A systematic selection of the non-aqueous phase in a bacterial two liquid phase bioreactor treating α-pinene

A systematic evaluation of the selection criteria of non-aqueous phases in two liquid phase bioreactors (TLPBs), also named two-phase partitioning bioreactors (TPPBs), was carried out using the biodegradation of α-pinene by Pseudomonas fluorescens NCIMB 11671 as a model process. A preliminary solvent screening was thus carried out among the most common non-aqueous phases reported in literature for volatile organic contaminants biodegradation in TLPBs: silicon oil, paraffin oil, hexadecane, diethyl sebacate, dibutyl-phtalate, FC 40, 1,1,1,3,5,5,5-heptamethyltrisiloxane (HMS), and 2,2,4,4,6,8,8-heptamethylnonane (HMN). FC 40, silicone oil, HMS, and HMN were first selected based on its biocompatibility, resistance to microbial attack, and α-pinene mass transport characteristics. FC 40, HMS, HMN, and silicone oil at 10% (v/v) enhanced α-pinene mass transport from the gas to the liquid phase by a factor of 3.8, 14.8, 11.4, and 8.6, respectively, compared to a single-phase aqueous system. FC 40 and HMN were finally compared for their ability to enhance α-pinene biodegradation in a mechanically agitated bioreactor. The use of FC 40 or HMN (both at 10% v/v) sustained non-steady state removal efficiencies (RE) and elimination capacities (EC) approximately 7 and 12 times higher than those achieved in the system without an organic phase, respectively. In addition, preliminary results showed that P fluorescens could uptake and mineralize α-pinene directly from the non aqueous phase.     

The Effect of Temperature on the Bioventing of Soil Contaminated with Toluene and Decane

The effect of temperature on evaporation and biodegradation rates during soil bioventing (SBV) was studied for a mixture of toluene and decane in bench-scale soil columns at a continuous air flow and consecutively at two different flow rates. The effect of temperature on SBV was monitored by GC headspace analysis of contaminant, CO₂ and O₂ concentrations in the soil gas over time. Separation of evaporation and biodegradation processes into three different phases based on their rates was used together with Q₁₀ and E₁₀ (values that give the factor by which biodegradation and evaporation rates increase when the temperature is raised by 10 degrees) to compare quantitatively the removal kinetics at 10 and 20°C. Adsorption of toluene and decane onto soil (a phase partitioning process) at 20 and 10°C was described with linear Freundlich isotherms. A temperature decrease from 20 to 10°C resulted in an increase of soil-air partitioning coefficients by a factor of 1.8 and of 2.1 for toluene and decane, respectively. The mean Q₁₀ value for the biodegradation of toluene was found to be 2.2 for a temperature rise from 10 to 20°C. A toluene content in the soil gas above 75% of the saturation concentration inhibited biodegradation at both temperatures. The SBV efficiency was dependent on temperature with respect to remediation time. SBV at 20°C resulted in a 99.8% and a 98.7% reduction of toluene and decane initial concentrations, respectively. To reach similar results at 10°C, about 1.6 times as much time and 1.4 times as much air were required; however, at both temperatures the total amounts of biodegraded hydrocarbons were approximately the same. The evaporation-to-biodegradation ratios at 20°C were 82.5:17.5 for toluene and 16:84 for decane, whereas at 10 °C they were 71:29 and 2:98, respectively. A comparison of Q₁₀ values showed that, except during the initial phase of SBV, only a modest decrease in biodegradation rates should be expected after a decrease in temperature from 20 to 10°C. Flow rate reduction had a significant impact on the toluene evaporation rate at a higher temperature, whereas for decane this rate was only slightly affected by temperature. In contrast to decane, the ratio between toluene vapor pressures at 20 and 10°C may be used to predict the removal of toluene by evaporation during the above-mentioned phases of SBV, when evaporation is important.     

