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Fermentation of Glucose and Xylose to Hydrogen in the Presence of Long Chain Fatty Acids

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Fermentation of Glucose and Xylose to Hydrogen in the Presence of Long Chain Fatty Acids University of Windsor Scholarship at UWindsor Electronic Theses and Dissertations Theses, Dissertations, and Major Papers 2009 Fermentation of Glucose and Xylose to Hydrogen in the Presence of Long Chain Fatty Acids Stephen Reaume University of Windsor Follow this and additional works at: https://scholar.uwindsor.ca/etd Recommended Citation Reaume, Stephen, "Fermentation of Glucose and Xylose to Hydrogen in the Presence of Long Chain Fatty Acids" (2009). Electronic Theses and Dissertations. 92. https://scholar.uwindsor.ca/etd/92 This online database contains the full-text of PhD dissertations and Masters’ theses of University of Windsor students from 1954 forward. These documents are made available for personal study and research purposes only, in accordance with the Canadian Copyright Act and the Creative Commons license—CC BY-NC-ND (Attribution, Non-Commercial, No Derivative Works). Under this license, works must always be attributed to the copyright holder (original author), cannot be used for any commercial purposes, and may not be altered. Any other use would require the permission of the copyright holder. Students may inquire about withdrawing their dissertation and/or thesis from this database. For additional inquiries, please contact the repository administrator via email ([email protected]) or by telephone at 519-253-3000ext. 3208. Fermentation of Glucose and Xylose to Hydrogen in the Presence of Long Chain Fatty Acids by Stephen Reaume A Thesis Submitted to the Faculty of Graduate Studies through the Environmental Engineering Program in Partial Fulfillment of the Requirements for the Degree of Master of Applied Science at the University of Windsor Windsor, Ontario, Canada 2009 © 2009 Stephen Reaume AUTHORS DECLARATION OF ORIGINALITY I hereby certify that I am the sole author of this thesis and that no part of this thesis has been published or submitted for publication. I certify that, to the best of my knowledge, my thesis does not infringe upon anyone’s copyright nor violate any proprietary rights and that any ideas, techniques, quotations, or any other material from the work of other people included in my thesis, published or otherwise, are fully acknowledged in accordance with the standard referencing practices. Furthermore, to the extent that I have included copyrighted material that surpasses the bounds of fair dealing within the meaning of the Canada Copyright Act, I certify that I have obtained a written permission from the copyright owner(s) to include such material(s) in my thesis and have included copies of such copyright clearances to my appendix. I declare that this is a true copy of my thesis, including any final revisions, as approved by my thesis committee and the Graduate Studies office, and that this thesis has not been submitted for a higher degree to any other University or Institution. iii Abstract Hydrogen is a clean, efficient and versatile energy source which makes it a suitable alternative to fossil fuels. Mixed anaerobic cultures has the potential to produce hydrogen in a sustainable way in methanogenic bacteria can be inhibited. Batch studies were performed to assess the fermentation of glucose and xylose individually and together to observe if the sugar mixture is effective in hydrogen fermentation. Experiments were performed using a variety of LCFAs in order to inhibit methanogens so hydrogen can be collected. The highest amount of hydrogen produced took place in cultures fed LA plus xylose, glucose and the 50%/50% sugar mixture with yields of 2.13 ±0.05, 2.46 ±0.19 and 2.32 ±0.17 mol H 2/mol sugar, respectively. The maximum yields generated on a mol hydrogen per mass of sugar was 13.65, 14.20 and 14.08 mmol H 2/g sugar for the respective sugars fermented. The final results showed that the ratio of the two different sugars did not have a significant difference in the hydrogen yield. iv DEDICATION I dedicate this thesis to my loving family, John, Mary and Mark Reaume v Acknowledgements I wish to truly thank my advisor, Dr. Jerald A. Lalman for his leadership, wisdom and guidance throughout this thesis. Without his advice, support and selfless contributions this study would not have been able to be conducted or completed. I would like to extend my gratitude to Dr. A. A. Asfour and Dr. P. Vacratsis for their valuable suggestions as my committee members. Thanks to all my close friends who have helped me along in my studies, including special thanks to Srimanta Ray for his invaluable advice, Noori Saady and SubbaRao Chaganti for their help in the lab and Paul Huggard for all his help and together all their help throughout this thesis was instrumental. I would