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Bacterial Dehalogenation of Mixtures of 1,2-Dichloroethane, 1,2

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Bacterial Dehalogenation of Mixtures of 1,2-Dichloroethane, 1,2 Louisiana State University LSU Digital Commons LSU Master's Theses Graduate School 2012 Bacterial Dehalogenation of Mixtures of 1,2-Dichloroethane, 1,2-Dichloropropane, and 1,1,2-Trichloroethane by Dehalogenimonas Jacob Laddye Dillehay Louisiana State University and Agricultural and Mechanical College, [email protected] Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_theses Part of the Civil and Environmental Engineering Commons Recommended Citation Dillehay, Jacob Laddye, "Bacterial Dehalogenation of Mixtures of 1,2-Dichloroethane, 1,2-Dichloropropane, and 1,1,2-Trichloroethane by Dehalogenimonas" (2012). LSU Master's Theses. 4099. https://digitalcommons.lsu.edu/gradschool_theses/4099 This Thesis is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Master's Theses by an authorized graduate school editor of LSU Digital Commons. For more information, please contact [email protected]. BACTERIAL DEHALOGENATION OF MIXTURES OF 1,2-DICHLOROETHANE, 1,2- DICHLOROPROPANE, and 1,1,2-TRICHLOROETHANE BY DEHALOGENIMONAS A Thesis Submitted to the Graduate Faculty of the Louisiana State University and Agricultural and Mechanical College in partial fulfillment of the requirements for the degree of Masters of Science in The Department of Civil and Environmental Engineering by Jacob Laddye Dillehay B.S., Louisiana State University, 2010 December 2012 ACKNOWLEDGEMENTS First, I would like to thank Dr. Bill Moe. His taking me on as an undergraduate student worker and later as a graduate student helped lay the foundation for a productive and successful career. His ability to play devil’s advocate has taught me to approach a problem from multiple angles. Furthermore, his professional criticism of both my research and technical writing has allowed me to develop into a more critical data analyst and a better technical communicator. I would also like to thank Dr. Fred Rainey and Dr. David Constant for their contribution as members of my graduate committee. Secondly, I would like to thank all my fellow lab workers. None of this research would have been possible without their help. To Dr. Kim Bowman, thank you for your tireless work shoulder deep in the anaerobic chamber. You set an extraordinary example that I have followed throughout my career as a graduate student. Your patience in training me and constant attention to detail kept me on the correct path and has allowed me to finish my degree in a timely manner. Andrew has been a great friend both in and out of the lab. His visits while I was trapped behind the GC made those twelve hour, measurements sessions fly by. I would like to especially thank my fiancé, Mea, for her unwavering support. None of this would have been possible without her partnership and unconditional love. Her patience and ability to listen made conducting the research and writing this thesis a more enjoyable process. Lastly, I would like to thank the Governor’s Biotechnology Initiative of the Louisiana Board of Regents and NPC Services, Inc. for their financial support of my research. ii TABLE OF CONTENTS ACKNOWLEDGEMENTS ...................................................................................................................... ii TABLE OF CONTENTS ......................................................................................................................... iii LIST OF TABLES ................................................................................................................................... iv LIST OF FIGURES .................................................................................................................................. v ABSTRACT ............................................................................................................................................ vii CHAPTER 1: INTRODUCTION AND LITERATURE REVIEW ......................................................... 1 1.1 Literature Review .......................................................................................................................... 1 1.1.1 Prevalence of Chlorinated Solvents .............................................................................................. 1 1.1.2 Microbial Solvent Mixture Degradation ....................................................................................... 2 1.2 Thesis Organization ...................................................................................................................... 8 CHAPTER 2: MATERIALS AND METHODS ....................................................................................... 