A dynamical model for phase transitions of lipids induced by calcium ions

Lipid membranes exist essentially in two different phases. A phase transition can be triggered off either by changing the temperature or, isothermally, by varying an external factor such as ionic concentration, pH, organic solvents, etc. Since the isothermal transition may be induced at physiological temperature, it may play an important regulatory role in diverse cellular functions. Based on the Landau-Ginsburg theory, the thermotropic transitions of lipids has been described by a number of models. In the present work a dynamical model for an isothermal phase transition of phospholipids induced by ionic binding is proposed. The properties of the model show that by ionic binding, phospholipids may form spatial heterogeneous distributions of lipids in fluid and crystalline phases. This heterogeneity possibly being the cause of membrane instabilities, which favour enhanced vesicle fusions observed in the presence of Ca2+.     

Estimation of Biodegradation Potential of Xenobiotic Organic Chemicals

A method is described to estimate the biodegradation potential of soluble, insoluble, and unknown organic chemicals. The method consists of two stages: (i) generation of a microbial inoculum in a bench scale semicontinuous activated sludge system during which microorganisms are acclimated to test material and the removal of dissolved organic carbon is monitored and (ii) biodegradability testing (CO₂ evolution) in a defined minimal medium containing the test material as the sole carbon and energy source and a dilute bacterial inoculum obtained from the supernatant of homogenized activated sludge generated in the semicontinuous activated sludge system. Removal and biodegradation are measured using nonspecific methods, at initial concentrations of 5 to 10 mg of dissolved organic carbon per liter. Biodegradability data are accurately described by a nonlinear computer model which allows the rate and extent of biodegradation for different compounds to be compared and statistically examined. The evaluation of data generated in the combined removability-biodegradability system allows the biodegradation potential of a variety of xenobiotic organic chemicals to be estimated.     

Evaluation of the interaction between biodegradation and sorption of phenanthrene in soil-slurry systems

This work develops and utilizes a non-steady-state model for evaluating the interactions between sorption and biodegradation of hydrophobic organic compounds in soil-slurry systems. The model includes sorption/desorption of a target compound, its utilization by microorganisms as a primary substrate existing in the dissolved phase, and/or the sorbed phase in biomass and soil, oxygen transfer, and oxygen utilization as an electron acceptor. Biodegradation tests with phenanthrene were conducted in liquid and soil-slurry systems. The soil-slurry tests were performed with very different mass transfer rates: fast mass transfer in a flask test at 150 rpm, and slow mass transfer in a roller-bottle test at 2 rpm. The results of liquid tests indicate that biodegradation of the soil-soluble organic fraction did not significantly enhance the biodegradation rate. In the slurry tests, phenanthrene was degraded more rapidly than in liquid tests, but at a similar rate in both slurry systems. Modeling analyses with several hypotheses indicate that a model without biodegradation of compound sorbed to the soil was not able to account for the rapid degradation of phenanthrene, particularly in the roller-bottle slurry test. The model with sorbed-phase biodegradation and the same biokinetic parameters, but unique mass transfer coefficients, simulated the experimental data in both slurry tests most successfully. Reduced mass transfer resistance to bacteria attached to the soil is the most likely phenomenon accounting for rapid sorbed-phase biodegradation.     

Mass transfer and hydrocarbon biodegradation of aged soil in slurry phase

Addition of toluene into slurry phase laboratory microcosm is proposed in order to increase desorption rate of hydrocarbons and as an alternative to improve bioavailability of hydrocarbon in aged soils. Our studies showed that toluene has a positive effect on desorption of total petroleum hydrocarbons (TPH). Addition of 14,000 mg toluene/kg of soil, in highly polluted soil, increased the consumption rate of hydrocarbons three times in comparison to control without solvent. In 30 days the initial TPH concentration in soil, 292,000 mg/kg, diminished 45%. Although toluene was able to dissolve complex organic compounds such as asphaltene fraction, it probably yielded a highly toxic toluene-hydrocarbons phase. The inhibitory effect of toluene-TPH was also studied. A substrate inhibition model was used: the k(m) and k(i) constants were 57 and 490 mg TPH/L liquid phase, respectively. Experimental data were well described when the proposed model included sequential desorption and biodegradation phenomena. Damk?hler number evaluation showed that rate of mass transfer was the limiting step in overall biodegradation in nonsolvent control. When high concentration of toluene was added, then bioreaction was the limiting step, but inhibitory effect should be considered. However, toluene addition at low concentrations facilitates the biodegradation of aromatic compounds.     

