Biodegradation of phenol and m-cresol in a batch and fed batch operated internal loop airlift bioreactor by indigenous mixed microbial culture predominantly Pseudomonas sp

An internal loop airlift reactor (ILALR) is developed and studied for biodegradation of phenol/m-cresol as single and dual substrate systems under batch and fed batch operation using an indigenous mixed microbial strain, predominantly Pseudomonas sp. The results showed that the culture could degrade phenol/m-cresol completely at a maximum concentration of 600mgl(-1) and 400mgl(-1), respectively. Batch ILALR study has revealed that phenol has been preferentially degraded by the microbial culture rather than m-cresol probably owing to the toxic effect of the later. Sum kinetic model evaluated the interaction between the phenol/m-cresol in dual substrate system, which resulted in a high coefficient of determination (R(2)) value >0.98). The fed batch results showed that the strain was able to degrade phenol/m-cresol with maximum individual concentrations 600mgl(-1) each in 26h and 37h, respectively. Moreover for fed batch operation, degradation rates increased with increase in feed concentration without any lag in the degradation profile.     

Apparent zero-order kinetics of phenol biodegradation by substrate-inhibited microbes at low substrate concentrations

The reaction kinetics for phenol biodegradation at low substrate concentrations can be estimated based on the analysis of changes in the dissolved oxygen concentration in the bulk liquid during biodegradation. The measured oxygen concentration changes with an interesting behavior as biodegradation proceeds. The oxygen concentration in the bulk liquid decreases rapidly in the early stages of degradation and subsequently decreases linearly and then rapidly recovers to the initial saturated level. Taking into account the oxygen transfer rate between gas and liquid phases and oxygen consumption rate by microbes, the change in the dissolved oxygen concentration can be simulated with an unsteady state mass balance equation and three kinetic models for the rate of phenol metabolism: a substrate-inhibited model; a zero-order model; and a combined model. In the combined model, it is assumed that, at phenol concentrations above 10 mg/L, the degradation rate is expressed by a substrate-inhibited model; whereas at concentrations below 10 mg/L the zero-order model is applied. It was found that the characteristics of the change in the dissolved oxygen concentration, especially the rapid increase at the end of degradation, can only be described by the combined kinetic model. This result suggests that conventional Haldane-type kinetics would be unsuitable for estimating the phenol consumption rate at low phenol concentrations, in particular, at concentrations less than 10 mg/L. (c) 1996 John Wiley & Sons, Inc.     

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.     

Degradation of phenol by aerobic granules and isolated yeast Candida tropicalis

Aerobic granules effectively degrade phenol at high concentrations. This work cultivated aerobic granules that can degrade phenol at a constant rate of 49 mg-phenol/g x VSS/h up to 1,000 mg/L of phenol. Fluorescent staining and confocal laser scanning microscopy (CLSM) tests demonstrated that an active biomass was accumulated at the granule outer layer. A strain with maximum ability to degrade phenol and a high tolerance to phenol toxicity isolated from the granules was identified as Candida tropicalis via 18S rRNA sequencing. This strain degrades phenol at a maximum rate of 390 mg-phenol/g x VSS/h at pH 6 and 30 degrees C, whereas inhibitory effects existed at concentrations >1,000 mg/L. The Haldane kinetic model elucidates the growth and phenol biodegradation kinetics of the C. tropicalis. The fluorescence in situ hybridization (FISH) and CLSM test suggested that the Candida strain was primarily distributed throughout the surface layer of granule; hence, achieving a near constant reaction rate over a wide range of phenol concentration. The mass transfer barrier provided by granule matrix did not determine the reaction rates for the present phenol-degrading granule.     

