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Protein and Lipid Characterization of Acheta Domesticus, Bombyx Mori, and Locusta Migratoria Dry Flours

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Protein and Lipid Characterization of Acheta Domesticus, Bombyx Mori, and Locusta Migratoria Dry Flours Graduate Theses, Dissertations, and Problem Reports 2018 Protein and Lipid Characterization of Acheta domesticus, Bombyx mori, and Locusta migratoria Dry Flours Emily N. Brogan Follow this and additional works at: https://researchrepository.wvu.edu/etd Part of the Food Chemistry Commons, and the Food Microbiology Commons Recommended Citation Brogan, Emily N., "Protein and Lipid Characterization of Acheta domesticus, Bombyx mori, and Locusta migratoria Dry Flours" (2018). Graduate Theses, Dissertations, and Problem Reports. 7498. https://researchrepository.wvu.edu/etd/7498 This Thesis is protected by copyright and/or related rights. It has been brought to you by the The Research Repository @ WVU with permission from the rights-holder(s). You are free to use this Thesis in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you must obtain permission from the rights-holder(s) directly, unless additional rights are indicated by a Creative Commons license in the record and/ or on the work itself. This Thesis has been accepted for inclusion in WVU Graduate Theses, Dissertations, and Problem Reports collection by an authorized administrator of The Research Repository @ WVU. For more information, please contact [email protected]. Protein and Lipid Characterization of Acheta domesticus, Bombyx mori, and Locusta migratoria Dry Flours Emily N. Brogan Thesis submitted to the Davis College of Agriculture, Natural Resources and Design at West Virginia University in partial fulfilment of the requirements for the degree of Master of Science in Nutritional and Food Science Jacek Jaczynski Ph.D., chair Kristen Matak Ph. D Yong-Lak Park, Ph. D Division of Animal and Nutritional Sciences Morgantown, West Virginia 2018 Keywords: Entomophagy, Insects, Nutrition, Food Supply, Protein, Lipid, Amino Acids Copyright 2018 Emily Brogan ABSTRACT Protein and Lipid Characterization of Acheta domesticus, Bombyx mori, and Locusta migratoria Dry Flours Emily N. Brogan Cricket (Acheta domesticus), silkworm pupae (Bombyx mori), and locust (Locusta migratoria) dry flours were obtained to examine the biochemical properties and composition of the flours. This study aimed to characterize the protein and lipid components of three insect species utilizing proximate composition analyses, amino acid composition analysis, protein solubility, SDS-PAGE, fatty acid composition analysis, and thin layer chromatography to determine lipid classes, and lipid extraction efficiency. Kjeldahl determined the cricket, locust, and silkworm flours contained 72.0%, 53.1%, and 71.2% protein, respectively. All proximate composition analyses were significantly (p<0.05) different between species. Amino acid composition analysis revealed that the flours contained 3.6-3.9% and 0.90-1.5% of lysine and methionine, respectively. Many countries have cereal grains and legumes as a staple in their diet that contain these limiting amino acids. Essential amino acids were 22% of total amino acids. Protein solubility revealed that the three species studied were most soluble in alkaline environments with highest protein solubility occurring at pH 13 at 66% solubility in silkworm. Lowest solubility occurred in more acidic conditions between pH 4-5. SDS-PAGE revealed five major protein fractions with estimated molecular weights of 27 (cuticle proteins), 41 (arginine kinase), which has been identified as an allergen in shrimp, 42 (actin), 71 (hemocyanin), and 220 (myosin) kDa. Soxhlet extraction determined cricket, silkworm, and locust flours contained 15.4%, 33.3%, and 11.4% lipid, respectively. The only omega-3 fatty acid found in the flours was α-linolenic acid. Silkworm flour contained the most α-linolenic acid at 33.3% of total fatty acids, followed by locust (13.7%), and cricket (0.6%). TLC resolved lipids for typical lipid classes. The four major lipid classes noted were triacylglycerol, free fatty acid, cholesterol, and phospholipid. Lipid extraction efficiency found that the organic solvents chloroform and methanol had the highest lipid extraction yield in both cricket (69.3%) and locust (93.0%). Methyl-tertiary-butyl ether (MTBE) extracted lipids most efficiently in the silkworm. Proteins and lipids found in insects need to be isolated to further or commercial use. DEDICATION AND ACKNOWLEDGMENTS I want to thank the Davis College at West Virginia University for funding my research. I would also like to thank my graduate committee Dr. Jacek Jaczynski, Dr. Kristen Matak, and Dr. Yong- Lak Park for sharing their knowledge with me. I could not have completed this research without the help of the students and faculty in