Thermal decomposition kinetics of wheat straw treated by Phanerochaete chrysosporium

Combining biological pretreatment with thermal processing may offer an alternative strategy for efficient conversion of lignocellulosic biomass into fuels and chemicals. The thermal decomposition kinetics of biologically pretreated wheat straw by Phanerochaete chrysosporium was investigated in this study using thermogravimetry (TG) - deconvoluted thermogravimetry (DTG) techniques and the Friedman method. This study revealed that biological pretreatment reduced the thermal degradation temperature of the biomass significantly. Relying on the thermal behavior of the biologically pretreated wheat straw, we proposed two biomass degradation phases during the biological degradation of wheat straw. The first phase of biodegradation (within 10 days of biological pretreatment) improved the efficiency of pyrolysis by reducing the temperature demand. In the second phase (after 10 days), although the efficiency of pyrolysis displayed the similar trend as the first phase, it showed a significant increase in activation energy demand. This process is greatly influenced by the residual lignin and cellulose ratios in the biomass. These experimental results will be useful in developing a biological pretreatment based thermochemical conversion process for lignocellulosic biomass.     

Biodegradability continuum and biodegradation kinetics of natural organic matter described by the beta distribution

We followed a long-term (up to 503 days) microbial mineralization of dissolved organic carbon (DOC) from lake water in a bioassay and described the kinetics of biodegradation with a new model based on a reactivity continuum approach. The biodegradability of DOC was expressed as the probability of biodegradation, which was assumed to follow a beta distribution. We compared the performance of our beta model to five earlier models: the simplest first order kinetic model, two G models, the power model and the gamma model. The simplest first order kinetic model described the decreasing microbial mineralization of DOC poorly (r ² = 0.73), but the other models explained the observed kinetics of biodegradation well (r ² > 0.95). When we assessed the extrapolation power of models beyond the length of the bioassay by reducing the amount of data, the predictive power of the G models was poor. Instead, the beta model predicted the biodegradation kinetics consistently and correctly based on even only three observations in time. The beta model provided also long-term predictions (up to 5,000 years) along the observed long-term mineralization trajectory of organic carbon in sediments. Additionally, the beta model formulated the biodegradability continuum of DOC, which was skewed towards low biodegradability. During the bioassay, the skew towards low biodegradability increased as the most biodegradable parts of DOC were consumed. The beta model describes the biodegradability continuum quantitatively and can predict biodegradation in a realistic manner, thus, improving our understanding about the biodegradability and the role of natural organic matter in the environment.     

A Process-Based Mathematical Model on Methane Production with Emission Indices for Control

In this paper, a process-based mathematical model is developed for the production of methane through biodegradation. It is a three-dimensional model given by ordinary differential equations. The results of the analysis of the model are interpreted through three emission indices, which are introduced for the first time. The estimation of either one or all of them can interpret the feasibility of the equilibrium and the long-term emission tendency of methane. The vulnerability of the methane production process with respect to soil temperature effects in methanogenic phase has been discussed and a feasible condition within a specified temperature range has defined for the nonvulnerability of the methane production process and also it has shown that under the same condition, zero-emission process of methane will be nonvulnerable with respect to the soil temperature effects in methanogenic phase. Lastly, condition for zero emission of methane is also obtained and it is interpreted through the emission indices.     

