George P. Prpich

ENHANCED BIODEGRADATION OF PHENOL BY A MICROBIAL CONSORTIUM IN A SOLID-LIQUID TWO PHASE PARTITIONING BIOREACTOR

Two phase partitioning bioreactors (TPPBs) operate by partitioning toxic substrates to or from an aqueous, cell-containing phase by means of second immiscible phase. Uptake of toxic substrates by the second phase effectively reduces their concentration within the aqueous phase to sub-inhibitory levels, and transfer of molecules between the phases to maintain equilibrium results in the continual feeding of substrate based on the metabolic demand of the microorganisms. Conventionally, a single pure species of microorganism, and a pure organic solvent, have been used in TPPBs. The present work has demonstrated the benefits of using a mixed microbial population for the degradation of phenol in a TPPB that uses solid polymer beads (comprised of ethylene vinyl acetate, or EVA) as the second phase. Polymer modification via an increase in vinyl acetate concentration was also shown to increase phenol uptake. Microbial consortia were isolated from three biological sources and, based on an evaluation of their kinetic performance, a superior consortium was chosen that offered improved degradation when compared to a pure strain of Pseudomonas putida ATCC 11172. The new microbial consortium used within a TPPB was capable of degrading high concentrations of phenol (≈2000mgl−1), with decreased lag time (10h) and increased specific rate of phenol degradation (0.71g phenolg−1cellh). Investigation of the four-member consortium showed that it consisted of two Pseudomonas sp., and two Acinetobacter sp., and tests conducted upon the individual isolates, as well as paired organisms, confirmed the synergistic benefit of their existence within the consortium. The enhanced effects of the use of a microbial consortium now offer improved degradation of phenol, and open the possibility of the degradation of multiple toxic substrates via a polymer-mediated TPPB system.

Remediation of PAH contaminated soils: application of a solid-liquid two-phase partitioning bioreactor.

The feasibility of a two-step treatment process has been assessed at laboratory scale for the remediation of soil contaminated with a model mixture of polycyclic aromatic hydrocarbons (PAHs) (phenanthrene, pyrene, and fluoranthene). The initial step of the process involved contacting contaminated soil with thermoplastic, polymeric pellets (polyurethane). The ability of three different mobilizing agents (water, surfactant (Biosolve) and isopropyl alcohol) to enhance recovery of PAHs from soil was investigated and the results were compared to the recovery of PAHs from dry soil. The presence of isopropyl alcohol had the greatest impact on PAH recovery with approximately 80% of the original mass of PAHs in the soil being absorbed by the polymer pellets in 48 h. The second stage of the suggested treatment involved regeneration of the PAH loaded polymers via PAH biodegradation, which was carried out in a solid-liquid two-phase partitioning bioreactor. In addition to the PAH containing polymer pellets, the bioreactor contained a microbial consortium that was pre-selected for its ability to degrade the model PAHs and after a 14 d period approximately 78%, 62% and 36% of phenanthrene, pyrene, and fluoranthene, respectively, had been desorbed from the polymer and degraded. The rate of phenanthrene degradation was shown to be limited by mass transfer of phenanthrene from the polymer pellets. In case of pyrene and fluoranthene a combination of mass transfer and biodegradation rate might have been limiting.

Polymer development for enhanced delivery of phenol in a solid-liquid two-phase partitioning bioreactor.

Two‐Phase Partitioning Bioreactors (TPPBs) have traditionally been used to partition toxic concentrations of xenobiotics from a cell‐containing aqueous phase by means of an immiscible organic solvent and to deliver these substrates back to the cells on the basis of metabolic demand and the maintenance of thermodynamic equilibrium between the phases. A limitation of TPPBs, which use organic liquid solvents, is the possibility that the solvent can be bioavailable, and this has therefore limited organic liquid TPPBs to the use of pure strains of microbes. Solid polymer beads have recently been introduced as a replacement for liquid organic solvents, offering similar absorption properties but with the capability to be used with mixed microbial populations. The present work was aimed at identifying a polymer with a greater capacity for and more rapid uptake and release of phenol for use as the second phase in a mixed culture TPPB. Polarity and hydrogen bonding capabilities between polymer and phenol were considered in the screening and selection process of candidate polymers. Hytrel (a copolymer of poly(butylene terephthalate) and butylene ether glycol terephthalate) polymer beads, offered improved capacity (19 mg phenol/g polymer at a fixed initial phenol concentration of 2000 mg/L) and a greater diffusivity (1.54 × 10−7 cm2/s) when compared to the capacity and diffusivity of previously used EVA (ethylene vinyl acetate) beads (12.4 mg phenol/g polymer and 3.73 × 10−9 cm2/s, respectively). Hytrel polymer beads were then used in a TPPB for the investigation of various substrate feeding strategies (fed‐batch, bead replacement, and concentrated spikes of phenol), with rapid and complete phenol degradation shown in all cases.