9 2.1 Chemicals ...................................................................................................................................... 9 2.2 Medium Preparation ...................................................................................................................... 9 2.3 Binary and Tertiary Mixture Experimental Design .................................................................... 10 2.4 Analytical Methods ..................................................................................................................... 11 CHAPTER 3: RESULTS AND DISCUSSION ...................................................................................... 13 3.1 Binary Mixture of 1,2-Dichloroethane (1,2-DCA) and 1,2-Dichloropropane (1,2-DCP) .......... 13 3.2 Binary Mixture of 1,2-Dichloroethane (1,2-DCA) and 1,1,2-Trichloroethane (1,1,2-TCA) ...... 16 3.2.1 Further Experiments on the Binary Mixture of 1,2-Dichloroethane ........................................... 19 3.3 Binary Mixture of 1,2-Dichloropropane (1,2-DCP) and 1,1,2-Trichlorethane (1,2-TCA) ......... 24 3.4 Tertiary Mixture of 1,2-Dichlorethane (1,2-DCA), 1,2-Dichloropropane (1,2-DCP), and 1,1,2-Trichloroethane (1,1,2-TCA) ............................................................................................. 28 3.5 Discussion and Implications ....................................................................................................... 32 CHAPTER 4: OVERALL CONCLUSIONS AND RECOMMENDATIONS FOR FUTURE WORK 36 4.1 Conclusions and Implications ..................................................................................................... 36 4.2 Future Work ................................................................................................................................ 37 REFERENCES ....................................................................................................................................... 39 APPENDIX: INDIVIDUAL BOTTLE GRAPHS .................................................................................. 43 VITA .................................................................................................................................................... 47 iii LIST OF TABLES Table 1.1 Chlorosolvent concentrations measured in groundwater from well W-1024-1 at the PPI site from which D. lykanthroporepellens BL-DC-9T was isolated (Bowman et al., 2006) ....................................................................................................................7 Table 1.2 Contaminant concentrations measured in groundwater used for isolation of Dehalogenimonas strain IP3-3 (Bowman et al., 2012)…………………………………………………………………………………………........7 Table 3.1 Thermodynamics of reduction half reactions for the chlorinated compounds of concern (data from De Wildeman and Verstraete,2003)… …….…………………..…………...33 iv LIST OF FIGURES Figure 3.1. Experimentally measured 1,2-DCA (closed diamonds), 1,2-DCP (closed squares), ethene (open diamonds), and propene (open squares) as a function of time in serum bottles inoculated with strain IP3-3 (left) or D. lykanthroporepellens BL-DC-9T (right)………………………………………………………………………………………….…15 Figure 3.2. Experimentally measured 1,2-TCA (closed circles), 1,2-DCA (closed diamonds),vinyl chloride (open circles), and ethene (open diamonds) as a function of time in serum bottles inoculated with strain IP3-3 (left) or D. lykanthroporepellens BL-DC-9T (right)...18 Figure 3.3. Experimentally measured 1,2-TCA (closed circles), 1,2-DCA (closed diamonds), vinyl chloride (open circles), and ethene (open diamonds) as a function of time in serum bottles inoculated with strain IP3-3 (left) or D. lykanthroporepellens BL-DC-9T (right)………..21 Figure 3.4. Experimentally measured 1,1,2-TCA, 1,2-DCA, vinyl chloride, and ethene as a function of time in serum bottles inoculated with strain IP3-3 (top) or D. lykanthroporepellens BL-DC-9T (bottom). Inoculum was grown on 1,2-DCA rather than 1,2-DCP. ……………………………………………………………………………...…….23 Figure 3.5. Experimentally measured 1,2-TCA (closed circles), 1,2-DCA (closed diamonds), 1,2-DCP (closed squares), vinyl chloride (open circles), ethene (open diamonds), and propene (open squares) as a function of time in serum bottles inoculated with strain IP3-3 (left) or D. lykanthroporepellens BL-DC-9T (right)……………..………………. ……..………26 Figure 3.6. Experimentally measured 1,2-TCA (closed circles), 1,2-DCA (closed diamonds), 1,2-DCP (closed squares), vinyl chloride (open circles), ethene (open diamonds), and propene (open squares) as a function of time in serum bottles inoculated with strain IP3-3 (left) or D. lykanthroporepellens BL-DC-9T (right)…… …………. …………..…….………....31 Figure A.1. Individual bottle analysis of experimentally measured 1,2-DCA (closed diamonds), 1,2-DCP (closed squares), ethene (open diamonds), and propene (open squares) as
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