Mathematical model for a three-phase fluidized bed biofilm reactor in wastewater treatment

A mathematical model for a three phase fluidized bed bioreactor (TFBBR) was proposed to describe oxygen utilization rate, biomass concentration and the removal efficiency of Chemical Oxygen Demand (COD) in wastewater treatment. The model consisted of the biofilm model to describe the oxygen uptake rate and the hydraulic model to describe flow characteristics to cause the oxygen distribution in the reactor. The biofilm model represented the oxygen uptake rate by individual bioparticle and the hydrodynamics of fluids presented an axial dispersion flow with back mixing in the liquid phase and a plug flow in the gas phase. The difference of settling velocity along the column height due to the distributions of size and number of bioparticle was considered. The proposed model was able to predict the biomass concentration and the dissolved oxygen concentration along the column height. The removal efficiency of COD was calculated based on the oxygen consumption amounts that were obtained from the dissolved oxygen concentration. The predicted oxygen concentration by the proposed model agreed reasonably well with experimental measurement in a TFBBR. The effects of various operating parameters on the oxygen concentration were simulated based on the proposed model. The media size and media density affected the performance of a TFBBR. The dissolved oxygen concentration was significantly affected by the superficial liquid velocity but the removal efficiency of COD was significantly affected by the superficial gas velocity. An erratum to this article can be found online at .     

Kinetics of phenol degradation using Pseudomonas putida MTCC 1194

Pseudomonas putida (MTCC 1194) has been used to degrade phenol in water in the concentration range 100–1000?ppm. The inhibition effects of phenol as substrate have become predominant above the concentration of 500?ppm (5.31?mmoles/dm³). The optimum temperature and initial pH required for maximum phenol biodegradation were 30?°C and 7.00 respectively. From the degradation data the activation energy (E _a ) was found to be equal to 13.8?kcal/g mole substrate reacted. The most suitable inoculum age and volume for highest phenol degradation were 12?hrs and 7% v/v respectively. Surfactants had negligible effect on phenol biodegradation process for this microorganism. Monod model has been used to interpret the free cell data on phenol biodegradation. The kinetic parameters have been estimated upto initial concentration of 5.31?mmoles/dm³. μ _max and K _S gradually increased with higher concentration of phenol. However, beyond the phenol concentration of 5.31?mmoles/dm³, the inhibition became prominant. The μ _max has been to be a strong function of initial phenol concentration. The simulated and the experimental phenol degradation profiles have good correspondence with each other.     

Mathematical model for evaluation of mass transfer limitations in phenol biodegradation by immobilized Pseudomonas putida

A mathematical model is proposed to analyze the mass transfer limitations in phenol biodegradation using Pseudomonas putida immobilized in calcium alginate. The model takes into account internal and external mass transfer limitations, substrate inhibition kinetics and the dependence of the effective diffusivity of phenol in alginate gel on cell concentration. The model is validated with the experimental data from batch fermentation. The effect of various operating conditions such as initial phenol concentration, initial cell loading, alginate gel loading on the biodegradation of phenol is experimentally demonstrated. Phenol degradation time is found to decrease initially and reach stationary value with increase in cell loading as well as gel loading. The model predicts these trends reasonably well and shows the presence of external mass transfer limitations. A new concept of effectiveness factor is introduced to analyze the overall performance of batch fermentation.     

Macro-kinetic investigation on phenol uptake from air by biofiltration: Influence of superficial gas flow rate and inlet pollutant concentration

The macro-kinetic behavior of phenol removal from a synthetic exhaust gas was investigated theoretically as well as experimentally by means of two identical continuously operating laboratory-scale biological filter bed columns. A mixture of peat and glass beads was used as filter material. After sterilization it was inoculated with a pure strain of Pseudomonas putida, as employed in previous experimental studies. To determine the influence of the superficial gas flow rate on biofilter performance and to evaluate the phenol concentration profiles along the column, two series of continuous tests were carried out varying either the inlet phenol concentration, up to 1650 mg . m(-3), or the superficial gas flow rate, from 30 to 460 m(3) . m(-2) . h(-1). The elimination capacity of the biofilter is proved by a maximum volumetric phenol removal rate of 0.73 kg . m(-3) . h(-1). The experimental results are consistent with a biofilm model incorporating first-order substrate elimination kinetics. The model may be considered a useful tool in scaling-up a biofiltration system. Furthermore, the deodorization capacity of the biofilter was investigated, at inlet phenol concentrations up to 280 mg . m(-3) and superficial gas flow rates ranging from 30 to 92 m(3) . m(-2) . h(-1). The deodorization of the gas was achieved at a maximum inlet phenol concentration of about 255 mg . m(-3), operating at a superficial gas flow rate of 30 m(3) . m(-2) . h(-1). (c) 1996 John Wiley & Sons, Inc.     