the Animal and Nutritional Sciences division at the university. I want to specifically thank Derek Warren, Casey Showman, Emily Nadenbousch, and Alleda Rose for their commitment to helping me succeed in the lab. I would like to thank Sarah Beamer for her guidance prior to her retirement as I learned how to navigate the lab, as well as Susan Slider for helping me troubleshoot in the lab and obtain my materials. Lastly, I would like to thank my family and fiancé for their unending support during my time at West Virginia University iv TABLE OF CONTENTS Abstract ........................................................................................................................................... ii Dedication and Acknowledgments ................................................................................................ iv Table of Contents ............................................................................................................................ v List of Tables ................................................................................................................................. vi List of Figures ............................................................................................................................... vii Chapter I.......................................................................................................................................... 1 Introduction ................................................................................................................................. 1 Chapter II ........................................................................................................................................ 5 Review of Literature ................................................................................................................... 5 Insect Nutrient Profile ............................................................................................................. 5 Insects as a Source of High Quality Protein ........................................................................... 6 Characterization of Insect Protein ........................................................................................... 7 Insects as a Source of High Quality Lipid ............................................................................ 11 Characterization of Insect Lipid ............................................................................................ 11 Summary ............................................................................................................................... 13 Chapter III ................................................................................................................................. 18 Chapter IV ................................................................................................................................. 39 v LIST OF TABLES Chapter III Table 1: Table 1. Proximate composition (% dry weight basis) of Acheta domesticus (cricket), Bombyx mori (silkworm), and Locusta migratoria (locust). Table 2: Amino acid composition (mg/g) of Acheta domesticus (cricket), Bombyx mori (silkworm), and Locusta migratoria (locust) with adult DRI reference values for EAA (mg/kg/day). Table 3: Protein solubility of Acheta domesticus (cricket), Bombyx mori (silkworm), and Locusta migratoria (locust) flours as is in deionized double distilled water pH adjusted to pH1- 13 with HCl and NaOH. Chapter IV Table 1: Proximate composition (% dry weight basis) of Acheta domesticus (cricket), Bombyx mori (silkworm), and Locusta migratoria (locust). Table 2: Fatty acid composition (w/w%) of Acheta domesticus (cricket), Bombyx mori (silkworm), and Locusta migratoria (locust) using FAMEs analysis (w/w%) compared to the DRI for adult men ages 31-50 years old. Table 3: Lipid extraction efficiency in Acheta domesticus (cricket), Bombyx mori (silkworm), and Locusta migratoria (locust) using the five organic solvents, chloroform:methanol (2:1), chloroform, hexane, heaxane:isopropanol (3:2), and methyl-tertiary-butyl ether (MTBE). vi LIST OF FIGURES Chapter III Figure 1: Protein solubility of Acheta domesticus (cricket), Bombyx mori (silkworm), and Locusta migratoria (locust) flours pH adjusted pH 1-13. Figure 2: SDS-PAGE 4-20% Tris-HCl gel identification of five major protein fractions using literature in Acheta domesticus, Locusta migratoria, and Bombyx mori flours as is with protein concentrated at 8 mg/mL with a BSA 10-250 kDa reference protein. Chapter IV Figure 1: Acheta domesticus (cricket) extracted lipid (5 µL/mL) thin layer chromatography plate. Figure 2: Locusta migratoria (locust) extracted lipid (5 µL/mL) thin layer chromatography plate. Figure 3: Bombyx mori (silkworm) extracted lipid (5 µL/mL) thin layer chromatography plate. vii CHAPTER I Introduction The Food and
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