Expanded application of a two-phase partitioning bioreactor through strain development and new feeding strategies.

This research demonstrated the microbial treatment of concentrated phenol wastes using a two-phase partitioning bioreactor (TPPB). TPPBs are characterized by a cell-containing aqueous phase and an immiscible and biocompatible organic phase that partitions toxic substrates to the cells on the basis of their metabolic demand and the thermodynamic equilibrium of the system. Process limitations imposed by the capability of wild-type Pseudomonas putida ATCC 11172 to utilize long chain alcohols were addressed by strain modification (transposon mutagenesis) to eliminate this undesirable biochemical characteristic, enabling use of a range of previously bioavailable organics as delivery solvents. Degradation of phenol in a system with the modified strain as catalyst and industrial grade Adol 85 NF (primarily oleyl alcohol) as the solvent was demonstrated, with the system ultimately degrading 36 g of phenol within 38 h. Volumetric phenol consumption rates by wild type P. putida ATCC 11172 and the genetically modified derivative revealed equivalent phenol degrading capabilities (0.49 g/L x h vs 0.47 g/L x h respectively, in paired fermentations), with the latter presenting a more efficient remediation option due to decreased solvent losses arising from the modified strain's forced inability to consume the delivery solvent as a substrate. Two feeding strategies and system configurations were evaluated to expand practical applications of TPPB technology. The ability to operate with a lower solvent ratio over extended periods revealed potential for long-term application of TPPB to the treatment of large masses of phenol while minimizing solvent costs. Repeated recovery of 99% of phenol from concentrated phenol solutions and subsequent treatment within a TPPB scheme demonstrated applicability of the approach to the remediation of highly contaminated "effluents" as well as large masses of bulk phenol. Operation of the TPPB system in a dispersed manner, rather than as two distinct phases, resulted in volumetric consumption rates similar to those previously achieved only in systems operated with enriched air.