Title Page
Abstract
Contents
Chapter 1. Introduction 8
1.1. Deterioration of vacuum-packed meat and its monitoring 8
1.2. Application of receptor-embedded nanodiscs as sensing material 10
Chapter 2. Materials and methods 14
2.1. Production of receptor-embedded nanodisc 14
2.1.1. Purification of olfactory receptors 14
2.1.2. Purification of membrane scaffold protein 15
2.1.3. SDS-PAGE analysis for receptors 16
2.1.4. Assembly and purification of receptor-embedded nanodiscs 16
2.1.5. Western blot for nanodiscs 16
2.2. Characterization of receptor-embedded nanodiscs 17
2.2.1. Dynamic light scattering 17
2.2.2. Tryptophan queching assay 17
2.3. Preparation and physicochemical analyses of vacuum-packed meat 18
2.3.1. Preparation of vacuum-packed meat 18
2.3.2. Total aerobic bacteria enumeration 18
2.3.3. pH measurement 18
2.3.4. Volatile basic nitrogen quantification 18
2.3.5. TBARS assay 19
2.4. Monitoring of quality of vacuum-packed meat with receptor-embedded nanodiscs 19
2.4.1. vacuum-packed meat sample preparation 19
2.4.2. Tryptophan quenching assay 20
2.4.3. Principal Component Analysis of tryptophan quenching assay results 20
Chapter 3. Results and discussion 21
3.1. Production of receptor-embedded nanodiscs 21
3.2. Characterization of receptor-embedded nanodiscs 26
3.3. Physicochemical analysis of vacuum-packed meat 30
3.4. Monitoring of quality of vacuum-packed meat with receptor-embedded nanodiscs 33
Chapter 4. Conclusion 36
Bibliography 37
초록 42
Table 3.1. Quality traits of vacuum-packed meat stored for 70 days 32
Figure 1.1. Responses of TAAR13c and TAAR13d nanodiscs to their ligands 11
Figure 1.2. Structure of receptor embedded nanodiscs 13
Figure 3.1. Coomassie blue staining result of receptors 23
Figure 3.2. Western blot results of receptors 24
Figure 3.3. Western blot results of receptor-embedded nanodiscs 25
Figure 3.4. Dynamic light scattering results of receptor-embedded nanodiscs 28
Figure 3.5. Tryptophan quenching assay results of receptor-embedded nanodiscs with their ligands 29
Figure 3.6. Physicochemical assay results of vacuum-packed meat by storage duration 31
Figure 3.7. Tryptophan quenching assay results of receptor-embedded nanodiscs with vacuum-packed meat 34
Figure 3.8. Pattern change visualization by principal component analysis 35