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Tropomyosin) Using Food Processing Methods Adeseye Lasekan University of Maine, Adeseye.Lasekan@Gmail.Com The University of Maine DigitalCommons@UMaine Electronic Theses and Dissertations Fogler Library Summer 8-11-2017 Attenuating the Antibody Reactivity of the Shrimp Major Allergen (Tropomyosin) using Food Processing Methods Adeseye Lasekan University of Maine, [email protected] Follow this and additional works at: http://digitalcommons.library.umaine.edu/etd Part of the Food Chemistry Commons, Food Processing Commons, and the Other Food Science Commons Recommended Citation Lasekan, Adeseye, "Attenuating the Antibody Reactivity of the Shrimp Major Allergen (Tropomyosin) using Food Processing Methods" (2017). Electronic Theses and Dissertations. 2732. http://digitalcommons.library.umaine.edu/etd/2732 This Open-Access Dissertation is brought to you for free and open access by DigitalCommons@UMaine. It has been accepted for inclusion in Electronic Theses and Dissertations by an authorized administrator of DigitalCommons@UMaine. For more information, please contact [email protected]. ATTENUATING THE ANTIBODY REACTIVITY OF THE SHRIMP MAJOR ALLERGEN (TROPOMYOSIN) USING FOOD PROCESSING METHODS By Adeseye O. Lasekan B.Sc. Federal University of Agriculture Abeokuta, 2007 M.S. Universiti Putra Malaysia, 2013 A DISSERTATION Submitted in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy (in Food and Nutrition Sciences) The Graduate School The University of Maine August 2017 Advisory Committee: Balunkeswar Nayak, Assistant Professor of Food Processing, Advisor Soheila Maleki, Lead Scientist, USDA-ARS, Southern Regional Research Center Dorothy Klimis-Zacas, Professor of Clinical Nutrition Eric Gallandt, Professor of Weed Ecology Denise Skonberg, Associate Professor of Food Science Vivian Wu, Research Leader, USDA-ARS, Western Regional Research Center © 2017 Adeseye O. Lasekan All Rights Reserved ii ATTENUATING THE ANTIBODY REACTIVITY OF THE SHRIMP MAJOR ALLERGEN (TROPOMYOSIN) USING FOOD PROCESSING METHODS By Adeseye Lasekan Dissertation Advisor: Dr. Balunkeswar Nayak An abstract of the Dissertation Presented in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy (in Food and Nutrition Sciences) August 2017 Shrimp is one of the major causes of adverse allergic reactions in both adults and children. Consumers with shrimp allergy have an immune system that is hypersensitive towards certain proteins in shrimp. The immune system of these consumers produces an antibody, immunoglobulin E (IgE), which binds to specific regions on these proteins and activates reactions leading to adverse symptoms. Most shrimp allergic consumers have IgE that binds to a shrimp myofibrillar protein known as tropomyosin. Other than avoidance of shrimp, structural modification of shrimp tropomyosin using food processing methods could influence the capacity of this allergenic protein to bind IgE. Thus, the objectives of this study were to evaluate the IgE binding capacity of tropomyosin extracted from whole shrimp following: 1) different methods of heat processing of shrimp, 2) exposure of shrimp muscle to high acid condition, 3) treatment with three microbial proteases, and 4) treatment with microbial and plant proteases. In study 1, the IgE binding of tropomyosin remained unchanged following boiling, steaming, baking, microwave roasting, grilling and frying of whole shrimp in comparison with tropomyosin from raw shrimp. However, high pressure steaming significantly reduced IgE binding capacity especially at lower concentration of antigen. In study 2, treatment of whole shrimp with vinegar reduced the solubility of tropomyosin and thus the IgE binding capacity of tropomyosin in the soluble extract was reduced. However, immunochemical analysis of the insoluble protein fraction indicated retention of significant IgE binding by tropomyosin following exposure to vinegar. In study 3, after treatment of whole shrimp with three microbial proteases, an alkaline protease significantly reduced (> 80%) the IgE reactivity of tropomyosin. Treatment of whole shrimp with alkaline protease and two plant proteases in study 4 revealed that, the plant proteases did not significantly reduce the IgE reactivity of tropomyosin. In summary, treatment of whole shrimp with enzymes that can cleave the IgE binding sites of tropomyosin could be the first step in the development of a hypoallergenic shrimp product. However, an in vivo assay like the cell mediator release assay is necessary to confirm the reduced immunoreactivity of tropomyosin following treatment with alkaline protease. DEDICATION I dedicate this dissertation to my parents and siblings for their love, support, encouragement and prayers. iii ACKNOWLEDGEMENTS First, I want to give thanks to God, the creator of all things, for the gift of life without which