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Managing Exoelectrogenic Microbial Community Development Through Bioprocess Control for Conversion of Biomass-Derived Streams

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Managing Exoelectrogenic Microbial Community Development Through Bioprocess Control for Conversion of Biomass-Derived Streams University of Tennessee, Knoxville TRACE: Tennessee Research and Creative Exchange Doctoral Dissertations Graduate School 8-2017 Managing exoelectrogenic microbial community development through bioprocess control for conversion of biomass-derived streams Alex James Lewis University of Tennessee, Knoxville, [email protected] Follow this and additional works at: https://trace.tennessee.edu/utk_graddiss Part of the Biological Engineering Commons, Biotechnology Commons, Environmental Microbiology and Microbial Ecology Commons, and the Systems Biology Commons Recommended Citation Lewis, Alex James, "Managing exoelectrogenic microbial community development through bioprocess control for conversion of biomass-derived streams. " PhD diss., University of Tennessee, 2017. https://trace.tennessee.edu/utk_graddiss/4699 This Dissertation is brought to you for free and open access by the Graduate School at TRACE: Tennessee Research and Creative Exchange. It has been accepted for inclusion in Doctoral Dissertations by an authorized administrator of TRACE: Tennessee Research and Creative Exchange. For more information, please contact [email protected]. To the Graduate Council: I am submitting herewith a dissertation written by Alex James Lewis entitled "Managing exoelectrogenic microbial community development through bioprocess control for conversion of biomass-derived streams." I have examined the final electronic copy of this dissertation for form and content and recommend that it be accepted in partial fulfillment of the equirr ements for the degree of Doctor of Philosophy, with a major in Energy Science and Engineering. Abhijeet Borole, Major Professor We have read this dissertation and recommend its acceptance: Brian Davison, Terry Hazen, Cong Trinh Accepted for the Council: Dixie L. Thompson Vice Provost and Dean of the Graduate School (Original signatures are on file with official studentecor r ds.) Managing exoelectrogenic microbial community development through bioprocess control for conversion of biomass-derived streams: A Dissertation Presented for the Doctor of Philosophy Degree The University of Tennessee, Knoxville Alex James Lewis August 2017 ACKNOWLEDGEMENTS First, I want to sincerely thank my advisor Abhijeet Borole for his mentorship during my dissertation. His guidance and open-door policy were invaluable to my development as a scientist and research success. I would also like to thank my committee members for their participation in this process, and for providing guidance and critiques to improve the way I approached science. I want to thank the Bredesen Center, specifically Lee Riedinger, Wanda Davis, and Tracey Bucher for all their help over these 4 years, you guys are the best! Lastly, I want to thank my friends and family who supported me through this process, and all the long hours and weekend work days! Thanks for always being there. ii ABSTRACT Bioelectrochemical systems are an emerging technology capable of utilizing aqueous waste streams generated during biomass conversion of lignocellulosic feedstocks to produce valuable co-products and thus, have potential to be integrated into biorefineries. In a microbial electrolysis cell, organic compounds are converted to electrons, protons, and CO2 by fermentative and exoelectrogenic bacteria in the anode compartment. By having the ability to extract electrons from waste streams, these systems can treat water while also producing hydrogen, and thus can improve the efficiency of biomass to fuel production by minimizing external hydrogen requirement and enabling water recycle. The overall goal of this research is to understand how changes in the way the reactors are operated affect the performance of the system, and the structure of microbial community within when converting a biomass-derived stream (BOAP). This can enable the design of optimal community structure for waste stream conversion, which can lead to improved and stable performance of the system. An integrated approach was taken to test parameters such as flow-rate, recycle, organic loading rate, feeding regime, and electrode potential using a suite of electrochemical, metabolic and genomic techniques to unravel the biocomplexity of these systems and the impact on the reactor microbial communities. Faster flow-rates and recycle operation led to better conversion of BOAP, but efficiencies decreased as organic loading rates increased. Exposure to high concentrations during fed-batch feeding resulted in a substantial loss of electrons that was alleviated through continuous operation. Additionally, high loading/concentration conditions selected for different microbial species. Furthermore, exposing the microbial communities to different anode voltages provided evidence that the benefits of using more negative anode iii potentials to increase electrical efficiency can be capture through long-term enrichment without sacrificing substantial output. Lastly, the microbial community was characterized using deep sequencing techniques, revealing novel players directing a wide range of compounds to electrons. The resulting data was used to develop correlations that will serve as the foundation for operating these systems for commercial applications. iv TABLE OF CONTENTS INTRODUCTION .......................................................................................................................... 1 References ................................................................................................................................... 7 CHAPTER I Hydrogen production from switchgrass via an integrated pyrolysis–microbial electrolysis process ......................................................................................................................... 9 Abstract ..................................................................................................................................... 10 Introduction ............................................................................................................................... 11 Methods..................................................................................................................................... 13 Biomass pyrolysis ................................................................................................................. 13 Bio-oil separation and characterization ................................................................................ 14 MFC and MEC construction ................................................................................................. 15 Inoculation and operation ..................................................................................................... 16 Chronoamperometry - Hydrogen production ........................................................................ 17 COD analysis ........................................................................................................................ 18 DNA extraction and 16s rRNA analysis ............................................................................... 19 Calculation of Coulombic efficiency and other conversion efficiencies .............................. 20 Results and Discussion ............................................................................................................. 22 Bio-oil production and pyrolysis yields ................................................................................ 22 Bio-oil and aqueous phase characterization .......................................................................... 22 Microbial anode development ............................................................................................... 23 16s rRNA analysis ................................................................................................................ 24 Batch operation ..................................................................................................................... 26 v Continuous substrate addition ............................................................................................... 29 Conversion of individual components of boap ..................................................................... 29 Energy efficiency .................................................................................................................. 31 Improvement in treatment of complex feed streams in MEC ............................................... 31 Advancements in conversion efficiency and hydrogen yield ............................................... 35 Implications for biorefinery application ............................................................................... 36 Conclusion ................................................................................................................................ 37 References ................................................................................................................................. 38 Appendix ................................................................................................................................... 41 CHAPTER II Understanding the impact of flow rate and recycle on the conversion of a complex biorefinery stream using a flow-through microbial electrolysis cell ............................................ 47 Abstract ..................................................................................................................................... 48 Introduction ..............................................................................................................................
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