Microbial succession during a composting process as evaluated by denaturing gradient gel electrophoresis analysis

Microbial succession during a laboratory-scale composting process of garbage was analysed by denaturing gradient gel electrophoresis (DGGE) combined with measurement of physicochemical parameters such as temperature, pH, organic acids, total dissolved organic carbon and water-soluble humic substance. From the temperature changes, a rapid increase from 25 to 58 degrees C and then a gradual decrease, four phases were recognized in the process as follows; mesophilic (S), thermophilic (T), cooling (C) and maturing (M). The polymerase chain reaction-amplified 16S rDNA fragments with universal (907R) and eubacterial (341F with GC clamp) primers were subjected to DGGE analysis. Consequently, the DGGE band pattern changed during the composting process. The direct sequences from DGGE bands were related to those of known genera in the DNA database. The microbial succession determined by DGGE was summarized as follows: in the S phase some fermenting bacteria, such as lactobacillus, were present with the existing organic acids; in the T phase thermophilic bacillus appeared and, after the C phase, bacterial populations were more complex than in previous phases and the phylogenetic positions of those populations were relatively distant from strains so far in the DNA database. Thus, the DGGE method is useful to reveal microbial succession during a composting process.     

Two-liquid-phase bioreactors for enhanced degradation of hydrophobic/toxic compounds

Two-liquid-phase culture systems involve the addition of a water-immiscible, biocompatible and non-biodegradable solvent to enhance a biocatalytic process. Two-liquid-phase bioreactors have been used since the mid-seventies for the microbial and enzymatic bioconversion of hydrophobic/toxic substrates into products of commercial interest. The increasing popularity of bioremediation technologies suggests a new area of application for this type of bioreactor. The toxicity and the limited bioavailability of many pollutants are important obstacles that must first be overcome in order to improve biodegradation processes. Two-liquid-phase bioreactors have the potential to resolve both limitations of biotreatment technologies by the enhancement of the mass-transfer rate of compounds with low bioavailability, and by the controlled delivery of apolar toxic compounds. This technology can also be useful in accelerating the enrichment of microorganisms degrading problematic pollutants. In this paper, we discuss the application of two-liquid-phase bioreactors to enhance the biodegradation of toxic/poorly bioavailable contaminants. Important microbial mechanisms involved in this type of system are described. Uptake of the substrates can be achieved by microorganisms freely dispersed in the aqueous phase and/or bound at the interface between the aqueous and the immiscible phases. Production of surface-active compounds and adhesion abilities are microbial features involved in the process. General guidelines for the design of two-liquid-phase bioreactors for biodegradation purposes are presented. Solvent selection should be established on specific criteria, which depend on the characteristics of target compound(s) and the microorganism(s) implicated in the biodegradation process. The central importance of maximizing the interfacial surface area is highlighted. The potential of this approach as an alternative to current biotreatment technologies is also discussed.     

Influence of a nonaqueous phase liquid (NAPL) on biodegradation of phenanthrene

A series of batch reactor experiments was carried out to examine the effect of a nonaqueous phase liquid (NAPL) on the biodegradation of a hydrophobic solute. A mathematical program model that describes physical processes of solute solubilization and partitioning between the NAPL and aqueous phases as well as microbial degradation and oxygen utilization was used to analyze the test data. The model calculates the cumulative changes in concentration of substrate, cell mass, carbon dioxide, and dissolved oxygen as a function of time. The equations incorporate the effects of solute solubilization, partitioning, biodegradation, as well as oxygen availability. Hexadecane was used as the model NAPL and was not biodegraded in the timeframe of the experiments performed. The model solute was the polyaromatic hydrocarbon, phenanthrene. In agreement with several previous studies, experimental measurements showed that hexadecane increased rates of mineralization of 15 mg phenanthrene when present at low mass but decreased rates at high mass. Model results suggest that partitioning of the phenanthrene into the hexadecane phase limits bioavailability at high NAPL mass. Further the model suggests that mineralization rates were higher with the low NAPL mass because aqueous phenanthrene concentrations were higher in those treatments from ca. 20 to 40 h than in other treatments. Finally, experiments showed that the presence of hexadecane, at all masses tested, resulted in a lower cell yield, effectively increasing the amount of CO₂ produced during the experiment. Model results suggest that this is due to changes in phenanthrene metabolism that are induced by the presence of the hexadecane phase. Model studies aimed at increasing rates of biodegradation by modifying operating conditions are described along with practical approaches to implementing these modifications.     