Treatment of phenolics containing synthetic wastewater in an internal loop airlift bioreactor (ILALR) using indigenous mixed strain of Pseudomonas sp. under continuous mode of operation

The scope of this study is to evaluate the performance of internal loop airlift bioreactor (ILALR) in treating synthetic wastewater containing phenol and m-cresol, in single and multi component systems. The microbe utilized in the process was an indigenous mixed strain of Pseudomonas sp. isolated from a wastewater treatment plant. The reactor was operated at both lower and higher hydraulic retention times (HRTs) i.e., 4.1 and 8.3 h, respectively, by providing an inlet feed flow rate of 5 and 10 mL/min. Shock loading experiments were also performed up to a maximum concentration of 800 mg/L for phenol at 8.3 h HRT and 500 mg/L for m-cresol at 4.1 h HRT. The study showed complete degradation of both phenol and m-cresol, when they were degraded individually at a HRT of 8.3 h. Experiments with both phenol and m-cresol present as mixtures were performed based on the 2² full factorial design of experiments.     

The role of laccase and peroxidase of Lentinus (Panus) tigrinus fungus in biodegradation of high phenol concentrations in liquid medium

The possibility of the usage of Lentinus tigrinus fungus strain VKM F-3616D for biodegradation of high (up to 5%) phenol concentrations in liquid medium and the involvement of laccase and peroxidase in this process have been studied. L. tigrinus fungus was demonstrated to effectively digrade phenol with easy biomass separation from the liquid. Decrease in phenol concentration was accompanied by increased secretion level and laccase activity at the preliminary stages of biodegradation, while that of peroxidase was at the latest stages of biodegradation. These enzyme secretions in distinct ratios and consequences are necessary for effective phenol biodegradation. An effective approach for phenol concentration decrease in the waste water of smoking shops in meat-processing factories using L. tigrinus fungus was described.     

Biodegradation of phenol and 4-chlorophenol by the yeast <Emphasis Type="Italic">Candida tropicalis</Emphasis>

Biodegradation of phenol and 4-chlorophenol (4-cp) using a pure culture of Candida tropicalis was studied. The results showed that C. tropicalis could degrade 2,000 mg l⁻¹ phenol alone and 350 mg l⁻¹ 4-cp alone within 66 and 55 h, respectively. The capacity of the strain to degrade phenol was obviously higher than that to degrade 4-cp. In the dual-substrate system, 4-cp intensely inhibited phenol biodegradation. Phenol beyond 800 mg l⁻¹ could not be degraded in the presence of 350 mg l⁻¹ 4-cp. Comparatively, low-concentration phenol from 100 to 600 mg l⁻¹ supplied a sole carbon and energy source for C. tropicalis in the initial phase of biodegradation and accelerated the assimilation of 4-cp, which resulted in the fact that 4-cp biodegradation velocity was higher than that without phenol. And the capacity of C. tropicalis to degrade 4-cp was increased up to 420 mg l⁻¹ with the presence of 100–160 mg l⁻¹ phenol. In addition, the intrinsic kinetics of cell growth and substrate degradation were investigated with phenol and 4-cp as single and mixed substrates in batch cultures. The results illustrated that the models proposed adequately described the dynamic behaviors of biodegradation by C. tropicalis.     

