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Musca Domestica) As Materials for the Development of Multifunctional Nutraceuticals

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Musca Domestica) As Materials for the Development of Multifunctional Nutraceuticals The potential usefulness of the housefly larvae (Musca domestica) as materials for the development of multifunctional nutraceuticals 著者 李 虎軍 その他のタイトル 多機能性食品の開発におけるイエバエ幼虫の素材と しての潜在的有用性 学位授与年度 平成30年度 学位授与番号 17104情生工第334号 URL http://hdl.handle.net/10228/00006874 The potential usefulness of the housefly larvae (Musca domestica) as materials for the development of multifunctional nutraceuticals （多機能性食品の開発におけるイエバエ幼虫 の素材としての潜在的有用性） 李 虎軍 Hujun Li Contents Abstract ........................................................................................................................... 1 Abbreviations .................................................................................................................. 3 Chapter 1 Introduction .................................................................................................................... 5 1.1 Background ............................................................................................................. 6 1.1.1 Expectation on insects ..................................................................................... 6 1.1.2 Housefly larvae (Musca domestica) and housefly larvae bioconversion system for livestock manure ......................................................................................... 9 1.1.2.1 Housefly larvae ........................................................................................... 9 1.1.2.2 Housefly larvae bioconversion system ..................................................... 10 1.1.3 Bioactive compounds derived from natural sources....................................... 11 1.1.3.1 Antioxidants .............................................................................................. 12 1.1.3.2 Angiotensin-converting enzyme inhibitors ............................................... 12 1.1.3.3 Dipeptidyl peptidase-IV inhibitors ........................................................... 14 1.2 Utilities of housefly larvae ................................................................................... 15 1.3 Objectives ............................................................................................................. 16 Chapter 2 Evaluation of usefulness of housefly larvae based on amino acid analysis ............. 17 Abstract ........................................................................................................................... 18 2.1 Introduction .......................................................................................................... 18 2.2 Materials ............................................................................................................... 19 2.3 Methods ................................................................................................................ 19 2.3.1 Amino acid analysis of housefly larvae and fish meal .................................. 19 2.3.2 Comparison of amino acid composition between housefly larvae and fishmeal ...................................................................................................................... 19 2.3.3 Comparison of amino acid composition between housefly larvae and beef, chicken, pork, casein, and hen egg ............................................................... 20 2.3.4 Calculation of the degree of similarity between housefly larvae and other resource in amino acids ................................................................................ 20 2.4. Results ................................................................................................................. 21 2.4.1 Amino acid composition of housefly larvae .................................................. 21 2.4.2 Amino acid composition of housefly larvae and fishmeal ............................ 21 I 2.4.3 Amino acid composition of housefly larvae and other protein sources ......... 28 2.4.4 The degree of similarity between housefly larvae and other resources in amino acids .............................................................................................................. 33 2.5 Discussion ............................................................................................................. 34 2.6 Conclusion ............................................................................................................ 36 Chapter 3 Preparation of housefly larvae water extract (HLWE) with multifunctional activities ......................................................................................................................... 37 Abstract ....................................................................................................................... 38 3.1 Introduction .......................................................................................................... 38 3.2 Materials ............................................................................................................... 39 3.3 Methods ................................................................................................................ 39 3.3.1 Preparation of HLWE .................................................................................... 39 3.3.2 Molecular distribution of HLWE by SDS-PAGE .......................................... 40 3.3.3 Fractionation of HLWE by ultrafiltration ...................................................... 40 3.3.4 Confirmation of fraction of HLWE by tricine-SDS-PAGE ........................... 40 3.3.5 Gel filtration chromatography ....................................................................... 40 3.3.6 Amino acid analysis of HLWE and its fraction ............................................. 41 3.3.7 Biological activity assays of the HLWE and its fractions ............................. 41 3.3.7.1 Measurement of antioxidant activity ........................................................ 41 3.3.7.2 ACE inhibition assay ................................................................................ 42 3.3.7.3 DPP-IV inhibition assay ........................................................................... 42 3.3.8 Statistical analysis ......................................................................................... 43 3.4 Results .................................................................................................................. 43 3.4.1 Preparation of HLWE and molecular distribution of HLWE by SDS-PAGE 43 3.4.2 Fractionation of HLWE and confirmation of fractionation by tricine-SDS-PAGE ........................................................................................ 44 3.4.3 Determination of molecular distribution of HLWE and its fractions by gel filtration chromatography ............................................................................. 44 3.4.4 Amino acid composition of HLWE and its fractions ..................................... 45 3.4.5 Biological activities ....................................................................................... 47 3.4.5.1 Antioxidant activity of HLWE and its fraction ......................................... 47 3.4.5.2 ACE- inhibitory activity assay ................................................................... 50 3.4.5.3 DPP-IV-inhibitory activity assay .............................................................. 52 3.5 Discussion ............................................................................................................. 54 II 3.6 Conclusion ............................................................................................................ 57 Chapter 4 Summary ....................................................................................................................... 58 Acknowledgements ........................................................................................................ 61 References ...................................................................................................................... 62 III Abstract Many bioactive compounds have been studied for the prevention or treatment of various diseases. Oxidative stress has been confirmed to be related to many chronic diseases, including hypertension and diabetes mellitus. Hypertension is a major chronic adult disease affecting 40% population worldwide, and angiotensin-converting enzyme (ACE, EC 3.4.15.1) plays an important role in elevation in blood pressure. Type 2 diabetes mellitus (DM) is the most common form of diabetes, which account for approximately 90 to 95% of all the diagnosed cases, and dipeptidyl peptidase-IV (DPP-IV, EC 3.4.14.5) is involved in this symptom. Therefore, the development of materials which have antioxidant, ACE-inhibitory (antihypertension), and DPP-IV-inhibitory (antidiabetic) activities become important to the prevention or treatment of these chronic diseases. Although many chemically synthesized antioxidants, ACE inhibitors, and DPP-IV inhibitors have been developed, it is still unclear whether these chemically synthesized compounds are sufficiently safe for long-term dosing or whether these compounds may have side effects. Therefore, bioactive compounds derived from natural sources are needed. Bioactive compounds are usually derived from plants, aquatic animals, and mammals. Although many types of insects have been used as foods and protein sources, there are few reports on biological activities of insects. Insects are an abundant resource, that, though not well explored, is expected to be a solution to food security in the future. Among them, the housefly has a high reproductive ability and
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