Modeling of crude oil biodegradation using two phase partitioning bioreactor

In this work, crude oil biodegradation has been optimized in a solid‐liquid two phase partitioning bioreactor (TPPB) by applying a response surface methodology based d ‐optimal design. Three key factors including phase ratio, substrate concentration in solid organic phase, and sodium chloride concentration in aqueous phase were taken as independent variables, while the efficiency of the biodegradation of absorbed crude oil on polymer beads was considered to be the dependent variable. Commercial thermoplastic polyurethane (Desmopan®) was used as the solid phase in the TPPB. The designed experiments were carried out batch wise using a mixed acclimatized bacterial consortium. Optimum combinations of key factors with a statistically significant cubic model were used to maximize biodegradation in the TPPB. The validity of the model was successfully verified by the good agreement between the model‐predicted and experimental results. When applying the optimum parameters, gas chromatography‐mass spectrometry showed a significant reduction in n‐alkanes and low molecular weight polycyclic aromatic hydrocarbons. This consequently highlights the practical applicability of TPPB in crude oil biodegradation. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:797–805, 2014     

Toluene biodegradation rates in unsaturated soil systems versus liquid batches and their relevance to field conditions

Contaminant biodegradation in unsaturated soils may reduce the risks of vapor intrusion. However, the reported rates show large variability and are often derived from slurry experiments that are not representative of unsaturated conditions. Here, different laboratory setups are used to derive the biodegradation capacity of an unsaturated soil layer through which gaseous toluene migrates from the water table upwards. Experiments in static unsaturated soil microcosms at 6–30 % water-filled porosity (WFP) and unsaturated soil columns at 9, 14, and 27 % WFP were compared with liquid batches containing the same culture of Alicycliphilus denitrificans. The biodegradation rates for the liquid batches were orders of magnitude lower than for the other setups. Hence, liquid batches do not necessarily reflect optimal conditions for bacteria; either oxygen or toluene mass transfer at the cell scale or the absence of soil–water–air interfaces seemed to be limiting bacterial activity. For the column setup, the rates were limited by mass supply. The microcosm results could be described by apparent first-order biodegradation constants that increased with WFP or through a numerical model that included biodegradation as a first-order process taking place in the liquid phase only. The model liquid phase first-order rates varied between 6.25 and 20 h^?1 and were not related to the water content. Substrate availability was the primary factor limiting bioactivity, with evidence for physiological stress at the lowest water-filled porosity. The presented approach is useful to derive liquid phase biodegradation rates from experimental data and to include biodegradation in vapor intrusion models.     

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.     

A novel high-pressure liquid-liquid extraction process for downstream processing in biotechnology: extraction of cardiac glycosides

This investigation examines phase equilibrium phenomena that can be used to create two water-like solvents for liquid-liquid extraction in downstream processing in biotechnology: a completely miscible, binary liquid mixture of water and a hydrophilic organic solvent (e. g., an alcohol) reveals a liquid phase split, when it is pressurized with a "near-critical" gas (i.e., a substance which at ambient conditions is a gas, near its critical temperature). This phase split results in two hydrophilic liquid phases. Making use of this phenomenon in process development first requires research on the phase split phenomenon and, second, research on the feasibility of biomolecule extraction and separation. In this study, basic fluid phase equilibrium phenomena are briefly described. Then, experimental results are reported for the partitioning of small amounts of cardiac glycosides (digitoxin and digoxin) on coexisting liquid phases in the high-pressure, three-phase, vapor-liquid-liquid equilibrium of the ternary system of "near critical" CO(2) + water + 1-propanol, at 313 K and 333 K. Finally, a process for extraction and separation of the aforementioned glycosides by means of the high-pressure phase equilibrium phenomenon is discussed.     

