A perfluorocarbon-based oxygen delivery system to a membrane bioreactor

Abstract

The white rot fungus, Phanerochaete chrysosporium strain BKMF-1767 (ATCC 24725), produces the extracellular enzymes, Lignin peroxidase (LiP) and Manganese peroxidase (MnP), that constitute the major route for lignin degradation by this organism. LiP and MnP have also been shown to play a major role in aromatic pollutant degradation. Due to the need for continuous production of LiP and MnP, a fixed-film bioreactor, classified as a membrane gradostat reactor (MGR), was developed. The implementation of batch-reactor operational parameters to the MGR system was found to be ineffective, thus creating the need for further research to improve the operational aspects of the MGR system to optimise its capabilities for continuous and industrial-scale operations. The research undertaken in this study, provides information that can be used to classify the dissolved oxygen (DO) transport kinetics into immobilised fixed-films of P. chrysosporium. Operational limitations of the MGR relating to environmental stresses in the bioreactor during operation and to biofilm deterioration, including limitations of DO mass transport, oxidative stress, trace element accumulation and polysaccharide storage in the fungal biomass, were evaluated in single capillary MGR systems (SCMGRs). These conditions were identified as existing in the continuous MGR systems. From DO profiles, the oxygen consumption and flux into the biofilms, including the distribution of DO, was determined to be dependent on the immobilised biofilm’s age. Younger biofilms showed higher DO distribution than older biofilms even when aeration was directed to the extracapillary space (ECS) of the reactor against the biofilm’s surface. An increase in anaerobic zone thickness was observed to be increasing with an increase in biofilm thickness. Although, DO kinetic parameters were comparable with those obtained in submerged mycelia pellets, higher oxygen consumption values were observed in biofilms grown in the SCMGRs. The limitations of MGR were identified as: 1) poor DO distribution in immobilised biofilms because of b-glucan production and storage in the immobilised

biomass, resulting in ethanol production; 2) the peroxidation of lipids of the biofilms, which in turn will affect the long-term performance of the biomass caused by oxygenation and 3) trace element ion accumulation enhanced by b-glucan production. Furthermore, trace element ion accumulation was higher in the MGRs than in batch cultures using the same nutrient medium. The development of a perfluorocarbon (PFC) emulsion for the MGRs to counteract these limitations was investigated. The compatibility of the emulsion with oxygen-carrying capacity was shown with an improvement in biomass generation, LiP/MnP production and overall consumption of primary substrates, mainly glucose and ammonium tartrate, in batch cultures. The emulsions investigated were based on the addition of oxygen carriers: Perfluorooctyl bromide (PFOB), Bis-(Perfluorobutyl) ethene (PFBE) and Perfluoropropylamine (PFPA), using Pluronic F 68 (PF 68) as the surfactant. Concentrations of 10 to 30% (w/v) PFC and 8.5% (w/v) PF 68 were tested successfully in batch cultures. The emulsions containing 10% (w/v) PFCs resulted in improved biomass performance as opposed to emulsions with higher PFC oil concentrations. An emulsion containing 10% (w/v) PFOB was used to evaluate its efficacy in the SCMGRs, as the biomass yield and overall enzyme production were superior to PFPA and PFBE-based emulsions with similar oil concentrations. After successfully applying PFOB and PF 68 to the SCMGRs, the following results were obtained: 1) reduced ethanol production; 2) reduced trace element accumulation; 3) lower b-glucan production and 4) improved DO-penetration ratio in immobilised biofilms.