this accomplishment would not have been possible. I also want to thank my advisor, Dr. Balunkeswar Nayak for giving me this opportunity in 2013 to pursue a doctoral degree at the University of Maine and for his support throughout the period of this program. I would like to appreciate members of my advisory committee Dr. Dorothy Klimis-Zacas, Dr. Denise Skonberg, Dr. Eric Gallandt, Dr. Soheila Maleki and Dr. Vivian Wu for their guidance, moral support and commitment to making sure that I understand all the relevant aspects of my research. Thank you all for the knowledge imparted. I am grateful to the School of Food and Agriculture for providing me with the financial aid for my doctoral program as well as travel funds to attend IFT conferences. Thank you very much for your investment and I would go from here to live a life worthy of that investment. I would also like to thank USDA – National Institute of Food and Agriculture and the University of Maine Graduate Student Government for providing funding that was used for different aspects of my research. Not forgetting, all the instructors of the classes that I took here at the University of Maine, thank you all for the knowledge imparted. A special appreciation goes to Ms. Katherine Davis Dentici. She is one of first people I met here at the University of Maine and throughout these past four years she has selflessly helped me in my work in the lab in so many more ways than I can count. Thank you so much Kath. Not forgetting my current and past lab members especially Tamanna Ramesh and Amma Amponsah, thank you so much for helping me to believe in myself at those moments of self-doubt and for modelling hard work, perseverance and optimism. iv Bouhee Kang, Xue Du, Praveen Kumar Sappati, Dhriti Nayyar, Bhargavi Rane, Hanjuan Cao, Angela Silke, Dhafer and Weeam Alshaibani, thank you guys for your encouragement and friendship. I also want to appreciate members of Dr. Maleki’s lab for providing us with polyclonal antibodies which was used in the last phase of my study. All these would not have been possible without the prayers, love and sacrifice of my parents Dr. and Mrs. Lasekan. I am grateful for your endless support, guidance and encouragement and for imparting in me the value of education which you always describe as an asset that no one can take away from me. I also want to thank my siblings Dr. Olusiji Lasekan, Gbemisola Lasekan and the recent addition to our family, Elizabeth (my niece), thank you all for being a source of joy and inspiration. v TABLE OF CONTENTS DEDICATION .................................................................................................................. iii ACKNOWLEDGMENTS ................................................................................................. iv LIST OF TABLES ............................................................................................................ xii LIST OF FIGURES ......................................................................................................... xiii CHAPTER 1. INTRODUCTION ...........................................................................................................1 1.1. Background .......................................................................................................1 1.2. Objectives .........................................................................................................7 2. LITERATURE REVIEW ................................................................................................8 2.1. Prevalence of Food Allergies ............................................................................8 2.2. Mechanism of Immune-Mediated Allergic Reaction .....................................11 2.2.1. The Gastrointestinal Tract as an Immune Organ .............................11 2.2.2. GALT Effector Cells and their Function .........................................14 2.2.2.1. Granulocytes.....................................................................15 2.2.2.2. Monocytes.........................................................................17 2.2.2.3. Lymphocytes.....................................................................18 2.2.3. Mechanism of IgE-Mediated Allergic Reaction ..............................21 2.3. The Major Food Allergens ..............................................................................25 2.3.1. Peanut and Tree nuts ........................................................................25 2.3.2. Milk ..................................................................................................28 2.3.3. Egg ...................................................................................................29 2.3.4. Fish ...................................................................................................31
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