liter) and atrazine (150 mg/liter) simultaneously. With cyanuric acid, the major intermediate of atrazine degradation, as an N source, both the growth rate and the phenol degradation rate were similar to those measured with ammonia as an N source. With atrazine as an N source, the growth rate and the phenol degradation rate were reduced to approximately 35% of those obtained for cyanuric acid. This presents clear evidence that although the first three enzymes of the atrazine degradation pathway are constitutively present, either these enzymes or the uptake of atrazine is the bottleneck that diminishes the growth rate of Pseudomonas sp. strain ADP with atrazine as an N source. Whereas atrazine and cyanuric acid showed no significant toxic effect on the cells, phenol reduces growth and activates or induces typical membrane-adaptive responses known for the genus Pseudomonas. Therefore Pseudomonas sp. strain ADP is an ideal bacterium for the investigation of the regulatory interactions among several catabolic genes and stress response mechanisms during the simultaneous degradation of toxic phenolic compounds and a xenobiotic N source such as atrazine.     

Simultaneous Degradation of Atrazine and Phenol by Pseudomonas sp. Strain ADP: Effects of Toxicity and Adaptation

The strain Pseudomonas sp. strain ADP is able to degrade atrazine as a sole nitrogen source and therefore needs a single source for both carbon and energy for growth. In addition to the typical C source for Pseudomonas, Na₂-succinate, the strain can also grow with phenol as a carbon source. Phenol is oxidized to catechol by a multicomponent phenol hydroxylase. Catechol is degraded via the ortho pathway using catechol 1,2-dioxygenase. It was possible to stimulate the strain in order to degrade very high concentrations of phenol (1,000 mg/liter) and atrazine (150 mg/liter) simultaneously. With cyanuric acid, the major intermediate of atrazine degradation, as an N source, both the growth rate and the phenol degradation rate were similar to those measured with ammonia as an N source. With atrazine as an N source, the growth rate and the phenol degradation rate were reduced to ~35% of those obtained for cyanuric acid. This presents clear evidence that although the first three enzymes of the atrazine degradation pathway are constitutively present, either these enzymes or the uptake of atrazine is the bottleneck that diminishes the growth rate of Pseudomonas sp. strain ADP with atrazine as an N source. Whereas atrazine and cyanuric acid showed no significant toxic effect on the cells, phenol reduces growth and activates or induces typical membrane-adaptive responses known for the genus Pseudomonas. Therefore Pseudomonas sp. strain ADP is an ideal bacterium for the investigation of the regulatory interactions among several catabolic genes and stress response mechanisms during the simultaneous degradation of toxic phenolic compounds and a xenobiotic N source such as atrazine.     

Isolation and identification of Paracoccus sp. FD3 and evaluation of its formaldehyde degradation kinetics

A formaldehyde-degrading bacterium strain, FD3, was isolated from contaminated soil and identified as Paracoccus sp. based on partial 16S rRNA gene sequence analysis. In batch culture, the bacterium metabolized 5,000 and 8,000 mg/L formaldehyde completely within 16 and 18 h, respectively, at 30°C (pH 7.0) with agitation at 150 rpm. The degradation kinetics was found to follow a first-order model at all initial formaldehyde concentrations with regression values greater than 0.99. Formaldehyde degradation rates increased from 532.37 to 2283.04 mg/L/h as the initial concentration of formaldehyde was increased from 1,000 to 8,000 mg/L. The growth of strain FD3 on formaldehyde as a sole carbon and energy source was well described by the Luong model with a maximal specific growth rate of 0.1754/h, a half-saturation constant of 309.02 mg/L, and a maximum substrate concentration of 3875.53 mg/L. Due to its high tolerance and degradation capacity to formaldehyde, Paracoccus sp., FD3 is considered an excellent candidate for use in degrading formaldehyde in wastewaters.     

Oxygen diffusion and consumption in active aerobic granules of heterogeneous structure

The interior structure of aerobic granules is highly heterogeneous, hence, affecting the transport and reaction processes in the granules. The granule structure and the dissolved oxygen profiles were probed at the same granule in the current work for possible estimation of transport and kinetic parameters in the granule. With the tested granules fed by phenol or acetate as carbon source, most inflow oxygen was consumed by an active layer thickness of less than 125 μm on the granule surface. The confocal laser scanning microscopy scans also revealed a surface layer thickness of approximately 100 μm consisting of cells. The diffusivities of oxygen transport and the kinetic constant of oxygen consumption in the active layers only were evaluated. The theoretical models adopted in literature that ignored the contributions of the layered structure of aerobic granule could have overlooked the possible limitations on oxygen transport.