Simultaneous Biodegradation of Benzene, Toluene, and p-Xylene in a Two-Phase Partitioning Bioreactor: Concept Demonstration and Practical Application

In this work, a mixture of benzene, toluene, and p-xylene was simultaneously biodegraded by Pseudomonas sp. ATCC 55595 in a two-phase partitioning bioreactor. This bioreactor consisted of a 1-L cell-containing aqueous medium phase and a 500mL immiscible organic phase. The organic solvent systematically selected for use in the bioreactor was Adol 85 NF, an industrial-grade, biocompatible solvent. In the first of three experiments, the organic phase was loaded with 2.0 g of benzene, 10.15 g of toluene, and 2.1 g of p-xylene, which partitioned into the aqueous phase at concentrations of 25, 50, and 8 mg/L, respectively. The system ultimately biodegraded all of the substrates within 144 h. During the rapid growth phase of this fermentation, the cells were oxygen-limited. This fermentation was therefore repeated using an enriched air supply to remove the oxygen limitation. The use of enriched air ultimately reduced the length of the fermentation to 108 h, thereby improving the overall volumetric consumption rates. Finally, 500 mL of Adol were used to recover 2.0 g of benzene, 10.15 g of toluene, and 2.1 g of p-xylene from silica sand that was contaminated as part of a simulated soil "spill". The solvent washing procedure was able to recover greater than 99% of each compound from the contaminated soil. The Adol was then transferred to the two-phase bioreactor to permit biological treatment of the BTX contaminants. This process was repeated when theJnitial BTX load had been consumed almost to exhaustion, and the solvent was able to recover the contaminants at greater than 99% efficiency once again. The system was ultimately able to degrade 4.0 g of benzene, 20.2 g of toluene, and 4.2 g of p-xylene within 144 h. These results represent an unprecedented level of BTX degradation and illustrate a potential practical application for this novel biotechnology.