Paul R. Bienkowski

Pseudomonas putida B2: a tod-lux bioluminescent reporter for toluene and trichloroethylene co-metabolism.

A tod-lux transcriptional fusion bioluminescent reporter strain, Pseudomonas putida B2, was developed to permit on-line analysis of trichloroethylene (TCE) transformation by toluene dioxygenase (todC1C2BA) in Pseudomonas putida F1. Strain B2 was exposed to toluene in growing and resting cell bioluminescence assays. The growing cells showed a direct correlation between bioluminescence and toluene concentration, while resting cells showed reproducible bioluminescence with repeated toluene exposures. In addition, P. putida B2 was encapsulated in alginate beads and used in a packed bed flow-through differential volume reactor. The TCE feed into the differential volume reactor was constant at 20 mg L−1 and toluene was pulsed in square-wave perturbations at 10 mg L−1. The system showed a direct correlation between the expression of the tod operon (as monitored by light output) and the co-metabolism of TCE. Approximately 20% of the TCE and 50% of the toluene was removed at a flow rate of 0.4 ml min−1. This approach allowed the on-line monitoring of tod gene expression and its relation to TCE biotransformation.

Solubility of the antibiotic penicillin V in supercritical CO2

Abstract The solubility of the antibiotic penicillin V in C0 2 has been measured at temperature from 314.85 K to 334.85 K and pressures form 7.987 MPa to 28.02 MPa using a dynamic flow apparatus. The equilibrium mole fraction in this region ranged from 6.2 × 10 −5 to 5.7 × 10 −4 , which 9is about 10 6 to 10 8 times the ideal gas solubility. The vapor pressure of penicillin V, which has not been reported in the literature, was estimated from the solubility data so the latter could be correlated. Other unknown physical properties of penicillin V were estimated using group contribution methods. The experimental solubility data were correlated using a compressed gas model with the Peng-Robinson equation-of-state and a theoretical model based on Kirkwood-Buff solution theory. Solubility results are compared for this system and other complex biological molecules taken from the literature.

Biodegradation of chlorinated aliphatic hydrocarbon mixtures in a single-pass packed-bed reactor

Aliphatic chlorinated compounds, such as trichloroethylene (TCE) and tetrachloroethylene (PCE), are major contaminants of ground water. A single-pass packed-bed bioreactor was utilized to study the biodegradation of organic waste mixtures consisting of PCE, TCE, and other short-chain chlorinated organics. The bioreactor consisted of two 1960-mL glass columns joined in a series. One column was packed with sand containing a microbial consortia enriched from a contaminated site. The other column provided a reservoir for oxygen and a carbon source of methane/propane that was recirculated through the reactor. Sampling was accomplished by both direct headspace and liquid effluent concentration analyses. The reactor was operated in a single-pass mode. Greater than 99% degradation of trichloroethylene, approaching drinking water standards, was observed when the bioreactor residence time ranged from 1.9 to 3.2 d. Typically, when the reactor was pulse-fed with methane, propane, and air, 1 mol of TCE was degraded/110 mol of substrate utilized. Perturbation studies were performed to characterize reactor behavior. The systems degradation behavior was affected by providing different carbon sources, a pulse feeding regime, supplementing microbial biomass, and by altering flow rates.

Small angle neutron scattering studies of dilute supercritical solutions. I. Pure supercritical neon

Small angle neutron scattering (SANS) results are presented for states of neon close to the critical isochore and supercritical temperatures between 47 and 60 K. The net intensities were converted to absolute units of differential cross section per unit sample volume by comparison with precalibrated secondary standards. The values of the differential cross section at zero scattering vector are in excellent agreement with values calculated from an equation of state. Evidence of three‐body forces in these data are compared to that present in Krypton data reported earlier. It is concluded that small angle experiments of the sort reported here should be repeated for the heavier noble gases, for which effects from three‐body forces are anticipated to be larger.

Dynamic response of naphthalene biodegradation in a continuous flow soil slurry reactor

Periodic perturbations were used to evaluate the system stability and robustness of naphthalene biodegradation in a continuous flow stirred tank reactor (CSTR) containing a soil slurry. The experimental design involved perturbing the test system using a sinusoidal input either of naphthalene or non-naphthalene organic carbon at different frequencies during steady state operation of the reactors. The response of the test system was determined by using time series off-gas analysis for naphthalene liquid phase concentration and degradation, total viable cell counts, and gene probe analysis of naphthalene degradative genotype, and by batch mineralization assays.Naphthalene biodegradation rates were very high throughout the experimental run (95 to >99% removed) resulting in very low or undetectable levels of naphthalene in the off-gas and reactor effluent. Attempts to reduce the rate of naphthalene biotransformation by either reducing the reactor temperature from 20°C to 10°C or the dissolved oxygen level (>1 mg/L) were unsuccessful. Significant naphthalene biodegradation was observed at 4°C. While variable, the microbial community as measured by population densities was not significantly affected by temperature changes. In terms of naphthalene biotransformation, the system was able to adapt readily to all perturbations in the reactor.