like to thank Mr. Bill Middleton for his advice, valuable assistance and technical support during my entire thesis without which I would still be stuck in the lab trying to figure out the instruments. Lastly I would like to thank NSERC, OGS and the Civil and Environmental Engineering Department for the financial support throughout this study. vi Table of Contents AUTHORS DECLARATION OF ORIGINALITY ……………………………… iii ABSTRACT………………………………………………………………………. iv DEDICATION …………………………………………………………………… v ACKNOWLEDGEMENTS ……………………………………………………… vi LIST OF TABLES ……………………………………………………………….. xi LIST OF FIGURES ……………………………………………………………… xiii NOMENCLATURE ……………………………………………………………… xvii CHAPTER 1 GENERAL INTRODUCTION 1.1 Hydrogen Production Methods…………………………………... 1 1.2 Biological Methods………………………………………………. 3 1.3 Objectives………………………………………………………… 4 CHAPTER 2 LITERATURE REVIEW 2.1 Anaerobic Fermentation………………………………………….. 6 2.1.1 Hydrolysis……………………………………………….. 6 2.1.2 Acidogenesis…………………………………………….. 7 2.1.3 Acetogenesis…………………………………………….. 7 2.1.4 Methanogenesis …………………………………………. 8 2.2 Anaerobic Hydrogen Production………………………………… 9 2.3 Thermodynamics ………………………………………………… 11 2.4 Xylose Production from Lignocellulose …………........................ 14 2.4.1 Xylose versus Glucose Degradation …………………….. 15 2.5 Factors Affecting Hydrogen Production…………………………. 17 2.5.1 Nutrients…………………………………………………. 20 2.5.2 pH………………………………………………………… 21 vii 2.5.3 Temperature……………………………………………… 23 2.5.4 Hydrogen Partial Pressure ………………………………. 24 2.5.5 Substrate Source and Concentration …………………….. 26 2.6 Microbial Inhibition……………………………………………… 26 2.6.1 Heat Treatment ………………………………………….. 27 2.6.2 Chemical Addition and Aeration ……………………….. 27 2.6.3 Electric Current ………………………………………….. 28 2.6.4 Product Inhibition ……………………………………….. 28 2.6.5 Long Chain Fatty Acids Inhibition ………...……………. 28 2.6.6 LCFA Inhibition ………………………………………… 29 CHAPTER 3 MATERIALS AND METHODS 3.1 Experimental Plan………………………………………………... 31 3.2 Inoculum Source ………………………………………………… 32 3.3 Basal Medium …………………………………………………… 33 3.4 Analytical Methods………………………………………............ 33 3.5 Chemicals ……………………………………………………….. 35 3.6 LCFA Delivery, Culture Bottle Preparation and Reaction Time... 36 3.7 Batch Reactor Operation and Culture Acclimation ……………... 37 CHAPTER 4 FERMENTATION OF GLUCOSE TO HYDROGEN IN THE PRESENCE OF LINOLEIC (LA) AND OLEIC ACID (OA) 4.1 Experimental Results and Data Analysis ………………………... 40 4.1.1 Hydrogen and Methane Production……………………… 41 4.1.2 Statistical Optimization – Full Factorial Design (FFD) … 47 4.1.3 Sugar Degradation ………………………………………. 53 4.1.4 Change in pH Value …………………………………….. 54 4.2 Discussion ………………………………………………………. 56 4.3 Conclusions ……………………………………………………… 59 viii CHAPTER 5 FERMENTATION OF XYLOSE IN THE PRESENCE OF LONG CHAIN FATTY ACIDS (LCFAs) 5.1 Introduction …………………………………………………….... 61 5.2 Experimental Design and Analytical Methods ………………….. 63 5.3 Results …………………………………………………………… 63 5.3.1 Xylose Degradation in the Presence of LA, OA, SA, PA, MA, LAU ……………………………………………..... 63 5.3.1.1 Hydrogen and Methane ……………………........ 63 5.3.2 Degradation of Glucose and Glucose/Xylose Mixture in the Presence of LA …………………………………… 71 5.3.2.1 Hydrogen and Methane (75%/25%, 50%/50%, 25%/75% Xylose/Glucose, Glucose)…….......... 71 5.3.3 Volatile Fatty Acids (VFA) (Xylose, 50%/50% Xylose/Glucose, Glucose)……………………………… 76 5.3.4 Alcohols (Xylose, 50%/50% Xylose/Glucose, Glucose) 79 5.3.5 Sugar Degradation ……………………………………….. 81 5.3.6 Electron Balance ………………………………………… 82 5.4 Discussion of Results ……………………………………………. 84 5.5 Conclusions ……………………………………………………… 89 CHAPTER 6 CONCLUSIONS…………………………………………………... 90 CHAPTER 7 ENGINEERING SIGNIFICANCE AND FUTURE RECCOMENDATIONS …………………………………………. 92 REFERENCES …………………………………………………………………… 93 APPENDICES ix Appendix A: Hydrogen Methane and Carbon Dioxide Calibration Curves …... 102 Appendix B: Volatile Fatty Acid Calibration Curves …………………………. 103 Appendix C: Sugar Calibration Curves ……………………………………….. 104 Appendix D: Alcohol Calibration Curves ……………………………………... 105 Appendix E: Sample Calculations …………………………………………….. 107 Appendix F: VSS, TSS, and pH data from feed reactor ………………………. 109 VITA AUCTORIS ……………………………………………………………….. 110 x List of Tables Table 2.1: Products of acidification of glucose………………………………….. 11 Table 2.2: Biological half-reactions and free energy associated with the reaction………………………………………………………………... 14 Table 2.3: Acetogenic reactions …………………………………………………. 19 Table 2.4 Typical hydrogen yields for mixed anaerobic microbial populations… 22 Table 2.5 Effects of temperature on hydrogen gas yield for mixed anaerobic cultures………………………………………………………………… 24 Table 2.6 Gibb’s free energy changes under standard conditions for typical hydrogen-releasing and hydrogen-consuming reactions……………… 25 Table 3.1: Experimental design for glucose fermentation to hydrogen in the presence of OA and
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