生物降解对黑碳及土壤上苯酚脱附行为的影响

农业土壤和黑碳(BC)两种不同的吸附剂吸附苯酚平衡后分离,每组一部分不做处理,另一部分通过加入无酚灭菌溶液脱附平衡后分离,制备得到在不同吸附位点上吸附有苯酚的两类不同类型的4种吸附苯酚的吸附剂,研究了在不同Pseudomonasputida ATCC 11172菌密度条件下吸附在这4种吸附剂上的苯酚的脱附行为.结果表明,土壤及BC对苯酚的吸附均呈现明显的非线性,可用Freundlich模型描述.吸附态的苯酚能否被微生物利用取决于微生物及吸附剂的性质,BC具有发达的微孔结构,微孔小于假单胞菌细胞尺寸,导致假单胞菌无法直接利用吸附在BC上的苯酚;土壤基本无微孔结构,微生物较易与吸附的苯酚发生表面接触,直接利用吸附态苯酚.BC和土壤上的吸附态苯酚的脱附行为能用三元位点模型很好地描述,模型计算结果表明BC上的苯酚脱附主要受慢速脱附和极慢速脱附控制,微生物降解速率受脱附控制,降解可加速BC上的慢速脱附和极慢速脱附;土壤上的苯酚脱附主要受快速脱附控制,微生物降解不受脱附速率限制,对土壤上的脱附行为基本无影响.     

Phenol degradation by Ralstonia eutropha: colorimetric determination of 2-hydroxymuconate semialdehyde accumulation to control feed strategy in fed-batch fermentations.

Phenol biodegradation by Ralstonia eutropha was modeled in different culture modes to assess phenol feeding in biotechnological depollution processes. The substrate-inhibited growth of R. eutropha was described by the Haldane equation with a Ks of 2 mg/L, a Ki of 350 mg/L and a mumax of 0.41 h(-1). Furthermore, growth in several culture modes was characterized by the appearance of a yellow color, due to production of a metabolic intermediate of the phenol catabolic pathway, 2-hydroxymuconic semialdehyde (2-hms) which was directly correlated to the growth rate and/or the phenol-degradation rate, because these two parameters are coupled (as seen by the constant growth yield of 0.68 g biomass/g phenol whatever the phenol concentration). This correlation between color appearance and metabolic activity was used to develop a control procedure for optimal phenol degradation. A mass-balance equation modeling approach combined with a filtering step using an extended Kalman filter enabled state variables of the biological system to be simulated. A PI controller, using the estimation of the phenol concentration provided by the modeling step, was then built to maintain the phenol concentration at a constant set-point of 0.1 g/L which corresponded to a constant specific growth rate of 0.3 h(-1), close to the maximal specific growth value of the strain. This monitoring strategy, validated for two fed-batch cultures, could lead, in self-cycling fermentation systems, to a productivity of more than 19 kg of phenol consumed/m(3)/d which is the highest value reported to date in the literature. This system of monitoring metabolic activity also protected the bacterial culture against toxicity problems due to the transient accumulation of phenol.     

The role of laccase and peroxidase of <Emphasis Type="Italic">Lentinus (Panus) tigrinus</Emphasis> fungus in biodegradation of high phenol concentrations in liquid medium

The possibility of the usage of Lentinus tigrinus fungus strain VKM F-3616D for biodegradation of high (up to 5%) phenol concentrations in liquid medium and the involvement of laccase and peroxidase in this process have been studied. L. tigrinus fungus was demonstrated to effectively degrade phenol with easy biomass deletion from the liquid. Decrease in phenol concentration was accompanied by increased secretion level and laccase activity at the preliminary stages of biodegradation, while that of peroxidase was at the latest stages of biodegradation. These enzyme secretions in distinct ratios and consequences are necessary for effective phenol biodegradation. An effective approach for phenol concentration decrease in the waste water of smoking shops in meat-processing factories using L. tigrinus fungus was described.     

