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Effects of Temperature, Light Intensity and Quality, Carbon Dioxide, And Worcester Polytechnic Institute Digital WPI Doctoral Dissertations (All Dissertations, All Years) Electronic Theses and Dissertations 2014-01-24 Effects of Temperature, Light Intensity and Quality, Carbon Dioxide, and Culture Medium Nutrients on Growth and Lipid Production of Ettlia oleoabundans Ying Yang Worcester Polytechnic Institute Follow this and additional works at: https://digitalcommons.wpi.edu/etd-dissertations Repository Citation Yang, Y. (2014). Effects of Temperature, Light Intensity and Quality, Carbon Dioxide, and Culture Medium Nutrients on Growth and Lipid Production of Ettlia oleoabundans. Retrieved from https://digitalcommons.wpi.edu/etd-dissertations/42 This dissertation is brought to you for free and open access by Digital WPI. It has been accepted for inclusion in Doctoral Dissertations (All Dissertations, All Years) by an authorized administrator of Digital WPI. For more information, please contact [email protected]. Effects of Temperature, Light Intensity and Quality, Carbon Dioxide, and Culture Medium Nutrients on Growth and Lipid Production of Ettlia oleoabundans by Ying Yang A Dissertation Submitted to the Faculty of WORCESTER POLYTECHNIC INSTITUTE in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Biology and Biotechnology by December 2013 Approved by: Dr. Pamela Weathers, Advisor Dr. Robert Thompson, Committee Member Dr. Luis Vidali, Committee Member Dr. Reeta Rao, Committee Member “A journey of a thousand miles begins with a single step.” — Lao Tzu (604 BC – 531 BC) ii Abstract Ettlia oleoabundans, a freshwater green microalga, was grown under different environmental conditions to study its growth, lipid yield and quality for a better understanding of the fundamental physiology of this oleaginous species. E. oleoabundans showed steady increase in biomass under low temperature and low light intensity, and at high temperature lipid cell content significantly increased independent of nitrate depletion. Studies on light quality showed that red light treatment did not change the biomass concentration, but stimulated lipid yield especially oleic acid, the most desirable biodiesel precursor. Moreover, no photoreversibility in lipid production was observed when applying alternating short-term red and far-red lights, which left the phytochrome effect still an open question. In addition, carbon dioxide enrichment via an air sparging system significantly boosted exponential growth and increased carbon conversion efficiency. Finally, a practical study demonstrated the feasibility of growing E. oleoabundans for high lipid production using a diluted agricultural anaerobic waste effluent as the medium. Together, these studies showed the potential of E. oleoabundans as a promising high yield feedstock for the production of high quality biodiesel. iii Acknowledgements Many people have helped me in the past five years to complete this dissertation and finish graduate school. First of all, I would like to express my sincere gratitude to my advisor, Dr. Pamela Weathers, for her continuous encouragement and guidance during my entire stay at WPI. She taught me how to become an independent and critical thinker throughout my research. Besides, she is extremely approachable and always ready to help in many different ways beyond school. I greatly appreciate her valuable advices on my career development and other topics in daily life, like travel destinations and food recipes, etc. I would like to deeply thank other three members in my graduate committee, Drs. Robert Thompson, Reeta Prusty Rao, and Luis Vidali, for their intelligent contribution to this dissertation. Dr. Thompson offered many insights not only on this specific lab research, but also on a broader picture of renewable energy from an engineering perspective. Dr. Prusty Rao kindly provided lots of useful information for this project and taught me how to find a better work-life balance in graduate school. I would also thank Dr. Vidali for his valuable input on experimental design and future directions of this study. Additional thanks go to Dr. Jay Xu (Arkansas State University) for his guidance during my two-month summer research in Jonesboro, Arkansas in 2009. I would also like to thank Dr. Joseph Duffy for all his encouragement and support during the past five years. Of course this dissertation would not have been possible without the help of many other faculty, staff, graduate and undergraduate students in WPI. I sincerely thank Drs. Melissa Towler and Patrick Arsenault for their tremendous assistance with my project on a daily basis. Also, thanks go to Dr. Will Lin and Andrew Butler for their help with the forever challenging GCMS iv analyses. I owe great thanks to the current and previous lab members, Liwen Fei and Khanhvan Nguyen, for their continuous encouragement and technical support along the way. Together we survived graduate school. In addition, I am grateful to the WPI 2012 Alexander DiIorio Fellowship Award in Bioprocessing, the Biology and Biotechnology Department at WPI, the 2009 Grant-in-Aid of Research Award and 2012 & 2013 Lewin Poster Awards by the Phycological Society of America, and the Arkansas Bioscience Institute for their generous funding to make this piece of work possible. I would like to thank my closest friends for making my life full of laughter and love. It is always a great feeling to contact and visit them, no matter how far apart we are, or how long we have not seen each other, friendships are always with me to go through all the good and bad days during the past few years. Also, as a sports fan, I am grateful to my favorite teams and players for teaching me to be tough and never give up. Their fighting spirits and persistent efforts in pursuit of success are just inspiring little things to cherish in real life. Last but not the least, I owe a huge debt of gratitude to my dear parents. It is such a long journey to get to this stage of my life and my parents are always the rock behind me to keep me motivated every single day. I deeply thank them for their love and understanding. None of my academic and personal development would be possible without their unconditional support. v Table of Contents Abstract ........................................................................................................................................................ iii Acknowledgements ...................................................................................................................................... iv Table of Contents ......................................................................................................................................... vi List of Tables ............................................................................................................................................... ix List of Figures .............................................................................................................................................. xi Chapter 1: Introduction and Literature Review ............................................................................................ 1 1.1 Microalgae and oleaginous algal species ............................................................................................ 2 1.1.1 Ettlia oleoabundans ..................................................................................................................... 4 1.1.2 Other microalgal species .............................................................................................................. 7 1.2 Biosynthesis of lipids in microalgae ................................................................................................... 7 1.3 Triacylglycerides-derived FAMEs and biodiesel quality ................................................................. 12 1.4 Photosynthetic machinery and photoreceptors in microalgae ........................................................... 14 1.4.1 Red/far-red light absorbing photoreceptor: phytochrome .......................................................... 16 1.5 Cultivation conditions for microalgae ............................................................................................... 18 1.5.1 Light ........................................................................................................................................... 19 1.5.2 Carbon Dioxide .......................................................................................................................... 22 1.5.3 Temperature ............................................................................................................................... 25 1.5.4 Culture medium ......................................................................................................................... 27 1.6 Wastewater as an alternative nutrient medium ................................................................................. 29 1.7 Objectives and organizations of thesis .............................................................................................. 31 Chapter 2: At high Temperature Lipid Production in Ettlia oleoabundans Occurs before Nitrate Depletion .................................................................................................................................................................... 33 Abstract ................................................................................................................................................... 34 2.1 Introduction ......................................................................................................................................
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