Thesis (M.S.)--University of Wisconsin--Madison, 1981. / Typescript. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 53-57).

Thesis (M. Sc.)--University of Canterbury, 2009. / Typescript (photocopy). Includes bibliographical references (leaves 115-130). Also available via the World Wide Web.

The metabolism of white-rot fungi has many proposed biotechnological applications. Their unique capability to depolymerize and catabolize lignin, the most recalcitrant component of lignocellulosic biomass, could be instrumental to the sustainable production of fuels, chemical, and materials from waste biomass feedstocks. The non-specific, oxidative nature of this lignin-degrading metabolism of white-rot fungi renders them capable of degrading a wide range of complex refractory organic compounds beyond lignin, including emerging micropollutants such as pharmaceuticals and pesticides which current wastewater treatment processes were not designed to remove. However, harnessing these metabolic capabilities into engineered bioprocesses has proven to be challenging. Common bioreactor design strategies were developed for traditionally-used unicellular bacteria and yeasts and are not necessarily appropriate for the more complex, filamentous white-rot fungi. Due to a lack of specific engineering strategies and other knowledge gaps, the realization of white-rot fungal bioprocesses has been hampered by low process efficiencies and operational challenges. This dissertation aims to expand the engineering toolbox for harnessing the metabolism of white-rot fungi in bioprocesses. Specifically, it proposes the addition of Fenton chemistry as an avenue to unlock the biotechnological potential of white-rot fungi. The production of hydroxyl radicals through the Fenton reaction is generally understood to be part of the lignin-degrading machinery of white-rot fungi and the addition of Fenton chemistry has been shown to synergistically enhance lignin degradation by white-rot fungi. Overall, the research presented here aims to demonstrate that incorporating Fenton chemistry into white-rot fungal bioprocesses not only synergistically increases lignin degradation efficiency, but also offers a potential solution for the operational challenges that have prevented the implementation of white-rot fungal bioprocesses. This dissertation was guided by five objectives aimed at illustrating the utility of coupling Fenton chemistry and white-rot fungi in engineered bioprocesses. The first objective was to demonstrate, optimize, and uncover the underlying mechanisms driving the synergistic degradation of lignin by white-rot fungi and Fenton chemistry. Through this assessment, it was found that lignin degradation increased synergistically from 58.8% to 80.2% in the presence of Fenton chemistry at the optimum concentration. This work also showed that Fe(II)/Fe(III) cycling and the induction of auxiliary ligninolytic pathways mediate this synergistic interaction. The second objective was to elucidate how Fenton chemistry influences the regulating mechanisms of ligninolytic activity in white-rot fungi, specifically C:N ratio. This showed that C:N ratio significantly influences lignin degradation in the absence of Fenton, but that this effect is blunted in the presence of Fenton. The third objective was to investigate how Fenton chemistry modulates the relationship between the concentration of fungal biomass and the extent of lignin. In the absence of Fenton, fungal biomass concentration was strongly correlated to the extent of lignin degradation. While this was also the case in the presence of Fenton chemistry at very low fungal biomass concentrations, this relationship became uncoupled at sufficiently high fungal biomass concentrations. The fourth objective was to evaluate Fenton chemistry as a selective disinfectant to allow for the persistence or enrichment of white-rot fungi in non-sterile settings. The model competitor E. coli became completely inactivated within hours at the optimal concentration of Fenton reagents, whereas the white-rot fungus P. chrysosporium survived and grew. Lastly, the fifth objective was to demonstrate the long-term performance of a continuously-operated bioreactor which integrated Fenton chemistry and white-rot fungal metabolism. A rotating biological contactor (RBC) combined with a rotating cathode electro-Fenton was constructed and a kinetic model based on batch tests was successfully developed and validated. The reactors were operated for over 100 days and reached stable lignin degradation performance at ~55%. Analysis of the microbial ecology of these reactors showed the persistence of the inoculated P. chrysosporium within the biofilms, as well as the enrichment for other lignin-degrading fungi and bacteria with aromatic catabolism and iron-reduction capabilities. Overall, this research provides insight into the potential and practical implications of integrating Fenton chemistry with white-rot fungi in bioprocesses. The lignin-degrading metabolism of white-rot fungi has long been of interest for biotechnological purposes, but attempts to operationalize them have thus far been unsuccessful at scale. In order to consider scaling white-rot fungi to full-scale operations such as wastewater treatment plants, a better understanding and tighter controls on the growth, ligninolytic activity, and ecological interactions of white-rot fungi are needed. This work proposes Fenton chemistry as a synergetic actor, selective promoter and regulator of white-rot fungal biomass and their production of lignin degrading enzymes.