臺灣博碩士論文加值系統

摘要 I
Abstract IV
Acknowledgement VII
Table of Contents VIII
List of Tables XIII
List of Figures XIV
1. Introduction 1
2. Literature Review 3
2.1 Wine 3
2.1.1 Fruit wine 4
2.2 Mulberry 7
2.2.1 Mulberry wine 8
2.3 Aroma compound in wine 9
2.4 Organic acid 14
2.4.1 Citric acid 15
2.5 Pectin 17
2.5.1 Influence of pectin to wine fermentation 19
2.5.2 Pectinase enzyme 20
2.5.2.1 Polygalacturonase 21
2.6 Yeast and co-fermentation for wine production 23
2.6.1 Saccharomyces cerevisiae 25
2.6.2 Hanseniaspora uvarum 28
3. Materials and Methods 31
3.1 Material 31
3.1.1 Equipments and instruments 31
3.1.2 Chemical reagents and mediums 32
3.1.2.1 Reagents 32
3.1.2.2 Media for microorganisms 34
3.2 Pectinolytic isolation for mulberry wine fermentation 36
3.2.1 Primary isolation 36
3.2.2 Pectinolytic yeast isolation 36
3.2.3 Pectin degradation test 38
3.2.4 Polygalacturonase enzyme activity 38
3.3 Characteristic analysis of selected yeasts and S. cerevisiae P13 strains for using in wine fermentation 39
3.3.1 Carbohydrate utilization test 39
3.3.2 Ethanol tolerance test 40
3.3.3 Citric acid tolerance test 40
3.3.4 Sulfate salt tolerance test 40
3.4 Yeast strain identification 41
3.4.1 API 32 C 41
3.4.2 VITEK® 2 compact 41
3.4.3 Characterization of yeast by scanning electron microscope 45
3.4.4 Polymerase chain reaction (PCR) 45
3.5 Experimental design 47
3.5.1 Mulberry wine fermentation 47
3.5.2 Mulberry wine analysis 48
3.5.2.1 Microbial enumeration 48
3.5.2.2 Reducing sugar 51
3.5.2.3 Total sugar 51
3.5.2.4 pH 51
3.5.2.5 Total soluble solid (TSS) 52
3.5.2.6 Titratable acidity 52
3.5.2.7 Total anthocyanin content 52
3.5.2.8 Color measurement 53
3.5.2.9 Determination ethanol content 54
3.5.2.10 Determination methanol content 55
3.5.2.11 Analysis of volatile compounds 56
3.5.2.11.1 Head space solid phase microextraction (HS-SPME) 56
3.5.2.11.2 GC-MS optimization 56
3.5.2.12 Determination of citric acid content 59
3.6 Statistical analysis 60
4. Results and Discussions 61
4.1 Microbial screening of pectinolytic yeast 61
4.1.1 Determination of pectin degradation 62
4.1.2 Determination of polygalacturonase activity 63
4.2 Principal characteristic of isolated yeasts and S. cerevisiae P13 65
4.2.1 Profile of carbohydrate utilization 65
4.2.2 Performance of ethanol tolerance 67
4.2.3 Performance of citric acid tolerance 67
4.2.4 Performance of sulfate salt tolerance 69
4.3 Identification of selected yeast 71
4.3.1 Identification of API 32 71
4.3.2 Identification of VITEK®2 71
4.3.3 Determination of cell dimension and characteristic of isolated yeast 74
4.3.4 Identification with PCR 76
4.4 Analytical parameters of mulberry wine 78
4.4.1 The enumeration of yeast 78
4.4.2 Kinetics changes of sugar and soluble solid 80
4.4.3 pH and titratable acidity 84
4.4.4 Change of color properties and anthocyanin 87
4.4.5 Ethanol and methanol contents 89
4.4.6 Volatile compounds profiles 90
4.4.7 Total citric acid 93
5. Conclusions 95
6. References 97
7. Appendices 109
Appendix 1. Pectin degradation clear zone 109
Appendix 2. Standard curve D-galacturonic acid 110
Appendix 3. The miniaturized utilization test of APl 32 C 111
Appendix 4. The mineral composition API 32 C kit 112
Appendix 5. The colony morphology in WL medium 113
Appendix 6. The standard calibration curve of ethanol 113
Appendix 7. The standard calibration curve of methanol 114
Appendix 8. Chromatogram of citric acid standard 114
Appendix 9. Chromatogram of volatile compound 115
8. Biosketch of Author 116
 

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