Biodegradation of a phenolic mixture in a solid-liquid two-phase partitioning bioreactor.

A solid–liquid two-phase partitioning bioreactor (TPPB) in which the non-aqueous phase consisted of polymer (HYTREL) beads was used to degrade a model mixture of phenols [phenol, o-cresol, and 4-chlorophenol (4CP)] by a microbial consortium. In one set of experiments, high concentrations (850xa0mg l−1 of each of the three substrates) were reduced to sub-inhibitory levels within 45xa0min by the addition of the polymer beads, followed by inoculation and rapid (8xa0h) consumption of the total phenolics loading. In a second set of experiments, the beneficial effect of using polymer beads to launch a fermentation inhibited by high substrate concentrations was demonstrated by adding 1,300 and 2,000xa0mg l−1 total substrates (equal concentrations of each phenolic) to a pre-inoculated bioreactor. At these levels, no cell growth and no degradation were observed; however, after adding polymer beads to the systems, the ensuing reduced substrate concentrations permitted complete destruction of the target molecules, demonstrating the essential role played by the polymer sequestering phase when applied to systems facing inhibitory substrate concentrations. In addition to establishing alternative modes of TPPB operation, the present work has demonstrated the differential partitioning of phenols in a mixture between the aqueous and polymeric phases. The polymeric phase was also observed to absorb a degradation intermediate (arising from the incomplete biodegradation of 4CP), which opens the possibility of using solid–liquid TPPBs during biosynthetic transformation to sequester metabolic byproducts.

Ex situ bioremediation of phenol contaminated soil using polymer beads

Polymer beads have been used to absorb high concentrations of phenol from soil decreasing the initial concentration of 2.3xa0gxa0kg−1 soil to 100xa0mgxa0kg−1 soil and achieving a phenol loading within the polymer beads of 27.5xa0mg phenolxa0g−1 beads. The phenol-loaded polymer beads were removed from the soil and placed in a bioreactor, which was then inoculated with a phenol-degrading microbial consortium. All of the phenol contained within the polymer beads was shown to desorb from the polymer matrix and was degraded by the microbial consortium. The beads were used again (twice) in a similar manner with no loss in performance.

On the Use, and Reuse, of Polymers for the Treatment of Hydrocarbon Contaminated Water Via a Solid-liquid Partitioning Bioreactor

Aqueous environments contaminated with diesel components pose a threat to the native biota due to the intrinsically toxic nature of the many hydrocarbon compounds present. In the event of diesel being released into an aqueous environment it is imperative that the contaminant is recovered in a rapid manner to ensure the safety of aquatic organisms as well as to maintain desired water quality. The research presented in this study investigates the potential of polymeric sorbents to recover diesel from a contaminated aqueous source. Thermoplastic materials, such as styrene butadiene derived polymers, were shown to substantially reduce diesel levels in excess of 98% with 90% of this recoverable fraction being removed in less than 30 min. Recyclable materials, such as used automobile tires, were shown to obtain similar results with added potential benefit including lower cost and reuse of a waste material. The polymeric sorbents were also biologically regenerated and this was accomplished in a solid–liquid two‐phase partitioning bioreactor, in which 65% of the initial diesel contamination was degraded within a 9 day period. The result of this work was the demonstration of a low cost, reusable remediation technology for the recovery, and destruction of diesel from aqueous environments.

Two-phase reactors applied to the removal of substituted phenols: comparison between liquid-liquid and liquid-solid systems

In this paper, a comparison is provided between liquid-liquid and liquid-solid partitioning systems applied to the removal of high concentrations of 4-nitrophenol. The target compound is a typical representative of substituted phenols found in many industrial effluents while the biomass was a mixed culture operating as a conventional Sequencing Batch Reactor and acclimatized to 4-nitrophenol as the sole carbon source. Both two-phase systems showed enhanced performance relative to the conventional single phase bioreactor and may be suitable for industrial application. The best results were obtained with the polymer Hytrel which is characterized by higher partition capability in comparison to the immiscible liquid solvent (2-undecanone) and to the polymer Tone™. A model of the two systems was formulated and applied to evaluate the relative magnitudes of the reaction, mass transfer and diffusion characteristic times. Kinetic parameters for the Haldane equation, diffusivity and mass transfer coefficients have been evaluated by data fitting of batch tests for liquid-liquid and liquid-solid two phase systems. Finally, preliminary results showed the feasibility of polymer regeneration to facilitate polymer reuse by an extended contact time with the biomass.