Biodegradation kinetics of naphthalene in nonaqueous phase liquid-water mixed batch systems: comparison of model predictions and experimental results

A model is formulated to describe dissolution of naphthalene from an insoluble nonaqueous phase liquid (NAPL) and its subsequent biodegradation in the aqueous phase in completely mixed batch reactors. The physicochemical processes of equilibrium partitioning and mass transfer of naphthalene between the NAPL and aqueous phases were incorporated into the model. Biodegradation kinetics were described by Monod's microbial growth kinetic model, modified to account for the inhibitory effects of 1,2-naphthoquinone formed during naphthalene degradation under certain conditions. System parameters and biokinetic coefficients pertinent to the NAPL-water systems were determined either by direct measurement or from nonlinear regression of the naphthalene mineralization profiles obtained from batch reactor tests with two-component NAPLs comprised of naphthalene and heptamethylnonane. The NAPLs contained substantial mass of naphthalene, and naphthalene biodegradation kinetics were evaluated over the time required for near complete depletion of naphthalene from the NAPL. Model predictions of naphthalene mineralization time profiles compared favorably to the general trends observed in the data obtained from laboratory experiments with the two-component NAPL, as well as with two coal tars obtained from the subsurface at contaminated sites and composed of many different PAHs (polycyclic aromatic hydrocarbon compounds). The effects of varying the NAPL mass and the naphthalene mole fractions in the NAPL are discussed. It was observed that the time to achieve a given percent removal of naphthalene does not change significantly with the initial mass of naphthalene in a fixed volume of the NAPL. Significant changes in the mineralization profiles are observed when the volume (and mass) of NAPL in the system is changed.     

Dual substrate biodegradation of a nonionic surfactant and pentachlorophenol by Sphingomonas chlorophenolica RA2

The simultaneous biodegradation of the nonionic surfactant Tween 20 (Tw20) and pentachlorophenol (PCP) by Sphingomonas chlorophenolica sp. Strain RA2 (RA2) was measured. As a sole substrate, Tw20 biodegradation was best described by the Contois kinetic model. During concurrent biodegradation of Tw20 and PCP, the biodegradation rates of Tw20 were not significantly affected by 50 or 100 mg/L PCP, but were significantly inhibited by 500 mg/L PCP. Decreases in cell yield in the presence of PCP suggest that PCP was acting as an uncoupler. Cultures were pre-grown on PCP or Tw20 before degradation of PCP to evaluate enzyme induction effects, and long lags before PCP biodegradation after growth on Tw20 occurred. Although biokinetic models could accurately describe some of the data sets of RA2 growth and Tw20 and PCP degradation, finding a single set of kinetic parameters that predicted all dual substrate tests was not achieved. The complicating factors to modeling PCP and Tw20 interactions are described and may be more widely applicable to the biodegradation of toxic organic compounds in the presence of a biodegradable surfactant.     

Model based evaluation of a contaminant plume development under aerobic and anaerobic conditions in 2D bench-scale tank experiments

The influence of transverse mixing on competitive aerobic and anaerobic biodegradation of a hydrocarbon plume was investigated using a two-dimensional, bench-scale flow-through laboratory tank experiment. In the first part of the experiment aerobic degradation of increasing toluene concentrations was carried out by the aerobic strain Pseudomonas putida F1. Successively, ethylbenzene (injected as a mixture of unlabeled and fully deuterium-labeled isotopologues) substituted toluene; nitrate was added as additional electron acceptor and the anaerobic denitrifying strain Aromatoleum aromaticum EbN1 was inoculated to study competitive degradation under aerobic / anaerobic conditions. The spatial distribution of anaerobic degradation was resolved by measurements of compound-specific stable isotope fractionation induced by the anaerobic strain as well as compound concentrations. A fully transient numerical reactive transport model was employed and calibrated using measurements of electron donors, acceptors and isotope fractionation. The aerobic phases of the experiment were successfully reproduced using a double Monod kinetic growth model and assuming an initial homogeneous distribution of P. putida F1. Investigation of the competitive degradation phase shows that the observed isotopic pattern cannot be explained by transverse mixing driven biodegradation only, but also depends on the inoculation process of the anaerobic strain. Transient concentrations of electron acceptors and donors are well reproduced by the model, showing its ability to simulate transient competitive biodegradation.