A novel membrane bioreactor for detoxifying industrial wastewater: I. Biodegradation of phenol in a synthetically concocted wastewater

A novel process has been used to biodegrade phenol present in an acidic (1 M HCI) and salty (5% w/w NaCl) synthetically bioreactor, in which the phenol present in the wastewater is separated from the inorganic components by means of a silicone rubber membrane. Transfer of the phenol from the wastewater and into a biological growth medium allows biodegradation to proceed under controlled conditions which are unaffected by the hostile inorganic composition of the wastewater. At a wastewater flow rate of 18 mL h(-1) (contact time 6 h), 98.5% of the phenol present in the wastewater at an inlet concentration of 1000 mg ( (-1) ) was degraded; at a contact time of 1.9 h, 65% of the phenol was degraded. Phenol degradation was accompanied by growth of a biofilm on the membrane tubes and by conversion of approximately 80% of the carbon entering the system to CO(2) carbon. Analysis of the transport of phenol across the membrane revealed that the major resistance to mass transfer arose in the diffusion of phenol across the silicone rubber membrane. A mathematical model was used to describe the transfer of phenol across the membrane and the subsequent diffusion and reaction of phenol in the biofilm attached to the membrane tube. This analysis showed that (a) the attached biofilm significantly lowers the mass transfer driving force for phenol across the membrane, and (b) oxygen concentration limits the phenol degradation rate in the biofilm. These conclusions from the model are consistent with the experimental results. (c) 1993 Wiley & Sons, Inc.     

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.     

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.     

Design of experiments in the biodegradation of phenol using immobilized Pseudomonas pictorum (NICM-2077) on activated carbon

Pseudomonas pictorum (NICM-2077) immobilized on various matrices are used to protect the microbes from confronting shock loads of concentrated phenol. The cells were immobilized in activated carbon and were used in biodegradation of phenol. Biodegradation of phenol using immobilized Activated Carbon matrix played an important role in reducing the toxicity of phenol. The degradation was carried and using the Box-Behnken model and analysis of variance have been applied to the experimental degradation studies. Response surface method with three levels of phenol concentration (0.200, 0.400, 0.600 g/l), activated carbon (0.5, 1.0, 1.5 g/l) and pH (7, 8, 9) were used in the identification of significant effects and interactions in the biodegradation studies. Phenol removal rate increases especially when the degradation medium was supplemented with utilizable carbon sources. A first-order polynomial regression model, which was used at first for analysis of the experiment had a significant lack of fit. Therefore, linear and quadratic terms were incorporated into the regression model through variable selection procedures. Effect of incubation phenol concentration, activated carbon and pH were significant. The predicted values using Box-Behnken model was found to be in close agreement with the experimental values, as indicated by the correlation coefficient values of 0.9463.     

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.     

Local overall volumetric gas-liquid mass transfer coefficients in gas-liquid-solid reversed flow jet loop bioreactor with a non-Newtonian fluid

The local overall volumetric gas-liquid mass transfer coefficients at the specified point in a gas-liquid-solid three-phase reversed flow jet loop bioreactor (JLB) with a non-Newtonian fluid was experimentally investigated by a transient gassing-in method. The effects of liquid jet flow rate, gas jet flow rate, particle density, particle diameter, solids loading, nozzle diameter and CMC concentration on the local overall volumetric gas-liquid mass transfer coefficient (K(L)a) profiles were discussed. It was observed that local overall K(L)a profiles in the three-phase reversed flow JLB with non-Newtonian fluid increased with the increase of gas jet flow rate, liquid jet flow rate, particle density and particle diameter, but decreased with the increase of the nozzle diameter and CMC concentration. The presence of solids at a low concentration increased the local overall K(L)a profiles, and the optimum of solids loading for a maximum profile of the local overall K(L)a was found to be 0.18x10(-3)m(3) corresponding to a solids volume fraction, varepsilon(S)=2.8%.