Production of ethanol from starch by co-immobilized Zymomonas mobilis -- Glucoamylase in a fluidized-bed reactor

The production of ethanol from starch was studied in a fluidized-bed reactor (FBR) using co-immobilizedZymomonas mobilis and glucoamylase. The FBR was a glass column of 2.54 cm in diameter and 120 cm in length. TheZ. mobilis and glucoamylase were co-immobilized within small uniform beads (1.2-2.5 mm diameter) of κ-carrageenan. The substrate for ethanol production was a soluble starch. Light steep water was used as the complex nutrient source. The experiments were performed at 35κC and pH range of 4.0-5.5. The substrate concentrations ranged from 40 to 185 g/L, and the feed rates from 10 to 37 mL/min. Under relaxed sterility conditions, the FBR was successfully operated for a period of 22 d, during which no contamination or structural failure of the biocatalyst beads was observed. Volumetric productivity as high as 38 g ethanol/(Lh), which was 74% of the maximum expected value, was obtained. Typical ethanol volumetric productivity was in the range of 15-20 g/(Lh). The average yield was 0.49 g ethanol/g substrate consumed, which was 90% of the theoretical yield. Very low levels of glucose were observed in the reactor, indicating that starch hydrolysis was the rate-limiting step.

Modeling infinite dilution activity coefficients of environmental pollutants in water using conformal solution theory

The fate of organic pollutants in the environment and in wastewater treatment processes is commonly modeled using a Henry`s law constant approach. By definition, Henry`s law constant is the product of a compound`s vapor pressure and infinite dilution activity coefficient. For many organic compounds in water solution, the infinite dilution activity coefficients are very large and are not adequately modeled by conventional methods such as UNIFAC. In this work, infinite dilution activity coefficients were determined for phenol, pyridine, aniline, p-toluidine, and o-toluidine in water by differential ebulliometry. An equation rigorously derived from conformal solution theory and van der Waals one-fluid mixing rules was used to model the temperature dependency of the infinite dilution activity coefficients. No corrections other than the introduction of two adjustable parameters were incorporated into the model to account for the strong interactions between molecules. Relationships derived from corresponding states theory were used to relate molecular parameters for size and energy interaction to the critical properties. Arithmetic mean combining rules and geometric mean combining rules were used to calculate size and interaction parameters, respectively.

Development of a Differential Volume Reactor System for Soil Biodegradation Studies

A bench scale experimental system was developed for the analysis of polycyclic aromatic hydrocarbon (PAH) degradation by mixed microbial cultures in PAH contaminated Manufactured Gas Plant (MGP) soils and on sand. The reactor system was chosen in order to provide a fundamental protocol capable for evaluating the performance of specific mixed microbial cultures on specific soil systems by elucidating the important system variables and their interactions. The reactor design and peripherals are described. A plug flow differential volume reactor (DVR) was used in order to remove performance effects related to reactor type, as opposed to system structure. This reactor system could be well represented mathematically. Methods were developed for on-line quantitative determination of PAH liquid phase concentrations. The mathematical models and experimental data are presented for the biodegradation of naphthalene on artificial and MGP soils.

Development of a systems analysis approach for resolving the structure of biodegrading soil systems.

An experimental and mathematical method is developed for the microbial systems analysis of polyaromatic hydrocarbon (PAH)-degrading mixed cultures in PAH-contaminated “town gas” soil systems. Frequency response is the primary experimental and data analysis tool used to probe the structure of these complicated systems. The objective is to provide a fundamental protocol for evaluating the performance of specific mixed microbial cultures on specific soil systems by elucidating the salient system variables and their interactions. Two well-described reactor systems, a constant volume stirred tank reactor (CSTR) and a plug flow differential volume reactor, are used in order to remove performance effects that are related to reactor type as opposed to system structure. These two reactor systems are well-defined systems that can be described mathematically and represent the two extremes of one potentially important system variable, macroscopic mass transfer. The experimental and mathematical structure of the protocol is described, experimental data is presented, and data analysis is demonstrated for the stripping, sorption, and biodegradation of napththalene.

A generalized quartic equation of state

A generalized quartic equation of state has been developed for nonpolar and polar fluids. The quartic equation of state needs only four fluid properties to predict physical and thermodynamic properties over a wide range of temperatures and pressures. These four properties are critical volume, critical temperature, acentric factor and dipole moment. The new equation of state models the repulsive forces between molecules correctly, unlike the cubic equations of state. One of the four roots obtained by solving the equation of state is always negative, making it possible to identify the remaining three roots like the roots of a cubic equation. The quartic equation of state shows superior performance compared with the Peng-Robinson equation of state.